Novel compounds for the diagnosis of TDP-43 proteinopathy

Abstract

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

Description

[Technical field]

[0001] FIELD OF THEINVENTION The present invention relates to compounds suitable for imaging TDP-43 (transactive 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 senile TDP-43 encephalopathy (LATE). The present invention also relates to methods for the preparation of the compounds, diagnostic compositions comprising the compounds, methods of using the compounds, kits comprising the compounds, and uses thereof. [Background technology]

[0002] 2. Background of the Invention Geriatric brain disorders, characterized by pathological aggregation of proteins (proteinopathies in the CNS) in the CNS and peripheral organs, are one of the leading causes of disability and death worldwide. The best-characterized protein that forms extracellular aggregates is amyloid beta (Aβ) in Alzheimer's disease (AD) and Aβ-related disorders. Other disease-related proteins prone to aggregation that cause neurodegeneration include, but are not limited to, tau, α-synuclein (a-syn), huntingtin, fused in sarcoma (FUS), dipeptide repeat proteins (DPRs) produced by atypical translation of C9orf72 repeat expansions, superoxide dismutase 1 (SOD1), and TDP-43. Diseases involving TDP-43 aggregates are collectively known as TDP-43 proteinopathies, and include, but are not limited to, amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration (FTLD), including frontotemporal dementia with TDP-43 pathology (FTLD-TDP, frontotemporal lobar degeneration with TDP-43 inclusions), and limbic-predominant late-life TDP-43 encephalopathy (LATE).

[0003] Introduction to TDP-43 Transactive response (TAR) DNA-binding protein 43 kDa (TDP-43) is a 414 amino acid protein encoded by the TARDBP gene (ALS10) on chromosome 1p36.2. TARDBP consists of six exons (exon 1 is noncoding; exons 2-6 code for proteins). TDP-43 belongs to the family of heterogeneous ribonucleoprotein (hnRNP) RNA-binding proteins (Wang et al., Trends in Molecular Medicine, Vol. 14, No. 11, 2008, 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) 1606-1609), namely, 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), a nuclear localization signal (NLS) that enables the NES to shuttle between the nucleus and the cytoplasm to transport bound mRNA, and a C-terminal glycine-rich domain 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) 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 usually primarily localized in the nucleus.In 2006, TDP-43 was identified as the protein that accumulates in the majority of cases of frontotemporal lobar degeneration (FTLD) with tau-negative, ubiquitin-positive inclusions (then called FTLD-TDP) and in most cases of amyotrophic lateral sclerosis (ALS) (Arai et al., Biochemical and Biophysical Research Communications, 351 (2006) 602-611; Neumann et al., Science, 314, (2006), 130-133).

[0004] Thirty-eight dominant-negative mutations in TDP-43, mainly located in the glycine-rich domain, have been identified in sporadic and familial ALS patients, as well as in hereditary FTD patients (K263E, N267S) (Figure 1; Lagier-Tourenne and Cleveland, Cell, 136, 2009, 1001-1004). TDP-43 is intrinsically 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), 285-296.).

[0005] TDP-43 in Neurodegeneration TDP-43 aggregates are associated with a number 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, chromosome 9p-linked dementia, corticobasal degeneration, frontotemporal lobar degeneration with ubiquitin-positive inclusions, argyrophilic grain disease, and Pick's disease), amyotrophic lateral sclerosis (sporadic ALS, with TARDBP mutations, and AN). G mutations), Alzheimer's disease (sporadic and familial), Down's syndrome, familial British dementia, polyglutamine diseases (Huntington's disease and SCA3), hippocampal sclerosis dementia and myopathies (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 shows several abnormal modifications, including hyperphosphorylation, ubiquitination, acetylation, and C-terminal fragmentation through proteolytic cleavage (Arai et al., Biochemical and Biophysical Research Communications, 351 (2006) 602-611; Neumann et al., Science, 314, (2006), 130-133; Neumann et al., Acta Neuropathol, (2009) 117:137-149; Hasegawa et al., Annals of Neurology, 2008, 64(1), 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 degenerated neurites (DNs), which are immunoreactive for TDP-43 as well as ubiquitin and p62, but negative for other neurodegenerative disease-associated proteins. Differences in the morphology of the inclusions and their tissue distribution are associated with specific mutations and / or clinical features. Four types of TDP-43 pathology by histological methods have been described so far (Mackenzie and Neumann, J. Neurochem., (2016), 138 (Suppl. 1), 54-70). FTLD-TDP type A cases are characterized by abundant short DNs and small, oval or semilunar NCIs, mainly in layer II of the neocortex (Fig. 2f in Mackenzie et al., J. Neurochem., 2016, 138 (Suppl. 1), 54-70). Cases of this condition usually present clinically with behavioral frontotemporal dementia (bvFTD) or nonfluent / agrammatic primary progressive aphasia (nfvPPA) and are associated with progranulin (GRN) mutations.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), 54-70, Fig. 2g). Most cases with concurrent FTD and ALS symptoms have been found to have FTLD-TDP type B pathology. Type C cases have abundant long and tortuous neurites, mainly in superficial cortical layers, with few or no NCIs (Mackenzie et al., J. Neurochem., 2016, 138 (Suppl. 1), 54-70, Fig. 2j). This pathology is particularly seen in cases presenting with svPPA (semantic primary progressive aphasia). FTLD-TDP type D shows abundant neuronal intranuclear inclusions (NIIs) in the neocortex and short DNs, with only rare NCIs (Fig. 2k in Mackenzie et al., J. Neurochem., 2016, 138 (Suppl. 1), 54-70). This pattern of pathology is seen only in cases with VCP mutations associated with inclusion body myositis.

[0007] TDP-43 in FTD Frontotemporal dementia (FTD) is a clinical term that covers a wide range of disorders based on the degeneration of the frontal and temporal lobes, a pathological feature called frontotemporal lobar degeneration (FTLD). FTD is the second most common cause of early degenerative dementia in the age group under 65 years (Le Ber, Revue Neurologique, 169 (2013), 811-819). FTD is represented 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. The diagnosis of these syndromes is complex, and a final conclusion can only be obtained through postmortem tissue analysis based on immunohistochemistry to detect aggregated proteins, and the description of the affected brain regions. With regard to pathological protein 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 pathology term describing FTD cases with TDP-43 pathology found primarily as cytoplasmic or neurite protein aggregates in neuronal and glial cells containing misfolded, insoluble, phosphorylated, and truncated TDP-43.

[0008] TDP-43 in ALS Amyotrophic lateral sclerosis (ALS) is an adult-onset neurodegenerative disorder characterized by early loss of upper and lower motor neurons. ALS progression is characterized by fatal paralysis and respiratory failure, with a disease course from illness to death of 1-5 years. In most cases of sporadic ALS, neuropathology is characterized by abnormal cytoplasmic accumulation of TDP-43 in neuronal and glial cells of the primary motor cortex, brainstem motor nuclei, spinal cord, and associated white matter tracts. ALS with dementia is associated with accumulation of TDP-43 in the supplementary motor area, neocortex, and hippocampus. The role of TDP-43 phosphorylation in ALS patients was investigated with the aid of phosphorylation-specific antibodies that tightly bind to nuclear and cytoplasmic TDP-43 inclusions. Amino acids S379, S403, S404, S409, and S410 have been identified as the major phosphorylation sites 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 of limbic-predominant senile TDP-43 encephalopathy (LATE) (LATE-NC) are defined by stereotypic TDP-43 proteinopathy in older adults with or without comorbid hippocampal sclerosis pathology. LATE-NC is a common TDP-43 proteinopathy associated with an amnesic dementia syndrome mimicking Alzheimer's dementia in retrospective autopsy studies. LATE is differentiated from frontotemporal lobar degeneration with TDP-43 pathology based on its epidemiological characteristics (LATE generally affects older subjects) and the relatively restricted neuroanatomical distribution of TDP-43 proteinopathy. No molecular-specific biomarkers of LATE exist. The discovery of the TDP-43 PET tracer may enable accurate and potentially earlier diagnosis, and monitoring of disease progression to facilitate long-term drug efficacy measurements in patients during clinical trials (including as a potential exclusion criterion for Alzheimer's disease clinical trials) as well as longitudinal studies of LATE clinical and pathological progression (Nelson et al., Brain, 2019, volume 142; issue 6, 1503 - 1527).

[0010] TDP-43 in AD and other diseases TDP-43 pathology occurs in up to 57% of brains of Alzheimer's disease patients (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., 2017 Jul;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 have cognitive impairment at death than TDP-43 negative controls. TDP-43 appears to be 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), then the hippocampus, limbic system, temporal lobes, and finally the fronto-striatum (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 features is insufficient, since clinical features may overlap with other diseases, especially in early stages.Therefore, there is an urgent need to develop biomarkers with sensitivity and specificity that allow differentiation between types of pathology within the FTD spectrum.Such tools will allow better detection and understanding of the specific types of pathology that cause neurodegeneration.Ultimately, this will lead to the development of diagnostic biomarkers that allow more efficient and accurate patient selection for long-term monitoring in clinical trials and to support the development of new therapeutic drugs for ALS and FTD.

[0012] Several approaches have aimed at developing biochemical biomarkers to distinguish different types of FTD pathology. Several studies have shown that TDP-43 concentrations are increased in the cerebrospinal fluid (CSF) of clinically defined FTD or FTD-MND populations, but there is significant overlap with controls or AD subjects, and it remains unclear whether this approach is 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 possible diagnostic tool for distinguishing FTLD-tau from FTLD-TDP is a low ratio of p-tau181 to tau in the CSF, below values ​​of 0.37 (Hu et al., Neurology., 2013;81(22):1945-1952). Another study showed that the level of phosphorylated tau in the CSF was 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] Alongside biochemical biomarkers, the development of imaging biomarkers would allow for early and specific detection of pathology in FTD and ALS. The ability to image TDP-43 deposition in the brain would be a considerable achievement in 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 shown direct binding to this target.

[0014] The development of TDP-43 specific PET tracers involves several challenges, including the low abundance and heterogeneous distribution of the target in the patient's brain, and the lack of a reference compound. To reduce background signal interference caused by non-specific off-target binding, and to reduce dosage requirements, TDP-43 imaging compounds should bind to the target with high affinity and selectivity. For imaging of TDP-43 aggregates associated with neurological disorders such as FTD and ALS, the imaging compound needs to penetrate and cross the blood-brain barrier to enter the relevant regions of the brain. To target intracellular amyloid-like inclusions such as TDP-43 aggregates, cell permeability is an additional requirement for the imaging compound. A further prerequisite to avoid compound accumulation in tissues, which may increase the risk of undesirable side effects, is rapid washout of the compound from the brain (or other target organs). [Prior art documents] [Patent documents]

[0015] [Patent Document 1] US 8,932,557 [Non-patent literature]

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

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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 the identification and differentiation of patients and patient groups with TDP-43 proteinopathies (e.g. FTD, FTLD-TDP, LATE, and ALS) and for the differentiation of TDP-43 proteinopathies from other proteinopathies. [Means for solving the problem]

[0018] Surprisingly, the inventors have discovered that compounds having formula (I) are capable of recognizing and binding to TDP-43 aggregates.

[0019] Summary of the Invention 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, Z 1 , Z 2 , Z 3 , and Z 4 are selected from CH and N, Z 1 If N, then Z 2 is CH; Z 2 If N, then Z 1 is CH; Z 3 If N, then Z 4 is CH; Z 4 If N, then Z 3 is CH; n is 1 or 2; R 1 is H or F; R 2 teeth a 5- or 6-membered carbocyclic ring optionally substituted with F, NH2, CN, and / or CH3; a 5- or 6-membered heterocycloalkyl ring, optionally substituted with F, NH2, CN, and / or CH3, containing one or more heteroatoms selected from N, O, and S; 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 is a 6-membered heteroaryl ring optionally substituted with F, NH2, CN, and / or CH3 and containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S. or a detectably labeled compound, stereoisomer, polymorph, racemic mixture, tautomer, pharma- ceutically acceptable salt, prodrug, hydrate, or solvate thereof, or mixtures thereof.

[0022] In another aspect, the present invention provides a diagnostic composition comprising a compound as defined in the present specification of formula (I) or a derivative thereof, and optionally at least one physiologically acceptable carrier, diluent, adjuvant and / or excipient, which can be used for imaging TDP-43 aggregates, in particular by positron emission tomography, or for diagnosing a disease, disorder or condition associated with TDP-43 aggregates, in particular by positron emission tomography.

[0023] In another aspect, the present invention provides a method comprising: A method of 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 the detection and optionally quantification of TDP-43 aggregates in tissue of a subject; · A method for diagnostic imaging of a subject's brain; Methods of collecting data for the diagnosis of a disease, disorder, or abnormality associated with TDP-43 aggregates or for the diagnosis of a TDP-43 proteinopathy: · A method for collecting data to determine a disease, disorder, or abnormality associated with TDP-43 aggregates, or a predisposition to a TDP-43 proteinopathy; A method for collecting data to monitor the progression of a disease, disorder, or abnormality associated with TDP-43 aggregates or to monitor 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 condition associated with TDP-43 aggregates to a pharmaceutical treatment. The present invention provides compounds according to the definition of compounds of formula (I) or derivatives thereof, which can be used in

[0024] In another aspect, the present invention provides compounds according to the definition of compounds of formula (I) or derivatives thereof, which can be used as biomarkers for TDP-43 aggregates or biomarkers for TDP-43 proteinopathies, as diagnostics or tools for TDP-43 proteinopathies, 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 compounds of formula (I) or derivatives thereof.

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

[0027] [Figure 1] Specific binding of increasing concentrations of [3H]-Compound 1 and [3H]-Compound 3 to recombinant TDP-43 aggregates (non-specific binding subtracted from total) using a one-site specific binding model in GraphPad Prism. [Diagram 2]Microautoradiography staining of pTDP-43 aggregates in FTLD-TDP brain sections with [3H]-Compound 1. Colocalization of argyrophilic granules (black dots, left panel) and fluorescent signal from pTDP-43 antibody staining (right panel) in frontal cortex brain sections from FTLD-TDP donors incubated with [3H]-Compound 1 (60 nM). Accumulation of argyrophilic granules (arrows) on pTDP-43 aggregates was found. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[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 may exist as racemates and racemic mixtures, stereoisomers (including diastereomeric mixtures and individual diastereomers, enantiomeric mixtures and single enantiomers, conformeric mixtures and single conformers), tautomers, atropisomers, 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. All pharma- ceutically acceptable salt forms, polymorphs, hydrates, solvates, prodrugs, and mixtures thereof are also included in the present invention. Unless otherwise specified, the term "compound of formula (I)" or "compound of the present invention" means "a compound of formula (I) or a detectably labeled compound thereof, stereoisomer, polymorph, racemic mixture, tautomer, pharma- ceutically 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 derivatives thereof, as well as isotopically labeled compounds (e.g., 18 F and 3 The term "compound of formula (I)" or "compound of the invention" means a compound as defined in any one of the embodiments mentioned below.

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

[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 pharma- ceutically acceptable solvent. Examples include C 1~4 Alcohols such as methanol or ethanol are included.

[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, for example formed from non-toxic inorganic or organic acids. For example, these conventional non-toxic salts include those derived from inorganic acids, such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like; and those 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. The 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, these salts can be prepared by reacting the free acid or free base form of these compounds with a stoichiometric amount of the appropriate acid or base 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. Lists of suitable salts can be found in Remington's Pharmaceutical Sciences, 18th issue, the disclosure of which is incorporated herein by reference. th ed., Mack Publishing Company, Easton, PA, 1990, page 1445. Typically, the pharma- ceutically acceptable salts are salts of amine residues in the compounds of the 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] In the present invention, a "diagnostic composition" is defined as a composition containing a compound of the invention in a form suitable for administration to a patient, eg, a mammal, such as a human.

[0036] "TDP-43 aggregates" refer to TDP-43-positive, multimer-rich aggregates of TDP-43. They can be found in intracellular deposits in a series of diseases called TDP-43 proteinopathies, particularly in amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), frontotemporal dementia (FTD), and limbic-predominant senile TDP-43 encephalopathy (LATE). TDP-43 aggregates can be found in the following morphologies: small, oval or crescent-shaped neuronal cytoplasmic inclusions (NCIs), lenticular neuronal intranuclear inclusions (NIIs), glial cytoplasmic inclusions (GCIs), degenerating neurites (DNs), and long, tortuous neurites. In pathological aggregates, TDP-43 often shows substantial increases in post-translational modifications, such as phosphorylation, ubiquitination, acetylation, SUMOylation, and proteolytic cleavage to generate C-terminal fragments.

[0037] The "preclinical state" of a disease is defined as a stage of the disease where disease-related changes at the molecular level have not yet given rise to an overt clinical picture in the patient.

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

[0039] The term "diagnosing" or "diagnosis" generally refers to the process or act of recognizing, determining, or concluding a disease or condition in a patient based on symptoms and signs and / or from the results of a diagnostic procedure. The method should be performed on multiple subjects. The average amount of compound (if any) bound to TDP-43 aggregates is then calculated and defined as a range.

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

[0041] A "healthy control subject" or "healthy subject" is an individual who does not show clinical evidence of a neurodegenerative disease. This individual meets the following criteria: Healthy male and female subjects with no clinically relevant findings upon 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. No clinical signs or symptoms of current neurological deficits, such as cognitive impairment or motor deficits must be met.

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

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

[0044] The term "predict" or "prediction" generally refers to the advance prognosis, forecast, or prognosis of a disease or condition in a patient who does not have the disease, disorder, or condition. For example, a prediction of a disease, disorder, or condition in a patient can indicate the likelihood, probability, or risk that the patient will suffer from the disease, disorder, or condition, for example, within a certain time period or by a certain age.

[0045] Detectable labels include suitable isotopes, such as radioisotopes, in particular positron- or gamma-emitters; 2 H, 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, more preferably 3 H and 18 F, most preferably 18 Examples include F.

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

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

[0048] The term "heterocycle" refers to a stable 5- or 6-membered heterocycle, including, but not limited to, any 5- or 6-membered saturated or unsaturated heterocycle. Unsaturated heterocycles include, but are not limited to, aromatic rings. Heterocycles contain one or more heteroatoms (e.g., 1 or 2 heteroatoms) selected from N, O, and S. The heteroatom is preferably N or O, more preferably N. Examples of 5- or 6-membered heterocycles include, for example, pyrrolyl, pyrrolidinyl, furanyl, tetrahydrofuranyl, thiophenyl, imidazolidinyl, pyrazolidinyl, imidazolyl, pyrazolyl, oxathiolidinyl, isoxthiolidinyl, oxathiolyl, isoxathiolyl, thiazolidinyl, isothiazolidinyl, thiazolyl, and isothiazolyl.

[0049] The term "5- or 6-membered heterocycloalkyl ring" is not particularly limited and includes any 5- or 6-membered saturated or unsaturated heterocyclic ring that is not an aromatic ring. The heterocycloalkyl ring contains one or more (e.g., 1 or 2) heteroatoms selected from N, O, and S. The heteroatoms are preferably N or O, more preferably N. Examples include, for example, pyrrolidinyl, tetrahydrofuranyl, imidazolidinyl, pyrazolidinyl, oxathiolidinyl, and isoxathiolidinyl.

[0050] The term "5-membered heteroaryl ring" is not particularly limited and includes any 5-membered aromatic heterocycle. The heteroaryl ring includes one or more (e.g., 1 or 2) heteroatoms selected from N, O, and S. The heteroatom is preferably N or O, more preferably N. Examples include, for example, pyrrolyl, furanyl, thiophenyl, imidazolyl, pyrazolyl, isoxathiolyl, thiazolidinyl, isothiazolidinyl, thiazolyl, and isothiazolyl. Pyrazolyl is preferred.

[0051] The term "6-membered heteroaryl ring" is not particularly limited and includes any 5-membered aromatic heterocycle. The heteroaryl ring contains one heteroatom selected from O and S, or two or more heteroatoms selected from O, N, and S. The heteroatom is preferably N or O, more preferably N. Examples include, for example, pyridinyl, pyridazinyl, pyrimidinyl, and pyrazinyl.

[0052] The term "leaving group" (LG) as used herein refers to any leaving group, meaning 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, 7, 71-83, Schemes 1, 2, 10, and 15). (Coenen, Fluorine-18 Labeling Methods: Features and Possibilities of Basic Reactions, (2006), in: Schubiger PA, Friebe M., Lehmann L., (eds), PET-Chemistry - The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, pp. 15-50, 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 sulfonates and C 6~10 More preferably, the leaving group (LG) is mesylate, tosylate, or nosylate. Even more preferably, the leaving group (LG) is mesylate.

[0053] As used herein, the term "detecting" encompasses quantitative and / or qualitative detection.

[0054] Detailed Description of the Invention Various embodiments of the present invention are described herein, it being recognized that features defined in each embodiment may be combined with other defined features to realize further embodiments of the present invention.

[0055] It will be understood that all definitions given with respect to formula (I) apply to all its subgenera, including formulas (Ia), (Ib), (Ic), (Id), (Ie), (If), (I*), (IH), (I*-H), (IF), (I*-F), (II), (II*), (IC), (I*-C), (III), (III*), (IV), and (IV*).

[0056] All provisions given with respect to formula (IH) or (IF) apply to (I*-H) or (I*-F) respectively.

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

[0058] [ka]

[0059] (In the formula, Z 1 , Z 2 , Z 3 , and Z 4 are selected from CH and N, Z 1 If N, then Z 2 is CH; Z 2 If N, then Z 1 is CH; Z 3 If N, then Z 4 is CH; Z 4 If N, then Z 3 is CH; n is 1 or 2; R 1 is H or F; R 2teeth a 5- or 6-membered carbocyclic ring optionally substituted with F, NH2, CN, and / or CH3; a 5- or 6-membered heterocycloalkyl ring, optionally substituted with F, NH2, CN, and / or CH3, containing one or more heteroatoms selected from N, O, and S; 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 is a 6-membered heteroaryl ring optionally substituted with F, NH2, CN, and / or CH3 and containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S. or a detectably labeled compound, stereoisomer, polymorph, racemic mixture, tautomer, pharma- ceutically acceptable salt, prodrug, hydrate, or solvate thereof, or mixtures thereof.

[0060] ring

[0061] [ka]

[0062] teeth

[0063] [ka]

[0064] It can be preferably selected from:

[0065] A preferred embodiment of the compound having formula (I) is

[0066] [ka]

[0067] It is.

[0068] In one embodiment of the first aspect, the present invention provides a compound of formula (I*);

[0069] [ka]

[0070] (In the formula, n is 1 or 2; R 1 is H or F; R 2 is a 5- or 6-membered carbocyclic ring or a 5- or 6-membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S, optionally substituted with F, NH2, and / or CH3; R 6 is H) or a detectably labeled compound, stereoisomer, polymorph, racemic mixture, tautomer, pharma- ceutically acceptable salt, prodrug, hydrate, or solvate thereof, or mixtures thereof.

[0071] 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.

[0072] The present invention relates to 1 is H or F. In a preferred embodiment, R 1 is H. In another preferred embodiment, R 1 is F.

[0073] The present invention relates to 2 but a 5- or 6-membered carbocyclic ring, optionally substituted with F, NH2, CN, and / or CH3, preferably the optional substituents being F, NH2, and / or CN; a 5- or 6-membered heterocycloalkyl ring, optionally substituted with F, NH2, CN, and / or CH3, containing one or more heteroatoms selected from N, O, and S, preferably the optional substituents being F, NH2, and / or CN; a 5-membered heteroaryl ring, optionally substituted with F, NH2, CN, and / or CH3, containing one or more heteroatoms selected from N, O, and S, preferably the optional substituents being CN and / or CH3; or This relates to compounds of formula (I) which are 6-membered heteroaryl rings optionally substituted with F, NH2, CN, and / or CH3, preferably the optional substituents being CN and / or CH3, and containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S.

[0074] Preferably, R 2 teeth a 5- or 6-membered carbocyclic 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 one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S.

[0075] R 2 Preferred examples of rings are given in the "Definitions" section above. Preferably, R 2 is phenyl or pyrazolyl, either of which may be optionally substituted with F, NH2, CN, and / or CH3, e.g., F, NH2, and / or CH3. In a preferred embodiment, phenyl may be optionally substituted with F, NH2, and / or CN. In a preferred embodiment, pyrazolyl may be optionally substituted with CH3.

[0076] R 2 The ring may be optionally substituted at any available position with F, NH2, CN, and / or CH3 (e.g., F, NH2, and / or CH3). Preferably, the 5- or 6-membered carbocyclic or heterocyclic ring is substituted with one or more of F, NH2, CN, and / or CH3 (e.g., F, NH2, and / or CH3).

[0077] In one embodiment, the present invention provides a method for the preparation of a compound comprising the steps of: 2 is a 5- or 6-membered carbocyclic ring or a 5- or 6-membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S, optionally substituted with F, NH2, and / or CH3. 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 F, NH2, and / or CH3. 6 is H.

[0078] In a preferred embodiment, the present invention comprises: R 2 but

[0079] [ka]

[0080] (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 8 is H, R 3 is CN and R 4 is NH2 and R 7 is H and R8 is H, R 3 is H and R 4 is NH2 and R 7 is H and R 8 is CN, or R 3 is H and R 4 is NH2 and R 7 is H and R 8 is F.

[0081] In another preferred embodiment, the present invention comprises: R 2 but

[0082] [ka]

[0083] (wherein X is N and R 5 is CH3).

[0084] In another preferred embodiment, the present invention comprises: R 2 but

[0085] [ka]

[0086] (wherein X is N and R 5 is H.

[0087] In one embodiment, the compound of formula (I) is 1 is F;R 2 but

[0088] [ka]

[0089] (wherein X is N and R 5 is CH3 or H, preferably CH3).

[0090] In one embodiment, the compound of formula (I) is 1 is H;R 2 but

[0091] [ka]

[0092] (wherein X is N and R 5 is CH3 or H, preferably CH3).

[0093] In one embodiment, the compound of formula (I) is 1 is F;R 2 but

[0094] [ka]

[0095] (wherein X is N and R 5 is CH3 or H, preferably CH3).

[0096] In one embodiment, the compound of formula (I) is 1 is H;R 2 but

[0097] [ka]

[0098] (wherein X is N and R 5 is CH3 or H, preferably CH3).

[0099] In one embodiment, the compound of formula (I) is1 is H;R 2 but

[0100] [ka]

[0101] (In the formula, R 3 is F and R 4 is -NH2, and R 7 is H and R 8 is H).

[0102] In one embodiment, the compound of formula (I) is 1 is F;R 2 but

[0103] [ka]

[0104] (In the formula, R 3 is F and R 4 is -NH2, and R 7 is H and R 8 is H).

[0105] In one embodiment, the compound of formula (I) is 1 is H;R 2 but

[0106] [ka]

[0107] (In the formula, R 3 is NH2 and R 4 is F and R 7 is H and R 8 is H).

[0108] In one embodiment, the compound of formula (I) is1 is F;R 2 but

[0109] [ka]

[0110] (In the formula, R 3 is NH2 and R 4 is F and R 7 is H and R 8 is H).

[0111] In one embodiment, the compound of formula (I) is 1 is H;R 2 but

[0112] [ka]

[0113] (In the formula, R 3 is CN and R 4 is NH2 and R 7 is H and R 8 is H).

[0114] In one embodiment, the compound of formula (I) is 1 is F;R 2 but

[0115] [ka]

[0116] (In the formula, R 3 is CN and R 4 is NH2 and R 7 is H and R 8 is H).

[0117] In one embodiment, the compound of formula (I) is1 is H;R 2 but

[0118] [ka]

[0119] (In the formula, R 3 is H and R 4 is NH2 and R 7 is H and R 8 is CN).

[0120] In one embodiment, the compound of formula (I) is 1 is F;R 2 but

[0121] [ka]

[0122] (In the formula, R 3 is H and R 4 is NH2 and R 7 is H and R 8 is CN).

[0123] In one embodiment, the compound of formula (I) is 1 is H;R 2 but

[0124] [ka]

[0125] (In the formula, R 3 is H and R 4 is NH2 and R 7 is H and R 8 is F).

[0126] In one embodiment, the compound of formula (I) is1 is F;R 2 but

[0127] [ka]

[0128] (In the formula, R 3 is H and R 4 is NH2 and R 7 is H and R 8 is F).

[0129] In all the above compounds having formula (I*), R 6 is H.

[0130] Preferred compounds of formula (I) include

[0131] [ka] [ka]

[0132] Examples include:

[0133] According to one embodiment, preferred compounds of formula (I) are

[0134] [ka]

[0135] can be selected from:

[0136] In one embodiment, preferred compounds of formula (I) are the following stereoisomers:

[0137] [ka]

[0138] can be selected from:

[0139] 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.

[0140] The type of detectable label is not particularly 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 identical to the natural amount of each isotope.Furthermore, the amount used should allow its detection by the detection method selected.

[0141] 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 F (detectable label). Any reference to "H" 1 H (stable) or 3 H (a detectable label, also called tritium, represented herein as "T").

[0142] Generally, the isotopic variants of the compounds of the present invention can be prepared by conventional procedures, for example, by exemplary methods, or by the preparation methods described in the following examples and preparations, using the appropriate isotopic variants of suitable reagents that are commercially available or prepared by known synthesis techniques.Radioisotopes, particularly positron emitters or gamma emitters, can be incorporated into the compounds of the present invention by methods that are common in the field of organic synthesis.Usually, they are introduced using starting materials that are appropriately labeled.Exemplary methods for introducing detectable labels are described, for example, in US 8,932,557, which is incorporated herein by reference.

[0143] 18 F can be attached at any position 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 18 It is also of special interest because of its potential use as an analytical standard during the manufacture, quality control, release, and clinical use of the F analogues.

[0144] In the compounds having formula (I), 18 F is, for example, R 2 as an F substituent in, or R 1 Preferably, it can be present as R 1 It exists as.

[0145] 3 When H is used as a detectable label, it is represented by -CT3 (T is -CT3) at any position where a CH3 group can be attached. 3 H). 3 Substitution with radioisotopes such as H can provide certain diagnostic advantages afforded by increased metabolic stability, for example by reduced defluorination, increased in vivo half-life, or reduced dosage requirements, while maintaining or improving the intrinsic efficacy of the compound.

[0146] In one embodiment, the present invention relates to a method for producing a cyclohexyl ether (CVD) having at least one hydrogen (H) and at least one tritium ( 3 H), wherein the tritium atom is replaced with a detectable label selected from the group consisting of: 3 3H) is detectably labeled with tritium ( 3 H), having the formula (I) 3 3H) is preferably defined as a compound detectably labeled with tritium ( 3 H), having the formula (I) 3 3H) is more preferably defined as a compound detectably labeled with tritium ( 3 H), having the formula (I) 3 H) detectably labeled compounds are further preferably defined.

[0147] In one embodiment, the present invention provides a compound of formula (I) having the formula (IT). In particular, the present invention provides a compound of formula (IT);

[0148] [ka]

[0149] (In the formula, n, Z 1 , Z 2 , Z 3 , Z 4 , R 1 , and R 2 is as defined herein for compounds of formula (I), 3 H), or a stereoisomer, polymorph, racemic mixture, tautomer, pharma- ceutically acceptable salt, prodrug, hydrate, or solvate thereof, or a mixture thereof. 1 , Z 2 , Z 3 , Z 4 In another embodiment, at least one of R 2is substituted with at least one CT3, or R 2 At least one hydrogen atom in is replaced by T.

[0150] In one embodiment, the present invention provides a compound of formula (I*) having the formula (I*-T). In particular, the present invention provides a compound of formula (I*-T);

[0151] [ka]

[0152] (In the formula, n, R 1 , R 2 , and R 6 is as defined herein for compounds of formula (I*) 3 H) or a stereoisomer, polymorph, racemic mixture, tautomer, pharma- ceutically acceptable salt, prodrug, hydrate, or solvate thereof, or a mixture thereof.

[0153] In a preferred embodiment, the present invention comprises: n is 1 or 2; preferably, n is 1; R 1 is H or F; (With respect to formula (IT), Z 1 , Z 2 , Z 3 , and Z 4 is as defined above; with respect to formula (I*-T), R 6 is T or H); T 3 H; R 2 but

[0154] [ka]

[0155] (In the formula, R 3 is F and R 4 is -NH2, and R 7 and R8 at least one of is T and, if applicable, the other is H; Preferably, R 7 is T;R 8 is T); Or, R 2 but

[0156] [ka]

[0157] (wherein X is N and R 5 is tritiated -CH3(CT3)

[0158] In one embodiment, the present invention provides R 1 is H; (With respect to formula (IT), Z 1 , Z 2 , Z 3 , and Z 4 is as defined above; with respect to formula (I*-T), R 6 is T); R 2 but

[0159] [ka]

[0160] (In the formula, R 3 is F;R 4 is -NH2; R 7 and R 8 is T and, if applicable, the other is H).

[0161] Preferably, R 7 is T;R 8 is T.

[0162] In one embodiment, the present invention provides R 1 is F; (With respect to formula (IT), Z 1 , Z 2 , Z 3 , and Z 4 is as defined above; with respect to formula (I*-T), R 6 is H); R 2 but

[0163] [ka]

[0164] (wherein X is N and R 5 is tritiated -CH3(CT3)

[0165] The tritium ( 3 Preferred detectably labeled compounds in H are

[0166] [ka]

[0167] wherein T is 3 (meaning H).

[0168] More preferably, the tritium (I) of the formula (IT) or (I*-T) according to the present invention is 3 H) the detectably labeled compound is

[0169] [ka]

[0170] and the like, wherein T is 3 (meaning H).

[0171] More preferably, the tritium ( 3 H) The detectably labeled compound is a stereoisomer.

[0172] [ka]

[0173] (Wherein, T is 3 H).

[0174] In one embodiment, the present invention provides a compound of formula (IF) 18 A compound detectably labeled with F;

[0175] [ka]

[0176] or a stereoisomer, polymorph, racemic mixture, tautomer, pharma- ceutically acceptable salt, prodrug, hydrate, or solvate thereof, or a mixture thereof; 1 , Z 2 , Z 3 , Z 4 , R 1 , and R 2 is as defined herein for compounds of formula (I), and at least one F is 18 It's F.

[0177] In one embodiment, the present invention relates to a compound of formula (I*-F)

[0178] [ka]

[0179] (where n, R 1 , R 2 , and R 6 is as defined herein for compounds of formula (I*) 18The present invention provides a compound detectably labeled with F, or a stereoisomer, polymorph, racemic mixture, tautomer, pharma- ceutically acceptable salt, prodrug, hydrate, or solvate thereof, or a mixture thereof.

[0180] In a preferred embodiment, the present invention provides a 1 but 18 The compound of formula (IF) wherein F is a detectable label.

[0181] In a further preferred embodiment, n is 1 or 2; R 1 teeth 18 F (detectable label); R 2 teeth

[0182] [ka]

[0183] (wherein X is N and R 5 is CH3 or H; or R 2 teeth

[0184] [ka]

[0185] (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 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, or R 3 is H and R 4 is NH2 and R 7 is H and R 8 is F).

[0186] In a preferred embodiment, the present invention comprises: n is 1 or 2; R 1 but 18 F (detectable label); R 2 but

[0187] [ka]

[0188] (wherein X is N and R 5 is CH3; R 6 is H).

[0189] In a preferred embodiment, n is 1. In another preferred embodiment, n is 2.

[0190] The compound of the present invention, 18 Preferred compounds detectably labeled with F are

[0191] [ka]

[0192] can be selected from:

[0193] More preferably, the compound of formula (IF) 18 Compounds detectably labeled with F are

[0194] [ka]

[0195] It is.

[0196] More preferably, the compound according to the invention is of formula (IF) 18 Compounds detectably labeled with F may be used in the detection of stereoisomers.

[0197] [ka]

[0198] It is possible.

[0199] 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.

[0200] The compounds of the present invention are particularly suitable for imaging TDP-43 aggregates. Imaging can be performed in mammals, particularly humans. Imaging is preferably in vitro imaging, ex vivo imaging, or in vivo imaging. More preferably, imaging is in vivo imaging. Even more preferably, imaging is brain imaging. Imaging can be eye / retina imaging, or central nervous system tissue imaging.

[0201] The compounds of the present invention are particularly suitable for use in diagnostic methods.The diagnostic method can be carried out on mammals, particularly humans.The tissue of interest on which the diagnostic method is carried out can be the brain, tissue of the central nervous system, tissue of the eye (e.g. retinal tissue), or other tissues or body fluids such as cerebrospinal fluid (CSF).The tissue is preferably brain tissue.

[0202] In the present invention, a "diagnostic composition" is defined as a composition comprising one or more compounds of the present invention in a form suitable for administration to a patient (e.g., a mammal such as a human) and suitable for use in diagnosing a particular disease, disorder, or abnormality in a tissue. In one embodiment, the diagnostic composition comprises a detectably labeled compound of the present invention as described above, and optionally at least one physiologically acceptable carrier, diluent, adjuvant, and / or excipient.

[0203] Preferred detectably labeled compounds of the present invention are compounds of formula (IT) or (IF).

[0204] The diagnostic composition is suitable for use in the diagnosis of a disease, disorder, or disorder associated with TDP-43 aggregates or a TDP-43 proteinopathy, as defined below. Preferably, the diagnostic composition further comprises a physiologically acceptable excipient, carrier, diluent, or adjuvant. Administration is preferably as defined below. More preferably, it is performed by injection of the composition as an aqueous solution. The diagnostic composition may optionally comprise further components such as a buffer; a pharma-ceutically acceptable solubilizer (e.g., cyclodextrin, or a surfactant such as Pluronic, Tween, or phospholipid); and a pharma-ceutically acceptable stabilizer or antioxidant (e.g., ascorbic acid, gentisic acid, or p-aminobenzoic acid). The dose of the compound of the present invention will vary depending on the exact compound administered, the patient's weight, and other variables that will be apparent to a physician skilled in the art.

[0205] Although the compounds of the invention can be administered alone, they are preferably formulated as diagnostic compositions in accordance with standard pharmaceutical practice. Thus, diagnostic compositions comprising a diagnostically effective amount of a compound of the invention in combination with a pharma- ceutically acceptable carrier, diluent, adjuvant, and / or excipient are part of the invention. Preferred pharma- ceutically acceptable carriers, diluents, adjuvants, and / or excipients are those that are physiologically compatible with the diagnostic compositions of the invention.

[0206] Pharmaceutically acceptable excipients are well known in the pharmaceutical art, see, for example, Remington's Pharmaceutical Sciences, 18 th Ed. (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.

[0207] 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, monohydric alcohols such as ethanol, isopropanol, and the like, polyhydric alcohols such as glycols, edible oils such as soybean oil, coconut oil, olive oil, safflower oil, cottonseed oil, and the like, oily esters such as ethyl oleate, isopropyl myristate, and the like, binders, adjuvants, solubilizers, thickeners, stabilizers, disintegrants, glidants, lubricants, buffers, emulsifiers, wetting agents, suspending agents, sweeteners, colorants, flavors, coating agents, preservatives, antioxidants, processing agents, drug delivery modifiers and drug delivery enhancers, such as calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, glucose, hydroxypropyl-β-cyclodextrin, polyvinylpyrrolidone, low melting point waxes, and ion exchange resins.

[0208] Routes of administration (delivery) of the compounds of the present invention include, but are not limited to, one or more of intravenous, gastrointestinal, intraspinal, intraperitoneal, intramuscular, oral (e.g., as tablets, capsules, or ingestible liquids), topical, mucosal (e.g., as nasal sprays or aerosols for inhalation), intranasal, parenteral (e.g., in 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 of administration (delivery) of the compounds of the present invention is parenteral.

[0209] The compounds of the invention (e.g., detectably labeled compounds, e.g., detectably labeled 3 H or 18 When a compound having 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 using infusion 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 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.

[0210] A physician will typically determine the actual dosage that will be most suitable for an individual patient. 3 H or 18 The dosage of the compound having F) will vary depending on the exact compound administered, the patient's weight, the size and type of sample, and other variables that will be apparent to a physician skilled in the art. In general, the dosage may preferably range from 0.001 μg / kg to 10 μg / kg, preferably 0.01 μg / kg to 1.0 μg / kg. The radiation dose may be, for example, 100 to 600 MBq, more preferably 150 to 450 MBq.

[0211] 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 disorders associated with TDP-43 aggregates. The compounds of the invention are particularly suitable for positron emission tomography imaging of TDP-43 aggregates.

[0212] The compounds of the invention disclosed herein are useful in treating diseases, disorders, or disorders associated with TDP-43 aggregates, or TDP-43 proteinopathies, such as 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, chromosome 9p-linked, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) (frontotemporal dementia T Frontotemporal lobar degeneration with DP-43 or ubiquitin-positive TDP-43 inclusions (FTLD-TDP)), argyrophilic grain disease, 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. sporadic ALS, those with TARDBP mutations, those with angiogenin (ANG) mutations), Alexander disease (AxD), limbic-predominant senile 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 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), and 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), It is particularly suitable for use in the diagnosis of a disease, disorder, or abnormality selected from, but not limited to, myopathy, traumatic brain injury (TBI), dementia with Lewy bodies (DLB), and Parkinson's disease (PD), preferably the disease, disorder, or abnormality associated with TDP-43 aggregates or 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 late-life TDP-43 encephalopathy (LATE).

[0213] In one embodiment, the TDP-43 aggregate-associated disease, disorder, or condition, or TDP-43 proteinopathy, is amyotrophic lateral sclerosis (ALS).

[0214] In one embodiment, the diagnosis of a disease, disorder, or abnormality associated with TDP-43 aggregates or a TDP-43 proteinopathy is Alzheimer's disease (AD).

[0215] 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 lobar dementia TDP-43 or frontotemporal lobar degeneration with TDP-43 inclusions (FTLD-TDP).

[0216] In one embodiment, the diagnosis of a disease, disorder, or condition associated with TDP-43 aggregates or TDP-43 proteinopathy is limbic-predominant late-life TDP-43 encephalopathy (LATE).

[0217] Methods and Uses In a third aspect, the present invention relates to the methods and uses listed below: A method of 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 the detection and optionally quantification of TDP-43 aggregates in tissue of a subject; · A method for diagnostic imaging of a subject's brain; 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; Methods of collecting data for the diagnosis of a disease, disorder, or abnormality associated with TDP-43 aggregates or for the diagnosis of a TDP-43 proteinopathy: · A method for collecting data to determine a disease, disorder, or abnormality associated with TDP-43 aggregates, or a predisposition to a TDP-43 proteinopathy; A method for collecting data to monitor the progression of a disease, disorder, or abnormality associated with TDP-43 aggregates or to monitor the progression of a TDP-43 proteinopathy in a patient; A method of collecting data to predict the responsiveness of a patient suffering from a disease, disorder, or condition associated with TDP-43 aggregates to a pharmaceutical treatment; Use of the compounds of the invention as biomarkers for TDP-43 aggregates or for TDP-43 proteinopathies, the use of the compounds of the invention as diagnostic agents or tools for TDP-43 proteinopathies, Use of the compounds of the invention as in vitro analytical standards or in vitro screening tools Regarding.

[0218] Any compound of the invention (e.g., a compound of formula (I), (IT), or (IF)) can be used in the methods summarized above. Preferably, the compound is a detectably labeled compound (e.g., a detectably labeled 3 H or 18 F).

[0219] The methods of the invention may include the step of contacting a sample, a particular body part or area suspected of containing TDP-43 aggregates with a compound of the invention.

[0220] The body is preferably a mammalian body, more preferably a human body, and includes the whole body or partial body regions / parts of a patient suspected of containing TDP-43 aggregates.

[0221] The sample may be selected from tissues or body fluids suspected of containing TDP-43 aggregates, which are samples 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 may be obtained from a mammal, more preferably a human. Preferably, the sample is an in vitro sample derived from a patient.

[0222] An in vitro sample or a particular body part or area obtained from a patient can be contacted with a compound of the invention by direct incubation.

[0223] In the in vivo method, a specific body part or region can be contacted with a compound of the present invention by administering an effective amount of the compound of the present invention to a patient, the effective amount being an amount suitable to allow the presence or absence of TDP-43 aggregates in the specific body part or region to be determined using a selected analytical technique.

[0224] The step of binding the compound of the invention to the TDP-43 aggregates includes allowing sufficient time for this 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 skill in the art by routine experimentation. For in vitro methods, the amount of time depends on the sample or the particular body part or body area, 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 compound of the invention to reach the particular body part or body area suspected of containing TDP-43 aggregates. The amount of time should not be too long to avoid washout and / or metabolism of the compound of the invention. The period can range, for example, from about 0 minutes to about 240 minutes (which is the duration of the PET scan during initial compound characterization (NHP PET and later FiH testing)).

[0225] The method of detecting the compound of the present invention bound to TDP-43 aggregates is not particularly limited, and depends in particular on the detectable level, the type of sample, the specific body part or body area, 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 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 in a sample or in the body. The imaging system is a system that shows an image of the detectably bound label, such as a radioisotope, especially a positron emitter or gamma emitter, as present in the tested sample, the tested specific body part, or the tested body area. Preferably, the compound of the present invention bound to TDP-43 aggregates is detected by an imaging device, such as a PET or SPECT spacer. The amount of compound bound to TDP-43 can be determined by visual or quantitative analysis, for example using PET scan images.

[0226] In one embodiment, the presence or absence of a compound of the invention bound to TDP-43 aggregates in a sample or in a particular body part or region can be correlated with the presence or absence of TDP-43 aggregates. The correlation can be qualitative or quantitative. In a preferred embodiment, this step comprises the following steps: - quantifying the amount of the compound of the invention 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 region; and - an optional step of comparing the amount of compound bound to TDP-43 aggregates in the sample or in a specific body part or region with a normal control value in a healthy control subject. Includes.

[0227] The amount of compound bound to TDP-43 aggregates can be determined by any suitable method, with a preferred method being positron emission tomography (PET).

[0228] In another embodiment, the presence or absence of a compound of the 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. The correlation can be qualitative or quantitative. In a preferred embodiment, this step comprises the following steps: - quantifying the amount of the compound of the invention 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 region; and - an optional step of comparing the amount of compound bound to TDP-43 aggregates in the sample or in a specific body part or region with a normal control value in a healthy control subject. Includes.

[0229] In any of the methods disclosed herein, steps (a)-(c) and, if present, optional step (d) can be repeated at least once. Repeating the steps is particularly useful in methods that collect data to monitor progression and to predict responsiveness. In these methods, it may be convenient to monitor the patient over time and repeat the steps after a certain period of time. The time interval before repeating the steps can be determined by the physician depending on the severity of the disease, disorder, or abnormality associated with TDP-43 aggregates or TDP-43 proteinopathy.

[0230] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease 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 the TDP-43 aggregates; and (c) detecting a compound bound to the TDP-43 aggregates. The present invention relates to a method for detecting a neurological disease, neurological disorder, or neurological abnormality associated with TDP-43 aggregates in a subject, comprising:

[0231] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease 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 disclosed herein; (b) binding the compound to the TDP-43 aggregates; and (c) detecting and optionally quantifying the compound bound to the TDP-43 aggregates using positron emission tomography. The present invention relates to a method (e.g., an in vivo or in vitro method) for detecting and optionally quantifying TDP-43 aggregates in a tissue of a subject, comprising:

[0232] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease 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 the 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 for diagnostic imaging of the brain of a subject, comprising:

[0233] 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 described above, in particular by PET.

[0234] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease 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 the TDP-43 aggregates; and (c) detecting a compound bound to the TDP-43 aggregates. The present invention relates to a method for imaging TDP-43 aggregates in a sample or a patient, in particular in the brain or in a sample taken from the brain of a patient, comprising:

[0235] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease 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 the TDP-43 aggregates; and (c) detecting the compound bound to the TDP-43 aggregates in the brain of the subject. The present invention relates to a method for imaging a disease, disorder, or abnormality associated with TDP-43 aggregates in a subject, comprising:

[0236] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease 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 the TDP-43 aggregates; and (c) detecting a compound bound to the TDP-43 aggregates. The present invention relates to a method for imaging a disease, disorder, or abnormality associated with TDP-43 aggregates in a subject, comprising:

[0237] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease 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 the TDP-43 aggregates; (c) detecting a compound bound to the TDP-43 aggregate; and (d) generating an image representing the location and / or amount of compound bound to the TDP-43 aggregates. The present invention relates to a method for imaging a disease, disorder, or abnormality associated with TDP-43 aggregates in a subject, comprising:

[0238] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease 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 the TDP-43 aggregates; and (c) detecting the compound bound to the TDP-43 aggregates by collecting positron emission tomography (PET) images of the tissue of the subject. The present invention relates to a method for positron emission tomography (PET) imaging of TDP-43 aggregates in tissue of a subject, comprising:

[0239] Preferably, the tissue is central nervous system (CNS) tissue, eye tissue, or brain tissue. More preferably, the tissue is brain tissue.

[0240] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease comprising the steps of: (a) contacting a sample, a specific body part or a body area suspected of containing TDP-43 aggregates with a compound of the invention, or a diagnostic composition comprising a compound of the invention, as disclosed herein; (b) binding the compound to the TDP-43 aggregates; and (c) detecting compounds bound 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 for imaging TDP-43 aggregates in a sample or a patient, comprising:

[0241] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease comprising the steps of: (a) contacting an in vitro sample suspected of containing TDP-43 aggregates with a compound of the invention, or a diagnostic composition comprising a compound of the invention, as disclosed herein; (b) binding the compound to the 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 for imaging TDP-43 aggregates in an in vitro patient sample, comprising:

[0242] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease comprising the steps of: (a) contacting a sample, or a specific body part or region, suspected of containing TDP-43 aggregates, with a compound of the invention, preferably a compound of formula (IT) or formula (IF), as disclosed herein; or with a diagnostic composition comprising a compound of the invention, preferably a compound of formula (IT) or formula (IF); (b) binding the compound to the TDP-43 aggregates; and (c) detecting the compound bound to the 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 for imaging TDP-43 aggregates in a patient or in a specific body part or region of a patient, comprising:

[0243] The step of imaging the sample, the patient, or a particular body part or region of the patient with an imaging system includes detecting the compound of the present invention bound to the TDP-43 aggregates using the imaging system disclosed herein. Detecting the compound of the present invention bound to the TDP-43 aggregates by imaging allows for identifying the distribution of TDP-43 aggregates in the sample, the patient, or a particular body part or region tested. PET imaging should be performed when the compound has entered the tissue and the compound has bound to the TDP-43 aggregates.

[0244] Determine 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, a particular body part or a body region suspected of containing TDP-43 aggregates using the compounds of the present invention.

[0245] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease comprising the steps of: (a) contacting a sample, a specific body part or a body area suspected of containing TDP-43 aggregates with a compound of the invention, or a diagnostic composition comprising a compound of the invention, as disclosed herein; (b) binding a compound of the invention to TDP-43 aggregates; (c) detecting a compound of the invention bound to TDP-43 aggregates; (d) quantifying the amount of the compound of the invention bound to the TDP-43 aggregates; and (e) An optional step of calculating the amount of TDP-43 aggregates in a sample, a particular body part or a body region. The present invention provides a method for determining the amount of TDP-43 aggregates in a sample, a specific body part or a body region suspected of containing TDP-43 aggregates, comprising:

[0246] At least one radiolabeled atom (e.g. 3 H, 2 H, or 18 When a detectably labeled compound of the invention, including F), binds to TDP-43 aggregates, a radioactive signal is observed.

[0247] Diagnose: In one embodiment, the present invention provides a method for producing a method for the treatment of a disease comprising the steps of: (a) detecting a compound of the invention bound to TDP-43 aggregates; and (b) correlating the presence or absence of a compound of the invention bound to TDP-43 aggregates with a disease, disorder, or condition associated with TDP-43 aggregates, or a TDP-43 proteinopathy. The present invention relates to a method for diagnosing a disease, disorder, or abnormality associated with TDP-43 aggregates, or a TDP-43 proteinopathy, comprising:

[0248] Preferably, the method for diagnosing a disease, disorder or abnormality associated with TDP-43 aggregates or a TDP-43 proteinopathy comprises the steps of: (a) contacting a sample, a specific body part or a body area suspected of containing TDP-43 aggregates with a compound of the invention, or a diagnostic composition comprising a compound of the invention, as disclosed herein; (b) binding a compound of the invention to TDP-43 aggregates; (c) detecting a compound of the invention bound to TDP-43 aggregates; and (d) correlating the presence or absence of a compound of the invention bound to TDP-43 aggregates with a disease, disorder, or condition associated with TDP-43 aggregates, or a TDP-43 proteinopathy. Includes.

[0249] In one embodiment, the present invention provides a method for producing a method for the treatment of a disease comprising the steps of: (a) contacting a sample or a specific body part or region suspected of containing TDP-43 aggregates with a compound of the invention, or a diagnostic composition comprising a compound of the invention, as disclosed herein; (b) binding a compound of the invention to TDP-43 aggregates; (c) detecting a compound of the invention bound to TDP-43 aggregates; and (d) an optional step of 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 a sample or in a particular body part or region. The present invention relates to a method for collecting data for the diagnosis of a disease, disorder, or abnormality associated with TDP-43 aggregates, or a TDP-43 proteinopathy, comprising:

[0250] After contacting the sample or a particular body part or region with the compound of the present invention, the compound binds to the 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 one of ordinary skill in the art through routine experimentation. The compound bound to the TDP-43 aggregates can then be detected by any suitable method. The particular method selected will depend on the detectable level selected. Examples of possible 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 magnetic resonance imaging (MRI). Fluorescent and / or nuclear imaging techniques can be used to monitor and / or visualize the distribution of the detectably labeled compound within the sample or a particular body part or region.

[0251] The optional 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 in a specific body part or body region, as mentioned above, comprises the following steps: - quantifying the amount of 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 region; and - an optional step of comparing the amount of compound bound to TDP-43 aggregates in the sample or in a specific body part or region with a normal control value in a healthy control subject. Includes.

[0252] The amount of compound bound to TDP-43 aggregates can be compared to normal control values ​​established in samples from healthy subjects or in specific body parts or regions, 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.

[0253] If the amount of compound bound to TDP-43 aggregates is greater than the normal control value defined herein, it can be predicted that the patient is afflicted with or has a high probability of being afflicted with a TDP-43 aggregate-associated disease, disorder, or condition, or a TDP-43 proteinopathy.

[0254] Determine predisposing factors: A further aspect of the invention relates to a method of collecting data to determine a predisposition to a disease, disorder or condition associated with TDP-43 aggregates or a TDP-43 proteinopathy, the method comprising the steps of: (a) contacting a sample or a specific body part or region suspected of containing TDP-43 aggregates with a compound of the invention, or a diagnostic composition comprising a compound of the invention, as disclosed herein; (b) binding a compound of the invention to TDP-43 aggregates; (c) detecting a compound of the invention bound to TDP-43 aggregates; and (d) an optional step of 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 a sample or in a particular body part or region. Includes.

[0255] The optional 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 in a specific body part or body region, as mentioned above, comprises the following steps: - quantifying the amount of 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 region; and - an optional step of comparing the amount of compound bound to TDP-43 aggregates in the sample or in a specific body part or region with a normal control value in a healthy control subject. Includes.

[0256] If the amount of compound bound to TDP-43 aggregates is higher than the normal control value of a healthy subject / reference subject, this indicates that the patient / subject is suffering from or at risk of developing a TDP-43 aggregate-associated disease, disorder, or condition. In particular, if the amount of compound bound to TDP-43 aggregates is higher than would be expected in an individual who does not show clinical evidence of a neurodegenerative disease, the patient may be considered to have a predisposition to a TDP-43 aggregate-associated disease, disorder, or condition, or a TDP-43 proteinopathy.

[0257] Monitor disease progression: In one embodiment, the present invention relates to a method for monitoring the progression of a disease, disorder or abnormality associated with TDP-43 aggregates or a TDP-43 proteinopathy in a patient. Typically, the patient is undergoing or has been treated for a disease, disorder or abnormality associated with TDP-43 aggregates or a TDP-43 proteinopathy. In particular, the treatment may include administration of an anti-TDP-43 drug.

[0258] A method for collecting data to monitor the progression of a disease, disorder, or abnormality associated with TDP-43 aggregates or to monitor the progression of a TDP-43 proteinopathy in a patient, comprising the steps of: (a) contacting a sample, specific body part or body area suspected of containing TDP-43 aggregates with a compound of the present invention; (b) binding a compound of the invention to TDP-43 aggregates; (c) detecting a compound of the invention bound to TDP-43 aggregates; (d) an optional step of 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 a sample or in a particular body part or region; and (e) the optional step of repeating steps (a) to (c) and, if present, optional step (d), at least once. Includes.

[0259] Steps (a)-(c), and optional step (d), if present, can 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, these steps should be repeated until no further progression of the disease is observed in the patient.

[0260] The optional 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 in a specific body part or body region, as mentioned above, comprises the following steps: - quantifying the amount of 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 region; and - an optional step of comparing the amount of compound bound to TDP-43 aggregates in the sample or in a specific body part or region with a normal control value in a healthy control subject. Includes.

[0261] In methods of monitoring progression over time, the amount of the compound of the invention bound to TDP-43 aggregates may optionally be compared at various time points during treatment, such as 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 the compound of the invention bound to TDP-43 aggregates may indicate that the disease is not progressing.

[0262] Predict responsiveness: In one embodiment, the present invention relates to a method for predicting the responsiveness of a patient suffering from a TDP-43 aggregate-associated disease, disorder or disorder or suffering from a TDP-43 proteinopathy to treatment of said TDP-43 aggregate-associated disease, disorder or disorder or TDP-43 proteinopathy.

[0263] The method can be used to predict the most suitable treatment for a patient. In particular, the treatment can include administration of an anti-TDP-43 drug.

[0264] A method for predicting the responsiveness of a patient suffering from a TDP-43 aggregate-associated disease, disorder, or disorder, or suffering from a TDP-43 proteinopathy, to a treatment for said TDP-43 aggregate-associated disease, disorder, or disorder, or TDP-43 proteinopathy, comprising the steps of: (a) contacting a sample, a specific body part or a body area suspected of containing TDP-43 aggregates with a compound of the invention, or a diagnostic composition comprising a compound of the invention, as disclosed herein; (b) binding a compound of the invention to TDP-43 aggregates; (c) detecting a compound of the invention bound to TDP-43 aggregates; (d) an optional step of 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 a sample or in a particular body part or region; and (e) the optional step of repeating steps (a) to (c) and, if present, optional step (d), at least once. may include.

[0265] Typically, the patient is undergoing / has undergone treatment for a disease, disorder, or condition associated with TDP-43 aggregates or a TDP-43 proteinopathy. In particular, the treatment may include the administration of a medicament suitable for treating the disease, disorder, or condition associated with TDP-43 aggregates.

[0266] This method allows prediction of a patient's responsiveness to a particular treatment. In one embodiment, responsiveness can be estimated, for example, by repeating steps (a)-(c) and optional step (d), if present, and monitoring the amount of the compound of the present invention bound to the TDP-43 aggregates over a period of time while the patient is undergoing treatment for a disease, disorder, or condition associated with TDP-43 aggregates or a TDP-43 proteinopathy. If the amount of the compound of the present invention bound to the TDP-43 aggregates decreases over time, the patient may be considered to be responsive to the treatment. If the amount of the compound bound to the TDP-43 aggregates remains essentially constant or increases over time, the patient may be considered to be non-responsive to the treatment.

[0267] Alternatively, responsiveness can be estimated by determining the amount of the compound of the invention bound to the TDP-43 aggregates. The amount of the compound bound to the 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 may refer to a control value of a healthy control subject. Alternatively, the control value may refer to a control value of a subject known to be responsive to a particular treatment, or the control value may refer to a control value of a subject known to be non-responsive to a particular treatment. The result regarding responsiveness can be "responsive" to a particular treatment, "non-responsive" to a particular treatment, or "undetermined responsiveness" to a particular treatment. Response to treatment can be different for each patient.

[0268] The optional 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 in a specific body part or body region, as mentioned above, comprises the following steps: - quantifying the amount of 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 region; and - an optional step of comparing the amount of compound bound to TDP-43 aggregates in the sample or in a specific body part or region with a normal control value in a healthy control subject. Includes.

[0269] A 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 show clinical evidence of a neurodegenerative disease.

[0270] In any of the methods summarized above, if the amount of compound bound to TDP-43 aggregates is greater than the normal control value, it can be predicted that the patient is suffering from or has a high probability of suffering from a disease, disorder, or condition associated with TDP-43 aggregates, or a TDP-43 proteinopathy.

[0271] Any compound of the present invention can be used in the methods summarized above. Preferably, the detectably labeled compounds of the present invention disclosed herein can be used in the methods summarized above.

[0272] In a fourth aspect, the present invention relates to the use of the compounds of the present invention as diagnostic agents or diagnostic 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 these cases, the compounds of the present invention may be detectably labeled compounds or may contain non-radioactive isotopes.

[0273] In another embodiment, the present invention further relates to the use of the compounds of the present invention as defined herein, more particularly the detectably labeled compounds of the present invention, as diagnostic biomarkers allowing more efficient and accurate patient selection, for example for long-term monitoring in clinical trials or to aid in the development of new therapeutics for the treatment of TDP-43 proteinopathies. In another embodiment, the present invention further relates to the use of the compounds of the present invention as defined herein, more particularly the detectably labeled compounds of the present invention, as biomarkers for TDP-43 aggregates or for TDP-43 proteinopathies.

[0274] In another embodiment, the compounds of the present invention can be used for research, in particular as analytical tools or reference molecules. The compounds can also be used to detect TDP-43 aggregates in vitro or in vivo. 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 described herein.

[0275] Kit of Parts In a fifth aspect, the present invention further relates to a kit for use in one or more methods of the present invention, comprising a compound of the present invention as described herein. Typically, the kit comprises a container holding the compound of the present invention and instructions for using 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)).

[0276] 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.

[0277] The dose of the detectably labeled compound 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 that will be apparent to a physician skilled in the art. In general, the dose may preferably range from 0.001 μg / kg to 10 μg / kg, preferably 0.01 μg / kg to 1.0 μg / kg. The radiation dose may be, for example, 100 to 600 MBq, more preferably 150 to 450 MBq.

[0278] In particular, these kits may be useful for carrying out the methods of the present invention (e.g., but not limited to, imaging, diagnostic, and monitoring methods) for diagnosing, for example, diseases, disorders, or disorders associated with TDP-43 aggregates, or TDP-43 proteinopathies. These kits may include all components necessary to carry out the methods provided herein. Typically, each component is stored separately in one full package. Suitable additional components to be included in the kit include, for example, buffers, detectable dyes, laboratory equipment, reaction vessels, instructions, etc. The instructions can be modified to suit the particular method in which the kit is to be used.

[0279] The present invention further relates to a kit for the preparation of the detectably labeled compounds of the present invention, in particular where the detectable label is a radioisotope. The kit thus comprises a precursor of the detectably labeled compound of formula (I) and a labeling agent which reacts with the precursor to introduce a detectable (e.g. radioactive) label. Preferred precursors include compounds of formula (II), (III) and (IV). The labeling agent which reacts with the precursor is 18 F or 3 The labeling agent can be an agent that introduces a detectable (e.g., radioactive) label, such as H. 18 F-can be a fluorinating agent.

[0280] Methods for preparing the compounds of the present invention In a sixth aspect, the present invention further relates to a process for preparing a compound of formula (I).

[0281] Non-radioactive isotopic compounds: In one embodiment, the present invention provides a method for producing a method for the treatment of a disease comprising the steps of: Compound of formula (II) and R 9 and reacting with a compound of formula (I):

[0282] [ka]

[0283] (In the formula, n, R 1 , R 2 , Z 1 , Z 2 , Z 3 , and Z 4 is as defined above; R 9 teeth a 5- or 6-membered carbocyclic ring optionally substituted with F, NH2, CN, and / or CH3; a 5- or 6-membered heterocycloalkyl ring, optionally substituted with F, NH2, CN, and / or CH3, containing one or more heteroatoms selected from N, O, and S; 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 one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S; wherein the 5- or 6-membered carbocyclic ring, the 5- or 6-membered heterocycloalkyl ring, the 5-membered heteroaryl ring, or the 6-membered heteroaryl ring is substituted with Br or I. The present invention relates to a process for preparing a compound of formula (I) above, comprising:

[0284] In a preferred embodiment, the compound having formula (II) has R 9 together with a compound of formula (IC), R 9 teeth

[0285] [ka]

[0286] (wherein X is N and R 5 is CH3 or H; Hal is Br or I, preferably Hal is Br; Or, R 9 teeth

[0287] [ka]

[0288] (wherein Hal is Br or I, preferably Hal is Br; R 3 is F and R 4 is NH2 and R 7 is H and R 8 is H; R 3is 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, or R 3 is H and R 4 is NH2 and R 7 is H and R 8 is F).

[0289] In one embodiment, the present invention relates to a process for preparing a compound of formula (I*) above, comprising the steps of: Formula (II*)

[0290] [ka]

[0291] (In the formula, n is 1 or 2; R 1 is H or F; R 6 is H; R 9 is a 5- or 6-membered carbocyclic ring or a 5- or 6-membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S, which may be optionally substituted with F, NH2, and / or CH3 and which is substituted with Br or I, and 9 By reacting with 2 obtaining a compound of formula (I*), The present invention relates to a method comprising the steps of:

[0292] A preferred embodiment of the method comprises: Formula (II)

[0293] [ka]

[0294] (In the formula, n is 1 or 2; R 1 is H or F; R 6 is H; R 9 but

[0295] [ka]

[0296] (wherein X is N and R 5 is CH3; Hal is Br or I, preferably Hal is Br; Or, R 9 but

[0297] [ka]

[0298] (In the formula, R 3 is F;R 4 is -NH2; R 7 is H;R 8 is H; Hal is Br or I, preferably Hal is Br; R 9 and reacting with the compound of formula (IC) (In the formula, n is 1 or 2; R 1 is H or F; R 6 is H; R 2 teeth

[0299] [ka]

[0300] (In the formula, R 3 is F and R 4 is -NH2, R 7 is H and R 8 is H); Or, R 2 teeth

[0301] [ka]

[0302] (wherein X is N and R 5 is CH3) A process for obtaining a compound of Includes.

[0303] A compound having the formula (II) and R 9 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.

[0304] In one embodiment, the present invention provides Preparation 1, a compound of formula (II) and R 9

[0305] [ka]

[0306] (wherein X is N and R 5 is CH3) with

[0307] [ka]

[0308] In another embodiment, the present invention relates to Preparation 2, a compound of formula (II) and R 9

[0309] [ka]

[0310] (wherein X is N and R 5 is CH3) with

[0311] [ka]

[0312] In another embodiment, the present invention relates to Preparation 3, a compound of formula (II) and R 9

[0313] [ka]

[0314] (In the formula, R 3 is F and R 4 is -NH2, R 7 is H and R 8 is H).

[0315] [ka]

[0316] Preferably, the method relates to a compound of the invention having formula (I) and a compound having formula (II) that does not have a detectable label.

[0317] Tritium (3 H) a compound detectably labeled with In one embodiment, the present invention relates to a compound having formula (I) in which at least one leaving group of the precursor of the compound is tritium ( 3 The present invention relates to a compound having formula (I), which comprises a step of radiolabeling a precursor of the compound having formula (I) with a radioisotope, the precursor being replaced with tritium (H). 3 H) provides a method for preparing a detectably labeled compound.

[0318] At least 1 to 3 hydrogen (H) are tritium ( 3 H), having the formula (I) 3 3H) is preferably defined as a compound detectably labeled with tritium ( 3 H), having the formula (I) 3 3H) is more preferably defined as a compound detectably labeled with tritium ( 3 H), having the formula (I) 3 H) detectably labeled compounds are further preferably defined.

[0319] In another embodiment, the present invention provides a process for preparing a compound of formula (I): Formula (III);

[0320] [ka]

[0321] (In the formula, Z 1 , Z 2 , Z 3 , and Z 4 are each selected from C-Br, CI, CH, and N, Z 1 If N, then Z 2 is C-Br, CI, or CH; Z 2 If N, then Z1 is C-Br, CI, or CH; Z 3 If N, then Z 4 is C-Br, CI, or CH; Z 4 If N, then Z 3 is C-Br, CI, or CH; n is 1 or 2; R 1 is H or F; R 10 teeth a 5- or 6-membered carbocyclic ring optionally substituted with Br, I, F, NH2, CN, and / or CH3; a 5- or 6-membered heterocycloalkyl ring optionally substituted with Br, I, F, NH2, CN, and / or CH3 and containing one or more heteroatoms selected from N, O, and S; a 5-membered heteroaryl ring optionally substituted with Br, I, 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 Br, I, F, NH2, CN, and / or CH3 and containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S; Z 1 , Z 2 , Z 3 , or Z 4 at least one of is selected from C-Br or CI; and / or R 10 contains Br or I, wherein at least one Br or I is replaced by CT or T; T is 3 H), or a stereoisomer, polymorph, racemic mixture, tautomer, pharma- ceutically acceptable salt, prodrug, hydrate, or solvate thereof, or a mixture thereof, is referred to as T (i.e. 3 H) radiolabeling step The present invention provides a method comprising:

[0322] In one embodiment, Br or I in C-Br or CI is replaced by T or CT3, preferably T.

[0323] In a preferred embodiment, Z 3 is C-Br or CI, where Br or I is replaced by T or CT3, preferably T.

[0324] In a further embodiment, R 10 contains Br or I, and Br or I is replaced by T or CT3.

[0325] In another embodiment, R 10 contains Br or I, and Br or I is replaced by T or CT. For example, R 10 teeth

[0326] [ka]

[0327] It is possible.

[0328] In another embodiment, the present invention provides a process for preparing a compound of formula (I*) comprising: Formula (III*):

[0329] [ka]

[0330] (In the formula, n is 1 or 2; R 1 is H or F; R 6 is Br or H; R 10 is a 5- or 6-membered carbocyclic ring or a 5- or 6-membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S, optionally substituted with F, Br, NH2, and / or CH3; wherein at least one proton of the 5- or 6-membered carbocyclic or heterocyclic ring is replaced by CT3; or At least one Br group is 3 H) to form a precursor compound having the formula 3 Radiolabeling with H The present invention provides a method comprising:

[0331] In another embodiment, the present invention provides a process for preparing a compound of formula (I*) comprising: Formula (IIIa*):

[0332] [ka]

[0333] (In the formula, n is 1 or 2; R 1 is H or F; R 6 is H; R 10 is a 5- or 6-membered carbocyclic ring or a 5- or 6-membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S, optionally substituted with F, NH2, and / or CH3, wherein at least one proton of the 5- or 6-membered carbocyclic or heterocyclic ring is replaced with CT3. 3 Radiolabeling with H The present invention provides a method comprising:

[0334] Preferably, the compound of formula (IIIa) contains one CT3 group.

[0335] In another embodiment, the present invention provides a process for preparing a compound of formula (I): Formula (IIIb*):

[0336] [ka]

[0337] (In the formula, n is 1 or 2; R 1 is H or F; R 6 is H or Br; R 10 is a 5- or 6-membered carbocyclic ring or a 5- or 6-membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S, optionally substituted with F, Br, NH2, and / or CH3; substituted with at least one Br; At least one Br group is 3 H) to form a precursor compound having the formula 3 Radiolabeling with H The present invention provides a method comprising:

[0338] Preferably, the compound of formula (IIIb*) comprises 1 to 3 3 H, and 16 3 H, and 21 3 More preferably, the compound of formula (IIIb) contains 2 or 3 or 16 H. 3 More preferably, the compound of formula (IIIb*) contains 16 3 Contains H.

[0339] In one embodiment, the present invention provides a process for preparing a compound of formula (I*-T): Formula (III*)

[0340] [ka]

[0341] (In the formula, n is 1 or 2; R 1 is H or F; R 10 teeth

[0342] [ka]

[0343] (In the formula, R 3 is F and R 4 is -NH2, R 12 and R 13 at least one of R is Br and, if applicable, the other is H; preferably, R 3 is F and R 4 is -NH2, R 12 is Br and R 13 is Br); R 6 is Br; Or, R 10 teeth

[0344] [ka]

[0345] (wherein X is N and R 11 is H); R 6 is H) 3 By radiolabeling with H, Formula (I*-T):

[0346] [ka]

[0347] (In the formula, n and R 1 is as defined above; T is 3 H; R 2 teeth

[0348] [ka]

[0349] (In the formula, R 3 is F and R 4 is -NH2, R 7 and R8 at least one of R 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 teeth

[0350] [ka]

[0351] (wherein X is N and R 5 is CT3); R 6 is H. The present invention provides a method comprising:

[0352] In another embodiment, the present invention relates to a compound of formula (III*):

[0353] [ka]

[0354] (In the formula, n is 1 or 2; R 1 is H or F; R 10 teeth

[0355] [ka]

[0356] (In the formula, R 3 is F;R 4 is -NH2; R 12 and R 13 at least one of R is Br and, if applicable, the other is H; preferably, R 3 is F and R 4 is -NH2, R 12is Br and R 13 is Br); R 6 is Br; Or, R 10 teeth

[0357] [ka]

[0358] (wherein X is N and R 11 is H); R 6 is H, Here, tritium (I*-T) 3 H) The detectably labeled compound is

[0359] [ka]

[0360] (In the formula, T is 3 H stands for; n is 1 or 2; R 1 is H or F; R 2 teeth

[0361] [ka]

[0362] (In the formula, R 3 is F and R 4 is -NH2, R 7 is T;R 8 is T); R 6 is T; Or, R 2 teeth

[0363] [ka]

[0364] (wherein X is N and R 5 is tritiated -CH3(CT3); R 6 is H) 3 The method comprises the step of radiolabeling a compound having the formula (IT) with H. 3 H) provides a method for preparing a detectably labeled compound.

[0365] Preferably, the tritium (III) compound according to the present invention has the formula (IT) 3 H) the detectably labeled compound is

[0366] [ka]

[0367] wherein T is 3 (meaning H).

[0368] Preferably, the precursor according to the invention has the formula (III)

[0369] [ka]

[0370] can be selected from:

[0371] 3 The methods used to introduce radioisotopes such as 3H are well known in the art, and one method is described below.

[0372] [ka]

[0373] 3The H radiolabeling agent can be tritium gas. The method can be carried out in the presence of a catalyst, such as palladium on carbon (Pd / C) or Lindlar's catalyst, a solvent, such as N,N-dimethylformamide (DMF), and a base, such as N,N-diisopropylethylamine (DIEA).

[0374] [ka]

[0375] Alternatively, the present invention relates to a method for treating 3 The present invention relates to a method of using a CT3 radiolabeled agent, which is H. The CT3 radiolabeled agent is ICT3 ( 3 H) of iodomethane. The process can be carried out in the presence of a solvent such as dimethylformamide (DMF) and a base such as cesium carbonate or sodium hydride.

[0376] Fluorine ( 18 F) A compound detectably labeled with: In one embodiment, the present invention provides a compound of formula (IV):

[0377] [ka]

[0378] (In the formula, n, Z 1 , Z 2 , Z 3 , Z 4 , and R 2 is as specified above, R 14 In the radiolabeling process, 18 is a leaving group that can be replaced by F) The precursor with radioisotope [ 18 The present invention relates to a method for radiolabeling a fluorine-containing compound comprising the step of radiolabeling the compound with fluorine (F). 18 F) A method for preparing a detectably labeled compound is provided.

[0379] In one embodiment, the present invention relates to a compound of formula (IV*):

[0380] [ka]

[0381] (In the formula, n is 1 or 2; R 6 is H; R 2 is a 5- or 6-membered carbocyclic ring or a 5- or 6-membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S, optionally substituted with F, NH2, and / or CH3; R 14 In the radiolabeling process, 18 is a leaving group that can be replaced by F) The precursor with radioisotope [ 18 The present invention relates to a method for radiolabeling a fluorine-containing compound comprising the step of radiolabeling the compound with fluorine (F). 18 F) A method for preparing a detectably labeled compound is provided.

[0382] In another embodiment, the present invention relates to a compound of formula (IV*)

[0383] [ka]

[0384] (In the formula, n is 1 or 2; R 6 is H; R 2 teeth

[0385] [ka]

[0386] (wherein X is N and R 5 is CH3) and; R 14 In the radiolabeling process, 18a precursor compound having a leaving group that can be replaced by F 18 By radiolabeling with F, Formula (I*-F)

[0387] [ka]

[0388] (In the formula, n, R 2 , and R 6 is as specified above, R 1 teeth 18 The present invention provides a method for preparing a compound detectably labeled with fluorine (18F) having the formula (I*-F), comprising the step of obtaining a compound having the formula (I*-F):

[0389] Fluorination is 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 can be carried out in the presence of an F-fluorinating agent.

[0390] Preferably, the leaving group (LG) is C 1~4 Alkyl sulfonate or C 6~10 More preferably, the leaving group (LG) is a mesylate, a tosylate, or a nosylate. Even more preferably, the leaving group (LG) is a mesylate.

[0391] 18 Suitable solvents for the F-fluorination step are known to those skilled in the art. The solvent may be selected, for example, from the group consisting of DMF, DMSO, acetonitrile, DMA, or mixtures thereof. Preferably, the solvent is acetonitrile or DMSO.

[0392] Preferably, fluorine ( 18 A method for preparing a detectably labeled compound 1 with F) is shown below, in which the leaving group (LG), in this case mesylate, of the precursor L1 is reacted with K[ 18 F] or [ 18 F]TBAF etc. 18 F-Fluorine ( 18 F) is replaced by a radiolabeling step.

[0393] [ka]

[0394] The compounds of the invention can be prepared by one of the general methods depicted in the following schemes, which are presented for illustrative purposes only and should not be construed as limiting.

[0395] In a seventh aspect, the present invention provides compounds of formulas (II), (III), and (IV) disclosed below: - Formula (II):

[0396] [ka]

[0397] (In the formula, R 1 , n, Z 1 , Z 2 , Z 3 , and Z 4 is as defined above; - Formula (III):

[0398] [ka]

[0399] (In the formula, R 1 , R 10 , n, Z 1 , Z2 , Z 3 , and Z 4 is as defined above; - Formula (IV):

[0400] [ka]

[0401] (In the formula, R 2 , n, Z 1 , Z 2 , Z 3 , and Z 4 is as specified above, R 14 is a leaving group LG) The present invention relates to a precursor compound of the above.

[0402] In one embodiment of the seventh aspect, the present invention relates to compounds of formula (II*), (III*), and (IV*) disclosed below: - Formula (II*):

[0403] [ka]

[0404] (In the formula, n is 1 or 2; R 1 is H or F; R 6 is H; - Formula (IIIa*):

[0405] [ka]

[0406] (In the formula, n is 1 or 2; R 1 is H or F; R 6 is H; R10 is a 5- or 6-membered carbocyclic ring or a 5- or 6-membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S, optionally substituted with F, NH2, and / or CH3. The present invention relates to a precursor compound of the above.

[0407] Preferably, R 10 teeth

[0408] [ka]

[0409] (wherein X is N and R 11 is H; R 6 is H). - Formula (IIIb*):

[0410] [ka]

[0411] (In the formula, n is 1 or 2; R 1 is H or F; R 6 is Br or H; R 10 is a 5- or 6-membered carbocyclic ring or a 5- or 6-membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S, optionally substituted with F, Br, NH2, and / or CH3; wherein at least one proton of a 5- or 6-membered carbocyclic or heterocyclic ring is replaced with at least one Br.

[0412] Preferably, R 10 teeth

[0413] [ka]

[0414] (In the formula, R 3 is F;R 4 is -NH2; R 12 and R 13 At least one of R is Br and, if applicable, the other is H; more preferably, R 3 is F and R 4 is -NH2, R 12 is Br and R 13 is Br; R 6 is Br). - Formula (IV*):

[0415] [ka]

[0416] (In the formula, n is 1 or 2; R 6 is H; R 2 is a 5- or 6-membered carbocyclic ring or a 5- or 6-membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S, optionally substituted with F, NH2, and / or CH3; R 14 is a leaving group LG).

[0417] Preferably, the leaving group (LG) is C 1~4 Alkyl sulfonate or C 6~10 More preferably, the leaving group (LG) is a mesylate, a tosylate, or a nosylate. Even more preferably, the leaving group (LG) is a mesylate.

[0418] R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 10, R 11 , R 12 , R 13 , R 14 , and n are as described above, including preferred embodiments.

[0419] Precursor compounds having formula (II), (III), (IV), (II*), (III*), or (IV*), or stereoisomers, polymorphs, racemic mixtures, tautomers, pharma- ceutically acceptable salts, prodrugs, hydrates, or solvates thereof, or mixtures of the above, are part of the present invention.

[0420] [Table 1]

[0421] General synthetic scheme: Synthetic scheme for the preparation of (R)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (compound 1)

[0422] [ka]

[0423] Synthetic scheme for the preparation of (R)-2-(6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (compound 2)

[0424] [ka]

[0425] Synthetic scheme for the preparation of (5-(3-amino-4-fluorophenyl)-2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (compound 3)

[0426] [ka]

[0427] [ 3 Synthetic scheme for the preparation of H] precursor compounds (R)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3 H. Synthetic scheme for the preparation of compound 1 (precursor)

[0428] [ka]

[0429] (R)-2-(6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3 H Synthetic scheme for the preparation of compound 2 (precursor)

[0430] [ka]

[0431] 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 H. Synthetic scheme for the preparation of compound 3 (precursor)

[0432] [ka]

[0433] 3H-labeled compounds can be prepared by catalytic dehalogenation with tritium gas from suitable precursor compounds containing halogen atoms (M. Saljoughian Synthesis (2002), 1781-1801) or by the synthesis of methyl iodide [ 3 H] (Y. Chen Chemistry 25 (2019):3405-3439). Preferably, 3 The solvent used in 3H-labeling is DMF or DMA, preferably the solvent is DMF.

[0434] 18 Synthetic scheme for the preparation of precursors for F labeling (S)-1-(5-(5-(1-methyl-1H-pyrazol-4-yl)-4-oxo-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)pyrrolidin-3-yl methanesulfonate( 18 F. Synthetic scheme for the preparation of compound 1 (precursor)

[0435] [ka]

[0436] (S)-1-(5-(5-(1-methyl-1H-pyrazol-4-yl)-4-oxo-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)piperidin-3-yl methanesulfonate( 18 F. Synthetic scheme for the preparation of compound 2 (precursor)

[0437] [ka]

[0438] The reaction is carried out in the presence of a fluorinating agent, and usually a solvent.

[0439] 18F-labeled compounds were synthesized using precursor compounds containing LG. 18 Reaction with F-fluorinating agent, LG 18 It can be prepared by replacing with F. 18 F-fluorination agent 18 Tetraalkylammonium salts of F (e.g. 18 F Tetra (C 1~6 alkyl) ammonium salts, for example 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 18 F, or Kryptofix

[0222] K 18 F. Preferably, 18 F-fluorination 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, 2853-2873; J. Fluorine Chem., 27 (1985):177-191; Coenen, Fluorine-18 Labeling Methods:Features and Possibilities of Basic Reactions, (2006), in: Schubiger PA, Friebe M., Lehmann L., (eds), PET-Chemistry - The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, pp. 15-50). Preferably, 18 The solvent used in the F-fluorination is DMF, DMSO, acetonitrile, DMA, or a mixture thereof, preferably the solvent is acetonitrile or DMSO.

[0440] In the above 18 Although the reactions are shown with F as the radioactive label, other radioactive labels may be introduced following similar procedures.

[0441] This invention is illustrated by the following examples which should not be construed as limiting. EXAMPLES

[0442] All reagents and solvents were obtained from commercial sources and used without further purification. 1 H NMR spectra were recorded on a Bruker DRX-400 MHz NMR spectrometer or a Bruker AV-400 MHz NMR spectrometer in deuterated solvents. Mass spectra (MS) were recorded on an Advion CMS mass spectrometer. Chromatography was performed using silica gel (Fluka: silica gel 60, 0.063-0.2 mm) and suitable 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.

[0443] Although some of the examples do not indicate that each compound is detectably labeled, a 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 will be appreciated that this can be readily prepared using starting materials containing F.

[0444] Example 1 Synthesis of (R)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (1) Compound 1

[0445] [ka]

[0446] 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 0-5°C. N-bromosuccinimide (8.35 g, 46.9 mmol) was added to the above solution in several portions 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 Na2SO4, 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 11.89 (s, 1H), 3.74 (t, 2H), 2.68 (t, 2H), 1.44 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0447] Step 2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) 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 h. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The resulting solid was recrystallized from ethanol to give compound C as a white solid (6.6 g, 71%). 1H NMR (500 MHz, DMSO-d6): δ 8.08 (s, 2H), 3.89 (t, 2H), 2.75 (t, 2H), 1.45 (s, 9H) LCMS (ESI) 270.3 m / z [M+H] + .

[0448] 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. To the above solution was added tert-butyl nitrite (4.3 mL, 36.4 mmol) and stirring was continued at -10°C for 1 h. To the above mixture was added copper(II) bromide (6.5 mL, 29.43 mmol) and 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 three times with ethyl acetate (300 mL) and water (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 silica gel (100-200 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90) to give 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), 1.50 (s, 9H). LCMS (ESI) 279 m / z [M+H-C4H8] + .

[0449] Step 4: Synthesis of tert-butyl 2-(6-fluoropyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (F1) Water (4.0 mL) and 1,4-dioxane (16 mL) were combined and the mixture was degassed by passing a stream of nitrogen through it. Then [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.245 g, 0.3 mmol), tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate D (1.0 g, 3.0 mmol), (6-fluoropyridin-3-yl)boronic acid E1 (0.507 g, 3.6 mmol), and cesium carbonate (2.9 g, 9.0 mmol) were added and the reaction mixture was heated in an oil bath at 80° C. for 12 hours. The reaction mixture was diluted twice with ethyl acetate (80 mL) and water (80 mL). The organic phase was separated, dried over Na2SO4, filtered and the solvent was removed under reduced pressure. The residue was purified by silica gel (100-200 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90→20 / 80) to give the title compound F1 as a white solid (0.5 g, 50%). 1 H NMR (500 MHz, DMSO-d6): δ 8.92 (d, 1H), 8.60 (m, 1H), 7.40 (dd, 1H), 4.09 (t, 2H), 3.17 (t, 2H), 1.50 (s, 9H). LCMS (ESI) 293.94 m / z [M+H-C4H8] + .

[0450] Step 5: Synthesis of (R)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 1) The title compound F1 (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 h using a CEM microwave. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (20 mL) and filtered on a Whatmann 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, Preparation 1, 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), 3.90 - 3.70 (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] + .

[0451] Step 6: Synthesis of (R)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (1) The commercially available reagent 4-bromo-1-methyl-1H-pyrazole 8 (0.57 g, 3.54 mmol), the title compound Preparation 1 (0.375 g, 1.18 mmol), copper(I) iodide (0.022 g, 0.1157 mmol), N,N'-dimethylethylenediamine (0.2 mL, 2.0 mmol), and potassium carbonate (0.49 g, 3.55 mmol) were suspended in 1,4-dioxane (10 mL). The reaction mixture was heated in an oil bath at about 120°C for 48 hours. The reaction mixture was diluted with ethyl acetate (100 mL) and washed with water (40 mL) and brine (40 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The resulting residue was purified by silica gel (180 g, 100-200 mesh) column chromatography using a dichloromethane / methanol gradient (100 / 0→99 / 1→98 / 2→97.5 / 2.5) to give a mixture of the title compound 1 and the title compound, Preparation 1. The mixture was further purified on a preparative TLC plate using dichloromethane / methanol (93 / 7) as the mobile phase to give the title compound 1 as a yellow solid (0.2 g, 39%). 1 H NMR (500 MHz, DMSO-d6): δ 8.75 (d, 1H), 8.07 (dd, 1H), 8.09 (dd, 1H), 8.04 (s, 1H), 7.68 (s, 1H), 6.64 (d, 1H), 5.47 (d, 1H), 4.07 (t, 2H), 3.83 (t, 2H), 3.72 (dd, 2H), 3.64 (dd, 1H), 3.50 (td, 1H), 3.21 (t, 2H), 2.24 (m, 2H). MS (ESI) 399.38 m / z [M+H] + HPLC: 99.87% [retention time: 5.87 min, Eclipse Plus C18].

[0452] Example 2 Synthesis of (R)-2-(6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one compound 2

[0453] [ka]

[0454] 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 0-5°C. N-bromosuccinimide (8.35 g, 46.9 mmol) was added to the above solution in several portions 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 Na2SO4, 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): 1 H NMR (500 MHz, DMSO-d6): δ 11.89 (s, 1H), 3.74 (t, 2H), 2.68 (t, 2H), 1.44 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0455] Step 2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) 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 h. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The resulting solid was recrystallized from ethanol to give compound C as a white solid (6.6 g, 71%). 1H NMR (500 MHz, DMSO-d6): δ 8.08 (s, 2H), 3.89 (t, 2H), 2.75 (t, 2H), 1.45 (s, 9H). LCMS (ESI) 270.3 m / z [M+H] + .

[0456] 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. To the above solution was added tert-butyl nitrite (4.3 mL, 36.4 mmol) and stirring was continued at -10°C for 1 h. To the above mixture was added copper(II) bromide (6.5 mL, 29.43 mmol) and 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 three times with ethyl acetate (300 mL) and water (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 silica gel (100-200 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90) to give 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), 1.50 (s, 9H). LCMS (ESI) 279 m / z [M+H-C4H8] + .

[0457] Step 4: Synthesis of tert-butyl 2-(6-fluoropyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (F1) Water (4.0 mL) and 1,4-dioxane (16 mL) were combined and the mixture was degassed by passing a stream of nitrogen through it. Then [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.245 g, 0.3 mmol), tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate D (1.0 g, 3.0 mmol), (6-fluoropyridin-3-yl)boronic acid E1 (0.507 g, 3.6 mmol), and cesium carbonate (2.9 g, 9.0 mmol) were added and the reaction mixture was heated in an oil bath at 80° C. for 12 hours. The reaction mixture was diluted twice with ethyl acetate (80 mL) and water (80 mL). The organic phase was separated, dried over Na2SO4, filtered and the solvent was removed under reduced pressure. The residue was purified by silica gel (100-200 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90→20 / 80) to give the title compound F1 as a white solid (0.5 g, 50%). 1 H NMR (500 MHz, DMSO-d6): δ 8.92 (d, 1H), 8.60 (m, 1H), 7.40 (dd, 1H), 4.09 (t, 2H), 3.17 (t, 2H), 1.50 (s, 9H). LCMS (ESI) 293.94 m / z [M+H-C4H8] + .

[0458] Step 5: Synthesis of (R)-2-(6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 2) The title compound F1 (0.55 g, 1.5 mmol), (R)-3-fluoropiperidine hydrochloride (0.328 g, 2.4 mmol), DIPEA (0.79 mL, 4.5 mmol) were suspended in n-butanol (6.6 mL) using a microwave vial. The sealed vial was then heated at 160° C. for 45 min using a CEM microwave. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (20 mL) and filtered on a Whatmann 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, Preparation 2, as a green solid (0.33 g, 63%). 1 H NMR (500 MHz, DMSO-d6): δ 8.7 (d, 1H), 8.02 (dd, 1H), 7.81 (s, 1H), 6.98 (d, 1H), 4.80 (d, 1H), 4.10 - 4.06 (m, 1H), 3.94 - 3.90 (m, 1H), 3.76 - 3.67 (m, 1H), 3.51 - 3.48 (m, 2H), 3.44 - 3.99 (m, 1H), 3.94 - 3.90 (m, 1H), 2.98 (t, 2H), 1.97 - 1.86 (m, 2H), 1.77 - 1.74 (m, 1H), 1.57 - 1.54 (m, 1H). LCMS: 95% [retention time: 7.0 min, Hypersil BDS C18], (ESI) 332.9 m / z [M+H] + .

[0459] Step 6: Synthesis of (R)-2-(6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (2) The commercially available reagent 4-bromo-1-methyl-1H-pyrazole 8 (0.218 g, 1.35 mmol), the title compound Preparation 2 (0.18 g, 0.54 mmol), copper(I) iodide (0.01 g, 0.05 mmol), N,N'-dimethylethylenediamine (0.095 mL, 0.9 mmol), and potassium carbonate (0.186 g, 1.35 mmol) were suspended in 1,4-dioxane (6 mL). The reaction mixture was heated in an oil bath at about 120° C. for 36 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by column chromatography on silica gel (180 g, 100-200 mesh) using a dichloromethane / methanol gradient (100 / 0→99 / 1→98 / 2→97.5 / 2.5) to give a mixture of the title compound 2 and the title compound Preparation 2. The mixture was further purified on a preparative TLC plate using dichloromethane / methanol (93 / 7) as the mobile phase to give the title compound 2 as a yellow solid (0.065 g, 29%). 1 H NMR (500 MHz, DMSO-d6): δ 8.73 (d, 1H), 8.05 - 8.02 (m, 2H), 7.67 (s, 1H), 8.04 (s, 1H), 6.99 (d, 1H), 4.81 (d, 1H), 4.10 - 4.05 (m, 3H), 4.00 - 3.90 (m, 1H), 3.83 (s, 3H), 3.45 - 3.39 (m, 1H), 3.21 (t, 2H), 1.97 - 1.87 (m, 2H), 1.80 - 1.73 (m, 1H), 1.60 - 1.53 (m, 1H). MS (ESI) 413.35 m / z [M+H] + HPLC: 97.4% [retention time: 6.89 min, Eclipse Plus C18].

[0460] Example 3 Synthesis of (5-(3-amino-4-fluorophenyl)-2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one compound 3

[0461] [ka]

[0462] 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 0-5°C. N-bromosuccinimide (8.35 g, 46.9 mmol) was added to the above solution in several portions 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 Na2SO4, 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 11.89 (s, 1H), 3.74 (t, 2H), 2.68 (t, 2H), 1.44 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0463] Step 2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) 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 h. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The resulting solid was recrystallized from ethanol to give compound C as a white solid (6.6 g, 71%). 1 H NMR (500 MHz, DMSO-d6): δ 8.08 (s, 2H), 3.89 (t, 2H), 2.75 (t, 2H), 1.45 (s, 9H). LCMS (ESI) 270.3 m / z [M+H]+ .

[0464] 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. To the above solution was added tert-butyl nitrite (4.3 mL, 36.4 mmol) and stirring was continued at -10°C for 1 h. To the above mixture was added copper(II) bromide (6.5 mL, 29.43 mmol) and 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 three times with ethyl acetate (300 mL) and water (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 silica gel (100-200 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90) to give 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), 1.50 (s, 9H). LCMS (ESI) 279 m / z [M+H-C4H8] + .

[0465] Step 4: Synthesis of 5-bromo-2-(pyrrolidin-1-yl)pyridine (F2) 5-Bromo-2-fluoropyridine E2 (1.5 g, 8.5 mmol) and pyrrolidine (3.6 g, 50.62 mmol) were dissolved in ethanol (8 mL) using a microwave vial. The sealed vial was then heated at 120° C. for 1 h using a CEM microwave. The reaction mixture was concentrated under reduced pressure to give the crude product. The resulting crude product was suspended in water (15 mL) and filtered on a Whatmann filter paper. The solid was washed with water (2×15 mL) and dried under reduced pressure to give the title compound F2 as an off-white solid (1.9 g, 98%). 1H NMR (500 MHz, DMSO-d6): δ 8.10 (d, 1H), 7.61 (dd, 1H), 6.42 (d, 1H), 3.34 - 3.33 (m, 4H), 1.94 - 1.91 (m, 4H). LCMS (ESI) 228.99 m / z [M+H] + .

[0466] Step 5: Synthesis of 2-(pyrrolidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (G1) 1,4-Dioxane (25 mL) was degassed by passing a stream of nitrogen through the mixture. Then [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.683 g, 0.83 mmol), 5-bromo-2-(pyrrolidin-1-yl)pyridine F2 (1.4 g, 4.2 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (4.25 g, 16.73 mmol), and potassium acetate (1.64 g, 16.73 mmol) were added and the reaction mixture was heated in an oil bath at 100°C for 3 hours. After completion of the reaction, the solvent was removed from the reaction mixture under reduced pressure. The residue was washed three times with n-hexane (100 mL). The reaction mixture was diluted twice with ethyl acetate (80 mL) and water (80 mL). The organic phase was dried over Na2SO4, filtered, and the solvent was removed under reduced pressure to give a pale yellow sticky solid G1 (5.7 g, crude product). The solid was used in the next step without further purification. LCMS (ESI) 275.22 m / z [M+H] + .

[0467] Step 6: Synthesis of tert-butyl 4-oxo-2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (H1) Water (10 mL) and 1,4-dioxane (40 mL) were combined and the mixture was degassed by passing a stream of nitrogen through it.[1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.342 g, 0.4 mmol), tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate D (1.4 g, 4.2 mmol), 2-(pyrrolidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine G1 (1.73 g, 6.3 mmol), and cesium carbonate (4.1 g, 12.6 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was diluted twice with ethyl acetate (80 mL) and water (80 mL). The organic phase was separated, dried over Na2SO4, filtered and the solvent was removed under reduced pressure. The residue was purified by basic silica gel (60-120 mesh) column chromatography using an ethyl acetate / n-hexane gradient (0 / 100→10 / 90→20 / 80→30 / 70→40 / 60) to give the title compound H1 as a pale yellow solid (0.8 g, 48%). 1 H NMR (500 MHz, DMSO-d6): δ 8.77 (d, 1H), 7.97 (dd, 1H), 6.40 (d, 1H), 4.14 (t, 2H), 3.54 (s, 4H), 3.10 (t, 2H), 2.04 (m, 4H), 1.57 (s, 9H). LCMS (APCI) 401.26 m / z [M+H] + .

[0468] Step 7: Synthesis of 2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 3) The title compound H1 (0.34 g, 0.99 mmol) was dissolved in DCM (8 mL) and cooled to 0° C. with stirring in an ice bath. 4M HCl in 1,4-dioxane (4.0 mL) was added to the above solution and stirring was continued at room temperature for 4 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 solution, the precipitated solid was filtered and dried to give the title compound, Preparation 3, as a pale yellow solid (0.25 g, 83%). 1 H NMR (500 MHz, DMSO-d6): δ 8.70 (d, 1H), 8.00 (dd, 1H), 7.80 (s, 1H), 6.55 (d, 1H), 3.50 - 3.40 (m, 6H), 2.97 (t, 2H), 1.97 - 1.92 (m, 4H). LCMS (ESI) 301.08 m / z [M+H] + .

[0469] Step 8: Synthesis of 5-(3-amino-4-fluorophenyl)-2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (3) The commercially available reagent 5-bromo-2-fluoroaniline (0.095 g, 0.5 mmol), the title compound Preparation 3 (0.05 g, 0.166 mmol), copper(I) iodide (0.003 g, 0.0166 mmol), N,N'-dimethylethylenediamine (0.03 mL, 0.28 mmol), and potassium carbonate (0.046 g, 0.33 mmol) were suspended in 1,4-dioxane (3 mL). The reaction mixture was heated in an oil bath at about 120° C. for 24 hours. The reaction mixture was concentrated under reduced pressure to give the crude product. The resulting residue was purified by silica gel (80 g, 100-200 mesh) column chromatography using a dichloromethane / methanol gradient (100 / 0→98 / 2) to give the title compound 3 as a yellow solid (80 mg, 73.4%). 1H NMR (500 MHz, DMSO-d6): δ 8.73 (d, 1H), 8.04 (dd, 1H), 7.02 - 6.98 (dd, 1H), 6.76 - 6.74 (dd, 1H), 6.58 (d, 1H), 6.50 - 6.48 (m, 1H), 5.24 (d, 1H), 3.98 (t, 2H), 3.47 (s, 4H), 3.17 (t, 2H), 1.97 (s, 4H). LCMS: 96.7% [retention time: 8.6 min, Hypersil BDS C18], (ESI) 410 m / z [M+H] + .

[0470] Example 4 (R)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3 H Synthesis of compound 1 (precursor)

[0471] [ka]

[0472] Step 1: Synthesis of tert-butyl 2-(6-fluoropyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (F1) Water (2.2 mL) and 1,4-dioxane (9.5 mL) were combined and the mixture was degassed by sonication while passing a stream of argon through it. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.019 g, 0.022 mmol), tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate D (0.15 g, 0.45 mmol), (6-fluoropyridin-3-yl)boronic acid (0.078 g, 0.55 mmol), and cesium carbonate (0.319 g, 0.9 mmol) were then added and the reaction mixture was heated in a sand bath at about 85° C. for 5 hours. The reaction mixture was diluted twice with ethyl acetate (200 mL) and water (80 mL). The organic phase was separated, dried over Na2SO4, filtered and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica (50 g HP-Ultra) using a Biotage Isolera system with an ethyl acetate / n-heptane gradient (5 / 95→10 / 90→20 / 80→30 / 70→40 / 60) to give the title compound F1 as a white solid (0.225 g, 71%). 1 H NMR (400 MHz, DMSO-d6): δ 8.92 (d, 1H), 8.60 (ddd, 1H), 7.41 (dd, 1H), 4.10 (t, 2H), 3.17 (t, 2H), 1.50 (s, 9H). LCMS (ESI) 293.94 m / z [M+H-C4H8] + .

[0473] Step 2: Synthesis of (R)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 1) The title compound F1 (0.1125 g, 0.3225 mmol), (R)-3-fluoropyrrolidine hydrochloride (0.122 g, 0.97 mmol), and N,N'-diisopropylethylamine (0.25 mL, 1.45 mmol) were suspended in n-butanol (10 mL) using a microwave vial. The sealed vial was then heated at 160°C for 1 h using a Biotage Initiator microwave. The reaction mixture was diluted twice with ethyl acetate / methanol (200 mL; 9 / 1) and washed with water (80 mL) and brine (80 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica (25 g HP-Ultra) using a Biotage Isolera system using a dichloromethane / methanol gradient (100 / 0→95 / 5→93 / 7→90 / 10) to give the title compound Preparation 1 as an off-white solid (0.151 g, 73%). 1 H NMR (400 MHz, DMSO-d6): δ 8.73 (d, 1H), 8.06 (dd, 1H), 7.81 (s, 1H), 6.64 (d, 1H), 5.48 (d, 1H), 3.85 - 3.61 (m, 4H), 3.50 (t, 2H), 2.99 (t, 2H), 2.32 - 2.17 (m, 2H). LCMS (ESI) 319.00 m / z [M+H] + .

[0474] Step 3: Synthesis of 4-bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole (E3) The commercially available reagent 4-bromo-1H-pyrazole (1.47 g, 10 mmol) was dissolved in acetonitrile (10 mL) and 3,4-dihydro-2H-pyran (2.39 mL, 26 mmol) was added. After addition of trifluoroacetic acid (0.05 mL, 0.69 mmol), the reaction mixture was heated at 90° C. for 2 h using a Biotage Initiator microwave. The reaction mixture was diluted with ethyl acetate (200 mL) and washed with water (80 mL) and brine (80 mL). The organic phase was separated, dried over Na2SO4, filtered and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica (100 g HP-Ultra) using a Biotage Isolera system using an ethyl acetate / n-heptane gradient (5 / 95→10 / 90→20 / 80→30 / 70→40 / 60) to give a pale yellow liquid. The title compound was further purified by silica (40 g Interchim) chromatography using a Biotage Isolera system using an ethyl acetate / n-heptane gradient (5 / 95→10 / 90→20 / 80→30 / 70) to afford the title compound E3 as a colorless liquid (0.989 g, 42%). 1 H NMR (400 MHz, CDCl3): δ 7.56 (s, 1H), 7.43 (s, 1H), 5.29 - 5.26 (m, 1H), 3.99 - 3.94 (m, 1H), 3.65 - 3.58 (m, 1H), 2.02 - 1.92 (m, 3H), 1.64 - 1.52 (m, 3H).

[0475] Step 4: Synthesis of 2-(6-((R)-3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (G2) The title compound Preparation 1 (0.05 g, 0.157 mmol), title compound E3 (0.109 g, 0.472 mmol), copper(I) iodide (0.003 g, 0.0157 mmol), N,N'-dimethylethylenediamine (0.025 mL, 0.27 mmol), and potassium carbonate (0.043 g, 0.314 mmol) were suspended in 1,4-dioxane (9 mL). The reaction mixture was heated in a sand bath at about 120° C. for 24 hours. The reaction mixture was diluted with ethyl acetate (100 mL) and washed with water (40 mL) and brine (40 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The residue was purified by silica (25 g HP-Ultra) chromatography using a Biotage Isolera system using a dichloromethane / methanol gradient (100 / 0→95 / 5→95 / 15) to give a mixture of the title compound G2 and the title compound Preparation 1. The mixture was further purified on a preparative TLC plate using dichloromethane / methanol (90 / 10) as the mobile phase to give the title compound G2 as a yellow solid (0.0384 g, 52%). 1 H NMR (400 MHz, DMSO-d6): δ 8.76 (d, 1H), 8.19 (s, 1H), 8.09 (dd, 1H), 7.81 (s, 1H), 6.65 (d, 1H), 5.48 (d, 1H), 5.41 - 5.38 (m, 1H), 4.10 (t, 2H), 3.95 - 3.91 (m, 1H), 3.87 - 3-61 (m, 4H), 3.55 - 3.48 (m, 1H), 3.23 (t, 2H), 2.34 - 2.26 (m, 2H), 2.10 - 2.05 (m, 1H), 1.97 - 1.89 (m, 2H), 1.72 - 1.64 (m, 1H), 1.57 - 1.52 (m, 2H). LCMS (ESI) 469.15 m / z [M+H] +

[0476] Step 5: Synthesis of (R)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (H2) The title compound G2 (0.0384 g, 0.082 mmol) was dissolved in dichloromethane (4 mL) and trifluoroacetic acid (2 mL) was added. The reaction mixture was stirred at room temperature for 18 h. The solvent was evaporated under reduced pressure, the residue was dissolved in methanol (5 mL) and the solvent was evaporated under reduced pressure. The residue was dissolved in ethyl acetate (80 mL) and saturated NaHCO3 (30 mL) was added. The organic phase was separated, washed with brine (30 mL), dried over Na2SO4, filtered and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica (25 g HP Ultra) using a Biotage Isolera system using a dichloromethane / methanol gradient (100 / 0 -> 95 / 5 -> 90 / 10 -> 80 / 20) to give the title compound H2 as a yellow solid (0.0188 g, 60%). 1 H NMR (400 MHz, DMSO-d6): δ 12.77 (br-s, 1H), 8.76 (d, 1H), 8.08 (dd, 1H), 8.03 - 7.76 (br-m, 2H), 6.65 (d, 1H), 5.49 (d, 1H), 4.10 (t, LCMS (ESI) 385.17 m / z [M+H] + .

[0477] Example 5 (R)-2-(6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3 H Synthesis of compound 2 (precursor)

[0478] [ka]

[0479] 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 0-5°C. N-bromosuccinimide (8.35 g, 46.9 mmol) was added to the above solution in several portions 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 Na2SO4, 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 11.89 (s, 1H), 3.74 (t, 2H), 2.68 (t, 2H), 1.44 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0480] Step 2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) 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 h. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The resulting solid was recrystallized from ethanol to give compound C as a white solid (6.6 g, 71%). 1 H NMR (500 MHz, DMSO-d6): δ 8.10 (s, 2H), 3.89 (t, 2H), 2.76 (t, 2H), 1.45 (s, 9H). LCMS (ESI) 270.3 m / z [M+H] + .

[0481] 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. To the above solution was added tert-butyl nitrite (4.3 mL, 36.4 mmol) and stirring was continued at -10°C for 1 h. To the above mixture was added copper(II) bromide (6.5 mL, 29.43 mmol) and 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 three times with ethyl acetate (300 mL) and water (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 silica gel (100-200 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90) to give 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), 1.50 (s, 9H). LCMS (ESI) 279 m / z [M+H-C4H8] + .

[0482] Step 4: Synthesis of tert-butyl 2-(6-fluoropyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (F1) Water (4.0 mL) and 1,4-dioxane (16 mL) were combined and the mixture was degassed by passing a stream of nitrogen through it. Then [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.245 g, 0.3 mmol), tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate D (1.0 g, 3.0 mmol), (6-fluoropyridin-3-yl)boronic acid (0.507 g, 3.6 mmol), and cesium carbonate (2.9 g, 9.0 mmol) were added and the reaction mixture was heated in an oil bath at 80° C. for 12 hours. The reaction mixture was diluted twice with ethyl acetate (80 mL) and water (80 mL). The organic phase was separated, dried over Na2SO4, filtered and the solvent was removed under reduced pressure. The residue was purified by silica gel (100-200 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90→20 / 80) to give the title compound F1 as a white solid (0.5 g, 50%). 1 H NMR (500 MHz, DMSO-d6): δ 8.92 (d, 1H), 8.60 (m, 1H), 7.40 (dd, 1H), 4.09 (t, 2H), 3.17 (t, 2H), 1.50 (s, 9H). LCMS (ESI) 293.94 m / z [M+H-C4H8] + .

[0483] Step 5: Synthesis of (R)-2-(6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 2) The title compound F1 (0.55 g, 1.5 mmol), (R)-3-fluoropiperidine hydrochloride (0.328 g, 2.4 mmol), and N,N-diisopropylethylamine (DIPEA) (0.79 mL, 4.5 mmol) were suspended in n-butanol (6.6 mL) using a microwave vial. The sealed vial was then heated at 160° C. for 45 minutes using a CEM microwave. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (20 mL) and filtered on a Whatmann 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, Preparation 2, as a green solid (0.33 g, 63%). 1 H NMR (500 MHz, DMSO-d6): δ 8.70 (d, 1H), 8.02 (dd, 1H), 7.81 (s, 1H), 6.98 (d, 1H), 4.80 (d, 1H), 4.10 - 4.06 (m, 1H), 3.94 - 3.90 (m, 1H), 3.76 - 3.67 (m, 1H), 3.51 - 3.48 (m, 2H), 3.44 - 3.99 (m, 1H), 3.94 - 3.90 (m, 1H), 2.98 (t, 2H), 1.97 - 1.86 (m, 2H), 1.77 - 1.74 (m, 1H), 1.57 - 1.54 (m, 1H). LCMS: 95% [retention time: 7.0 min, Hypersil BDS C18], (ESI) 332.9 m / z [M+H] + .

[0484] Step 6: Synthesis of (R)-2-(6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (G3) 4-Iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (0.194 g, 0.6 mmol), the title compound Preparation 2 (0.1 g, 0.3 mmol), copper(I) iodide (0.03 g, 0.15 mmol), N,N'-dimethylethylenediamine (0.03 mL, 0.3 mmol), and potassium carbonate (0.08 g, 0.6 mmol) were suspended in 1,4-dioxane (10 mL). The reaction mixture was heated in an oil bath at about 120° C. for 24 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by column chromatography on basic silica gel (100 g, 100-200 mesh) using a dichloromethane / methanol gradient (100 / 0→98 / 2→97 / 3) to afford the title compound G3 as a yellow solid (0.12 g, 75%). 1 H NMR (500 MHz, DMSO-d6): δ 8.73 (d, 1H), 8.22 (s, 1H), 8.05 (dd, 1H), 7.83 (s, 1H), 6.99 (d, 1H), 5.40 (s, 2H), 4.81 (d, 1H), 4.10 - 4.05 (m, 3H), 3.94 (d, 1H), 3.76 - 3.68 (m, 1H), 3.54 (t, 2H), 3.42 (t, 1H), 3.22 (t, 2H), 1.99 - 1.94 (m, 2H), 1.89 - 1.75 (m, 1H), 1.56 - 1.53 (m, 1H), 0.85 (t, 2H), -30.39 (s, 9H). MS (ESI) 529.3 m / z [M+H] + .

[0485] Step 7: Synthesis of (R)-2-(6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (H3) The title compound G3 (0.12 g, 0.23 mmol) was dissolved in dichloromethane (4 mL) and cooled to 0° C. with stirring in an ice bath. 4M HCl in 1,4-dioxane (2.0 mL) was added to the above solution and stirring was continued at room temperature for 4 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 solution and the precipitated solid was filtered. The solid was further purified by silica gel (100 g, 100-200 mesh) column chromatography using dichloromethane / methanol gradient (100 / 0→98 / 2→97 / 3) to give the title compound H3 as a yellow solid (0.06 g, 55%). 1 H NMR (500 MHz, DMSO-d6): δ 12.79 (s, 1H), 8.73 (d, 1H), 8.04 (dd, 2H), 7.79 (d, 1H), 6.99 (d, 1H), 4.88 - 4.72 (m, 1H), 4.13 - 4.06 (m, 3H), 3.97 - 3.91 (m, 1H), 3.77 - 3.66 (m, 1H), 3.45 - 3.40 (m, 1H), 3.21 (t, 2H), 1.98 - 1.87 (m, 2H), 1.79 - 1.72 (m, 1H), 1.59 - 1.53 (m, 1H). MS (ESI) 399.22 m / z [M+H] + HPLC: 98.2% [retention time: 7.65 min, Hypersil BDS C18].

[0486] Example 6 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 H Synthesis of compound 3 (precursor)

[0487] [ka]

[0488] 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 0-5°C. N-bromosuccinimide (8.35 g, 46.9 mmol) was added to the above solution in several portions 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 Na2SO4, 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 8.10 (s, 2H), 3.89 (t, 2H), 2.76 (t, 2H), 1.45 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0489] Step 2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) 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 h. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The resulting solid was recrystallized from ethanol to give compound C as a white solid (6.6 g, 71%). 1 H NMR (500 MHz, DMSO-d6): δ 8.10 (s, 2H), 3.89 (t, 2H), 2.76 (t, 2H), 1.45 (s, 9H). LCMS (ESI) 270.3 m / z [M+H] + .

[0490] 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. To the above solution was added tert-butyl nitrite (4.3 mL, 36.4 mmol) and stirring was continued at -10°C for 1 h. To the above mixture was added copper(II) bromide (6.5 mL, 29.43 mmol) and 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 three times with ethyl acetate (300 mL) and water (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 silica gel (100-200 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90) to give 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), 1.50 (s, 9H). LCMS (ESI) 279 m / z [M+H-C4H8] + .

[0491] Step 4: Synthesis of tert-butyl 4-oxo-2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (E4) Water (10 mL) and 1,4-dioxane (40 mL) were combined and the mixture was degassed by passing a stream of nitrogen through it.[1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.342 g, 0.4 mmol), tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate D (1.4 g, 4.2 mmol), 2-(pyrrolidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (1.73 g, 6.3 mmol), and cesium carbonate (4.1 g, 12.6 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was diluted twice with ethyl acetate (80 mL) and water (80 mL). The organic phase was separated, dried over Na2SO4, filtered and the solvent was removed under reduced pressure. The residue was purified by basic silica gel (60-120 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90→20 / 80→30 / 70→40 / 60) to give the title compound E4 as a pale yellow solid (0.8 g, 48%). 1 H NMR (500 MHz, DMSO-d6): δ 8.77 (d, 1H), 7.97 (dd, 1H), 6.40 (d, 1H), 4.14 (t, 2H), 3.54 (s, 4H), 3.10 (t, 2H), 2.04 (m, 4H), 1.57 (s, 9H). LCMS (APCI) 401.26 m / z [M+H] + .

[0492] Step 5: Synthesis of 2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (F3) The title compound E4 (0.34 g, 0.99 mmol) was dissolved in dichloromethane (8 mL) and cooled to 0° C. with stirring in an ice bath. 4M HCl in 1,4-dioxane (4.0 mL) was added to the above solution and stirring was continued at room temperature for 4 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 solution, and the precipitated solid was filtered and dried to obtain the title compound F3 as a pale yellow solid (0.25 g, 83%). 1 H NMR (500 MHz, DMSO-d6): δ 8.70 (d, 1H), 8.00 (dd, 1H), 7.80 (s, 1H), 6.55 (d, 1H), 3.50 - 3.40 (m, 6H), 2.97 (t, 2H), 1.97 - 1.92 (m, 4H). LCMS (ESI) 301.08 m / z [M+H] + .

[0493] Step 6: Synthesis of 5-(3-amino-4-fluorophenyl)-2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (G4) Commercially available reagent 5-bromo-2-fluoroaniline (0.095 g, 0.5 mmol), title compound F3 (0.05 g, 0.166 mmol), copper(I) iodide (0.003 g, 0.0166 mmol), N,N'-dimethylethylenediamine (0.03 mL, 0.28 mmol), and potassium carbonate (0.046 g, 0.33 mmol) were suspended in 1,4-dioxane (3 mL). The reaction mixture was heated in an oil bath at about 120° C. for 24 hours. The reaction mixture was concentrated under reduced pressure to give the crude product. The resulting residue was purified by silica gel (80 g, 100-200 mesh) column chromatography using a dichloromethane / methanol gradient (100 / 0→98 / 2) to give the title compound G4 as a yellow solid (80 mg, 73%). 1H NMR (500 MHz, DMSO-d6): δ 8.73 (d, 1H), 8.04 (dd, 1H), 7.02 - 6.98 (dd, 1H), 6.76 - 6.74 (dd, 1H), 6.58 (d, 1H), 6.50 - 6.48 (m, 1H), 5.24 (d, 1H), 3.98 (t, 2H), 3.47 (s, 4H), 3.17 (t, 2H), 1.97 (s, 4H). LCMS: 96.7% [retention time: 8.6 min, Hypersil BDS C18], (ESI) 410 m / z [M+H] + .

[0494] Step 7: 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 (H4) The title compound G4 (90 g, 0.22 mmol) was dissolved in chloroform (15 mL) and cooled to 0-5° C. N-bromosuccinimide (0.15 g, 0.44 mmol) was added to the above solution in several portions and stirring was continued at 5-20° C. for 1 h. The reaction mixture was evaporated under reduced pressure. The resulting residue was purified by silica gel (60 g, 100-200 mesh) column chromatography using a dichloromethane / methanol gradient (100 / 0→99 / 1→98 / 2) to give compound H4 as a yellow solid (0.035 g, 25%). 1 H NMR (500 MHz, DMSO-d6): δ 8.72 (d, 1H), 8.31 (d, 1H), 7.59 (d, 1H), 5.74 (s, 2H), 3.89 (t, 2H), 3.73 (t, 4H), 3.30 - 3.39 (m, 2H), 1.92 - 1.89(m, 4H). LCMS: 97% [retention time: 12.84 min, Hypersil BDS C18], (ESI) 645.6 m / z [M+2H] + .

[0495] Example 7 (S)-1-(5-(5-(1-methyl-1H-pyrazol-4-yl)-4-oxo-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)pyrrolidin-3-yl methanesulfonate( 18 F. Synthesis of precursor of compound 1)

[0496] [ka]

[0497] 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 0-5°C. N-bromosuccinimide (8.35 g, 46.9 mmol) was added to the above solution in several portions 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 Na2SO4, 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 11.89 (s, 1H), 3.74 (t, 2H), 2.68 (t, 2H), 1.44 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0498] Step 2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) 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 h. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The resulting solid was recrystallized from ethanol to give compound C as a white solid (6.6 g, 71%). 1 H NMR (500 MHz, DMSO-d6): δ 8.10 (s, 2H), 3.89 (t, 2H), 2.76 (t, 2H), 1.45 (s, 9H). LCMS (ESI) 270.3 m / z [M+H] + .

[0499] 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. To the above solution was added tert-butyl nitrite (4.3 mL, 36.4 mmol) and stirring was continued at -10°C for 1 h. To the above mixture was added copper(II) bromide (6.5 mL, 29.43 mmol) and 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 three times with ethyl acetate (300 mL) and water (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 silica gel (100-200 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90) to give 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), 1.50 (s, 9H). LCMS (ESI) 279 m / z [M+H-C4H8] + .

[0500] Step 4: Synthesis of tert-butyl 4-oxo-2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (F1) Water (10 mL) and 1,4-dioxane (40 mL) were combined and the mixture was degassed by passing a stream of nitrogen through it. Then [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.342 g, 0.4 mmol), tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate D (1.4 g, 4.2 mmol), 6-fluoropyridine-3-boronic acid (1.73 g, 6.3 mmol), and cesium carbonate (4.1 g, 12.6 mmol) were added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was diluted twice with ethyl acetate (80 mL) and water (80 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The residue was purified by basic silica gel (60-120 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90→20 / 80→30 / 70→40 / 60) to afford the title compound F1 as a pale yellow solid (0.8 g, 48%). 1 H NMR (500 MHz, DMSO-d6): δ 8.77 (d, 1H), 7.97 (dd, 1H), 6.40 (d, 1H), 4.14 (t, 2H), 3.54 (s, 4H), 3.10 (t, 2H), 2.04 (m, 4H), 1.57 (s, 9H). LCMS (APCI) 401.26 m / z [M+H] + .

[0501] Step 5: Synthesis of 2-(6-fluoropyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (G5) The title compound F1 (1.14 g, 3.2 mmol) was dissolved in dichloromethane (20 mL) and cooled to 0° C. with stirring in an ice bath. 4M HCl in 1,4-dioxane (10 mL) was added to the above solution and stirring was continued at room temperature for 16 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 solution, the precipitated solid was filtered and dried to give the title compound G5 as a white solid (0.66 g, 80%) without further purification. LCMS (ESI) 250.15 m / z [M+H] +

[0502] Step 6: Synthesis of (S)-2-(6-(3-hydroxypyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (H6) The title compound G5 (0.66 g, 2.65 mmol), (S)-pyrrolidin-3-ol hydrochloride (0.49 g, 3.97 mmol), diisopropylethylamine (1.41 mL, 7.9 mmol) were suspended in ethanol (7.3 mL) using a microwave vial. The sealed vial was then heated at 120° C. for 1.5 h using a CEM microwave. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (20 mL) and filtered on a Whatmann 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 H6 as a pale yellow solid (0.7 g, 83%). 1 H NMR (500 MHz, DMSO-d6): δ 8.70 (d, 1H), 8.00 (dd, 1H), 7.78 (s, 1H), 6.55 (d, 1H), 5.02 (d, 1H), 4.41 (d, 1H), 3.56 - 3.47 (m, 5H), 2.97 (t, 2H), 2.06 - 1.98 (m, 2H). LCMS (ESI) 317.15 m / z [M+H] + .

[0503] Step 7: Synthesis of (S)-2-(6-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (I1) The title compound H6 (0.95 g, 3.0 mmol) and imidazole (1.84 g, 27 mmol) were dissolved in dimethylformamide (25 mL) with stirring in an ice bath at 0° C. tert-Butyldimethylsilyl chloride (1.81 g, 12 mmol) was added and stirring was continued at room temperature for 16 h. The reaction mixture was quenched with ice-cold water (100 mL) and the precipitated solid was filtered on a Whatmann filter paper and dried to give the title compound I1 as a pale yellow solid (1.0 g, 78%). 1 H NMR (500 MHz, DMSO-d6): δ 8.70 (d, 1H), 8.01 (dd, 1H), 7.80 (s, 1H), 6.58 (d, 1H), 4.59 (s, 1H), 3.66 - 3.62 (m, 2H), 3.57 - 3.47 (m, 3H), 2.97 (t, 2H), 2.10 (dd, 1H), 2.11 - 1.91 (m, 2H), 0.82 (s, 9H), 0.09 (d, 6H). LCMS: (ESI) 637.44 m / z [M+H] + .

[0504] Step 8: Synthesis of (S)-2-(6-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)pyridin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (J1) The title compound I1 (1.02 g, 2.4 mmol), 4-bromo-1-methyl-1H-pyrazole (1.14 g, 7.1 mmol), copper(I) iodide (0.053 g, 0.28 mmol), N,N'-dimethylethylenediamine (0.42 mL, 4.1 mmol), and potassium carbonate (0.98 g, 7.1 mmol) were suspended in 1,4-dioxane (70 mL). The reaction mixture was heated in an oil bath at about 120 °C for 24 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by column chromatography on basic silica gel (150 g, 100-200 mesh) using a dichloromethane / methanol gradient (100 / 0 → 99 / 1 → 98 / 2 → 97 / 3 → 95 / 5) to give the title compound J1 as a yellow solid (660 mg, 54%). 1 H NMR (500 MHz, DMSO-d6): δ 8.69 (d, 1H), 7.99 (dd, 2H), 7.63 (s, 1H), 6.56 (d, 1H), 4.55 (s, 1H), 4.02 (t, 2H), 3.79 (s, 3H), 3.62 - 3.47(m, 3H), 3.32 (d, 1H), 3.16 (s, 2H), 2.07 (t, 1H), 1.88 (s, 1H), 0.82 (s, 9H), 0.06 (d, 6H). LCMS: (ESI) 511.37 m / z [M+H] + .

[0505] Step 9: Synthesis of (S)-2-(6-(3-hydroxypyrrolidin-1-yl)pyridin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (K1) The title compound J1 (0.65 g, 1.27 mmol) was dissolved in dichloromethane (13 mL) and cooled to 0° C. with stirring in an ice bath. 4M HCl in 1,4-dioxane (6.5 mL) was added to the above solution and stirring was continued at room temperature for 4 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 solution, and the precipitated solid was filtered and dried to obtain the title compound K1 as a pale yellow solid (0.5 g, 99%).1 H NMR (500 MHz, DMSO-d6): δ 8.69 (d, 1H), 7.99 (dd, 2H), 7.63 (s, 1H), 6.53 (d, 1H), 4.99 (s, 1H), 4.37 (s, 1H), 4.02 (t, 2H), 3.79 (s, 3H), 3.51 (dd, 3H), 3.16 (t, 2H), 2.00 (q, 1H), 1.89 (s, 1H). LCMS (ESI) 397.39 m / z [M+H] +

[0506] Step 10: Synthesis of (S)-1-(5-(5-(1-methyl-1H-pyrazol-4-yl)-4-oxo-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)pyrrolidin-3-yl methanesulfonate (L1) The title compound K1 (0.68 g, 1.7 mmol) and triethylamine (14.3 mL, 103 mmol) were suspended in dichloromethane (90 mL) with stirring. The mixture was cooled to 0° C. in an ice bath. Methanesulfonyl chloride (4.0 mL, 51.4 mmol) was added and stirring was continued at room temperature for 24 h. The reaction mixture was quenched with cold water (150 mL) and extracted twice with dichloromethane (150 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 (400 g, 230-400 mesh) using a dichloromethane / methanol gradient (100 / 0→99 / 1→98 / 2) to afford the title compound L1 as a pale yellow solid (0.1 g, 12.3%). 1H NMR (500 MHz, DMSO-d6): δ 8.76 (d, 1H), 8.09 (dd, 1H), 8.04 (s, 1H), 7.68 (s, 1H), 6.65 (d, 1H), 5.45 (s, 1H), 4.07 (t, 2H), 3.83 LCMS (ESI) 475.3 m / z [M+H] + HPLC: 96.9% [retention time: 6.08 min, Eclipse plus C18].

[0507] Example 8 (S)-1-(5-(5-(1-methyl-1H-pyrazol-4-yl)-4-oxo-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)piperidin-3-yl methanesulfonate( 18 F. Synthesis of precursor of compound 2

[0508] [ka]

[0509] 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 0-5°C. N-bromosuccinimide (8.35 g, 46.9 mmol) was added to the above solution in several portions 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 Na2SO4, filtered and the solvent was removed under reduced pressure to give the title compound B as a white solid (10 g, 73%). 1H NMR (500 MHz, DMSO-d6): δ 11.89 (s, 1H), 3.74 (t, 2H), 2.68 (t, 2H), 1.44 (s, 9H). LCMS (ESI) 292.04 m / z [M+H] + .

[0510] Step 2: Synthesis of tert-butyl 2-amino-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (C) 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 h. The reaction mixture was diluted twice with ethyl acetate (500 mL) and water (200 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The resulting solid was recrystallized from ethanol to give compound C as a white solid (6.6 g, 71%). 1 H NMR (500 MHz, DMSO-d6): δ 8.10 (s, 2H), 3.89 (t, 2H), 2.76 (t, 2H), 1.45 (s, 9H). LCMS (ESI) 270.3 m / z [M+H] + .

[0511] 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. To the above solution was added tert-butyl nitrite (4.3 mL, 36.4 mmol) and stirring was continued at -10°C for 1 h. To the above mixture was added copper(II) bromide (6.5 mL, 29.43 mmol) and 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 three times with ethyl acetate (300 mL) and water (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 silica gel (100-200 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90) to give 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), 1.50 (s, 9H). LCMS (ESI) 279 m / z [M+H-C4H8] + .

[0512] Step 4: Synthesis of tert-butyl 4-oxo-2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate (F1) Water (10 mL) and 1,4-dioxane (40 mL) were combined and the mixture was degassed by passing a stream of nitrogen through it. Then [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.342 g, 0.4 mmol), tert-butyl 2-bromo-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate D (1.4 g, 4.2 mmol), 6-fluoropyridine-3-boronic acid (1.73 g, 6.3 mmol), and cesium carbonate (4.1 g, 12.6 mmol) were added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was diluted twice with ethyl acetate (80 mL) and water (80 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The residue was purified by basic silica gel (60-120 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→10 / 90→20 / 80→30 / 70→40 / 60) to afford the title compound F1 as a pale yellow solid (0.8 g, 48%). 1 H NMR (500 MHz, DMSO-d6): δ 8.77 (d, 1H), 7.97 (dd, 1H), 6.40 (d, 1H), 4.14 (t, 2H), 3.54 (s, 4H), 3.10 (t, 2H), 2.04 (m, 4H), 1.57 (s, 9H). LCMS (APCI) 401.26 m / z [M+H] + .

[0513] Step 5: Synthesis of 2-(6-fluoropyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (G5) The title compound F1 (1.14 g, 3.2 mmol) was dissolved in dichloromethane (20 mL) and cooled to 0° C. with stirring in an ice bath. 4M HCl in 1,4-dioxane (10 mL) was added to the above solution and stirring was continued at room temperature for 16 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 solution, the precipitated solid was filtered and dried without further purification to give the title compound G5 as a white solid (0.66 g, 80%). LCMS (ESI) 250.15 m / z [M+H] + .

[0514] Step 6: Synthesis of (S)-2-(6-(3-hydroxypiperidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (H7) The title compound G5 (0.65 g, 2.6 mmol), (S)-piperidin-3-ol hydrochloride (1.07 g, 7.8 mmol), and diisopropylethylamine (2.7 mL, 15.7 mmol) were suspended in ethanol (21 mL) using a microwave vial. The sealed vial was then heated at 150° C. for 2 h using a CEM microwave. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (20 mL) and filtered on a Whatmann 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 H7 as a pale yellow solid (0.75 g, 87%). 1 H NMR (500 MHz, DMSO-d6): δ 8.68 (d, 1H), 7.99 (dd, 1H), 7.79 (s, 1H), 6.90 (d, 1H), 4.90 (d, 1H), 4.23 - 3.99 (m, 2H), 3.53 - 3.47 LCMS (ESI) 331.1 m / z [M+H] + .

[0515] Step 7: Synthesis of (S)-2-(6-(3-((tert-butyldimethylsilyl)oxy)piperidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (I2) The title compound H7 (0.2 g, 0.6 mmol) and imidazole (0.74 g, 10.9 mmol) were dissolved in dimethylformamide (15 mL) with stirring in an ice bath at 0° C. tert-Butyldimethylsilyl chloride (1.81 g, 4.9 mmol) was added and stirring was continued at room temperature for 24 h. The reaction mixture was quenched with ice-cold water (100 mL) and the precipitated solid was filtered on a Whatmann filter paper and dried to give the title compound I2 as a pale yellow solid (0.23 g, 86%). 1 H NMR (500 MHz, DMSO-d6): δ 8.68 (d, 1H), 7.99 (dd, 1H), 7.80 (s, 1H), 6.90 (d, 1H), 4.09 (d, 1H), 3.87 - 3.85 (m, 1H), 3.75 - 3.70 (m, 1H), 3.5 - 3.47 (m, 2H), 3.2 - 3.17 (m, 1H), 2.98 (t, 2H), 1.88 - 1.87 (m, 1H), 1.77 - 1.75 (m, 1H), 1.55 - 1.43 (m, 2H), 0.82 (s, 9H), 0.07 (d, 6H). LCMS: (ESI) 445.38 m / z [M+H] + .

[0516] Step 8: Synthesis of (S)-2-(6-(3-((tert-butyldimethylsilyl)oxy)piperidin-1-yl)pyridin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (J2) The title compound I2 (0.24 g, 0.54 mmol), 4-iodo-1-methyl-1H-pyrazole 11 (0.34 g, 1.6 mmol), copper(I) iodide (0.051 g, 0.27 mmol), N,N'-dimethylethylenediamine (0.056 mL, 0.54 mmol), and potassium carbonate (0.15 g, 1.1 mmol) were suspended in 1,4-dioxane (25 mL). The reaction mixture was heated in an oil bath at about 120 °C for 24 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by column chromatography on basic silica gel (150 g, 100-200 mesh) using a dichloromethane / methanol gradient (100 / 0 → 99 / 1 → 98 / 2 → 97 / 3) to give the title compound J2 as a yellow solid (0.21 g, 74%). 1 H NMR (500 MHz, DMSO-d6): δ 8.70 (d, 1H), 8.04 (s, 1H), 8.01 (dd, 1H), 7.67 (s, 1H), 6.91 (d, 1H), 4.10 - 4.05 (m, 3H), 3.90 - 3.80 (m, 4H), 3.76 - 3.71 (m, 1H), 3.37 -3.34 (m, 1H), 3.23 - 3.18 (m, 3H), 1.90 - 1.76 (m, 2H), 1.55 - 1.41 (m, 1H), 1.88 (s, 1H), 0.82 (s, 9H), 0.07 (d, 6H). LCMS: (ESI) 525.34 m / z [M+H] + .

[0517] Step 9: Synthesis of (S)-2-(6-(3-hydroxypiperidin-1-yl)pyridin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (K2) The title compound J2 (0.21 g, 0.4 mmol) was dissolved in dichloromethane (8.4 mL) and cooled to 0° C. with stirring in an ice bath. 4M HCl in 1,4-dioxane (4.2 mL) was added to the above solution and stirring was continued at room temperature 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 to pH 8-9 with saturated aqueous sodium bicarbonate solution, and the precipitated solid was filtered and dried to obtain the title compound K2 as a pale yellow solid (0.16 g, 97%). 1 H NMR (500 MHz, DMSO-d6): δ 8.71 (d, 1H), 8.04 - 8.00 (m, 2H), 7.67 (s, 1H), 6.92 (d, 1H), 4.90 (d, 1H), 4.20 (d, 1H), 4.08 - 4.00 (m, 3H), 3.83 (s, 3H), 3.53 -3.50 (m, 1H), 3.20 - 3.18 (m, 4H), 2.97 - 2.93 (m, 1H), 1.90 (s, 1H), 1.75 (s, 1H), 1.43 (s, 2H). LCMS (ESI) 411.23 m / z [M+H] + .

[0518] Step 10: Synthesis of (S)-1-(5-(5-(1-methyl-1H-pyrazol-4-yl)-4-oxo-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)piperidin-3-yl methanesulfonate (L2) The title compound K2 (0.5 g, 1.2 mmol) and triethylamine (6.9 mL, 48.7 mmol) were suspended in dichloromethane (80 mL) with stirring. The mixture was cooled to 0° C. in an ice bath. Methanesulfonyl chloride (2.9 mL, 24.4 mmol) was added and stirring was continued at room temperature for 24 h. The reaction mixture was quenched with cold water (250 mL) and extracted twice with dichloromethane (250 mL). The organic phase was separated, dried over Na2SO4, filtered and the solvent was removed under reduced pressure. The residue obtained was purified by washing twice with 60% ethyl acetate in hexane (50 mL) and 60% ethyl acetate in hexane (50 mL). The solid obtained was further purified by precipitation method by dissolving in dichloromethane (25 mL) and precipitation using n-hexane (40 mL). The solid was filtered and dried to give the title compound L2 as a yellow solid (0.133 g, 22%). 1 H NMR (500 MHz, DMSO-d6): δ 8.75 (d, 1H), 8.07 (dd, 1H), 8.04 (s, 1H), 7.68 (s, 1H), 6.98 (d, 1H), 4.80 - 4.76 (m, 1H), 4.07 (t, 2H), 4.00 (dd, 1H), 3.89 - 3.87 (m, 1H), 3.83 (s, 3H), 3.73 - 3.60 (m, 2H), 3.23 (s, 3H), 3.20 (d, 2H), 3.10 (s, 1H), 2.07 - 2.01 (m, 1H), 1.89 - 1.86 (m, 1H), 1.79 - 1.74 (m, 1H), 1.65 - 1.55 (m, 1H). LCMS (ESI) 489.37 m / z [M+H] + . HPLC: 86.7% [retention time: 7.11 min, Eclipse plus C18].

[0519] Example 9 Tritiated (R)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3 H Synthesis of Compound 1)

[0520] [ka]

[0521] T is 3 It means H.

[0522] The tritium vessel was charged with the title compound H2 (0.001 g, 0.002 mmol), followed by sodium hydride (0.001 g, 0.006 mmol), dimethylformamide (0.1 mL), and iodomethane [ 3 [H] (0.1 mL, 50 mCi) was added. The vessel was sealed and the solution was stirred at room temperature for 3 h. The reaction mixture was evaporated under reduced pressure. The material was purified on a silica gel column. The mobile phase was removed under reduced pressure and the product was redissolved in 0.05% TFA in water / acetonitrile. The material was further purified by reverse phase HPLC. The mobile phase was removed under reduced pressure and the product was redissolved in ethanol and 3 H-Compound 1 (6 mCi, SA 75.83 Ci / mmol, purity 99%) was obtained.

[0523] Example 10 Tritiated (R)-2-(6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-5-(1-methyl-1H-pyrazol-4-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3 Synthesis of H-compound 2)

[0524] [ka]

[0525] T is 3 It means H.

[0526] The tritium vessel was charged with the title compound H3 (0.001 g, 0.002 mmol), followed by sodium hydride (0.001 g, 0.006 mmol), dimethylformamide (0.1 mL), and iodomethane [ 3[H] (0.1 mL, 50 mCi) was added. The vessel was sealed and the solution was stirred at room temperature for 4 h. The reaction mixture was evaporated under reduced pressure. The material was purified on a silica gel column. The mobile phase was removed under reduced pressure and the product was redissolved in 0.05% TFA in water / acetonitrile. The material was further purified by reverse phase HPLC. The mobile phase was removed under reduced pressure and the product was redissolved in ethanol and 3 H-Compound 2 (6.5 mCi, SA 81.5 Ci / mmol, purity 99%) was obtained.

[0527] Example 11 Tritiated (5-(3-amino-4-fluorophenyl)-2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3 Synthesis of H-compound 3)

[0528] [ka]

[0529] T is 3 It means H.

[0530] A tritium vessel was charged with 10% Pd on carbon (0.002 g, 0.005 mmol), followed by a solution of H4 (0.002 g, 0.003 mmol) in dimethylformamide (3 mL) and N,N-diisopropylethylamine (DIEA) (5 μL). The vessel was attached to a tritium line and pressurized to 0.5 atm with tritium gas at -200 °C. The solution was stirred at room temperature for 3 h, then cooled to -200 °C and excess gas was removed. The reaction mixture was evaporated under reduced pressure. The material was purified on a silica gel column. The mobile phase was removed under reduced pressure and the product was redissolved in 0.05% TFA in water / acetonitrile. The material was further purified by reverse phase HPLC. The mobile phase was removed under reduced pressure and the product was redissolved in ethanol to give 1.0 mL of 100% TFA. 3 H-Compound 3 (2.2 mCi, SA 55.8 Ci / mmol, purity 99%) was obtained.

[0531] Example 12 Fluorinated (5-(3-amino-4-fluorophenyl)-2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 18 F. Synthesis of Compound 1)

[0532] [ka]

[0533] Drying 18 A solution of (S)-1-(5-(5-(1-methyl-1H-pyrazol-4-yl)-4-oxo-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)pyrrolidin-3-yl methanesulfonate L1 (1 mg in 1 mL of anhydrous DMSO) was added to the microwave vial containing [F] fluoride and the vessel was heated at 110° C. for 15 min. The reaction vessel was cooled to 40° C. and the mixture was diluted with HPLC buffer (4 mL). The resulting mixture was passed through a Sep-Pak® Alumina-N Light cartridge. The crude material was purified by semi-preparative HPLC and the collected purified fractions were passed through a Strata® C18-E cartridge. The final product, 18 F-Compound 1 was filtered through a sterile filter onto the final product vial inside the dispensing hot cell. The final product vial was assayed and a sample was removed for QC testing. The identity of the product was confirmed by 19 F-Confirmed by co-injection with a sample of the reference compound.

[0534] Synthesis of the building block (S)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 4)

[0535] [ka]

[0536] Step 1: Synthesis of (S)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 4) Compound F1 (0.8 g, 2.29 mmol), (S)-3-fluoropyrrolidine hydrochloride (0.575 g, 4.6 mmol), and DIEA (1.6 mL, 1.18 g, 9.16 mmol) were suspended in n-butanol (15 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 160° C. for 2 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (20 mL) and filtered on a Whatmann 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, Preparation 4, as an off-white solid (0.7 g, 96%). 1 H NMR (400 MHz, DMSO-d6): δ 8.73 (d, 1H), 8.05 (dd, 1H), 7.80 (s, 1H), 6.63 (d, 1H), 5.47 (m, 1H), 3.73 (m, 3H), 3.5(m, 3H), 2.98 (t, 2H), 2.23 (m, 2H). LCMS (ESI) 319.00 m / z [M+H] + .

[0537] Synthesis of the building block (S)-2-(4-(3-fluoropyrrolidin-1-yl)phenyl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 5)

[0538] [ka]

[0539] Step 1: Synthesis of tert-butyl 2-(4-chlorophenyl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate To a degassed solution of N,N-dimethylformamide (12.5 mL) was added compound D (0.5 g, 1.5 mmol), 4-chlorophenylboronic acid (1.17 g, 7.5 mmol), cesium carbonate (0.975 g, 3.0 mmol), copper(I) chloride (0.15 g, 1.5 mmol), palladium acetate (0.017 g, 0.075 mmol), and 1,1'-bis(diphenylphosphino)ferrocene (0.083 g, 0.15 mmol). The reaction mixture was heated in a sand bath at about 90° C. for 5 h. The reaction mixture was diluted with ethyl acetate (200 mL) and water (120 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The residue was purified by silica gel (100-200 mesh) column chromatography using ethyl acetate / n-hexane gradient (0 / 100→3 / 97→5 / 95) to afford the title compound as a white solid (0.23 g, 42%). 1 H NMR (500 MHz DMSO-d6): δ 8.05 (d, 2H), 7.62 (d, 2H), 4.08 (t, 2H), 3.15 (t, 2H), 1.49 (s, 9H). LCMS (ESI) 308.8 m / z [M+H-Boc] + .

[0540] Step 2: Synthesis of (S)-2-(4-(3-fluoropyrrolidin-1-yl)phenyl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 5) To the degassed 1,4-dioxane solution (10 mL) was added the title compound from step 1 above (0.1 g, 0.275 mmol), (S)-3-fluoropyrrolidine hydrochloride (0.07 g, 0.55 mmol), palladium(II) acetate (0.006 g, 0.027 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (0.026 g, 0.054 mmol), and cesium carbonate (0.36 g, 1.1 mmol). The reaction mixture was heated in a sand bath at about 120° C. for 16 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel (100-200 mesh) column chromatography using an ethyl acetate / n-hexane gradient (0 / 100→10 / 90→15 / 85→20 / 80) to give Boc-Preparation 5 as a yellow solid (0.03 g, 26%) and Preparation 5 as a yellow solid (0.03 g, 34%). Boc-Preparation 5: 1 H NMR (500 MHz DMSO-d6): δ 7.81 (d, 2H), 6.64 (d, 2H), 5.45 (d, 1H), 4.0 (t, 2H), 3.49 (m, 4H), 3.03 (t, 2H), 2.2 (m, 2H), 1.45 (s, 9H). LCMS (ESI) 418 m / z [M+H] + Preparation 5: 1 H NMR (500 MHz DMSO-d6): δ 7.84 (dd, 2H), 6.66 (d, 2H), 5.48 (d, 1H), 3.56 (m, 5H), 3.41 (q, 1H), 2.96 (t, 2H), 2.23 (m, 2H). LCMS (ESI) 317.9 m / z [M+H] + .

[0541] Synthesis of the building block (R)-2-(4-(3-fluoropyrrolidin-1-yl)phenyl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 6)

[0542] [ka]

[0543] Step 1: Synthesis of tert-butyl (R)-2-(4-(3-fluoropyrrolidin-1-yl)phenyl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate To a degassed toluene solution (5 mL) was added the title compound of step 1 of the synthesis of building block Preparation 5 (0.04 g, 0.1 mmol), (R)-3-fluoropyrrolidine hydrochloride (0.028 g, 0.21 mmol), tris(dibenzylideneacetone)dipalladium(0) (0.01 g, 0.01 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.012 g, 0.02 mmol), and cesium carbonate (0.1 g, 0.3 mmol). The reaction mixture was heated in a sand bath at about 100° C. for 8 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel (100-200 mesh) column chromatography using an ethyl acetate / n-hexane gradient (0 / 100→5 / 95→10 / 90) to give the title compound as a yellow solid (0.015 g, 33%). 1 H NMR (500 MHz DMSO-d6): δ 7.85 (d, 2H), 6.68 (d, 2H), 5.49 (d, 1H), 4.04 (t, 2H), 3.59 (m, 3H), 3.43 (m, 1H), 3.07 (t, 2H), 2.28 (m, 2H), 1.49 (s, 9H). LCMS (ESI) 362.0 m / z [M+H-Boc] + .

[0544] Step 2: Synthesis of (R)-2-(4-(3-fluoropyrrolidin-1-yl)phenyl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 6) The title compound from step 1 above (0.04 g, 0.096 mmol) was dissolved in CH2Cl2 (2 mL) and cooled to 0°C with stirring in an ice bath. 4M HCl in 1,4-dioxane (0.5 mL) was then added and the reaction mixture was stirred at room temperature for 4 hours. 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 to give Preparation 6 as a pale yellow solid (0.022 g, 73%). 1 H 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) 317.38 [M+H] + .

[0545] (Examples 13 to 28) Following the coupling procedures reported for the preparation of Example 1 utilizing the building blocks and halogen derivatives shown in Table 1, the following compounds were prepared.

[0546] [Table 2A]

[0547] [Table 2B]

[0548] [Table 2C]

[0549] [Table 2D]

[0550] [Table 2E]

[0551] Synthesis of the building block (S)-2-(6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 7)

[0552] [ka]

[0553] Step 1: (S)-2-(6-(3-fluoropiperidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 7) Compound F1 (0.2 g, 0.57 mmol), (S)-3-fluoropiperidine hydrochloride (0.158 g, 1.177 mmol), and DIEA (0.4 mL, 0.29 g, 2.29 mmol) were suspended in n-butanol (10 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 170° C. for 2.5 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (20 mL) and filtered on a Whatmann 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, Preparation 7, as a pale yellow solid (0.16 g, 84%). 1 H NMR (400 MHz, DMSO-d6): δ 8.7 (d, 1H), 8.02 (dd, 1H), 7.81 (s, 1H), 6.98 (d, 1H), 4.8 (m, 1H), 4.08 (m, 1H), 3.92 (td, 1H), 3.72(qd, 1H), 3.49 (td, 2H), 3.42 (t, 1H), 2.98 (t, 2H), 1.92 (m, 2H), 1.76 (tt, 1H).1.56 (m, 1H). LCMS (ESI) 333.1 m / z [M+H] + .

[0554] (Examples 29 to 33) Following the coupling procedures reported in Example 1 utilizing the building blocks and halogen derivatives shown in Table 2, the following compounds were prepared.

[0555] [Table 3A]

[0556] [Table 3B]

[0557] [Table 3C]

[0558] Synthesis of the building block 2-(5-(pyrrolidin-1-yl)pyridin-2-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 8)

[0559] [ka]

[0560] Step 1: Synthesis of tert-butyl 2-(5-chloropyridin-2-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate To a degassed solution of N,N-dimethylformamide (9.2 mL) was added compound D (0.3 g, 0.9 mmol), (5-chloropyridin-2-yl)boronic acid (0.43 g, 1.8 mmol), cesium carbonate (0.588 g, 1.62 mmol), copper(I) chloride (0.089 g, 0.9 mmol), palladium(II) acetate (0.01 g, 0.046 mmol), and 1,1'-bis(diphenylphosphino)ferrocene (0.05 g, 0.092 mmol). The reaction mixture was heated in a sand bath at about 85° C. for 5 hours. The reaction mixture was diluted with ethyl acetate (200 mL) and water (80 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica (50 g HP-Ultra) using a Biotage Isolera system using an ethyl acetate / n-heptane gradient (5 / 95→10 / 90→20 / 80→30 / 70→40 / 60) to afford the title compound as a white solid (0.237 g, 72%). 1 H NMR (400 MHz, DMSO-d6): δ 8.79 (d, 1H), 8.20-8.15 (m, 2H), 4.10 (t, 2H), 3.17 (t, 2H), 1.50 (s, 9H). LCMS (ESI) 309.94 / 311.96 m / z [M + H-C4H8] + .

[0561] Step 2: Synthesis of 2-(5-(pyrrolidin-1-yl)pyridin-2-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 8) To a degassed 1,4-dioxane solution (10 mL) was added the title compound from step 1 above (0.170 g, 0.464 mmol), pyrrolidine (0.497 g, 0.7 mmol), 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (0.058 g, 0.093 mmol), tris(dibenzylideneacetone)dipalladium(0) (0.0425 g, 0.0464 mmol), and cesium carbonate (0.452 g, 1.39 mmol). The reaction mixture was heated at 120 °C for 3 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by chromatography on silica (100-200 mesh) using a MeOH / CH2Cl2 gradient (1 / 99 to 2 / 98) to afford the title compound, Preparation 8, as a yellow solid (0.07 g, 50%). 1 H NMR (500 MHz, DMSO-d6): δ 8.01 (d, 1H), 7.93 (d, 1H), 7.77 (s, 1H), 7.0 (dd, 1H), 3.49 (t, 2H), 3.36 (t, 4H), 2.97 (t, 2H), 1.99 (m, 4H). LCMS (ESI) 301.13 m / z [M+H] + .

[0562] (Example 34) Following the coupling procedures reported in Example 1 using the building blocks and halogen derivatives shown in Table 3, the following compounds were prepared.

[0563] [Table 4]

[0564] Synthesis of the building block (R)-2-(5-(3-fluoropyrrolidin-1-yl)pyrazin-2-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 9)

[0565] [ka]

[0566] Step 1: Synthesis of (R)-2-bromo-5-(3-fluoropyrrolidin-1-yl)pyrazine Commercially available 2,5-dibromopyrazine (0.2 g, 8.4 mmol), (R)-3-fluoropyrrolidine hydrochloride (2.6 g, 21.0 mmol), and Et3N (6.0 mL, 4.35 g, 43 mmol) were suspended in n-butanol (20 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 120° C. for 3 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (70 mL) and filtered on a Whatmann filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound as an off-white solid (1.9 g, 93%). 1 H NMR (400 MHz DMSO-d6): δ 8.31 (S,1H), 7.65 (d,1H), 5.40 (dt,1H), 3.83 (q,1H), 3.68 (m,1H), 3.58 (td,2H), 2.44 (m,1H), 2.17 (m,1H). LCMS (ESI) 245.70 / 247.7 m / z [M+H] + .

[0567] Step 2: Synthesis of (R)-2-(3-fluoropyrrolidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazine The title compound from step 1 above (1.9 g, 7.75 mmol) was dissolved in 1,4-dioxane (40 mL) and the mixture was degassed by passing a stream of nitrogen through it. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.63 g, 0.77 mmol), bispinacolatodiboron (3.9 g, 15.5 mmol), and potassium acetate (1.52 g, 15.5 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was washed three times with n-hexane (100 mL) and concentrated under reduced pressure to give the title compound as a brown solid (5 g, quantitative). LCMS (ESI) 294.28 m / z [M+H] + .

[0568] Step 3: Synthesis of tert-butyl (R)-2-(5-(3-fluoropyrrolidin-1-yl)pyrazin-2-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate Compound D (0.5 g, 1.5 mmol) was dissolved in 1,4-dioxane (25 mL) and the solution was degassed by a stream of nitrogen. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.12 g, 0.15 mmol), the title compound from step 2 above (1.9 g, 6.6 mmol), and tripotassium phosphate (1.0 g, 4.5 mmol) were then added and the reaction mixture was heated in an oil bath at 80° C. for 5 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel (basic, 100-200 mesh) column chromatography using an ethyl acetate / n-hexane gradient (0 / 100→5 / 95→15 / 85→20 / 80→30 / 70) to give the title compound as a pale yellow solid (0.25 g, 41%). 1 H NMR (500 MHz DMSO-d6): δ 8.81 (S,1H), 8.14 (s,1H), 5.51 (d,1H), 4.06 (t,2H), 3.82 (m,3H), 3.58 (m,1H), 3.11 (t,2H), 2.24 (m,2H), 1.47 (s,9H). LCMS (ESI) 363.9 m / z [M+H] + .

[0569] Step 4: Synthesis of (R)-2-(5-(3-fluoropyrrolidin-1-yl)pyrazin-2-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 9) The title compound from step 3 above (0.25 g, 0.59 mmol) was dissolved in CH2Cl2 (10 mL) and the mixture was cooled to 0° C. in an ice bath. 4M HCl in 1,4-dioxane (2.5 mL) was then added and stirring was continued at room temperature 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 to give the title compound, Preparation 9, as a pale yellow solid (0.17 g, 89%). 1 H NMR (500 MHz DMSO-d6): δ 8.79 (S, 1H), 8.12 (s, 1H), 7.83 (s, 1H), 5.51 (d, 1H), 3.81 (m, 3H), 3.57 (m, 1H), 3.15 (td, 2H), 3.00 (t, 2H), 2.25 (m, 2H). LCMS (ESI) 319.9 m / z [M+H] + .

[0570] Synthesis of the building block (S)-2-(5-(3-fluoropyrrolidin-1-yl)pyrazin-2-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 10)

[0571] [ka]

[0572] Step 1: Synthesis of (S)-2-bromo-5-(3-fluoropyrrolidin-1-yl)pyrazine Commercially available 2,5-dibromopyrazine (2.0 g, 8.4 mmol), (R)-3-fluoropyrrolidine hydrochloride (2.6 g, 21.0 mmol), and Et3N (6.0 mL, 4.35 g, 43 mmol) were suspended in n-butanol (20 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 120° C. for 3 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (70 mL) and filtered on a Whatmann filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound as an off-white solid (1.9 g, 95%). 1 H NMR (500 MHz DMSO-d6): δ 8.21 (S, 1H), 7.64 (s, 1H), 5.47 (m, 1H), 3.66 (m, 3H), 3.44 (dt, 1H), 2.22 (m, 2H). LCMS (ESI) 246.05 / 248.05 m / z [M+H] + .

[0573] Step 2: Synthesis of (S)-2-(3-fluoropyrrolidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazine The title compound from step 1 above (0.97 g, 3.95 mmol) was dissolved in 1,4-dioxane (20 mL) and the mixture was degassed by passing a stream of nitrogen through it. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.323 g, 0.39 mmol), bispinacolatodiboron (2.0 g, 7.9 mmol), and potassium acetate (0.78 g, 7.9 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was washed three times with n-hexane (100 mL) and concentrated under reduced pressure to give the title compound as a brown solid (1.4 g, quantitative). LCMS (ESI) 294.16 m / z [M+H] + .

[0574] Step 3: Synthesis of tert-butyl (S)-2-(5-(3-fluoropyrrolidin-1-yl)pyrazin-2-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate Compound D (1.0 g, 3.0 mmol) was dissolved in 1,4-dioxane (50 mL) and the solution was degassed by a stream of nitrogen. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.24 g, 0.3 mmol), the title compound from step 2 above (2.28 g, 7.8 mmol), and tripotassium phosphate (1.9 g, 9.0 mmol) were then added and the reaction mixture was heated in an oil bath at 80° C. for 5 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel (basic, 100-200 mesh) column chromatography using an ethyl acetate / n-hexane gradient (0 / 100→15 / 85→20 / 80→25 / 75) to give the title compound as a pale yellow solid (0.13 g, 10%). 1 H NMR (400 MHz DMSO-d6): δ 8.81 (S, 1H), 8.15 (s, 1H), 5.52 (d, 1H), 4.06 (t, 2H), 3.82 (m, 3H), 3.58 (m, 1H), 3.11 (t, 2H), 2.23 (m, 2H), 1.49 (s, 9H). LCMS (ESI) 420.1 m / z [M+H] + .

[0575] Step 4: Synthesis of (S)-2-(5-(3-fluoropyrrolidin-1-yl)pyrazin-2-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 10) The title compound from step 3 above (0.13 g, 0.31 mmol) was dissolved in CH2Cl2 (6.5 mL) and the mixture was cooled to 0° C. in an ice bath. 4M HCl in 1,4-dioxane (1.3 mL) was then added and stirring was continued at room temperature 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 to give the title compound, Preparation 10, as a pale yellow solid (0.07 g, 71%). 1 H NMR (500 MHz DMSO-d6): δ 8.79 (d, 1H), 8.12 (d, 1H), 7.83 (s, 1H), 5.51 (d, 1H), 3.80 (m, 3H), 3.54 (m, 3H), 3.00 (t, 2H), 2.21 (m, 2H). LCMS (ESI) 319.95 m / z [M+H] + .

[0576] Synthesis of the building block 2-(5-(4-fluoropiperidin-1-yl)pyrazin-2-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 11)

[0577] [ka]

[0578] Step 1: Synthesis of 2-bromo-5-(4-fluoropiperidin-1-yl)pyrazine Commercially available 2,5-dibromopyrazine (2.0 g, 8.4 mmol), 4-fluoropiperidine hydrochloride (2.8 g, 21.0 mmol), and Et3N (6.0 mL, 4.2 g, 42 mmol) were suspended in n-butanol (20 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 120° C. for 3 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (70 mL) and filtered on a Whatmann filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound as an off-white solid (2.15 g, 98%). 1H NMR (500 MHz DMSO-d6): δ 8.21 (d, 1H), 4.9 (m, 1H), 3.72 (m, 2H), 3.56 (m, 2H), 1.93 (m, 2H) 1.72 (m, 2H). LCMS (ESI) 260.15 / 262.15 m / z [M+H] + .

[0579] Step 2: Synthesis of 2-(4-fluoropiperidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazine The title compound from step 1 above (2.2 g, 8.5 mmol) was dissolved in 1,4-dioxane (44 mL) and the mixture was degassed by passing a stream of nitrogen through it. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.704 g, 0.85 mmol), bispinacolatodiboron (4.2 g, 16.6 mmol), and potassium acetate (1.6 g, 16.6 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was washed three times with n-hexane (100 mL) and concentrated under reduced pressure to give the title compound as a brown solid (4.3 g, quantitative). LCMS (ESI) 308.15 m / z [M+H] + .

[0580] Step 3: Synthesis of tert-butyl 2-(5-(4-fluoropiperidin-1-yl)pyrazin-2-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate Compound D (0.2 g, 0.6 mmol) was dissolved in 1,4-dioxane (10 mL) and the solution was degassed by a stream of nitrogen. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.049 g, 0.06 mmol), the title compound from step 2 above (0.6 g, 1.98 mmol), and tripotassium phosphate (0.382 g, 1.8 mmol) were then added and the reaction mixture was heated in an oil bath at 80° C. for 8 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel (basic, 100-200 mesh) column chromatography using an ethyl acetate / n-hexane gradient (0 / 100→15 / 85→20 / 80) to give the title compound as a pale yellow solid (0.19 g, 73%). 1 H NMR (500 MHz DMSO-d6): δ 8.79 (S, 1H), 8.48 (s, 1H), 4.95 (d, 1H), 4.06 (t, 2H), 3.93 (s, 1H), 3.82 (m, 4H), 3.11 (t, 2H), 1.97 (m, 2H), 1.79 (d, 2H), 1.49 (s, 9H). LCMS (ESI) 434.35 m / z [M+H] + .

[0581] Step 4: Synthesis of 2-(5-(4-fluoropiperidin-1-yl)pyrazin-2-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 11) The title compound from step 3 above (0.19 g, 0.438 mmol) was dissolved in CH2Cl2 (9.5 mL) and the mixture was cooled to 0° C. in an ice bath. 4M HCl in 1,4-dioxane (1.9 mL) was then added and stirring was continued at room temperature 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 to give the title compound, Preparation 11, as a pale yellow solid (0.12 g, 82%). 1H NMR (500 MHz DMSO-d6): δ 8.77 (d, 1H), 8.46 (t, 1H), 7.85 (s, 1H), 4.95 (m, 1H), 3.86 (t, 2H), 3.75 (m, 2H), 3.50 (td, 2H), 3.00 (t, 2H), 1.97 (m, 2H), 1.78 (d, 2H). LCMS (ESI) 334.3 m / z [M+H] + .

[0582] Synthesis of the building block (S)-2-(5-(3-fluoropiperidin-1-yl)pyrazin-2-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 12)

[0583] [ka]

[0584] Step 1: Synthesis of (S)-2-bromo-5-(3-fluoropiperidin-1-yl)pyrazine Commercially available 2,5-dibromopyrazine (1.0 g, 4.2 mmol), (S)-3-fluoropiperidine hydrochloride (1.4 g, 10.5 mmol), and Et3N (3.0 mL, 2.1 g, 21 mmol) were suspended in n-butanol (10 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 120° C. for 3 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (70 mL) and filtered on a Whatmann filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound as an off-white solid (0.98 g, 95%). 1 H NMR (500 MHz DMSO-d6): δ 8.19 (d, 2H), 4.80 (m, 1H), 4.00 (m, 1H), 3.83 (dt, 1H), 3.60 (qd, 1H), 3.35 (d, 1H), 1.90 (m, 2H), 1.74 (m, 1H), 1.55 (m, 1H). LCMS (ESI) 260.05 / 262.05m / z [M+H]+ .

[0585] Step 2: Synthesis of (S)-2-(3-fluoropiperidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazine The title compound from step 1 above (2.2 g, 8.46 mmol) was dissolved in 1,4-dioxane (44 mL) and the mixture was degassed by passing a stream of nitrogen through it. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.7 g, 0.846 mmol), bispinacolatodiboron (4.2 g, 16.6 mmol), and potassium acetate (1.6 g, 16.6 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was washed three times with n-hexane (100 mL) and concentrated under reduced pressure to give the title compound as a brown solid (4.3 g, quantitative). LCMS (ESI) 308.15 m / z [M+H] + .

[0586] Step 3: Synthesis of tert-butyl (S)-2-(5-(3-fluoropiperidin-1-yl)pyrazin-2-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate Compound D (0.2 g, 0.6 mmol) was dissolved in 1,4-dioxane (10 mL) and the solution was degassed by a stream of nitrogen. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.049 g, 0.06 mmol), the title compound from step 2 above (0.6 g, 1.98 mmol), and tripotassium phosphate (0.382 g, 1.8 mmol) were then added and the reaction mixture was heated in an oil bath at 80° C. for 8 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel (basic, 100-200 mesh) column chromatography using an ethyl acetate / n-hexane gradient (0 / 100→15 / 85→20 / 80) to give the title compound as a pale yellow solid (0.19 g, 73%). 1H NMR (500 MHz DMSO-d6): δ 8.79 (S, 1H), 8.48 (s, 1H), 4.95 (d, 1H), 4.06 (t, 2H), 3.93 (s, 1H), 3.82 (m, 4H), 3.11 (t, 2H), 1.97 (m, 2H), 1.79 (d, 2H), 1.49 (s, 9H). LCMS (ESI) 434.35 m / z [M+H] + .

[0587] Step 4: Synthesis of (S)-2-(5-(3-fluoropiperidin-1-yl)pyrazin-2-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 12) The title compound from step 3 above (0.19 g, 0.438 mmol) was dissolved in CH2Cl2 (9.5 mL) and the mixture was cooled to 0° C. in an ice bath. 4M HCl in 1,4-dioxane (1.9 mL) was then added and stirring was continued at room temperature 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 to give the title compound, Preparation 12, as a pale yellow solid (0.12 g, 82%). 1 H NMR (500 MHz DMSO-d6): δ 8.77 (d, 1H), 8.46 (t, 1H), 7.85 (s, 1H), 4.95 (m, 1H), 3.86 (t, 2H), 3.75 (m, 2H), 3.50 (td, 2H), 3.00 (t, 2H), 1.97 (m, 2H), 1.78 (d, 2H). LCMS (ESI) 334.3 m / z [M+H] + .

[0588] (Examples 35 to 41) Following the coupling procedures reported in Example 1 using the building blocks and halogen derivatives shown in Table 4, the following compounds were prepared.

[0589] [Table 5A]

[0590] [Table 5B]

[0591] Synthesis of the building block 2-(2-(pyrrolidin-1-yl)pyrimidin-5-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 13)

[0592] [ka]

[0593] Step 1: Synthesis of 5-bromo-2-(pyrrolidin-1-yl)pyrimidine Commercially available 2-iodo-5-bromopyrimidine (1.1 g, 3.8 mmol) and pyrrolidine (3 ml, 2.7 g, 3.8 mmol) were suspended in n-butanol (6 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 120° C. for 1 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (70 mL) and filtered on a Whatmann filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound as an off-white solid (0.88 g, quantitative). 1 H NMR (500 MHz DMSO-d6): δ 1 H NMR (500 MHz DMSO-d6): δ 8.29 (s, 2H), 3.53 (m, 4H), 2.0 (m, 4H).

[0594] Step 2: Synthesis of 2-(pyrrolidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine The title compound from step 1 above (2.5 g, 10.9 mmol) was dissolved in 1,4-dioxane (18 mL) and the mixture was degassed by passing a stream of nitrogen through it. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.89 g, 1.09 mmol), bispinacolatodiboron (5.56 g, 22 mmol), and potassium acetate (2.16 g, 22 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 3 h. The reaction mixture was washed three times with n-hexane (100 mL) and concentrated under reduced pressure to give the title compound as a brown solid (5.0 g, quantitative). LCMS (ESI) 276.1 m / z [M+H] + .

[0595] Step 3: Synthesis of tert-butyl 4-oxo-2-(2-(pyrrolidin-1-yl)pyrimidin-5-yl)-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate Compound D (0.7 g, 2.1 mmol) was dissolved in 1,4-dioxane (25 mL) and the solution was degassed by a stream of nitrogen. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.17 g, 0.2 mmol), the title compound from step 2 above (1.0 g, 3.6 mmol), and cesium carbonate (2.05 g, 6.3 mmol) were then added and the reaction mixture was heated in an oil bath at 95-100 °C for 5 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel (basic, 100-200 mesh) column chromatography using an ethyl acetate / n-hexane gradient (0 / 100 → 10 / 90 → 20 / 80 → 30 / 70 → 40 / 60) to give the title compound as a pale yellow solid (0.32 g, 20%). 1 H NMR (500 MHz DMSO-d6): δ 8.86 (s, 2H), 4.15 (t, 2H), 3.66 (m, 4H), 3.12 (t, 2H), 2.04 (m, 4H), 1.57(s, 9H), 1.55 (s, 2H). LCMS (ESI) 402.28 m / z [M+H] + .

[0596] Step 4: Synthesis of 2-(2-(pyrrolidin-1-yl)pyrimidin-5-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 13) The title compound from step 3 above (0.32 g, 0.8 mmol) was dissolved in CH2Cl2 (10 mL) and the mixture was cooled to 0° C. in an ice bath. 4M HCl in 1,4-dioxane (3 mL) was then added and stirring was continued at room temperature for 4 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 to give the title compound, Preparation 13, as a pale yellow solid (0.2 g, 83%). 1 H NMR (400 MHz DMSO-d6): δ 8.90 (s, 2H), 7.85 (s, 1H), 3.57 (t, 4H), 3.49 (dt, 2H), 2.99 (t, 2H), 1.96 (m, 4H). LCMS (ESI) 302.14 m / z [M+H] + .

[0597] Synthesis of the building block (R)-2-(2-(3-fluoropyrrolidin-1-yl)pyrimidin-5-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 14)

[0598] [ka]

[0599] Step 1: Synthesis of (R)-5-bromo-2-(3-fluoropyrrolidin-1-yl)pyrimidine Commercially available 2-iodo-5-bromopyrimidine (2.0 g, 7.02 mmol), (R)-3-fluoropyrrolidine hydrochloride (1.76 g, 1.4 mmol), and Et3N (3.9 mL, 2.84 g, 2.8 mmol) were suspended in ethanol (12 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 120° C. for 1 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (70 mL) and filtered on a Whatmann filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound as an off-white solid (1.65 g, 97%). 1 H NMR (400 MHz DMSO-d6): δ 8.47 (s, 1H), 5.43 (m, 1H), 3.68 (m, 2H), 3.46 (dt, 1H), 2.18 (m, 2H). LCMS (ESI) 245.70 / 247.75 m / z [M+H] + .

[0600] Step 2: Synthesis of (R)-2-(3-fluoropyrrolidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine The title compound from step 1 above (1.65 g, 6.7 mmol) was dissolved in 1,4-dioxane (33 mL) and the mixture was degassed by passing a stream of nitrogen through it. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.89 g, 1.09 mmol), bispinacolatodiboron (5.56 g, 22 mmol), and potassium acetate (2.16 g, 22 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was washed three times with n-hexane (100 mL) and concentrated under reduced pressure to give the title compound as a brown solid (5.0 g, quantitative). LCMS (ESI) 294.16 m / z [M+H] + .

[0601] Step 3: Synthesis of tert-butyl (R)-2-(2-(3-fluoropyrrolidin-1-yl)pyrimidin-5-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate Compound D (0.5 g, 1.5 mmol) was dissolved in 1,4-dioxane (25 mL) and the solution was degassed by a stream of nitrogen. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.122 g, 0.15 mmol), the title compound from step 2 above (1.8 g, 6.3 mmol), and cesium carbonate (1.4 g, 4.5 mmol) were then added and the reaction mixture was heated in an oil bath at 100°C for 5 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel (basic, 100-200 mesh) column chromatography using an ethyl acetate / n-hexane gradient (0 / 100→10 / 90→20 / 80→30 / 70) to give the title compound as a pale yellow solid (0.24 g, 38%). 1 H NMR (500 MHz DMSO-d6): δ 8.97 (d, 2H), 5.47 (m, 1H), 4.07 (m, 2H), 3.90 (m, 2H), 3.75 (ddd, 1H), 3.59 (td, 1H), 3.16 (t, 1H), 3.11 (t, 2H), 2.23 (m, 2H), 1.49 (s, 9H). LCMS (ESI) 428.8 m / z [M+H] +

[0602] Step 4: Synthesis of (R)-2-(2-(3-fluoropyrrolidin-1-yl)pyrimidin-5-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 14) The title compound from step 3 above (0.24 g, 0.5 mmol) was dissolved in CH2Cl2 (5 mL) and the mixture was cooled to 0° C. in an ice bath. 4M HCl in 1,4-dioxane (2.5 mL) was then added and stirring was continued at room temperature 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 to give the title compound, Preparation 14, as a pale yellow solid (0.135 g, 74%). 1 H NMR (500 MHz DMSO-d6): δ 8.94 (d, 2H), 7.86 (s, 1H), 5.47 (d, 1H), 3.89 (m, 2H), 3.74 (ddd, 1H), 3.58 (td, 1H), 3.50 (td, 2H), 3.00 (t, 2H), 2.24 (m, 2H). LCMS (ESI) 320 m / z [M+H] + .

[0603] Synthesis of the building block (S)-2-(2-(3-fluoropyrrolidin-1-yl)pyrimidin-5-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 15)

[0604] [ka]

[0605] Step 1: Synthesis of (S)-5-bromo-2-(3-fluoropyrrolidin-1-yl)pyrimidine Commercially available 2-iodo-5-bromopyrimidine (1.5 g, 5.26 mmol), (S)-3-fluoropyrrolidine hydrochloride (1.32 g, 1.05 mmol), and Et3N (3.0 mL, 2.13 g, 2.1 mmol) were suspended in ethanol (9 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 120° C. for 1 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (70 mL) and filtered on a Whatmann filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound as an off-white solid (1.25 g, 97%). 1 H NMR (500 MHz DMSO-d6): δ 8.47 (s, 2H), 5.43 (m, 1H), 3.68 (m, 3H), 3.46 (m, 1H), 2.20 (m, 2H). LCMS (ESI) 246.06 / 247.91 m / z [M+H] + .

[0606] Step 2: Synthesis of (S)-2-(3-fluoropyrrolidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine The title compound from step 1 above (1.3 g, 5.3 mmol) was dissolved in 1,4-dioxane (26 mL) and the mixture was degassed by passing a stream of nitrogen through it. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.4 g, 0.53 mmol), bispinacolatodiboron (2.8 g, 10.6 mmol), and potassium acetate (1 g, 10.6 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was washed three times with n-hexane (100 mL) and concentrated under reduced pressure to give the title compound as a brown solid (4.0 g, quantitative). LCMS (ESI) 294.04 m / z [M+H] + .

[0607] Step 3: Synthesis of tert-butyl (S)-2-(2-(3-fluoropyrrolidin-1-yl)pyrimidin-5-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate Compound D (0.5 g, 1.5 mmol) was dissolved in 1,4-dioxane (25 mL) and the solution was degassed by a stream of nitrogen. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.122 g, 0.15 mmol), the title compound from step 2 above (2.0 g, 6.9 mmol), and cesium carbonate (1.4 g, 4.5 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel (basic, 100-200 mesh) column chromatography using an ethyl acetate / n-hexane gradient (0 / 100→10 / 90→20 / 80) to give the title compound as a pale yellow solid (0.27 g, 42%). 1 H NMR (400 MHz DMSO-d6): δ 8.97 (d, 2H), 5.47 (m, 1H), 4.06 (t, 2H), 3.19 (m, 2H), 3.75 (m, 2H), 3.59 (m, 1H), 3.11 (t, 2H), 2.22 (m, 2H), 1.49 (s, 9H). LCMS (ESI) 420.36 m / z [M+H] + .

[0608] Step 4: Synthesis of (S)-2-(2-(3-fluoropyrrolidin-1-yl)pyrimidin-5-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 15) The title compound from step 3 above (0.27 g, 0.62 mmol) was dissolved in CH2Cl2 (3 mL) and the mixture was cooled to 0°C in an ice bath. 4M HCl in 1,4-dioxane (2.5 mL) was then added and stirring was continued at room temperature for 2 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 to give the title compound, Preparation 15, as a pale yellow solid (0.18 g, 87%). 1 H NMR (500 MHz DMSO-d6): δ 8.94 (d, 2H), 7.86 (s, 1H), 5.47 (d, 1H), 3.89 (m, 2H), 3.75 (m, 1H), 3.58 (td, 1H), 3.50 (td, 2H), 3.00 (t, 2H), 2.21 (m, 2H),1.19 (d, 2H). LCMS (ESI) 320.26 m / z [M+H] + .

[0609] Synthesis of the building block 2-(5-(4-fluoropiperidin-1-yl)pyrazin-2-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 16)

[0610] [ka]

[0611] Step 1: Synthesis of 5-bromo-2-(4-fluoropiperidin-1-yl)pyrimidine)pyrazine Commercially available 2-iodo-5-bromopyrimidine (2.0 g, 7.02 mmol), 4-fluoropiperidine hydrochloride (1.47 g, 10.5 mmol), and DIEA (3.4 mL, 2.64 g, 2.1 mmol) were suspended in ethanol (20 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 120° C. for 1.5 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (70 mL) and filtered on a Whatmann filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound as an off-white solid (1.77 g, 97%). 1 H NMR (400 MHz DMSO-d6): δ 8.44 (s, 2H), 4.91 (m, 1H), 3.86 (m, 2H), 3.71 (m, 2H), 1.89 (m, 2H), 1.69 (m, 2H).

[0612] Step 2: Synthesis of 2-(4-fluoropiperidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine The title compound from step 1 above (1.77 g, 8.8 mmol) was dissolved in 1,4-dioxane (36 mL) and the mixture was degassed by passing a stream of nitrogen through it. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.55 g, 0.68 mmol), bispinacolatodiboron (3.4 g, 13.6 mmol), and potassium acetate (1.3 g, 13.6 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was washed three times with n-hexane (100 mL) and concentrated under reduced pressure to give the title compound as a brown solid (5.0 g, quantitative). LCMS (ESI) 307.15 m / z [M+H] + .

[0613] Step 3: Synthesis of tert-butyl 2-(2-(4-fluoropiperidin-1-yl)pyrimidin-5-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate Compound D (0.2 g, 0.6 mmol) was dissolved in 1,4-dioxane (10 mL) and the solution was degassed by a stream of nitrogen. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.05 g, 0.06 mmol), the title compound from step 2 above (0.885 g, 2.88 mmol), and cesium carbonate (0.585 g, 1.8 mmol) were then added and the reaction mixture was heated in an oil bath at 100°C for 8 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel (basic, 100-200 mesh) column chromatography using an ethyl acetate / n-hexane gradient (0 / 100→10 / 90→20 / 80) to give the title compound as a pale yellow solid (0.09 g, 34%). 1 H NMR (400 MHz DMSO-d6): δ 8.95 (s, 2H), 4.96 (dt, 1H), 4.06 (t, 2H), 3.94 (m, 5H), 3.11 (t, 2H), 1.94 (m, 2H) 1.75 (m, 2H), 1.49 (s, 9H), 1.27 (s, 1H), 1.07 (s, 4H). LCMS (ESI) 378.3 m / z [M+H-Boc] + .

[0614] Step 4: Synthesis of 2-(2-(4-fluoropiperidin-1-yl)pyrimidin-5-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 16) The title compound from step 3 above (0.09 g, 0.2 mmol) was dissolved in CH2Cl2 (4.5 mL) and the mixture was cooled to 0° C. in an ice bath. 4M HCl in 1,4-dioxane (0.9 mL) was then added and stirring was continued at room temperature for 2 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 to give the title compound, Preparation 16, as a pale yellow solid (0.064 g, 92%). 1H NMR (500 MHz DMSO-d6): δ 8.92 (s, 2H), 7.87 (s, 1H), 4.95 (m, 1H), 3.93 (m, 4H), 3.50 (td, 2H), 3.0 (t, 2H) 1.94 (qt, 2H), 1.75 (m, 2H). LCMS (ESI) 334.25 m / z [M+H] + .

[0615] Synthesis of the building block (S)-2-(2-(3-fluoropiperidin-1-yl)pyrimidin-5-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 17)

[0616] [ka]

[0617] Step 1: Synthesis of (S)-5-bromo-2-(3-fluoropiperidin-1-yl)pyrimidine Commercially available 2-iodo-5-bromopyrimidine (1.5 g, 5.28 mmol), (S)-3-fluoropiperidine hydrochloride (1.1 g, 7.92 mmol), and DIEA (2.6 mL, 1.98 g, 15.8 mmol) were suspended in ethanol (37 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 120° C. for 1.5 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (70 mL) and filtered on a Whatmann filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound as an off-white solid (1.35 g, 99%). 1 H NMR (500 MHz DMSO-d6): δ 8.44 (s, 1H), 4.77 (m, 1H), 4.23 (m, 1H), 4.05 (dt, 1H), 3.64 (qd, 1H), 3.40 (t, 1H), 1.89 (m, 2H), 1.70 (m, 1H), 1.52 (m, 1H). LCMS (ESI) 259.7 / 261.75 m / z [M+H] + .

[0618] Step 2: Synthesis of (S)-2-(3-fluoropiperidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine The title compound from step 1 above (1.35 g, 5.23 mmol) was dissolved in 1,4-dioxane (27 mL) and the mixture was degassed by passing a stream of nitrogen through it. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.42 g, 0.52 mmol), bispinacolatodiboron (2.6 g, 10. mmol), and potassium acetate (1 g, 10.4 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 5 h. The reaction mixture was washed three times with n-hexane (100 mL) and concentrated under reduced pressure to give the title compound as a brown solid (5.0 g, quantitative). LCMS (ESI) 308.28 m / z [M+H] + .

[0619] Step 3: Synthesis of tert-butyl (S)-2-(2-(3-fluoropiperidin-1-yl)pyrimidin-5-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate Compound D (1.0 g, 3 mmol) was dissolved in 1,4-dioxane (15 mL) and the solution was degassed by a stream of nitrogen. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.245 g, 0.3 mmol), the title compound from step 2 above (3.2 g, 12.6 mmol), and cesium carbonate (2.9 g, 9 mmol) were then added and the reaction mixture was heated in an oil bath at 100°C for 5 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel (basic, 100-200 mesh) column chromatography using an ethyl acetate / n-hexane gradient (0 / 100→10 / 90→20 / 80) to give the title compound as a pale yellow solid (0.98 g, 75%). 1H NMR (400 MHz DMSO-d6): δ 8.94 (s, 1H), 4.84 (m, 1H), 4.49 (m, 1H), 4.31 (td, 1H), 4.06 (t, 2H), 3.71 (dd, 1H) 3.47 (m, 1H), 3.11 (t, 2H), 1.92 (m, 2H), 1.73 (m, 1H), 1.60 (m, 1H), 1.49 (s, 9H). LCMS (ESI) 434.0 m / z [M+H] + .

[0620] Step 4: Synthesis of (S)-2-(2-(3-fluoropiperidin-1-yl)pyrimidin-5-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 17) The title compound from step 3 above (0.98 g, 2.26 mmol) was dissolved in CH2Cl2 (20 mL) and the mixture was cooled to 0° C. in an ice bath. 4M HCl in 1,4-dioxane (9.8 mL) was then added and stirring was continued at room temperature for 2 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 to give the title compound, Preparation 17, as a pale yellow solid (0.67 g, 89%). 1 H NMR (400 MHz DMSO-d6): δ 8.91 (s, 1H), 7.86 (s, 1H), 4.84 (m, 1H), 4.45 (m, 1H), 4.26 (m, 1H), 3.73 (m, 1H) 3.48 (m, 3H), 3.00 (t, 1H), 1.91 (m, 2H), 1.75 (m, 1H), 1.57 (m, 1H). LCMS (ESI) 334.0 m / z [M+H] + .

[0621] Synthesis of the building block (S)-2-(2-(3-fluoropiperidin-1-yl)pyrimidin-5-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 18)

[0622] [ka]

[0623] Step 1: Synthesis of (R)-5-bromo-2-(3-fluoropiperidin-1-yl)pyrimidine Commercially available 2-iodo-5-bromopyrimidine (1 g, 3.52 mmol), (R)-3-fluoropiperidine hydrochloride (0.73 g, 5.28 mmol), and DIEA (1.7 mL, 1.3 g, 5.28 mmol) were suspended in ethanol (25 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 120° C. for 1 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (70 mL) and filtered on a Whatmann filter paper. The solid was washed with water (20 mL) and dried under reduced pressure to give the title compound as an off-white solid (0.9 g, 99%). 1 H NMR (500 MHz DMSO-d6): δ 8.44 (s, 2H), 4.77 (m, 1H), 4.23 (m, 1H), 4.05 (td, 1H), 3.64 (qd, 1H), 3.40 (m, 1H), 1.88 (m, 2H), 1.70 (qd, 1H), 1.53 (m, 1H). LCMS (ESI) 260.00 / 261.96 m / z [M+H] + .

[0624] Step 2: Synthesis of (R)-2-(3-fluoropiperidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine The title compound from step 1 above (0.9 g, 3.48 mmol) was dissolved in 1,4-dioxane (18 mL) and the mixture was degassed by passing a stream of nitrogen through it. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.28 g, 0.34 mmol), bispinacolatodiboron (1.7 g, 6.97 mmol), and potassium acetate (0.68 g, 6.97 mmol) were then added and the reaction mixture was heated in an oil bath at 100° C. for 5 hours. The reaction mixture was washed three times with n-hexane (100 mL) and concentrated under reduced pressure to give the title compound as a brown solid (2.7 g, quantitative). LCMS (ESI) 308.44 [M+H] + .

[0625] Step 3: Synthesis of tert-butyl (R)-2-(2-(3-fluoropiperidin-1-yl)pyrimidin-5-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-carboxylate Compound D (0.5 g, 1.5 mmol) was dissolved in 1,4-dioxane (25 mL) and the solution was degassed by a stream of nitrogen. [1,1'-bis(diphenyl-phosphino)ferrocene]-dichloropalladium(II) complex with dichloromethane (0.12 g, 0.15 mmol), the title compound from step 2 above (1.8 g, 6 mmol), and cesium carbonate (1.46 g, 4.5 mmol) were then added and the reaction mixture was heated in an oil bath at 100°C for 5 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel (basic, 100-200 mesh) column chromatography using an ethyl acetate / n-hexane gradient (05 / 95→15 / 85→30 / 70) to give the title compound as a pale yellow solid (0.474 g, 73%). 1H NMR (400 MHz DMSO-d6): δ 8.94 (s, 2H), 4.84 (m, 1H), 4.49 (m, 1H), 4.31 (m, 1H), 4.06 (t, 2H), 3.71 (dd, 1H), 3.47 (m, 1H), 3.11 (t, 2H), 1.92 (m, 2H), 1.73 (m, 1H), 1.60 (m, 1H), 1.49 (s, 9H). LCMS (ESI) 434.1 [M+H] + .

[0626] Step 4: Synthesis of (R)-2-(2-(3-fluoropiperidin-1-yl)pyrimidin-5-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one (Preparation 18) The title compound from step 3 above (0.47 g, 1.08 mmol) was dissolved in CH2Cl2 (10 mL) and the mixture was cooled to 0° C. in an ice bath. 4M HCl in 1,4-dioxane (4.7 mL) was then added and stirring was continued at room temperature for 2 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 to give the title compound, Preparation 18, as a pale yellow solid (0.3 g, 83%). 1 H NMR (400 MHz DMSO-d6): δ 7.85 (s, 1H), 4.83 (m, 1H), 4.45 (m, 1H), 4.26 (m, 1H), 3.72 (dd, 1H), 3.48 (m, 3H), 3.00 (t, 2H), 1.90 (m, 2H), 1.73 (m, 1H), 1.58 (dd, 1H). LCMS (ESI) 334.0 [M+H] + .

[0627] (Examples 42 to 50) Following the coupling procedures reported in Example 1 using the building blocks and halogen derivatives shown in Table 5, the following compounds were prepared.

[0628] [Table 6A]

[0629] [Table 6B]

[0630] [Table 6C]

[0631] (Example 51) (R)-2-(5-bromo-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(5-bromopyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3 H Synthesis scheme of compound 52 (precursor)

[0632] [ka]

[0633] Step 1: Synthesis of (R)-5-(5-bromopyridin-3-yl)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one Compound Preparation 1 (0.046 g, 0.144 mmol), 3,5-dibromopyridine (0.112 g, 0-472 mmol), copper(I) iodide (0.003 g, 0.0157 mmol), N,N'-dimethylethylenediamine (0.025 mL, 0.27 mmol), and potassium carbonate (0.043 g, 0.314 mmol) were suspended in 1,4-dioxane (9 mL). The reaction mixture was heated in a sand bath at about 120° C. for 24 hours. The reaction mixture was diluted twice with ethyl acetate (200 mL) and washed with water (80 mL) and brine (80 mL). The organic phase was separated, dried over Na2SO4, filtered, and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica (25 g HP-Ultra) using a Biotage Isolera system with CH2Cl2 / MeOH (100 / 0→98 / 2→93 / 7→96 / 4→95 / 5→90 / 10) to give 3 H The precursor of compound 52 was obtained as a yellow solid (0.0814 g). The title compound was further purified on a preparative TLC plate using CH2Cl2 / MeOH (95 / 5) as the mobile phase to give the title compound as a yellow solid (0.0677 g, 49%). 1 H NMR (400 MHz, DMSO-d6): δ 8.79 (d, 1H), 8.68 (d, 1H), 8.59 (d, 1H), 8.18 (t, 1H), 8.10 (dd, 1H), 6.66 (d, 1H), 5.49 (d, 1H), 4.18 (t, 2H), 3.88-3.62 (m, 3H), 3.55-3.49 (m, 1H), 3.25 (t, 2H), 2.33-2.25 (m, 2H). LCMS (ESI) 474.01 / 475.97 m / z [M+H] +

[0634] Step 2: (R)-2-(5-bromo-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(5-bromopyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3 H Synthesis of compound 52 (precursor) The title compound from step 1 above (0.0677 g, 0.143 mmol) was dissolved in CH2Cl2 (3.5 mL) and acetonitrile (3.5 mL). The mixture was cooled to 0° C., N-bromosuccinimide (0.101 g, 0.574 mmol) was added and the reaction mixture was stirred at 0° C. for 3 h. The reaction mixture was diluted with ethyl acetate (80 mL) and washed with water / brine (40 mL; 1 / 1). The organic phase was separated, dried over Na2SO4, filtered and the solvent was removed under reduced pressure. The residue was purified by chromatography on silica (25 g Interchim) using a Biotage Isolera system using an ethyl acetate / n-heptane gradient (5 / 95→80 / 20→80 / 20) to give 3 H The precursor of compound 52 was obtained as a yellow solid. 3 The precursor of compound 52 was treated with methanol (3 mL), sonicated for 1 min, and the solvent was evaporated under reduced pressure to give 3 H The precursor of compound 52 was obtained as a yellow solid (0.0481 g, 60%). 1 H NMR (400 MHz, DMSO-d6): δ 8.77 (d, 1H), 8.68 (d, 1H), 8.60 (d, 1H), 8.37 (d, 1H), 8.18 (t, 1H), 5.45 (d, 1H), 4.19 (t, 2H), 4.07 (ddd, 1H), 3.98-3.84 (m, 3H), 3.26 (t, 2H), 2.28-2.07 (m, 2H). LCMS (ESI) 552.09 / 554.06 / 556.01 m / z [M+H] +

[0635] (Example 52) Tritiated (R)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 3 Synthesis of H-compound 52)

[0636] [ka]

[0637] Experimental procedure for exchange of halogens with tritium: The tritium reaction vessel was charged with 10% palladium on charcoal (0.5 mg), followed by 3 H A solution of compound 52 precursor (0.5 mg) in DMF (0.3 mL) was added followed by DIEA (0.005 mL). The vessel was attached to a tritium line and pressurized to 0.5 atm with tritium gas at -200 °C. The solution was stirred at room temperature for 2 h, cooled to -200 °C, and excess gas was removed. The reaction flask was rinsed with 4 x 1 mL methanol, each methanol wash passed through a Celite pad. The combined methanol was removed under reduced pressure. Crude yield: 30 mCi. The material was purified by HPLC and the mobile phase removed under reduced pressure. 3 H-Compound 52 was redissolved in absolute ethanol. Yield 15 mCi, purity >99%. Specific activity was determined by MS to be 47.8 Ci / mmol.

[0638] (Example 53) 2-Amino-3,5-dibromo-6-(2-(5-bromo-6-(pyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridin-5(4H)-yl)benzonitrile ( 3 H Synthesis of compound 16 (precursor)

[0639] [ka]

[0640] Compound 16 (0.15 g, 0.36 mmol) was dissolved in DMF (10 mL). The mixture was cooled to -50°C and N-bromosuccinimide (0.224 g, 1.26 mmol) was added in portions. The reaction mixture was allowed to warm to 0°C and then to room temperature over 1 h. The reaction mixture was diluted with ethyl acetate (80 mL) and water (100 mL). The organic phase was separated, dried over Na2SO4, filtered and the solvent was removed under reduced pressure. The residue was further purified by preparative TLC plate using CH2Cl2 / MeOH (98 / 2) as the mobile phase to give 3 H The precursor of compound 16 (0.033 g, 15%) was obtained.1 H NMR (500 MHz, DMSO-d6): δ 8.73 (d, 1H), 8.32 (d, 1H), 8.11 (s, 1H), 6.55 (d, 2H), 3.97 (m, 2H), 3.73 (s, 4H), 3.25 (m, 2H), 3.26 (m, 2H), 1.91 (s, 4H).

[0641] (Example 54) Tritium-labeled 2-amino-6-(4-oxo-2-(6-(pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-5(4H)-yl)benzonitrile ( 3 Synthesis scheme of H-compound 16)

[0642] [ka]

[0643] Experimental procedure for exchange of halogens with tritium: 3 H Compound 16 precursor (1.82 mg), 8.53 mg Pd / C (10% metal), and 10 μl DIEA were suspended in 0.4 ml DMF. The suspension was degassed three times with a high vacuum manifold and stirred under an atmosphere of tritium gas (5.9 Ci) at room temperature for 1 h. The pressure was started at 620 mbar and ended at 516 mbar at room temperature. The solvent was removed under reduced pressure and the exchange of the labile tritium was carried out by adding methanol / CHCl (0.3 mL, 1 / 1), stirring the solution, and removing the solvent again under reduced pressure. This process was repeated three times. Finally, the well-dried solid was extracted with methanol / CHCl (5 mL, 1 / 1) and the suspension was filtered through a 0.2 μm nylon membrane to obtain a clear yellow solution. The desired product was isolated from the HPLC solvent mixture by solid phase extraction. The HPLC solution was neutralized with aqueous NaHCO and the volume of the fractions was partially reduced on a rotary evaporator. The product was then extracted with a Phenomenex StrataX cartridge eluted with 5 ml of ethanol. 3H Compound 16 exhibited a radiochemical purity of >99% and the specific activity was determined to be 72.5 Ci / mmol by MS.

[0644] (Example 55) (S)-2-amino-3,5-dibromo-6-(2-(5-bromo-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridin-5(4H)-yl)benzonitrile ( 3 H Synthesis of compound 21 (precursor)

[0645] [ka]

[0646] Step 1: (S)-2-amino-3,5-dibromo-6-(2-(5-bromo-6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridin-5(4H)-yl)benzonitrile ( 3 H Synthesis of compound 21 (precursor) Compound 21 (0.08 g, 0.18 mmol) was dissolved in DMF (10 mL) and the solution was cooled to -50 °C. N-bromosuccinimide (0.131 g, 0.737 mmol) was added in portions and the reaction mixture was allowed to warm to 20 °C over 1 h. The reaction mixture was diluted with cold water (100 mL) and the precipitate was collected by filtration. The solid was purified by preparative TLC plate using CH2Cl2 / MeOH (98.5 / 1.5) as the mobile phase to give 3 H The precursor of compound 21 was obtained as a pale yellow solid (0.038 g, 32%). 1 H NMR (500 MHz DMSO-d6): δ 8.72 (d, 1H), 8.33 (d, 1H), 8.07 (s, 1H), 6.5 (s, 2H), 5.4 (d, 1H), 3.95 (m, 6H), 3.21 (m, 2H), 2.13 (m, 2H).

[0647] (Example 56) Tritium-labeled (S)-2-amino-6-(2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridin-5(4H)-yl)benzonitrile ( 3 H Synthesis scheme of compound 21)

[0648] [ka]

[0649] Experimental procedure for exchange of halogens with tritium: 3 H Compound 21 precursor (3.94 mg), 45.1 mg Lindlar catalyst (10% metal), and 20 μl DIEA were suspended in 0.5 ml DMF. The suspension was degassed three times with a high vacuum manifold and stirred under an atmosphere of tritium gas (8.7 Ci) at room temperature for 16 h. The pressure was started at 640 mbar and ended at 428 mbar at room temperature. The solvent was removed under reduced pressure and the exchange of the labile tritium was carried out by adding methanol / CHCl (0.3 mL, 1 / 1), stirring the solution, and removing the solvent again under reduced pressure. This process was repeated three times. Finally, the well-dried solid was extracted with methanol / CHCl (5 mL, 1 / 1) and the suspension was filtered through a 0.2 μm nylon membrane to give a clear yellow solution. The desired product was isolated from the HPLC solvent mixture by solid phase extraction. The HPLC solution was neutralized with aqueous NaHCO and the volume of the fractions was partially reduced on a rotary evaporator. The product was then extracted with a Phenomenex StrataX cartridge eluted with 5 ml of ethanol. 3 H Compound 21 exhibited a radiochemical purity of >99% and the specific activity was determined to be 64.4 Ci / mmol by MS.

[0650] (Example 57) (R)-1-(5-(5-(3-(bis(tert-butoxycarbonyl)amino)-4-fluorophenyl)-4-oxo-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)pyrrolidin-3-yl methanesulfonate( 18 F. Synthesis of compound 26 (precursor)

[0651] [ka]

[0652] Step 1: Synthesis of (R)-2-(6-(3-hydroxypyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one Compound F1 (0.4 g, 1.14 mmol), (R)-pyrrolidin-3-ol hydrochloride (0.424 g, 3.4 mmol), and DIEA (1.2 mL, 0.88 g, 6.8 mmol) were suspended in n-butanol (20 mL) using a microwave vial. The sealed vial was then heated in a CEM microwave at 160° C. for 2 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was suspended in water (20 mL) and filtered on a Whatmann 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 as an off-white solid (0.29 g, 80%). 1 H NMR (400 MHz, DMSO-d6): δ 8.7 (d, 1H), 8.01 (dd, 1H), 7.79 (s, 1H), 6.58 (d, 1H), 4.59 (s, 1H), 3.64 (dd, 1H), 3.52 (m, 4H), 3.3 (m, 1H), 2.98 (t, 2H), 2.02 (m, 2H), 0.86 (s, 9H), 0.2 (d, 2H). LCMS (ESI) 317.33 m / z [M+H] +

[0653] Step 2: Synthesis of (R)-2-(6-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one The title compound from step 1 above (0.33 g, 1.04 mmol) and imidazole (0.75 g, 10.4 mmol) were dissolved in DMF (12.5 mL). The reaction mixture was cooled to 0° C. in an ice bath and tert-butyldimethylsilyl chloride (0.8 g, 5.2 mmol) was added in portions. The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with cold water (150 mL) and the precipitate was collected by filtration and dried to give the title compound as a white solid (0.4 g, 83%). 1 H NMR (400 MHz, DMSO-d6): δ 8.7 (d, 1H), 8.0 (dd, 1H), 7.79 (s, 1H), 6.58 (d, 1H), 4.59 (s, 1H), 3.64 (dd, 1H), 3.52 (m, 4H), 3.33 (m, 1H), 2.98 (t, 1H), 2.02(m, 2H), 0.86 (s, 9H), 0.1 (d, 6H). LCMS (ESI) 431.41 m / z [M+H] +

[0654] Step 3: Synthesis of (R)-5-(3-amino-4-fluorophenyl)-2-(6-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one The title compound from step 2 above (0.4 g, 0.92 mmol), 5-bromo-2-fluoroaniline (0.44 g, 2.32 mmol), copper(I) iodide (0.089 g, 0.46 mmol), N,N'-dimethylethylenediamine (0.1 mL, 0.92 mmol), and potassium carbonate (0.52 g, 3.7 mmol) were suspended in 1,4-dioxane (40 mL). The reaction mixture was heated in a sand bath at about 120 °C for 24 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by silica gel (100-200 mesh) column chromatography using a MeOH / CHCl gradient (0 / 100→1 / 99→2 / 98) to afford the title compound as a pale yellow solid (0.45 g, 90%). 1 H NMR (500 MHz, DMSO-d6): δ 8.73 (d, 1H), 8.04 (dd, 1H), 7.0 (dd, 1H), 6.76 (dd, 1H), 6.6 (d, 1H), 6.49 (t, 1H), 5.22 (d, 1H), 4.59 (s, LCMS (ESI) 540.1 m / z [M+H] +

[0655] Step 4: Synthesis of tert-butyl (R)-(tert-butoxycarbonyl)(5-(2-(6-(3-((tert-butyldimethylsilyl)oxy)pyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridin-5(4H)-yl)-2-fluorophenyl)carbamate The title compound from step 3 above (0.45 g, 0.83 mmol), 4-(dimethylamino)pyridine (0.2 g, 1.6 mmol), and di-tert-butyl dicarbonate (0.86 mL, 0.81 g, 3.75 mmol) were suspended in CHCl (72 mL) at 0° C. The reaction mixture was then stirred at room temperature for 24 h. The reaction mixture was diluted with water (30 mL) and MeOH (10 mL). The organic phase was separated, dried over NaSO, filtered, and the solvent was removed under reduced pressure. The residue was purified by silica gel (basic, 100-200 mesh) chromatography using a MeOH / CHCl gradient (0 / 100→0.5 / 99.5) to afford the title compound as a pale yellow solid (0.28 g, 36%). 1 H NMR (500 MHz, DMSO-d6): δ 8.75 (d, 1H), 8.05 (m, 1H), 7.43 (m, 2H), 7.35 (t, 1H), 6.6 (d, 1H), 4.6 (s, 1H), 4.06 (t, 2H), 3.37 (m, 4H), 3.22 (t, 2H), 2.1-1.9 (m, 2H), 1.4 (s, 18H), 0.86 (s, 9H), 0.1 (d, 6H). LCMS (ESI) 740.57 m / z [M+H] +

[0656] Step 5: Synthesis of tert-butyl (R)-(tert-butoxycarbonyl)(2-fluoro-5-(2-(6-(3-hydroxypyrrolidin-1-yl)pyridin-3-yl)-4-oxo-6,7-dihydrothiazolo[5,4-c]pyridin-5(4H)-yl)phenyl)carbamate The title compound from step 4 above (0.28 g, 0.37 mmol) was dissolved in THF (6 mL) and cooled to -20°C with stirring in a dry ice bath. Tetrabutylammonium fluoride (0.196 g, 0.75 mmol) was then added and stirring was continued at room temperature for 2.5 h. The reaction mixture was diluted with water (20 mL) and extracted with 10% MeOH in CH2Cl2 (3x20 mL). The organic phase was separated, dried over Na2SO4, filtered and the solvent was removed under reduced pressure. The residue was purified by silica gel (basic, 100-200 mesh) chromatography using a MeOH / CH2Cl2 gradient (0 / 100 -> 1 / 99 -> 2 / 98) to give the title compound as a pale yellow solid (0.2 g, 87%). 1 H NMR (500 MHz, DMSO-d6): δ 8.70 (d, 1H), 8.0 (dd, 1H), 7.39 (m, 2H), 7.3 (d, 1H), 6.53 (d, 1H), 5.0 (d, 1H), 4.37 (s, 1H), 4.02 (t, 2H), 3.49 (m, 3H), 3.3 (m, 1H), 1.9 (m, 2H), 1.36 (s, 18H). LCMS (ESI) 626.48 m / z [M+H] + .

[0657] Step 6: (R)-1-(5-(5-(3-(bis(tert-butoxycarbonyl)amino)-4-fluorophenyl)-4-oxo-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)pyridin-2-yl)pyrrolidin-3-yl methanesulfonate( 18 F. Synthesis of compound 26 (precursor) The title compound from step 5 above (0.19 g, 0.30 mmol) was dissolved in pyridine (7 mL) and cooled to -50°C with stirring in a dry ice bath. Methanesulfonyl chloride (0.514 g, 4.5 mmol) was then added and stirring was continued at room temperature for 1.5 h. The reaction mixture was diluted with water (35 mL) and the precipitate was collected by filtration, washed with water (2x15 mL) and dried. 18 F The precursor of compound 26 was obtained as a pale yellow solid (0.2 g, 94%). 1H NMR (500 MHz, DMSO-d6): δ 8.77 (d, 1H), 8.1 (dd, 1H), 7.43 (m, 2H), 7.3 (t, 1H), 6.56 (d, 1H), 5.45 (s, 1H), 4.06 (t, 2H), 3.77 (m, 3H), 3.53 (m, 1H), 3.27 (s, 3H), 3.23 (t, 2H), 2.35 (d, 2H), 1.4 (s, 18H). LCMS (ESI) 704.3 m / z [M+H] + .

[0658] (Example 58) Fluorinated (S)-5-(3-amino-4-fluorophenyl)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-6,7-dihydrothiazolo[5,4-c]pyridin-4(5H)-one ( 18 F. Synthesis of compound 26)

[0659] [ka]

[0660] Drying 18 F]Tetrabutylammonium fluoride 18 F The precursor of compound 26 (1 mg in 0.7 mL) was added and the reaction mixture was heated at 100° C. for 10 min, followed by Boc deprotection with 1 M HCl at 90° C. for 8 min. The crude reaction mixture was diluted with HPLC mobile phase (MeOH / NH4OAc solution (20 mM, 55 / 45, v / v)), injected onto the HPLC, and purified by reversed-phase HPLC. The product fractions were collected, diluted with aqueous ascorbic acid, and passed through a tC18 cartridge. After washing the cartridge with aqueous ascorbic acid, the product was eluted with ethanol and the cartridge was rinsed with a saline formulation of ascorbic acid. The resulting mixture was passed through a sterile filter and further diluted with saline to give the final product. 18 F - Preparation of compound 26 was obtained. The identity of the product was confirmed by: 19 F-Determined by co-injection with a sample of the reference compound.

[0661] Biological assay description 1. General method Human brain material for these studies was obtained from Professor William Seeley at the Neurodegenerative Disease Brain Bank UCSF (funded by NIH grants P01AG019724 and P50AG023501, the Consortium for Frontotemporal Dementia Research, and the Tau Consortium) and Professor Tammaryn Lashley at the Queen Square Brain Bank for Neurological Disorders, UCL. All material was collected from donors, who provided written informed consent for the brain bank to perform brain autopsies and to use the material and clinical information for research purposes.

[0662] 1.1. Radioligands The specific radioactivities were 75.83 Ci / mmol (1.0 mCi / mL), 52.4 or 47.8 Ci / mmol (1.0 mCi / mL), and 72.5 Ci / mmol (1.0 mCi / mL), respectively. 3 H]-Compound 1, [ 3 H]-Compound 52, and [ 3 H]-Compound 16 was used in the assay described below.

[0663] 1.2. Preparation of recombinant TDP-43 aggregates The TDP-43 aggregation protocol (Shimonaka et al., 2016) was performed to generate TDP-43 aggregates. Highly pure protein preparations without degradation were used for the aggregation procedure.

[0664] 1.3. 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 (frontal or temporal cortex) samples were homogenized in homogenization-solubilization (HS) buffer at 1:4 (w / v) ratio using a tissue homogenizer (Precellys) with a CKmix homogenization tube at 4°C. The following sequence was used for homogenization: 3 cycles of 30 s at 5000 rpm with 15 s pause between each cycle. Homogenized samples were aliquoted and stored at -80°C in 1.5 mL low protein binding tubes.

[0665] Brain homogenates were thawed on ice and resuspended in HS buffer to obtain a final concentration of 2% Sarkosyl, 1 unit / μL Benzonase, and 1 mM MgCl2. Samples were then incubated at 37°C for 45 min with constant agitation 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 20,000g 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 one step. The pellet was then washed with phosphate-buffered saline (PBS) and centrifuged at 20,000g for 30 min at 4°C. The final pellet was resuspended in 200 μL of PBS and stored at -80°C (Sarkosyl-insoluble fraction). Samples were analyzed by immunoblotting under denaturing conditions.

[0666] 2. Assay 2.1 [ 3 Determination of the Kd of [H]-Compound 1 on recombinant TDP-43 aggregates by filter binding assay Recombinant FLAG-tagged TDP-43 aggregates (1.3 μM) were incubated with increasing radioligand concentrations from 0 to 150 nM [ 3H]-Compound 1 was incubated for 2 h at room temperature in a total reaction volume of 100 μL. Duplicate samples of 45 μL (corresponding to each concentration) were vacuum filtered onto UniFilter-96 GF / C plates. Filters were washed five times with ice-cold buffer (50 mM Tris pH 7.4) and then dried overnight. Signals were quantified on a Microbeta Trilux instrument (Perkin Elmer) after adding 30 μL of scintillation fluid to each well for 30 min. Nonspecific signals were determined with an excess of unlabeled Compound 1 (10 μM) and specific binding was calculated by subtracting nonspecific signals from total signals. Kd (dissociation constant) and R2 (a parameter that quantifies the goodness of fit, ranging from 0.0 to 1.0; the best curve fit is obtained with a value of 1.0) were obtained by fitting the specific binding data with nonlinear regression analysis using a one-site specific binding model in GraphPad Prism.

[0667] result: [ 3 The dissociation constant (Kd) of [H]-compound 1 was determined with recombinant TDP-43 aggregates in a filter binding assay. 3 [H]-Compound 1 showed strong specific binding resulting in a large dynamic range, with a Kd value of 15 nM for recombinant FLAG-tagged TDP-43 aggregates (Figure 1). Data from three independent experiments yielded an average Kd of 18.4 ± 4 nM.

[0668] [ 3 The dissociation constant (Kd) of [H]-compound 3 was determined with recombinant TDP-43 aggregates in a filter binding assay. 3 H]-Compound 3 exhibited strong specific binding yielding a large dynamic range and showed a Kd value of 24.3 nM for recombinant FLAG-tagged TDP-43 aggregates (Figure 1).

[0669] 2.2. [ 3 Microautoradiography staining with [H]-compound 1 All tissues were collected from donors, with each brain bank obtaining written informed consent for brain autopsy and use of material and clinical information for research purposes. 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 and mounted on glass slides. Sections were kept at -80°C until use.

[0670] Brain sections were immunostained using a commercial antibody specific for phosphorylated serine at amino acids 409 / 410 (anti-pTDP-43 pS409 / 410, Cosmobio, TIP-PTD-P02). Sections were fixed with 4% formaldehyde (Sigma, 252549) for 15 min at 4°C and subjected to three 5 min washes with 1X PBS (Dulbecco's Phosphate Buffered Saline, Sigma, D1408) at room temperature. Sections were then saturated and permeabilized in blocking buffer (PBS, 10% normal goat serum (NGS), 0.25% Triton X-100) for 1 h at room temperature and incubated overnight at 4°C with primary antibody against pTDP-43 (diluted 1 / 250 in PBS, 5% NGS, 0.25% Triton X-100). The next day, sections were subjected to three 5-min washes in 1X PBS and then incubated with secondary AlexaFluor647-labeled goat anti-rabbit antibody (Abcam, ab150079, 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.

[0671] For microautoradiography, 3[H]-Compound 1 was incubated on the sections at 60 nM in 50 mM Tris buffer pH 7.4 at room temperature for 45 min. The sections were then washed as follows: one 1 min wash in ice-cold 50 mM Tris-HCl pH 7.4 buffer, two 1 min washes in ice-cold 70% ethanol, one 1 min wash in ice-cold 50 mM Tris-HCl pH 7.4 buffer, and finally a short rinse in ice-cold distilled water. The sections were then dried under airflow for 1 h before being exposed to Ilford Nuclear Emulsion Type K5 (Agar Scientific, AGP9281) at 4°C in a light-tight slide storage box for 5 days. After 7 days, sections were developed by sequential immersion 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.

[0672] For image acquisition, sections were mounted on a Panoramic250 slide scanner (3DHistech) with a 40x objective that acquires brightfield and fluorescent images separately, or on a Panoramic Scan II (3DHistech) with a 20x objective and imaged using ProLong Gold Antifade reagent (Invitrogen P36930). Fluorescent and brightfield images were aligned using the Visiopharm image analysis software suite.

[0673] Incubated on human brain slices [ 3 Microautoradiography signals from [H]-compound 1 were detected in the form of accumulated argyrophilic granules that colocalized with immunofluorescence signals from the pTDP-43 antibody, indicating that [H]-compound 1 is a cytoplasmic marker for TDP-43 aggregates. 3 H]-suggesting target binding of compound 1. Brain slices from FTLD-TDP donors were [3 Incubation with [H]-compound 1 (60 nM) demonstrated colocalization with pTDP-43 aggregates (Figure 2). To our knowledge, this is the first compound to show targeted binding to TDP-43 aggregates on human brain sections by autoradiography techniques.

[0674] 2.3. Microradiometric binding competition assay for determination of binding affinity Human FTD sarkosyl-insoluble brain extracts were spotted onto microarray slides. The slides were then stained with 75 nM or 40 nM of 3 H]-Compound 52 ligand or [ 3 H]-Compound 16 ligand and 2 μM and 250 nM of Example Compound (non-radiolabeled). In some cases, non-radiolabeled Example Compound was further evaluated for a series of different concentrations varying from 0.24 nM to 2 μM for determination of inhibition constant (Ki). After incubation, slides were washed and scanned with a real-time autoradiography system (BeaQuant, ai4R). Quantification of the signal was performed using Beamage image analysis software (ai4R). Non-specific signal was determined with an excess of non-radiolabeled Compound 52 or Compound 16 (2 μM) and specific binding was calculated by subtracting the non-specific signal from the total signal. Composition was calculated as a percentage, where 0% was defined as specific binding in the presence of vehicle and 100% was defined as the value obtained in the presence of an excess of non-radiolabeled Compound 52 or Compound 16. K i Values ​​were calculated in Prism 7 from GraphPad by applying a nonlinear regression curve fit using a one-site specific binding model. 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, replicates from independent experiments or K i The average of the values ​​is reported.

[0675] result: The Example Compounds were used to measure the activity of TDP-43 aggregates derived from the brains of FTD patients. 3 H]-ligand ([ 3 H]-Compound 52 or [ 3 The results of the microradiometric binding competition assay for the example compounds tested are shown below in Table 6 as % competition at 2 μM and 250 nM. i The values ​​are also shown.

[0676] Examples 1 to 3 and 13 to 50 According to the above screening method, the following compounds were characterized:

[0677] [Table 7A]

[0678] [Table 7B]

[0679] [Table 7C]

[0680] [Table 7D]

Claims

1. Formula (I); 【Chemical 1】 (wherein, Z 1 , Z 2 , Z 3 , and Z 4 are each selected from CH and N, where Z 1 When N is Z 2 is CH; Z 2 When N is N, Z 1 is CH; Z 3 When N is N, Z 4 is CH; Z 4 When N is N, Z 3 is CH; n is 1 or 2; R 1 is H or F; R 2 is F, NH 2 , CN, and / or CH 3 a 5- or 6-membered carbocyclic ring optionally substituted with F, NH 2 , CN, and / or CH 3 optionally substituted with, and containing one or more heteroatoms selected from N, O, and S, a 5- or 6-membered heterocycloalkyl ring F, NH 2 , CN, and / or CH 3 optionally substituted with, and containing one or more heteroatoms selected from N, O, and S, a 5-membered heteroaryl ring, or F, NH 2 , CN, and / or CH 3 optionally substituted with, and selected from 6-membered heteroaryl rings containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S), or a detectably labeled compound thereof, stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate, or a mixture thereof.

2. Formula (Ia); 【Chemical 2】 (wherein, n is 1 or 2; R 1 is H or F; R 2 is F, NH 2 , CN, and / or CH 3 a 5- or 6-membered carbocyclic ring optionally substituted with F, NH 2 , CN, and / or CH 3 which may be optionally substituted with, and contains one or more heteroatoms selected from N, O, and S, a 5- or 6-membered heterocycloalkyl ring F, NH 2 , CN, and / or CH 3 optionally substituted with, and containing one or more heteroatoms selected from N, O, and S, a 5-membered heteroaryl ring, or F, NH 2 , CN, and / or CH 3 optionally substituted with, and is a 6-membered heteroaryl ring containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S), the compound according to claim 1, or a detectably labeled compound thereof, stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate, or a mixture thereof.

3. Formula (Ib); 【Chemical 3】 (wherein, n is 1 or 2; R 1 is H or F; R 2 is F, NH 2 , CN, and / or CH 3 a 5- or 6-membered carbocyclic ring optionally substituted with F, NH 2 , CN, and / or CH 3 which may be optionally substituted with 2 , 3 , and contains one or more heteroatoms selected from N, O, and S, and is a 5- or 6-membered heterocycloalkyl ring F, NH 2 , CN, and / or CH 3 optionally substituted with, and containing one or more heteroatoms selected from N, O, and S, a 5-membered heteroaryl ring, or F, NH 2 , CN, and / or CH 3 optionally substituted with, and is a 6-membered heteroaryl ring containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S), a compound according to claim 1, or a detectably labeled compound thereof, stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate, or a mixture thereof.

4. Formula (Ic); [Chemical Formula 4] (wherein, n is 1 or 2; R 1 is H or F; R 2 is F, NH 2 , CN, and / or CH 3 a 5- or 6-membered carbocyclic ring optionally substituted with F, NH 2 , CN, and / or CH 3 optionally substituted with, and containing one or more heteroatoms selected from N, O, and S, a 5- or 6-membered heterocycloalkyl ring F, NH 2 , CN, and / or CH 3 optionally substituted with, and containing one or more heteroatoms selected from N, O, and S, a 5-membered heteroaryl ring, or F, NH 2 , CN, and / or CH 3 optionally substituted with, and is a 6-membered heteroaryl ring containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S), the compound according to claim 1, or a detectably labeled compound thereof, stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate, or a mixture thereof.

5. Formula (Id); [Chemical Formula 5] (wherein, n is 1 or 2; R 1 is H or F; R 2 is F, NH 2 , CN, and / or CH 3 a 5- or 6-membered carbocyclic ring optionally substituted with F, NH 2 , CN, and / or CH 3 optionally substituted with 2 , 3 , CN, and / or CH, and containing one or more heteroatoms selected from N, O, and S, a 5- or 6-membered heterocycloalkyl ring F, NH 2 , CN, and / or CH 3 optionally substituted with, and containing one or more heteroatoms selected from N, O, and S, a 5-membered heteroaryl ring, or F, NH 2 , CN, and / or CH 3 optionally substituted with, and is a 6-membered heteroaryl ring containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S), the compound according to claim 1, or a detectably labeled compound thereof, stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate, or a mixture thereof.

6. Formula (Ie); 【Chemical Formula 6】 (wherein, n is 1 or 2; R 1 is H or F; R 2 is F, NH 2 , CN, and / or CH 3 a 5- or 6-membered carbocyclic ring optionally substituted with F, NH 2 , CN, and / or CH 3 optionally substituted with 2 , 3 , CN, and / or CH, and containing one or more heteroatoms selected from N, O, and S, a 5- or 6-membered heterocycloalkyl ring F, NH 2 , CN, and / or CH 3 optionally substituted with, and containing one or more heteroatoms selected from N, O, and S, a 5-membered heteroaryl ring, or F, NH 2 , CN, and / or CH 3 optionally substituted with, and is a 6-membered heteroaryl ring containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S), the compound according to claim 1, or a detectably labeled compound thereof, stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate, or a mixture thereof.

7. R 2 is [Chemical Formula 7] (wherein, R 3 is F, and R 4 is NH 2 and R 7 is H, and R 8 is H, R 3 is NH 2 and R 4 is F, and R 7 is H, and R 8 is H, R 3 is CN, and R 4 is NH 2 and R 7 is H, and R 8 is H, R 3 is H, and R 4 is NH 2 and R 7 is H, and R 8 is CN, or R 3 is H, and R 4 is NH 2 and R 7 is H, and R 8 is F); or, R 2 is 【Chemical 8】 (wherein X is N, and R 5 is CH 3 or H), the compound according to claim 1.

8. The compound according to claim 1, comprising a detectable label.

9. The detectable label is 3 H or 18 F, and the compound according to claim 8.

10. Formula (I-T); 【Chemical Formula 9】 (wherein, Z 1 、Z 2 、Z 3 、and Z 4 are each selected from CH and N, where Z 1 When N is N, Z 2 is CH; Z 2 When N is N, Z 1 is CH; Z 3 When N is N, Z 4 is CH; Z 4 When N is N, Z 3 is CH; n is 1 or 2; R 1 is H or F; R 2 is F, NH 2 , CN, and / or CH 3 a 5- or 6-membered carbocyclic ring optionally substituted with F, NH 2 , CN, and / or CH 3 optionally substituted with 2 and 3 , and containing one or more heteroatoms selected from N, O, and S, a 5- or 6-membered heterocycloalkyl ring F, NH 2 , CN, and / or CH 3 optionally substituted with 2 , 3 , and containing one or more heteroatoms selected from N, O, and S, a 5-membered heteroaryl ring, or F, NH 2 , CN, and / or CH 3 optionally substituted with, and selected from 6-membered heteroaryl rings containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S, here Z 1 、Z 2 、Z 3 、Z 4 at least one of them is selected from C-T; or, R 2 is replaced by at least one CT 3 or at least one hydrogen atom in R 2 is replaced by T), the compound according to claim 9, or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate thereof, or a mixture thereof.

11. Formula (I-Ta), (I-Tb), (I-Td), or (I-Te); 【Chemical 10】 (wherein, n is 1 or 2; R 1 is H or F; R 6 is T or H; T is 3 H; R 2 is 【Chemical 11】 (wherein, R 3 is F, and R 4 is NH 2 and R 7 and R 8 at least one of which is T, R 3 is CN, and R 4 is NH 2 and R 7 and R 8 at least one of them is T, R 4 is NH 2 and R 8 is -CN, and R 3 and R 7 at least one of them is T, or, R 4 is NH 2 and R 8 is F, and R 3 and R 7 at least one of them is T, and in the corresponding case, the other is H); or, R 2 is 【Chemical 12】 (wherein X is N, and R 5 is CT 3 ); R 6 The compound according to claim 10, or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate thereof, or a mixture thereof, having (wherein R is H).

12. 【Fig. 13】 (wherein T is 3 H), the compound according to claim 10, selected from

13. Formula (I-F); 【Chemical Formula 14】 (wherein, Z1, Z2, Z3, and Z4 are each independently selected from CH and N, where when Z1 is N, Z2 is CH; when Z2 is N, Z1 is CH; when Z3 is N, Z4 is CH; when Z4 is N, Z3 is CH; n is 1 or 2; R1 is H or F; R2 is a 5- or 6-membered carbocyclic ring optionally substituted with F, NH2, CN, and / or CH3, a 5- or 6-membered heterocycloalkyl ring optionally substituted with F, NH2, CN, and / or CH3 and containing one or more heteroatoms selected from N, O, and S, 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 Optionally substituted with F, NH₂, CN, and / or CH₃, selected from 6-membered heteroaryl rings containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S, wherein at least one F is 18 F, preferably, R 1 is 18 F), the compound according to claim 9, or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate thereof, or a mixture thereof.

14. 【Fig. 15】 The compound according to claim 13, which is

15. A diagnostic composition comprising the compound according to any one of claims 1 to 14 and optionally at least one physiologically acceptable carrier, diluent, adjuvant, and / or excipient.

16. The diagnostic composition according to claim 15 for use in a diagnostic method.

17. The diagnostic composition according to claim 15 for use in imaging TDP-43 aggregates, particularly by positron emission tomography.

18. The diagnostic composition according to claim 15 for use in the diagnosis of a disease, disorder, or abnormality associated with TDP-43 aggregates, particularly by positron emission tomography.

19. The following steps: (a) Administering the diagnostic composition according to claim 15 to a subject; (b) Binding a compound to TDP-43 aggregates; and (c) Detecting the compound bound to the TDP-43 aggregates A method for imaging a disease, disorder, or abnormality associated with TDP-43 aggregates in a subject, comprising the above steps.

20. The following steps: (d) Generating an image representing the position and / or amount of the compound bound to the TDP-43 aggregates The method for imaging a disease, disorder, or abnormality associated with TDP-43 aggregates according to claim 19, further comprising the above step.

21. The following steps: (a) Administering the diagnostic composition according to claim 15 to a subject; (b) Binding a compound to TDP-43 aggregates; and (c) Detecting the compound bound to the TDP-43 aggregates by collecting a positron emission tomography (PET) image of the subject's tissue A method for positron emission tomography (PET) imaging of TDP-43 aggregates in a subject's tissue, comprising the above steps.

22. The method for positron emission tomography (PET) imaging of TDP-43 aggregates in a subject's tissue according to claim 21, wherein the tissue is tissue of the central nervous system (CNS), eye tissue, or brain tissue, and preferably, the tissue is brain tissue.

23. The following steps: (a) Contacting a sample suspected of containing TDP-43 aggregates or a specific body part or body region with the diagnostic composition according to claim 15; (b) Binding a compound to TDP-43 aggregates; (c) Detecting the compound bound to the TDP-43 aggregates using positron emission tomography; and (d) Optionally, quantifying the amount of the compound bound to the TDP-43 aggregates A method for detecting and optionally quantifying TDP-43 aggregates in a subject's tissue, comprising the above steps.

24. The following steps: (a) Contacting a sample suspected of containing TDP-43 aggregates or a specific body part or body region with the diagnostic composition according to claim 15; (b) Binding a compound to TDP-43 aggregates; (c) Detecting the compound bound to the TDP-43 aggregates; and (d) Optionally, correlating the presence or absence of the compound bound to the TDP-43 aggregates with the presence or absence of the TDP-43 aggregates in the sample or the specific body part or body region A method of collecting data for the diagnosis of a disease, disorder, or abnormality associated with TDP-43 aggregates, or for the diagnosis of TDP-43 proteinopathy.

25. The following steps: (a) Contacting a sample suspected of containing TDP-43 aggregates, or a specific body part or body region, with the diagnostic composition according to claim 15; (b) Binding a compound to the TDP-43 aggregates; (c) Detecting the compound bound to the TDP-43 aggregates; and (d) Optionally, correlating the presence or absence of the compound bound to the TDP-43 aggregates in the sample or specific body part or body region with the presence or absence of the TDP-43 aggregates A method of collecting data for determining a predisposition to a disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy.

26. The following steps: (a) Contacting a sample, specific body part or body region suspected of containing TDP-43 aggregates with the diagnostic composition according to claim 15; (b) Binding a compound to the TDP-43 aggregates; (c) Detecting the compound bound to the TDP-43 aggregates; (d) Optionally, correlating the presence or absence of the compound bound to the TDP-43 aggregates in the sample or specific body part or body region with the presence or absence of the TDP-43 aggregates; and (e) Optionally, repeating steps (a) to (c), and optionally step (d) if present, at least once A method of collecting data for monitoring the progression of a disease, disorder, or abnormality associated with TDP-43 aggregates, or for monitoring the progression of TDP-43 proteinopathy, in a patient.

27. The following steps: (a) Contacting a sample, specific body part or body region suspected of containing TDP-43 aggregates with the diagnostic composition according to claim 15; (b) Binding a compound to the TDP-43 aggregates; (c) Detecting the compound bound to the TDP-43 aggregates; (d) Optionally, correlating the presence or absence of the compound bound to the TDP-43 aggregates in the sample or specific body part or body region with the presence or absence of the TDP-43 aggregates; and (e) Optionally, repeating steps (a) to (c), and optionally step (d) if present, at least once A method of collecting data for predicting the responsiveness of a patient suffering from a disease, disorder, or abnormality associated with TDP-43 aggregates, including **Claim 28** An optional 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 specific body part or body region is the following steps: - Quantifying the amount of the compound bound to TDP-43 aggregates; - Correlating the amount of the compound bound to TDP-43 aggregates with the amount of TDP-43 aggregates in a sample or a specific body part or body region; and - An optional step of comparing the amount of the compound bound to TDP-43 aggregates in a sample or a specific body part or body region with a normal control value in a healthy control subject The method according to claim 24, comprising the above steps. **Claim 29** A composition for use as a biomarker for TDP-43 aggregates or a biomarker for TDP-43 proteinopathy, comprising the compound according to any one of claims 1 to 14. **Claim 30** A composition for use as a diagnostic agent or diagnostic tool for TDP-43 proteinopathy, comprising the compound according to any one of claims 1 to 14. **Claim 31** A composition for use as a standard substance for in vitro analysis or an in vitro screening tool, comprising the compound according to any one of claims 1 to 14. **Claim 32** A disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is selected from frontotemporal dementia (FTD, such as sporadic or familial with or without motor neuron disease (MND), with progranulin (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valosin-containing protein (VCP) mutation, chromosome 9p-linked, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) (including frontotemporal dementia TDP-43 or frontotemporal lobar degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP)), argyrophilic grain disease, Pick's disease, semantic primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent primary progressive aphasia (e.g., nfvPPA), amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation), 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 syndrome, familial British dementia, polyglutamine disease (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 mutation of valosin-containing protein (VCP); and Paget's disease of bone and frontotemporal dementia), oculopharyngeal muscular dystrophy with rimmed vacuoles, myofibrillar myopathy with mutation of the myotilin (MYOT) gene or mutation of the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), and Parkinson's disease (PD), preferably, a disease, disorder, or abnormality associated with TDP-43 aggregates, or 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), the diagnostic composition according to claim 18.

33. A disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is frontotemporal dementia (FTD, e.g., sporadic or familial with or without motor neuron disease (MND), with progranulin (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valosin-containing protein (VCP) mutation, chromosome 9p-linked, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) (including frontotemporal lobar dementia TDP-43 or frontotemporal lobar degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP)), argentophilic grain disease, Pick's disease, semantic primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent primary progressive aphasia (e.g., nfvPPA), amyotrophic lateral sclerosis (ALS, e.g., sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation), 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 syndrome, familial British dementia, polyglutamine disease (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 valosin-containing protein (VCP) mutation; and Paget's disease of bone and frontotemporal dementia), oculopharyngeal muscular dystrophy with rimmed vacuoles, myofibrillar myopathy with mutation of the myotilin (MYOT) gene or mutation of the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), and Parkinson's disease (PD), preferably, the disease, disorder, or abnormality associated with TDP-43 aggregates, or 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), the method according to claim 19.

34. A disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is 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, chromosome 9p-linked, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) (including frontotemporal lobar degeneration with TDP-43 inclusions (FTLD-TDP) or ubiquitin-positive TDP-43 inclusions), argentophilic grain disease, Pick's disease, semantic primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent primary progressive aphasia (e.g., nfvPPA), amyotrophic lateral sclerosis (ALS, e.g., sporadic ALS, with TARDBP mutations, 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 syndrome, familial British dementia, polyglutamine disease (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); and 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), Lewy body dementia (DLB), and Parkinson's disease (PD), preferably, the disease, disorder, or abnormality associated with TDP-43 aggregates, or 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), the method according to claim 20.

35. A disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is frontotemporal dementia (FTD, such as sporadic or familial with or without motor neuron disease (MND), with progranulin (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valosin-containing protein (VCP) mutation, chromosome 9p-linked, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) (including frontotemporal lobar dementia TDP-43 or frontotemporal lobar degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP)), argyrophilic grain disease, Pick's disease, semantic primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent primary progressive aphasia (e.g., nfvPPA), amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation), 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 syndrome, familial British dementia, polyglutamine disease (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 mutation of valosin-containing protein (VCP); and Paget's disease of bone and frontotemporal dementia), oculopharyngeal muscular dystrophy with rimmed vacuoles, myofibrillar myopathy with mutation of the myotilin (MYOT) gene or mutation of the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), and Parkinson's disease (PD), preferably, the disease, disorder, or abnormality associated with TDP-43 aggregates, or 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), the method according to claim 21.

36. A disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is frontotemporal dementia (FTD, e.g., sporadic or familial with or without motor neuron disease (MND), with progranulin (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valosin-containing protein (VCP) mutation, chromosome 9p-linked, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) (including frontotemporal lobar degeneration with TDP-43 inclusions in frontotemporal dementia (FTLD-TDP)), argyrophilic grain disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent primary progressive aphasia (e.g., nfvPPA), amyotrophic lateral sclerosis (ALS, e.g., sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation), 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 syndrome, familial British dementia, polyglutamine disease (Huntington's disease and spinocerebellar ataxia type 3 (SCA3; also known as Machado-Joseph disease)), hippocampal sclerosis-related dementia and myopathy (sporadic inclusion body myositis, inclusion body myopathy with valosin-containing protein (VCP) mutation; and Paget's disease of bone and frontotemporal dementia), oculopharyngeal muscular dystrophy with rimmed vacuoles, myofibrillar myopathy with mutation of the myotilin (MYOT) gene or mutation of the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), and Parkinson's disease (PD), preferably, a disease, disorder, or abnormality associated with TDP-43 aggregates, or 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), the method according to claim 22.

37. A disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is frontotemporal dementia (FTD), such as sporadic or familial (with or without motor neuron disease (MND)), with progranulin (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valosin-containing protein (VCP) mutation, chromosome 9p-linked, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) (including frontotemporal lobar degeneration with TDP-43 (FTLD-TDP) or ubiquitin-positive TDP-43 inclusions), argyrophilic grain disease, Pick's disease, semantic variant primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent primary progressive aphasia (e.g., nfvPPA), amyotrophic lateral sclerosis (ALS), such as sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation, 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 syndrome, familial British dementia, polyglutamine disease (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 valosin-containing protein (VCP) mutation; and Paget's disease of bone and frontotemporal dementia), oculopharyngeal muscular dystrophy with rimmed vacuoles, myofibrillar myopathy with mutation of the myotilin (MYOT) gene or mutation of the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), and Parkinson's disease (PD), preferably, a disease, disorder, or abnormality associated with TDP-43 aggregates, or 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), the method according to claim 23.

38. A disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is frontotemporal dementia (FTD, such as sporadic or familial, with or without motor neuron disease (MND), with progranulin (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valosin-containing protein (VCP) mutation, chromosome 9p-linked, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) (including frontotemporal lobar dementia TDP-43 or frontotemporal lobar degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP)), argentophilic grain disease, Pick's disease, semantic primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent primary progressive aphasia (e.g., nfvPPA), amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation), 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 syndrome, familial British dementia, polyglutamine disease (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 valosin-containing protein (VCP) mutation; and Paget's disease of bone and frontotemporal dementia), oculopharyngeal muscular dystrophy with rimmed vacuoles, myofibrillar myopathy with mutation of the myotilin (MYOT) gene or mutation of the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), and Parkinson's disease (PD), preferably, the disease, disorder, or abnormality associated with TDP-43 aggregates, or 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), the method according to claim 24.

39. A disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is frontotemporal dementia (FTD, e.g., sporadic or familial with or without motor neuron disease (MND), with progranulin (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valosin-containing protein (VCP) mutation, chromosome 9p-linked, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) (including frontotemporal lobar dementia TDP-43 or frontotemporal lobar degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP)), argentophilic grain disease, Pick's disease, semantic primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent primary progressive aphasia (e.g., nfvPPA), amyotrophic lateral sclerosis (ALS, e.g., sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation), 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 syndrome, familial British dementia, polyglutamine disease (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 valosin-containing protein (VCP) mutation; and Paget's disease of bone and frontotemporal dementia), oculopharyngeal muscular dystrophy with rimmed vacuoles, myofibrillar myopathy with mutation of the myotilin (MYOT) gene or mutation of the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), and Parkinson's disease (PD), preferably, a disease, disorder, or abnormality associated with TDP-43 aggregates, or 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), the method according to claim 25.

40. A disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is frontotemporal dementia (FTD, such as sporadic or familial with or without motor neuron disease (MND), with progranulin (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valosin-containing protein (VCP) mutation, chromosome 9p-linked, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) (including frontotemporal lobar dementia TDP-43 or frontotemporal lobar degeneration with ubiquitin-positive TDP-43 inclusions (FTLD-TDP)), argyrophilic grain disease, Pick's disease, semantic primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent primary progressive aphasia (e.g., nfvPPA), amyotrophic lateral sclerosis (ALS, such as sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation), 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 syndrome, familial British dementia, polyglutamine disease (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 valosin-containing protein (VCP) mutation; and Paget's disease of bone and frontotemporal dementia), oculopharyngeal muscular dystrophy with rimmed vacuoles, myofibrillar myopathy with mutation of the myotilin (MYOT) gene or mutation of the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), and Parkinson's disease (PD), preferably, the disease, disorder, or abnormality associated with TDP-43 aggregates, or 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), the method according to claim 26.

41. A disease, disorder, or abnormality associated with TDP-43 aggregates, or TDP-43 proteinopathy, is frontotemporal dementia (FTD, e.g., sporadic or familial with or without motor neuron disease (MND), with progranulin (GRN) mutation, with C9orf72 mutation, with TARDBP mutation, with valosin-containing protein (VCP) mutation, chromosome 9p-linked, corticobasal degeneration, frontotemporal lobar degeneration (FTLD) (including frontotemporal lobar degeneration with TDP-43 (FTLD-TDP) or ubiquitin-positive TDP-43 inclusions), argyrophilic grain disease, Pick's disease, semantic primary progressive aphasia (svPPA), behavioral variant FTD (bvFTD), non-fluent primary progressive aphasia (e.g., nfvPPA), amyotrophic lateral sclerosis (ALS, e.g., sporadic ALS, with TARDBP mutation, with angiogenin (ANG) mutation), 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 syndrome, familial British dementia, polyglutamine disease (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 valosin-containing protein (VCP) mutation; and Paget's disease of bone and frontotemporal dementia), oculopharyngeal muscular dystrophy with rimmed vacuoles, myofibrillar myopathy with mutation of the myotilin (MYOT) gene or mutation of the gene encoding desmin (DES), traumatic brain injury (TBI), Lewy body dementia (DLB), and Parkinson's disease (PD), preferably, a disease, disorder, or abnormality associated with TDP-43 aggregates, or 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), the method according to claim 27.

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

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

44. The diagnostic composition 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 dementia with TDP-43 (FTLD-TDP) or frontotemporal lobar degeneration with TDP-43 inclusions.

45. The diagnostic composition 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).

46. Formula (II); 【Chemical Formula 16】 (Wherein, Z 1 、Z 2 、Z 3 、and Z 4 are each selected from C-Br, C-I, C-H, and N, where Z 1 When N is N, Z 2 is C-Br, C-I, or C-H; Z 2 When N is N, Z 1 is C-Br, C-I, or C-H; Z 3 When N is N, Z 4 is C-Br, C-I, or C-H; Z 4 When N is N, Z 3 is C-Br, C-I, or C-H; n is 1 or 2; R 1 a compound having (wherein R is H or F), or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate thereof, or a mixture thereof.

47. Formula (III); 【Chemical 17】 (Wherein, Z 1 、Z 2 、Z 3 、and Z 4 are each selected from C-Br, C-I, C-H, and N, where Z 1 When N is N, Z 2 is C-Br, C-I, or C-H; Z 2 When N is N, Z 1 is C-Br, C-I, or C-H; Z 3 When N is N, Z 4 is C-Br, C-I, or C-H; Z 4 When N is N, Z 3 is C-Br, C-I, or C-H; n is 1 or 2; R 1 is H or F; R 10 is Br, I, F, NH 2 , CN, and / or CH 3 a 5- or 6-membered carbocyclic ring optionally substituted with Br, I, F, NH 2 , CN, and / or CH 3 which may optionally be substituted with 2 , 3 , and contains one or more heteroatoms selected from N, O, and S, a 5- or 6-membered heterocycloalkyl ring Br, I, F, NH 2 , CN, and / or CH 3 optionally substituted with, and containing one or more heteroatoms selected from N, O, and S, a 5-membered heteroaryl ring, or Br, I, F, NH 2 , CN, and / or CH 3 which may be optionally substituted with 2 , 3 , and is a 6-membered heteroaryl ring containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S; Z 1 、Z 2 、Z 3 、 or Z 4 at least one of which is selected from C-Br or C-I, and / or, R 10 a compound having (wherein R is Br or I), or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate thereof, or a mixture thereof.

48. The following steps: Formula (III); 【Chemical Formula 18】 (Wherein, Z 1 、Z 2 、Z 3 、and Z 4 are each selected from C-Br, C-I, C-H, and N, where Z 1 When N is N, Z 2 is C-Br, C-I, or C-H; Z 2 When N is N, Z 1 is C-Br, C-I, or C-H; Z 3 When N is N, Z 4 is C-Br, C-I, or C-H; Z 4 When N is N, Z 3 is C-Br, C-I, or C-H; n is 1 or 2; R 1 is H or F; R 10 is Br, I, F, NH 2 , CN, and / or CH 3 a 5- or 6-membered carbocyclic ring optionally substituted with Br, I, F, NH 2 , CN, and / or CH 3 which may be optionally substituted with 2 , 3 , and contains one or more heteroatoms selected from N, O, and S, and is a 5- or 6-membered heterocycloalkyl ring Br, I, F, NH 2 , CN, and / or CH 3 optionally substituted with, and containing one or more heteroatoms selected from N, O, and S, a 5-membered heteroaryl ring, or Br, I, F, NH 2 , CN, and / or CH 3 which may be optionally substituted with 2 , 3 , Br, I, F, NH, CN, and / or CH, and is a 6-membered heteroaryl ring containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S; Z 1 、Z 2 、Z 3 、 or Z 4 at least one of which is selected from C-Br or C-I, and / or, R 10 contains Br or I, Here, at least one of Br or I is CT 3 or T is replaced, T is 3 a compound having), or a stereoisomer, polymorph, racemic mixture, tautomer, pharmaceutically acceptable salt, prodrug, hydrate, or solvate thereof, or a mixture thereof 3 the step of radiolabeling with H A method for preparing a compound having formula (I) as defined in claim 9, comprising.

49. Formula (IV): 【Chemical Formula 19】 (Wherein, n, Z 1 , Z 2 , Z 3 , Z 4 , and R 2 is as defined in claim 1; R 14 is a leaving group (LG)) A compound having.

50. The following steps: Formula (IV): 【Chemical 20】 (Wherein, Z 1 、Z 2 、Z 3 、and Z 4 are each selected from CH and N, where Z 1 When N is N, Z 2 is CH; Z 2 When N is N, Z 1 is CH; Z 3 When N is N, Z 4 is CH; Z 4 When N is N, Z 3 is CH; n is 1 or 2; R 2 is F, NH 2 , CN, and / or CH 3 a 5- or 6-membered carbocyclic ring optionally substituted with F, NH 2 , CN, and / or CH 3 optionally substituted with 2 , 3 , and containing one or more heteroatoms selected from N, O, and S, a 5- or 6-membered heterocycloalkyl ring F, NH 2 , CN, and / or CH 3 optionally substituted with, and containing one or more heteroatoms selected from N, O, and S, a 5-membered heteroaryl ring, or F, NH 2 , CN, and / or CH 3 which may be optionally substituted with 2 , 3 , CN, and / or CH, and is a 6-membered heteroaryl ring containing one heteroatom selected from O and S or two or more heteroatoms selected from O, N, and S; R 14 is a compound having a leaving group (which is replaced by F in the radiolabeling step) 18 and is subjected to a step of radiolabeling with F 18 in the radiolabeling step A method for preparing a compound having formula (I) as defined in claim 9, comprising.

51. A kit for preparing a radiopharmaceutical preparation, comprising a precursor of a detectably labeled compound as defined in any one of claims 8 to 14, which is a compound of formula (II) as defined in claim 46, a compound of formula (III) as defined in claim 47, or a compound of formula (IV) as defined in claim 49.

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