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WO2024126840A1 - Novel compounds for diagnosis - Google Patents

Novel compounds for diagnosis Download PDF

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Publication number
WO2024126840A1
WO2024126840A1 PCT/EP2023/086185 EP2023086185W WO2024126840A1 WO 2024126840 A1 WO2024126840 A1 WO 2024126840A1 EP 2023086185 W EP2023086185 W EP 2023086185W WO 2024126840 A1 WO2024126840 A1 WO 2024126840A1
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WO
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Prior art keywords
compound
c4alkyl
c4alkoxy
haloc1
alpha
Prior art date
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PCT/EP2023/086185
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French (fr)
Inventor
Jérôme Molette
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Ac Immune Sa
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Publication of WO2024126840A1 publication Critical patent/WO2024126840A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Definitions

  • the present invention relates to novel compounds of formula (I), or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, that can be employed in the imaging of alpha-synuclein aggregates and determining an amount thereof.
  • the compounds can be used for diagnosing a disease, disorder or abnormality associated with alpha-synuclein (a-synuclein, A-synuclein, aSynuclein, A-syn, a-syn, aSyn, a-syn) aggregates, such as Parkinson’s disease or multiple system atrophy, determining a predisposition to such a disease, disorder or abnormality, prognosing such a disease, disorder or abnormality, monitoring the evolution of the disease in a patient suffering from such a disease, disorder or abnormality, monitoring the progression of such a disease, disorder or abnormality and predicting responsiveness of a patient suffering from such a disease, disorder or abnormality to a treatment thereof.
  • the present invention also relates to processes for the preparation of the compounds and their precursors, diagnostic compositions comprising the compounds, methods of using the compounds, kits comprising the compounds and their uses thereof.
  • amyloid beta amyloid beta
  • Amyloid-like proteins that form mainly intracellular aggregates include, but are not limited to, Tau, alpha-synuclein, and huntingtin (HTT).
  • Diseases involving alpha-synuclein aggregates are generally listed as synucleinopathies (or alpha-synucleinopathies) and these include, but are not limited to Parkinson’s disease (PD) or multiple system atrophy (MSA).
  • PD Parkinson’s disease
  • MSA multiple system atrophy
  • Synucleinopathies with primarily neuronal aggregates include, but are not limited to, Parkinson's disease (sporadic, familial with SNCA (the gene encoding for the alpha-synuclein protein) mutations or SNCA gene duplication or triplication, familial with mutations in other genes than SNCA, pure autonomic failure and Lewy body dysphagia), SNCA duplication carrier, Lewy Body dementia (LBD), dementia with Lewy bodies (DLB) (“pure” Lewy body dementia), Parkinson’s disease dementia (PDD), diffuse Lewy body disease (DLBD), Alzheimer’s disease, sporadic Alzheimer’s disease, familial Alzheimer's disease with APP mutations, familial Alzheimer's disease with PS-1 , PS-2 or other mutations, familial British dementia, Lewy body variant of Alzheimer’s disease and normal aging in Down syndrome.
  • Parkinson's disease sporadic, familial with SNCA (the gene encoding for the alpha-synuclein protein
  • Synucleinopathies with neuronal and glial aggregates of alpha-synuclein include, but are not limited to, multiple system atrophy (MSA) (Shy-Drager syndrome, striatonigral degeneration and olivopontocerebellar atrophy).
  • MSA multiple system atrophy
  • alpha-synuclein-immunoreactive lesions are, but are not limited to, traumatic brain injury, chronic traumatic encephalopathy, dementia puglistica, tauopathies (Pick's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration and Niemann-Pick type C1 disease, frontotemporal dementia with Parkinsonism linked to chromosome 17), motor neuron disease, Huntington’s disease, amyotrophic lateral sclerosis (sporadic, familial and ALS-dementia complex of Guam), neuroaxonal dystrophy, neurodegeneration with brain iron accumulation type 1 (Hallervorden-Spatz syndrome), prion diseases, Creutzfeldt-Jakob disease, ataxia telangiectatica, Meige’s syndrome, subacute sclerosing panencephalitis, Gerstmann- Straussler-Scheinker disease, inclusion-body myositis, Gaucher disease,
  • Alpha-synuclein is a 140 amino acid natively unfolded protein (Iwai et al., Biochemistry 1995, 34(32), 10139-10145).
  • the sequence of alpha-synuclein can be divided into three main domains: 1 ) the N- terminal region comprising of residues 1-60, which contains the 11-mer amphipatic imperfect repeat residues with highly conserved hexamer (KTKEGV).
  • This region has been implicated in regulating alpha-synuclein binding to membranes and its internalization; 2) the hydrophobic Non Amyloid beta Component (NAC) domain spanning residues 61-95; which is essential for alpha-synuclein fibrillization; and 3) the C-terminal region spanning residues 96-140 which is highly acidic and prolinerich and has no distinct structural propensity.
  • Alpha-synuclein has been shown to undergo several posttranslational modifications, including truncations, phosphorylation, ubiquitination, oxidation and/or transglutaminase covalent cross linking (Fujiwara et al., Nat. Cell. Biol.
  • Tyr-125 residues can be phosphorylated by two Src family protein tyrosine kinases, c-Src and Fyn (Ellis et al., J. Biol. Chem.
  • Alpha-synuclein has proved to be an outstanding substrate for protein tyrosine kinase p72 syk (Syk) in vitro-, once it is extensively Tyr-phosphorylated by Syk or tyrosine kinases with similar specificity, it loses the ability to form oligomers, suggesting a putative anti- neurodegenerative role for these tyrosine kinases (Negro et al., FASEB J. 2002, 16(2), 210-212).
  • Alpha-synuclein can be Ser-phosphorylated by protein kinases CKI and CKII (Okochi et al., J. Biol. Chem. 2000, 275(1 ), 390-397).
  • Ser-129 is also phosphorylated by G-protein-coupled receptor protein kinases (Pronin et al., J. Biol. Chem. 2000, 275(34), 26515-26522). Extensive and selective phosphorylation of alpha-synuclein at Ser-129 is evident in synucleinopathy lesions, including Lewy bodies (Fujiwara et al., Nat. Cell. Biol. 2002, 4(2); 160-164). Other post-translational modifications in the carboxyl-terminal, including glycosylation on Ser-129 (McLean et al., Neurosci. Lett. 2002, 323(3), 219-223) and nitration on Tyr-125, -133, and -136 (Takahashi et al., Brain Res.
  • Abnormal protein aggregation appears to be a common feature in aging brain and in several neurodegenerative diseases (Trojanowski et al., 1998, Cell Death Differ. 1998, 5(10), 832-837, Koo et al., Proc. Natl. Acad. Sci. 1999, 96(18), 9989-9990, Hu et al., Chin. Sci. Bull. 2001 , 46, 1-3); although a clear role in the disease process remains to be defined.
  • alpha-synuclein (or some of its truncated forms) readily assembles into filaments resembling those isolated from the brain of patients with Lewy Body (LB) dementia and familiar PD (Crowther et al., FEBS Lett. 1998, 436(3), 309-312).
  • Alpha-synuclein and its mutated forms (A53T and A30P) have a random coil conformation and do not form significant secondary structures in aqueous solution at low concentrations; however, at higher concentrations they are prone to self-aggregate, producing amyloid fibrils (Wood et al., J. Biol. Chem. 1999, 274(28), 19509-19512).
  • Parkinson’s disease is the most common neurodegenerative motor disorder.
  • PD is mainly an idiopathic disease, although in at least 5% of the PD patients the pathology is linked to mutations in one or several specific genes.
  • alpha-synuclein gene A30P, E46K, H50Q, G51 D, A53T
  • duplications and triplications of the alpha-synuclein gene have been described in patients that developed PD, underlining the role of alpha-synuclein in PD pathogenesis (Lesage et al., Hum. Mol. Genet., 2009, 18, R48-59).
  • the pathogenesis of PD remains elusive. However, growing evidence suggests a role for the pathogenic folding of the alpha-synuclein protein that leads to the formation of amyloid-like fibrils. Indeed, the hallmarks of PD are the presence of intracellular alpha-synuclein aggregate structures called Lewy Bodies and neurites mainly in the nigral neurons, as well as the death of dopaminergic neurons in the substantia nigra and elsewhere.
  • Alpha-synuclein is a natively unfolded presynaptic protein that can misfold and aggregate into larger oligomeric and fibrillar forms which are linked to the pathogenesis of PD.
  • Lewy Bodies are diffusely distributed throughout the cortices of the brain and in addition to Lewy bodies and neurites, more threads and dot-like structures (Lewy dots) were found to be immunopositive for alpha-synuclein phosphorylated at Ser-129 (Outeiro et al., Mol. Neurodegener. 2019, 14, 5).
  • MSA multiple system atrophy
  • MSA is a rare and sporadic neurodegenerative disorder that manifests with rapidly progressive autonomic and motor dysfunction, as well as variable cognitive decline. Such disorders include Shy-Drager syndrome, striatonigral degeneration and olivopontocerebellar atrophy.
  • the disease can be clinically subclassified in parkinsonian (MSA-P) or cerebellar (MSA-C) variant, depending on the predominant motor phenotype (Fanciulli et al., N. Engl. J. Med. 2015; 372, 249-63).
  • GCIs glial cytoplasmic inclusions
  • Parkinson’s disease is largely clinical and depends on the presence of a specific set of symptoms and signs (the initial core feature being bradykinesia, rigidity, rest tremor and postural instability), the absence of atypical features, a slowly progressive course, and the response to a symptomatic drug therapy, mainly limited to a dopamine replacement therapy.
  • the accurate diagnosis requires sophisticated clinical skills and is open to a degree of subjectivity and error, as several other degenerative and non-degenerative diseases can mimic PD symptoms (multiple system atrophy (MSA), progressive supranuclear palsy (PSP), Alzheimer’s disease (AD), essential tremor, dystonic tremor), (Guideline No. 113: Diagnosis and pharmacological management of Parkinson’s disease, January 2010. SIGN).
  • MSA multiple system atrophy
  • PSP progressive supranuclear palsy
  • AD Alzheimer’s disease
  • AD essential tremor
  • dystonic tremor dystonic tremor
  • Computed tomography CT and conventional magnetic resonance imaging (MRI) brain scans of people with Parkinson’s disease (PD) usually appear normal.
  • CT computed tomography
  • MRI magnetic resonance imaging
  • PD Parkinson’s disease
  • Examples are ioflupane ( 123 l) (trade name DaTSCAN) and iometopane (Dopascan) for SPECT or fluorodeoxyglucose ( 18 F) ( 18 F-FDG) and dihydrotetrabenazine ( 11 C) ( 11 C-DTBZ) for PET,
  • a pattern of reduced dopaminergic activity in the basal ganglia can aid in diagnosing PD, particularly in the symptomatic stage (Brooks, J. Nucl. Med., 2010, 51 , 596-609; Redmond, Neuroscientist, 2002, 8, 457-88; Wood, Nat. Rev. Neurol., 2014, 10, 305).
  • biomarkers that have been investigated in different body fluids (cerebrospinal fluid (CSF), plasma, saliva) include alpha-synuclein levels but also DJ-1 , Tau and Abeta, as well as neurofilaments proteins, interleukins, osteopontin and hypocrontin (Schapira Curr. Opin. Neurol. 2013; 26(4):395-400), but so far none of these biomarkers alone or in combination can be used as a determinant diagnostic test. To our knowledge, no approved alpha-synuclein diagnostic agent is currently on the market despite a crucial need for Parkinson's disease research and drug development (Eberling et al., J Parkinsons Dis. 2013; 3(4):565-7).
  • alpha-synuclein deposition in the brain would be a huge achievement for alpha- synucleopathies research, including Parkinson’s disease (PD) and MSA research, diagnosis, and drug development.
  • PD Parkinson’s disease
  • MSA MSA research, diagnosis, and drug development.
  • the accumulation of aggregated alpha-synuclein in the brain is considered a key pathological hallmark of PD and MSA and can start many years before the appearance of the symptoms. Therefore, alpha-synuclein is a priority target for drug development given not only its likely contribution to neurodegeneration but also because it can offer the possibility to treat the disease while still in the asymptomatic or prodromal stages.
  • alpha-synuclein pathology could be useful as a biomarker to (i) detect the presence of the disease potentially in early stages, (ii) to evaluate disease progression and (iii) to be used as a pharmacodynamics tool for drug development.
  • the development of an alpha-synuclein PET imaging agent is considered nowadays key for an accurate diagnosis of synucleinopathies as well as to support the clinical development of therapeutics targeting alpha-synuclein, starting from the optimal selection of the trial population (Eberling, Dave and Frasier, J. Parkinson's Disease, 3, 565-567 (2013)).
  • alpha-synuclein imaging compounds should bind with high affinity and selectivity to their target.
  • imaging compounds For imaging of alpha-synuclein aggregates associated with neurological diseases such as Parkinson's Disease or multiple system atrophy, imaging compounds need to penetrate the blood brain barrier and pass into the relevant regions of the brain.
  • cell permeability is a further requirement of imaging compounds.
  • a further prerequisite in order to avoid unnecessary accumulation of the compound which may result in increased risk of unwanted side-effects is a fast compound wash-out from the brain (or other targeting organ).
  • WO 2011/128455 refers to specific compounds which are suitable for treating disorders associated with amyloid proteins or amyloid-like proteins.
  • US 2012/0302755 relates to certain imaging agents for detecting neurological dysfunction. Further compounds for the diagnosis of neurodegenerative disorders on the olfactory epithelium are discussed in WO 2012/037928.
  • WO 2010/063701 refers to a certain in vivo imaging agent for use in a method to determine the presence of, or susceptibility to, Parkinson's disease, wherein the in vivo imaging agent comprises an alpha-synuclein binder labelled with an in vivo imaging moiety, and wherein the in vivo imaging agent binds to alpha-synuclein with a binding affinity.
  • US 2014/0142089 relates to a method for preventing or treating a degenerative brain disease, the method comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising a specific compound, a pharmaceutically acceptable salt, an isomer, a solvate, a hydrate, and a combination thereof.
  • WO 2009/155017 describes aryl or heteroaryl substituted azabenzoxazole derivatives, which are stated to be useful as tracers in positron emission tomography (PET) imaging to study amyloid deposits in the brain in vivo to allow diagnosis of Alzheimer's disease.
  • PET positron emission tomography
  • WO 2016/033445 refers to a specific compound for imaging huntingtin protein.
  • WO 2017/153601 WO 2019/234243 and WO 2021/224489 refer to bicyclic compounds for imaging alpha-synuclein aggregates.
  • the present invention provides compounds that can be employed in diagnosing a disease, disorder or abnormality associated with alpha-synuclein aggregates, such as Parkinson's disease or MSA, prognosing such a disease, disorder or abnormality, and monitoring the progression of such a disease, disorder or abnormality.
  • the compounds should be suitable for determining a predisposition to such a disease, disorder or abnormality, monitoring the progression of the disease, disorder or abnormality, or predicting the responsiveness of a patient who is suffering from such a disease, disorder or abnormality to the treatment with a certain medicament.
  • the compounds should be suitable for positron emission tomography (PET) imaging of a disease, disorder or abnormality associated with alpha-synuclein aggregates and / or detecting and optionally quantifying alpha-synuclein aggregates.
  • PET positron emission tomography
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, halo C1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl; or
  • R 1 is halo, haloC1-C4alkyl, halo C1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl; or R 1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and R 4 and R 5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl.
  • J is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl ;
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo;
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl.
  • the invention is also directed to a compound having the following subformula (la) or a detestably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
  • R 1 and R 2 are defined above, R 3 is halo, or C1-C4alkyl; and q is 0, 1 or 2.
  • the present invention provides a diagnostic composition comprising a compound of formula (I), and optionally at least one pharmaceutically acceptable excipient, carrier, diluent and/or adjuvant.
  • the present invention provides a compound of formula (I), or a diagnostic composition as defined herein, which can be used in the imaging of alpha-synuclein aggregates.
  • the compound of formula (I), or the diagnostic composition can be for use in positron emission tomography imaging of alpha-synuclein aggregates.
  • the compound of formula (I) or the diagnostic composition, as defined herein can be for use for in vitro imaging, ex vivo imaging, or in vivo imaging, preferably the use is for in vivo imaging, more preferably the use is for brain imaging.
  • the compound of formula (I) or the diagnostic composition, as defined herein can be for use in diagnosis.
  • the present invention refers to a method of diagnosing a disease, disorder or abnormality associated with alpha-synuclein aggregates in a subject, the method comprising the steps:
  • the present invention refers to a method of positron emission tomography (PET) imaging of alpha-synuclein aggregates in a tissue of a subject, the method comprising the steps:
  • the present invention is directed to a method for the detection and optionally quantification of alpha-synuclein aggregates in a tissue of a subject, the method comprising the steps: (a) Bringing a sample or a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound of formula (I), or a diagnostic composition which comprises a compound of formula (I), as defined herein;
  • the present invention is also directed to a method of collecting data for the diagnosis of a disease, disorder or abnormality associated with alpha-synuclein aggregates wherein the method comprises the steps:
  • the present invention also refers to a method of collecting data for determining a predisposition to a disease, disorder or abnormality associated with alpha-synuclein aggregates, the method comprising the steps:
  • the present invention also relates to a method of collecting data for prognosing a disease, disorder or abnormality associated with alpha-synuclein aggregates, wherein the method comprises the steps: (a) Bringing a sample, a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound of the formula (I), or a diagnostic composition which comprises a compound of formula (I), as defined herein;
  • step (e) Optionally repeating steps (a) to (c) and, if present, optional step (d) at least one time.
  • the present invention is directed to a method of collecting data for monitoring the progression of a disease, disorder or abnormality associated with alpha-synuclein aggregates in a patient, the method comprising the steps:
  • step (e) Optionally repeating steps (a) to (c) and, if present, optional step (d) at least one time.
  • the present invention relates to a method of collecting data for predicting responsiveness of a patient suffering from a disease, disorder or abnormality associated with alpha- synuclein aggregates to a medicament, the method comprising the steps:
  • the invention is further directed to a compound of formula (lll-F): or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1F is a 4- to 6-membered heterocyclyl, which is optionally substituted with at least one C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably wherein R 1F is a 4- to 6-membered heterocyclyl; or
  • R 1F is C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably R 1F is C1-C4alkoxy, or C1-C4alkyl; or
  • R 1F is -NH-C3-C6cycloalkyl or Cs-Cacycloalkyl
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl ;
  • R 4 and R 5 are independently selected from H, and C1-C4alkyl
  • LG is a leaving group; and n is at least 1 (e.g., 1 , or 2 or 3), preferably 1.
  • the invention refers to a compound of formula (lll-F) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1F is a 4- to 6-membered heterocyclyl
  • R 1F is C1-C4alkoxy, or C1-C4alkyl
  • R 1F is -NH-C3-C6cycloalkyl or C3-C6cycloalkyl
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
  • LG is a leaving group; and n is at least 1 .
  • the invention refers to a compound of formula (lll-F’) (lll-F’) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
  • (A) is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or CrC4alkyl;
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, CrC4alkoxy, NR 4 R 5 , or C1-C4alkyl; or
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -Cg-Cgcycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 4 and R 5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl
  • R 2F is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from C1-C4alkoxy, and C1-C4alkyl;
  • LG is a leaving group
  • a further aspect of the invention relates to a compound of formula (l-F) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
  • (A ) is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or CrC4alkyl;
  • R 1F is a 4- to 6-membered heterocyclyl, which is optionally substituted with at least one C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably R 1F is a 4- to 6-membered heterocyclyl; or
  • R 1F is C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably R 1F is C1-C4alkoxy, or C1-C4alkyl; or
  • R 1F is -NH-C3-C6cycloalkyl or C3-C6cycloalkyl; and R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
  • R 4 and R 5 are independently selected from H, and C1-C4alkyl; and n is at least 1 (e.g., 1 , 2 or 3), preferably 1.
  • the invention is direct to a compound of formula (l-F) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
  • 6 ⁇ is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1F is a 4- to 6-membered heterocyclyi
  • R 1F is C1-C4alkoxy, or C1-C4alkyl
  • R 1F is -NH-C3-C6cycloalkyl or C3-C6cycloalkyl
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and n is at least 1 , preferably 1.
  • the invention is directed to a compound a compound of formula (l-F’): or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
  • ⁇ -2 is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1 is a 4- to 6-membered heterocyclyi which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl; or
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 4 and R 5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl; and R 2F is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from C1-C4alkoxy, and C1-C4alkyl.
  • the invention is further directed to a compound of formula (lll-H) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
  • (A) is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably R 1 is a 4- to 6- membered heterocyclyl which is optionally substituted with at least one halo; or
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably halo, haloC1-C4alkyl, haloC1-C4alkoxy, C-i-C 4 alkoxy, or C1-C4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 2 is a 5-membered or 6-membered heteroaryl, optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
  • R 4 and R 5 are independently selected from H, CrC 4 alkyl and haloC1-C4alkyl;
  • X is bromo, chloro or iodo; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; and with the proviso that the compound of formula (lll-H) comprises at least one X.
  • a compound of formula (lll-H) is defined or a stereoisomer, racemic mixture, pharmaceuticaliy acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo;
  • R 1 is halo, haloC1-C4alkyl, haloCi ⁇ alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or
  • -C3-C6cycloalkyl is optionally substituted with at least one halo
  • R2 is a 5-membered or 6-membered heteroaryl, optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
  • X is bromo, chloro or iodo; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; and with the proviso that the compound of formula (II l-H) comprises at least one X.
  • the invention is further directed to a compound of formula (l-H) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably R 1 is a 4- to 6- membered heterocyclyl which is optionally substituted with at least one halo; or
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -C 3 -C 6 cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C 3 -C 6 cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
  • R 4 and R 5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl; Y is D, CDs, T or CT 3 ; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; with the proviso that the compound of formula (l-H) comprises at least one D, CD 3 , T or CT 3 , wherein D is 2 H (deuterium) and T is 3 H (Tritium). In one embodiment, the compound of formula (l-H) comprises at least one T or CT 3 . In one embodiment, the compound of formula (l-H) comprises at least one D or CD 3 .
  • the invention relates to a compound of formula (l-H) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof,
  • CA is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloCrC4alkyl, haloC 1 -C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo;
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -C 3 -Cecycloalkyl, wherein the C3-C6cycloalkyl of -NH-C 3 -Cecycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
  • Y is D, CD 3 , T or CT 3 ; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; with the proviso that the compound of formula (l-H) comprises at least one D, CD 3 , T or CT 3 , wherein D is Deuterium and T is 3 H (Tritium).
  • the invention is further directed to a method of preparing a compound of formula (l-F) by reacting a compound of formula (lll-F) with a 18 F-fluorinating agent (e.g., K 18 F, Rb 18 F, Cs 18 F, Na 18 F, tetra(Ci-6alkyl)ammonium salt of 18 F, Kryptofix[222] 18 F, tetrabutylammonium [ 18 F]fluoride or any other suitable agents), so that the Leaving Group (LG) is replaced by 18 F.
  • a 18 F-fluorinating agent e.g., K 18 F, Rb 18 F, Cs 18 F, Na 18 F, tetra(Ci-6alkyl)ammonium salt of 18 F, Kryptofix[222] 18 F, tetrabutylammonium [ 18 F]fluoride or any other suitable agents
  • the invention is further directed to a method of preparing a compound of formula (l-H) by reacting the compound of formula (lll-H) with a 3 H radiolabelling agent (e.g., tritium gas or any other suitable agents), so that X is replaced by T or CT3.
  • a 3 H radiolabelling agent e.g., tritium gas or any other suitable agents
  • the invention is further directed to a method of preparing a compound of formula (l-H) by reacting the compound of formula (lll-H) with a 2 H radiolabelling agent comprising D, (e.g., D2O, D4-methanol or any other suitable agents) preferably in the presence of a catalyst like Pd/C, so that X is replaced by D or CD3 (D is deuterium).
  • a 2 H radiolabelling agent comprising D, (e.g., D2O, D4-methanol or any other suitable agents) preferably in the presence of a catalyst like Pd/C, so that X is replaced by D or CD3 (D is deuterium).
  • the invention is further directed to the use of the compound of formula (I) as an in vitro analytical reference or an in vitro screening tool.
  • the invention is further directed to a test kit for detection and/or diagnosis of a disease, disorder or abnormality associated with alpha-synuclein aggregates, wherein the test kit comprises at least one compound of formula (I) as defined herein, preferably at least one detectably labelled compound, more preferably at least one compound of formula (l-F) or (l-H).
  • the test kit comprises at least one compound of formula (I) as defined herein, preferably at least one detectably labelled compound, more preferably at least one compound of formula (l-F) or (l-H).
  • the invention is further directed to a kit for preparing a radiopharmaceutical preparation, wherein the kit comprises a sealed vial containing at least one compound of formula (lll-F) or (lll-H).
  • C1-C4alkyl refers to a saturated straight or branched hydrocarbon chain consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to four carbon atoms, and which is attached to the rest of the molecule by a single bond.
  • suitable alkyl groups having 1 to 4 carbon atoms include, but are not limited to, methyl, ethyl, propyl, isopropyl, 1 -methylethyl, n-butyl, t-butyl and isobutyl.
  • C1-C4alkoxy refers to a radical of the formula -ORa where Ra is a C1-C4alkyl radical as generally defined above. Examples of C1-C4alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, and isobutoxy.
  • halogenC1-C4alkyl or "haloC1-C4alkyl” refer to a C1-C4alkyl radical as defined above, substituted with one or more (e.g., 1 , 2 or 3, preferably 1 or 2, more preferably 1) halo radicals as defined below.
  • haloC1-C4alkyl include, but are not limited to, trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1 ,3-dibromopropan-2-yl, 3-bromo-2- fluoropropyl and 1 ,4,4-trifluorobutan-2-yl.
  • halogenC1-C4alkoxy refers to a C1-C4alkoxy radical as defined above, substituted with one or more (e.g., 1 , 2 or 3, preferably 1 or 2, more preferably 1 ) halo radicals as defined below.
  • haloC1-C4alkoxy include, but are not limited to, trifluoromethoxy, difluoromethoxy, fluoromethoxy, 2,2,2-trifluoroethoxy, 3,3,3-trifluoropropoxy, 4,4,4-trifluorobutoxy, 2,2-difluorobutoxy, and 4-bromobutoxy.
  • heterocyclyl refers to a stable 4- to 6-membered non-aromatic monocyclic ring radical which comprises 1 or 2 heteroatoms which are, e.g., selected from N, O or S.
  • the heterocyclyl group can be unsaturated or saturated.
  • the heterocyclyl radical may be bonded via a carbon atom or a heteroatom.
  • Examples include, but are not limited to, azetidinyl, oxetanyl, pyrrolidinyl, pyrrolidyl, tetrahydrofuryl, tetrahydrothienyl, piperidyl, piperazinyl, tetrahydropyranyl, or morpholinyl, preferably azetidinyl, pyrrolidinyl, or piperidyl, more preferably pyrrolidinyl.
  • heteroaryl refers to a 5- or 6-membered aromatic monocyclic ring, which comprises 1 , 2, or 3 heteroatoms independently selected from N, O and S.
  • the heteroaryl radical may be bonded via a carbon atom or heteroatom selected from N, O and S.
  • heteroaryl include, but are not limited to, thiopyranyl, dioxanyl, pyranyl, pyrazinyl, pyridazinyl, pyrimidyl, isothiazolyl, pyrazolyl, thiazolyl or pyridyl, with pyridyl, isothiazolyl, pyrazolyl, and thiazolyl being preferred.
  • Hal or “halogen” or “Halo” refers to F, Cl, Br, and I. With respect to diagnostic and pharmaceutical applications, F (e.g., 19 F and 18 F) is particularly preferred.
  • LG leaving group
  • LG is any leaving group and means an atom or group of atoms that can be replaced by another atom or group of atoms. Examples are given e.g. in Synthesis (1982), p. 85-125, table 2, Carey and Sundberg, Organische Synthese, (1995), page 279- 281 , table 5.8; or Netscher, Recent Res. Dev. Org. Chem., 2003, 7, 71-83, schemes 1 , 2, 10 and 15 and others).
  • the "leaving group” is selected from halogen, C1-C4 alkylsulfonate and C6-C10arylsulfonate, wherein the Ce-Cwarylsulfonate can be optionally substituted with -CH3 or -NO2.
  • the term “compound of the invention” refers to a compound of formula (I), or of subformulae thereof (e.g. (la), (l-F), (l-H*), (l-H)), or a detectably labelled compound, stereoisomer (including diastereomeric mixtures and individual diastereomer, enantiomeric mixture and single enantiomer, mixture of conformers and single conformer), racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof. It is understood that every reference to a compound of formula (I) also covers the subformulae thereof (e.g. (la), (l-F), (l-H*), (l-H)).
  • the compounds of the formulae (II l-F) and (I I l-H) will be referred to as the precursors of the compounds of the present invention.
  • Compounds of the present invention and their precursors having one or more optically active carbons can exist as racemates and racemic mixtures, stereoisomers (including diastereomeric mixtures and individual diastereomers, enantiomeric mixtures and single enantiomers, mixtures of conformers and single conformers), tautomers, atropoisomers, and rotamers. All isomeric forms are included in the present invention.
  • “Pharmaceutically acceptable salts” are defined as derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof.
  • Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like.
  • the pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids.
  • such conventional non-toxic salts include those derived from inorganic acids such as, but not limited to, hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as, but not limited to, acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, and the like.
  • inorganic acids such as, but not limited to, hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like
  • organic acids such as, but not limited to
  • the pharmaceutically acceptable salts of the compounds of the present invention and their precursors can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods.
  • such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two.
  • Organic solvents include, but are not limited to, nonaqueous media like ethers, ethyl acetate, ethanol, isopropanol, or acetonitrile. Lists of suitable salts can be found in Remington’s Pharmaceutical Sciences, 18 th ed., Mack Publishing Company, Easton, PA, 1990, p. 1445, the disclosure of which is hereby incorporated by reference.
  • “Pharmaceutically acceptable” is defined as those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit/risk ratio.
  • Solvates can be formed from the compound of the present invention and any suitable pharmaceutically acceptable solvent.
  • suitable pharmaceutically acceptable solvent examples include C1-4 alcohols (such as methanol or ethanol).
  • the patients or subjects in the present invention are typically animals, particularly mammals, more particularly humans.
  • Alpha-synuclein aggregates are multimeric beta-sheet rich assemblies of alpha-synuclein monomers that can form either soluble oligomers or soluble/insoluble protofibrils or mature fibrils which coalesce into intracellular deposits detected as a range of Lewy pathologies in Parkinson’s disease and other synucleinopathies.
  • Alpha-synuclein aggregates that are composing Lewy pathologies can be detected as having the following morphologies: Lewy bodies, Lewy neurites, premature Lewy bodies or pale bodies, perikaryal deposits with diffuse, granular, punctate or pleomorphic patterns.
  • alpha-synuclein aggregates are the major component of intracellular fibrillary inclusions detected in oligodendrocytes (also referred to as glial cytoplasmic inclusions) and in neuronal somata, axons and nuclei (referred to as neuronal cytoplasmic inclusions) that are the histological hallmarks of multiple system atrophy.
  • Alpha-synuclein aggregates in Lewy pathologies often display substantial increase in post-translational modifications such as phosphorylation, ubiquitination, nitration, and truncation.
  • Lewy bodies are abnormal aggregates of protein that develop inside nerve cells in Parkinson’s disease (PD), Lewy body dementia and other synucleinopathies. Lewy bodies appear as spherical masses that displace other cell components. Morphologically, Lewy bodies can be classified as being brainstem or cortical type. Classic brainstem Lewy bodies are eosinophilic cytoplasmic inclusions consisting of a dense core surrounded by a halo of 5-10-nm-wide radiating fibrils, the primary structural component of which is alpha-synuclein; cortical Lewy bodies differ by lacking a halo. The presence of Lewy bodies is a hallmark of Parkinson’s disease.
  • Lewy neurites are abnormal neuronal processes in diseased neurons, containing granular material, abnormal alpha-synuclein (a-syn) filaments similar to those found in Lewy bodies, dot-like, varicose structures and axonal spheroids. Like Lewy bodies, Lewy neurites are a feature of a- synucleinopathies such as dementia with Lewy bodies and Parkinson's disease.
  • Glial cytoplasmic inclusions are argyrophilic cytoplasmic aggregates in oligodentroglial cells composed of filamentous alpha-synuclein. Morphologically appear as triangles, half-moon or sickle shapes.
  • inclusions composed of alpha-synuclein filaments are detected in neurons in the cytoplasm or beneath the nuclear membrane termed neuronal cytoplasmic inclusions and neuronal nuclear inclusions respectively.
  • GCIs are recognized as the defining morphological feature of MSA; their widespread distribution is a criterion for the definite post-mortem neuropathological diagnosis of MSA.
  • the compounds of formula (I) can bind to alpha-synuclein aggregates.
  • the type of bonding with the compounds of formula (I) has not been elucidated and any type of bonding is covered by the present invention.
  • the wording "compound bound to the alpha-synuclein aggregates" and the like are used interchangeably herein and are not considered to be limited to any specific type of bonding.
  • the present invention relates to a compound of formula (I): or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC 1 -C4alkoxy, C1-C4alkoxy, or C1-C4alkyl, if substituted, ⁇ - ⁇ is substituted with 1 , 2 or 3, preferably 1 or 2, more preferably with 1 halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloCr C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo; or
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and R 4 and R 5 are independently selected from H, CrC4alkyl and haloC1-C4alkyl. Throughout the present application, R 4 and R 5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl, preferably at least one of R 4 and R 5 is not H.
  • the optional substituent of is preferably halo or C1-C4alkyl.
  • R 1 is pyridyl which can be substituted at any available position by R 1 , more preferably R 1 -V> is being as defined above.
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl (e.g., 1 to 3, preferably 1 or 2, more preferably 1 ).
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo (e.g., 1 to 3, preferably 1 or 2, more preferably 1 ).
  • R 1 is a 4- to 6-membered heterocyclyl which is substituted with at least one halo.
  • the heterocyclyl is substituted with at least one halo, more preferably with one or two halo, even more preferably with one halo.
  • R 1 is a 4- to 6- membered heterocyclyl which is unsubstituted.
  • R 1 is a 4- to 6-membered heterocyclyl selected from the following: wherein R 1a is F or H, preferably F. In another preferred embodiment R 1a is H.
  • R 1 is a 4- or 5-membered heterocyclyl selected from the following: wherein R 1a is F or H, preferably F. In another preferred embodiment R 1a is H.
  • R 1 is a 5-membered heterocyclyl which is:
  • R 1 is a 5-membered heterocyclyl which is: o .
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl. In an embodiment R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl.
  • R 1 is NR 4 R 5 , preferably wherein R 4 and R 5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl.
  • R 1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo.
  • F is preferably 19 F or 18 F.
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl, preferably haloC1-C4alkyl, or C1-C4alkyl.
  • R 2 is a 5-membered or 6-membered heteroaryl selected from the following: wherein
  • R 2a is independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
  • R 2b is selected from H, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and s is 0, 1 or 2 (preferably 0 or 1 , more preferably 0). In another preferred embodiment s is 1 .
  • R 2 is a 5-membered or 6-membered heteroaryl selected from the following: wherein
  • R 2b is selected from haloC1-C4alkyl, haloC1-C4alkoxy, alkoxy, H or C1-C4alkyl, preferably H, haloC1-C4alkyl (preferably wherein halo is F) or C1-C4alkyl, preferably H or C1-C4alkyl.
  • the invention provides a compound having the formula (la): or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein R 1 and R 2 are as defined above, and R 3 is halo, or C1-C4alkyl; and q is 0, 1 or 2.
  • the present invention provides a compound of formula (I), wherein the compound is selected from
  • the present invention provides a compound of formula (I), wherein the compound is selected from
  • the present invention provides a compound of formula (I) which is a detectably labelled compound.
  • the detectable label can be a radioisotope.
  • the compound of formula (I) comprises at least one radioisotope.
  • the detectable label is a radioisotope selected from 18 F, 2 H and 3 H. Most preferably, the radioisotope is selected from 18 F and 3 H.
  • the present invention provides a compound of formula (I) wherein R 1 is
  • the present invention provides a compound of formula (I) wherein R 1 is -O-CH 2 -CH 2 - 18 F.
  • the present invention provides a compound of formula (I) wherein R z is , wherein R 2b is -CH 2 -CH 2 - 18 F or -CH 2 -CH 2 -CH 2 - 18 F.
  • the present invention provides a compound of formula (I) wherein R 2 is
  • the present invention provides a compound of formula (I), wherein the compound is a detectably labelled compound of formula (l-F): or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl; which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1F is a 4- to 6-membered heterocyclyl, which is optionally substituted with at least one C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably a 4- to 6-membered heterocyclyl; or
  • R 1F is C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably C1-C4alkoxy, or C1-C4alkyl; or
  • R 1F is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl
  • R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
  • R 4 and R 5 are independently selected from H, and C1-C4alkyl; and n is at least 1 (e.g., 1, 2 or 3), preferably 1.
  • R 1F is -NH-C3-C6cycloalkyl, C3-C6cycloalkyl, C1-C4alkoxy, or C1-C4alkyl or heterocyclyl.
  • -R 1F -( 18 F) n is selected from the following: More preferably, -R 1F -( 18 F) n is selected from the following:
  • -R 1F -( 18 F) n is:
  • the present invention provides a compound of formula (I), wherein the compound is a detectably labelled compound of formula (l-F’): or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloCr
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 4 and R 5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl
  • R 2F is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from C1-C4alkoxy.
  • -R 2F -( 18 F) n is selected from the following
  • the detectably labelled compound of formula (l-F) or (l-F’) comprises at least one 18 F.
  • the detectably labelled compound of formula (l-F) or (l-F’) comprises one 18 F.
  • the present invention provides a compound of formula (I), wherein the compound is a detectably labelled compound of formula (l-H*) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof,
  • haloC1-C4alkyl is optionally substituted with at least one halo; haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or (XUalkyl;
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo; or
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or CrC4alkyl, preferably halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and R 4 and R 5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl; with the proviso that the compound of formula (l-H*) comprises at least one 2 H (deuterium “D”) or 3 H (Tritium “T”), preferably 1 , 2, or 3 D or T, even more preferably 2 or 3 D or T.
  • 2 H deuterium “D”
  • Tritium “T” Tritium “T”
  • the compound of formula (l-H*) comprises at least one 3 H (Tritium “T”), preferably 1 , 2, or 3 T, even more preferably 2 or 3 T.
  • the 3 H can be present as T or as -CT3.
  • the 2 H can be present as D or as -CD3.
  • the compound is a detectably labelled compound of formula (l-H) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloCrC 4 alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, CrC 4 alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo; or
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or CrC 4 alkyl, preferably halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and CrC 4 alkyl;
  • R 4 and R 5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl;
  • Y is D, CD 3 , T or CT 3 ; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; with the proviso that the compound of formula (l-H) comprises at least one D, CD 3 , T or CT 3 , wherein D is 2 H (Deuterium) and T is 3 H (Tritium).
  • the compound of formula (l-H) comprises at least one D or CD 3 .
  • the compound of formula (l-H) comprises at least one T or CT 3 .
  • deuterium or tritium can present at any available position at which a hydrogen is present.
  • deuterium or tritium can be present either directly bound to the 5-membered or 6-membered heteroaryl (such as in the form of D or T) or can be present in the haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl (such as in the form of CD 3 or CT 3 ).
  • deuterium or tritium can be, e.g., directly bound to the 4- to 6-membered heterocyclyl.
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl, and p is 1 , 2 or 3, preferably 1.
  • R 2 is a 5-membered or 6-membered heteroaryl selected from the following: wherein
  • R 2a is independently selected from T, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
  • R 2b is selected from H, T, haloC1-C4alkyl and C1-C4alkyl, wherein haloC1-C4alkyl and C1-C4alkyl optionally comprise one or more T (preferably R 2b is selected from T or CT3); and s is 0, 1 or 2 (preferably 0 or 1 , more preferably 0).
  • R 2a is -T, -OCH3, -CH 3 or -H; and R 2b is selected from -H, -T or -CT3.
  • R 2 is a 5-membered or 6-membered heteroaryl selected from the following: wherein
  • R 2b is selected from T, H or C1-C4alkyl (e.g., CT3), preferably T, H or C1-C4alkyl (e.g., CT 3 ).
  • the detectably labelled compound of formula (l-H*) or (l-H) comprises one, two or three T.
  • the detectably labelled compound of formula (l-H*) or (l-H) comprises one T.
  • the detectably labelled compound of formula (l-H*) or (l-H) comprises two T.
  • the detectably labelled compound of formula (l-H*) or (l-H) comprises three T such as -CT3.
  • R 2 is a 5-membered or 6-membered heteroaryl selected from the following: wherein
  • R 2a is independently selected from D, haloC1-C4alkyl, haloC1-C4alkoxy, CrC4alkoxy, and C1-C4alkyl;
  • R 2b is selected from H, D, haloC1-C4alkyl and C1-C4alkyl, wherein haloC1-C4alkyl and C1-C4alkyl optionally comprise one or more D (preferably R 2b is selected from D or CD3); and s is 0, 1 or 2 (preferably 0).
  • R 2a is -D, -OCH3, -CHsor -H; and R 2b is selected from -H, -D or -CD3.
  • R 2 is a 5-membered or 6-membered heteroaryl selected from the following: wherein
  • R 2b is selected from D, H or C1-C4alkyl (e.g., CD3).
  • the detectably labelled compound of formula (l-H*) or (l-H) comprises one, two or three D.
  • the detectably labelled compound of formula (l-H*) or (l-H) comprises one D.
  • the detectably labelled compound of formula (l-H*) or (l-H) comprises two D.
  • the detectably labelled compound of formula (l-H*) or (l-H) comprises three D such as -CD3.
  • the invention provides a detectably labelled compound of formula (l-H*) or (l-H) wherein 3 H Tritium (“T”) can be replaced by 2 H Deuterium (“D”).
  • the deuterated compound can be prepared by reacting a compound of formula (lll-H) with a 2 H radiolabelling agent.
  • the compounds of the present invention and their precursors can be detectably labelled.
  • the type of the label is not specifically limited and will depend on the detection method chosen. Examples of possible labels include isotopes such as radionuclides, positron emitters, and gamma emitters, preferably the detectable label is a radioisotope.
  • the radioisotope, positron emitter, or gamma emitter is to be present in an amount which is not identical to the natural amount of the respective radioisotope, positron emitter, or gamma emitter.
  • the employed amount should allow detection thereof by the chosen detection method.
  • suitable isotopes such as radionuclides, positron emitters and gamma emitters include 2 H, 3 H, 11 C, 13 N, 15 O, and 18 F, preferably 2 H, 3 H and 18 F.
  • 18 F-labelled compounds are particularly suitable for imaging applications such as PET.
  • the corresponding compounds which include fluorine having a natural 19 F isotope are also of particular interest as they can be used as analytical standards and references during manufacturing, quality control, release, and clinical use of their 18 F-analogs.
  • substitution with isotopes such as deuterium, i.e. 2 H or D may afford certain diagnostic and therapeutic advantages resulting from greater metabolic stability by reducing for example defluorination, increased in vivo half-life or reduced dosage requirements, while keeping or improving the original compound efficacy.
  • Isotopic variations of the compounds of the invention and their precursors can generally be prepared by conventional procedures such as by the illustrative methods or by the preparations described in the Examples and Preparative Examples hereafter using appropriate isotopic variations of suitable reagents, which are commercially available or prepared by known synthetic techniques.
  • Radionuclides, positron emitters and gamma emitters can be included into the compounds of the present invention and their precursors by methods which are usual in the field of organic synthesis. Typically, they will be introduced by using a correspondingly labelled starting material when the desired compound of the present invention and its precursor is prepared. Illustrative methods of introducing detectable labels are described, for instance, in US 2012/0302755.
  • the position at which the detectable label is to be attached to the compounds of the present invention and their precursors is not particularly limited.
  • the radionuclides, positron emitters and gamma emitters can be attached at any position where the corresponding non-emitting atom can also be attached.
  • 18 F can be attached at any position which is suitable for attaching F.
  • R 1 is substituted with 18 F.
  • 3 H can be attached at any available position at which H is present. If 2 H is employed as a detectable label it can be attached at any available position at which H is present.
  • the present invention relates further to a compound of formula (lll-F) that is a precursor of the compound of formula (l-F) (lll-F) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein fA') is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1F is a 4- to 6-membered heterocyclyl, which is optionally substituted with at least one C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably R 1F is a 4- to 6-membered heterocyclyl; or
  • R 1F is C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably R 1F is C1-C4alkoxy, or C1-C4alkyl; or
  • R 1F is -NH-C3-C6cycloalkyl or C3-C 6 cycloalkyl
  • R 2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
  • R 4 and R 5 are independently selected from H, and C1-C4alkyl
  • LG is a leaving group; and n is at least 1 (e.g., 1, 2 or 3; preferably 1 ).
  • (LG) n -R 1F is selected from the following: wherein x is at least 1 (e.g., 1 to 3), preferably 1 or 2, even more preferably 2.
  • (LG) n -R 1F is selected from the following: wherein x is 1 or 2, more preferably 2.
  • (LG) n -R 1F is :
  • the present invention relates further to a compound of formula (lll-F') that is a precursor of the compound of formula ( l-F’):
  • R 1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl; or
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or CrC4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C 3 -C 6 cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 4 and R 5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl ;
  • R 2F is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from C1-C4alkoxy, and C1-C4alkyl;
  • LG is a leaving group
  • -R 2F -( LG ) is selected from the following: wherein R 2b is -CH 2 -CH 2 -LG or -CH 2 -CH 2 -CH 2 -LG; or
  • the Leaving Group (LG) in (lll-F) or (lll-F’) is halogen, C1-C4 alkylsulfonate, C1-C4alkyl ammonium, or Ce-Cwarylsulfonate, wherein the Ce-C-ioarylsulfonate can be optionally substituted with -CH3 or -NO 2 . More preferably, the Leaving Group (LG) is bromo, chloro, iodo, C6-C4alkylsulfonate, or Ce-Cwarylsulfonate, wherein the Ce-Cwarylsulfonate can be optionally substituted with -CH3 or -NO2.
  • the Leaving Group (LG) is mesylate, tosylate or nosylate. Even more preferably, the Leaving Group (LG) is mesylate, or nosylate. More preferably the Leaving Group (LG) is mesylate.
  • the present invention relates to a compound of formula (lll-H), a precursor of the compound of formula (l-H): or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
  • ( A ) is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
  • R 1 is a 4- to 6-membered heterocyclyl which is' optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or C1-C4alkyl, preferably optionally substituted with at least one halo; or
  • R 1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR 4 R 5 , or CrC4alkyl, preferably halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
  • R 1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
  • R 2 is a 5-membered or 6-membered heteroaryl, optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and
  • R 4 and R 5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; and
  • X is bromo, chloro or iodo; with the proviso that the compound of formula (lll-H) comprises at least one X (e.g., 1 , 2 or 3 X, preferably 1 or 2 X).
  • halo is preferably F, 18 F or 19 F.
  • X is attached to the 6-membered heteroaryl of and or the 5-membered or 6-membered heteroaryl of R 2 . If halo is present as a substituent of and X is present, the X can be present in addition of halo.
  • R 2 is selected from the following: wherein
  • R 2a is independently selected from X, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; wherein haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkyl, or C1-C4alkoxy are optionally substituted with one or more X; s is 0, 1 or 2 (preferably 0 or 1 ); and
  • R 2b is selected from H, X, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl.
  • R 2 is selected from the following: wherein
  • R 2a is X
  • R 2b is selected from H, X, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl, preferably H, X, haloC1-C4alkyl, and C1-C4alkyl, more preferably X.
  • R 2b is selected from haloC1-C4alkyl, and haloC1-C4alkoxy; s is 0, 1 or 2 (preferably 0); and wherein C1-C4alkyl is optionally substituted by one or more X.
  • the detectably labelled compound of formula (lll-H) comprises one, two or three X.
  • the detectably labelled compound of formula (lll-H) comprises one or two X.
  • the detectably labelled compound of formula (lll-H) comprises two X, X is selected from bromo, chloro and iodo. In a preferred embodiment X is bromine.
  • the present invention relates further to a method for preparing a compound of formula (I), or of subformulae thereof (e.g. (I-F), (l-F’), (l-H*), (l-H)), and in particular a compound of formula (lll-F), (lll-F’), or (lll-H).
  • the present invention relates to a method for preparing a compound of formula (l-F) by reacting a compound of formula (lll-F) with a 18 F-fluorinating agent, so that LG is replaced by 18 F .
  • a compound of formula (lll-F) with a 18 F-fluorinating agent, so that LG is replaced by 18 F .
  • R 1F , R 2 , n, and LG are as defined herein above.
  • the present invention relates to a method for preparing a compound of formula (l-F’) by reacting a compound of formula (lll-F’) with a 18 F-fluorinating agent, so that LG is replaced by 18 F.
  • a compound of formula (lll-F’) with a 18 F-fluorinating agent, so that LG is replaced by 18 F.
  • R 1 , R 2F and LG are as defined herein above.
  • Suitable solvents for the 18 F-fluorination comprise DMF, DMSO, acetonitrile, DMA, or mixtures thereof, preferably acetonitrile or DMSO.
  • Suitable agents for the 18 F-fluorination are selected from K 18 F, Rb 18 F, Cs 18 F, Na 18 F, tetra(Ci-6alkyl)ammonium salt of 18 F, Kryptofix[222] 18 F and tetrabutylammonium [ 18 F]fluoride.
  • the present invention relates to a method of preparing a compound of formula (l-H) by reacting a compound of formula (lll-H) with a 3 H radiolabelling agent. wherein R 1 , R 2 , X, Y, m, and p are as defined herein above.
  • the present invention relates to a method of preparing a compound of formula (l-H*), by reacting a compound of formula (lll-H*) with a 3 H or D radiolabelling agent.
  • a compound of formula (lll-H*) with a 3 H or D radiolabelling agent.
  • R 1 , R 2 , X, m, and p are as defined herein above and wherein Y is D, CDs, T or CT3.
  • the 3 H radiolabelling agent can be tritium gas.
  • the method can be conducted in the presence of a catalyst such as palladium on carbon (Pd/C), a solvent such as dimethylformamide (DMF) and a base such as N,N-diisopropylethylamine (DIEA).
  • a catalyst such as palladium on carbon (Pd/C)
  • a solvent such as dimethylformamide (DMF)
  • DIEA N,N-diisopropylethylamine
  • the compound of formula (lll-H*) can be reacted with a 2 H radiolabelling agent comprising D (e.g., D2O, D4-methanol or any other suitable agents), preferably in the presence of a catalyst like Pd/C, so that X is replaced by D (D is deuterium, 2 H).
  • a 2 H radiolabelling agent comprising D (e.g., D2O, D4-methanol or any other suitable agents), preferably in the presence of a catalyst like Pd/C, so that X is replaced by D (D is deuterium, 2 H).
  • the present invention relates to a method for preparing a compound of formula (l-H*) by radiolabelling a compound of formula (lll-H*) (reacting the compound of formula (lll-H*)) with a 3 H radiolabelling agent like CT3 radiolabelling agent, wherein T is 3 H, so that X is replaced by CT3.
  • the CT3 radiolabelling agent can be ICT3 (derivative of iodomethane with 3 H).
  • the method can be conducted in the presence of a solvent such as dimethylformamide (DMF) and a base such as caesium carbonate or sodium hydride. Kits
  • the precursor compounds of the present invention can also be employed in kits for the preparation of radiopharmaceutical preparations. Due to the radioactive decay, the radiopharmaceuticals are usually prepared immediately before use.
  • the kit typically comprises a precursor of the compound of the present invention, and an agent which reacts with the precursor to introduce a radioactive label into the compound of the present invention.
  • the precursor of the compound of the present invention can, for example, be a compound having the formula (lll-F), (lll-H*) or (lll-H).
  • the agent can be an agent which introduces a radioactive label such as 18 F, 3 H, or D.
  • the kit of parts is a test kit for the detection and/or diagnosis of a disease, disorder or abnormality associated with alpha-synuclein aggregates, wherein the test kit comprises at least one precursor of the compound of the present invention (e.g. a compound having the formula (lll-F), (lll-H*) or (lll-H)).
  • the test kit comprises at least one precursor of the compound of the present invention (e.g. a compound having the formula (lll-F), (lll-H*) or (lll-H)).
  • the kit of parts is a kit for preparing a radiopharmaceutical preparation, wherein the kit comprises a sealed vial containing at least one precursor of the compound of the present invention (e.g. a compound having the formula (lll-F), (lll-H*) or (lll-H)).
  • a sealed vial containing at least one precursor of the compound of the present invention (e.g. a compound having the formula (lll-F), (lll-H*) or (lll-H)).
  • the kit is for use in the imaging of alpha-synuclein aggregates, wherein the imaging is preferably conducted by positron emission tomography, or is for use for in vitro imaging, ex vivo imaging, or in vivo imaging, preferably the use is for in vivo imaging. More preferably, the use is for brain imaging.
  • the compounds of the present invention are particularly suitable for imaging of alpha-synuclein aggregates including. With respect to alpha-synuclein protein, the compounds are particularly suitable for binding to various types of alpha-synuclein aggregates including, but not limited to, Lewy bodies and/or Lewy neurites.
  • the imaging can be conducted in mammals, preferably in humans.
  • the imaging is preferably in vitro imaging, ex vivo imaging, or in vivo imaging. More preferably the imaging is in vivo imaging: Even more preferably, the imaging is preferably brain imaging.
  • the imaging can also be eye/retinal imaging.
  • the compounds of the present invention are particularly suitable for use in diagnostics.
  • the diagnostics can be conducted for mammals, preferably for humans.
  • the tissue of interest on which the diagnostic is conducted can be brain, tissue of the central nervous system, tissue of the eye (such as retinal tissue), tissue of peripheral organs such as the gut or other tissues, or body fluids such as cerebrospinal fluid (CSF) or blood.
  • the tissue is preferably brain tissue.
  • the present invention provides a diagnostic composition
  • a diagnostic composition comprising a compound of the invention, and optionally at least one pharmaceutically acceptable excipient, carrier, diluent and/or adjuvant.
  • the compounds of the present invention are suitable for use in the diagnosis of diseases, disorders and abnormalities associated with alpha- synuclein aggregates.
  • the diagnostic composition which comprises a compound of the present invention is also suitable for use in the diagnosis of diseases, disorders and abnormalities associated with alpha-synuclein aggregates.
  • the compound of the present invention, or the diagnostic composition comprising a compound of the invention is suitable for use in imaging, such as in vitro imaging, ex vivo imaging, or in vivo imaging, preferably the use is for in vivo imaging, more preferably the use is for brain imaging. In particular, the use is in humans.
  • the compounds of the present invention or the diagnostic composition are particularly suitable for use in positron emission tomography imaging of alpha-synuclein aggregates.
  • the compounds of the present invention are suitable for use in the diagnosis of diseases, disorders or abnormalities associated with alpha-synuclein aggregates including, but not limited to, Lewy bodies and/or Lewy neurites, or a predisposition therefor, wherein the diseases, disorders or abnormalities are selected from (including, but not limited to) Parkinson's disease (sporadic, familial with alpha-synuclein mutations, familial with mutations other than alpha-synuclein, pure autonomic failure and Lewy body dysphagia), SNCA duplication carrier, dementia with Lewy bodies (“pure” Lewy body dementia), Alzheimer’s disease, sporadic Alzheimer’s disease, familial Alzheimer's disease with APP mutations, familial Alzheimer's disease with PS-1 , PS-2 or other mutations, familial British dementia, Lewy body variant of
  • the compounds of the present invention are suitable for use in the diagnosis of diseases, disorders or abnormalities associated with alpha-synuclein aggregates including, but not limited to, neuronal and glial aggregates of alpha synuclein, includingmultiple system atrophy (MSA) (Shy-Drager syndrome, striatonigral degeneration and olivopontocerebellar atrophy).
  • MSA multiple system atrophy
  • alpha-synuclein-immunoreactive lesions include traumatic brain injury, chronic traumatic encephalopathy, tauopathies (Pick's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration and Niemann-Pick type C1 disease), motor neuron disease, amyotrophic lateral sclerosis (sporadic, familial and ALS-dementia complex of Guam), neuroaxonal dystrophy, neurodegeneration with brain iron accumulation type 1 (Hallervorden-Spatz syndrome), prion diseases, ataxia telangiectatica, Meige’s syndrome, subacute sclerosing panencephalitis, Gaucher disease as well as other lysosomal storage disorders (including Kufor- Rakeb syndrome and Sanfilippo syndrome) and rapid eye movement (REM) sleep behavior disorder (Jellinger, Mov Disord 2003, 18 Suppl.
  • tauopathies Pick's disease, frontotemporal dementia, progressive supranucle
  • the compounds of the present invention are suitable for use in the diagnosis of Parkinson's disease, multiple system atrophy, dementia with Lewy bodies, Parkinson’s disease dementia, SNCA duplication carrier, or Alzheimer’s disease, more preferably Parkinson’s disease (PD) or multiple system atrophy (MSA).
  • Parkinson's disease multiple system atrophy
  • dementia with Lewy bodies dementia with Lewy bodies
  • Parkinson’s disease dementia dementia with Lewy bodies
  • SNCA duplication carrier or Alzheimer’s disease
  • PD Parkinson’s disease
  • MSA multiple system atrophy
  • the method comprises the steps of:
  • tissue of interest such as brain tissue, tissue of the central nervous system (CNS), tissue of the eye, tissue of peripheral organs or other tissues
  • body fluid such as cerebrospinal fluid (CSF) or blood
  • CSF cerebrospinal fluid
  • the compounds of the present invention can be used for imaging of alpha-synuclein aggregates in any sample or a specific body part or body area of a patient which is suspected to contain alpha- synuclein aggregates.
  • the compounds are able to pass the blood-brain barrier.
  • CNS central nervous system
  • eye such as retinal tissue
  • peripheral organs such as the gut or other tissues
  • body fluids such as cerebrospinal fluid (CSF) or blood.
  • the compounds of the present invention are preferably administered in the form of a diagnostic composition comprising the compound of the invention.
  • a "diagnostic composition” is defined in the present invention 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 which is suitable for use in the diagnosis of the specific disease, disorder or abnormality at issue.
  • a diagnostic composition further comprises a pharmaceutically acceptable excipient, carrier, diluent or adjuvant.
  • Administration is preferably carried out as defined below. More preferably by injection of the composition as an aqueous solution.
  • Such a composition may optionally contain further ingredients such as buffers; pharmaceutically acceptable solubilizers (e.g., cyclodextrins or surfactants such as Pluronic, Tween or phospholipids); and pharmaceutically acceptable stabilisers or antioxidants (such as ascorbic acid, gentisic acid or para-aminobenzoic acid).
  • pharmaceutically acceptable solubilizers e.g., cyclodextrins or surfactants such as Pluronic, Tween or phospholipids
  • pharmaceutically acceptable stabilisers or antioxidants such as ascorbic acid, gentisic acid or para-aminobenzoic acid.
  • the invention also provides a diagnostic composition which comprises a diagnostically effective amount of a compound of the present invention in admixture with, optionally, at least one pharmaceutically acceptable excipient, carrier, diluent or adjuvant.
  • compositions are well known in the pharmaceutical art, and are described, for example, in Remington's Pharmaceutical Sciences, 15 th Ed., Mack Publishing Co., New Jersey (1975).
  • the pharmaceutical excipient can be selected with regard to the intended route of administration and standard pharmaceutical practice.
  • the excipient must be acceptable in the sense of being not deleterious to the recipient thereof.
  • compositions of the present invention may comprise, for example, solvents such as monohydric alcohols such as ethanol, isopropanol and polyhydric alcohols such as glycols and edible oils such as soybean oil, coconut oil, olive oil, safflower oil, cottonseed oil, oily esters such as ethyl oleate, isopropyl myristate, binders, adjuvants, solubilizers, thickening agents, stabilizers, disintegrants, glidants, lubricating agents, buffering agents, emulsifiers, wetting agents, suspending agents, sweetening agents, colorants, flavors, coating agents, preservatives, antioxidants, processing agents, drug delivery modifiers and enhancers such as calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose
  • the routes for administration (delivery) of the compounds of the invention include, but are not limited to, one or more of: intravenous, gastrointestinal, intraspinal, intraperitoneal, intramuscular, oral (e. g. as a tablet, capsule, or as an ingestible solution), topical, mucosal (e. g. as a nasal spray or aerosol for inhalation), nasal, parenteral (e. g. by an injectable form), intrauterine, intraocular, intradermal, intracranial, intratracheal, intravaginal, intracerebroventricular, intracerebral, subcutaneous, ophthalmic (including intravitreal or intracameral), transdermal, rectal, buccal, epidural and sublingual.
  • the route of administration (delivery) of the compounds of the invention is intravenous.
  • the compounds can be administered orally in the form of tablets, capsules, ovules, elixirs, solutions or suspensions, which may contain flavoring or coloring agents, for immediate-, delayed-, modified-, sustained-, pulsed- or controlled-release applications.
  • the tablets may contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate and glycine, disintegrants such as starch (preferably corn, potato or tapioca starch), sodium starch glycolate, croscarmellose sodium and certain complex silicates, and granulation binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin and acacia. Additionally, lubricating agents such as magnesium stearate, stearic acid, glyceryl behenate and talc may be included. Solid compositions of a similar type may also be employed as fillers in gelatin capsules.
  • excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate and glycine
  • disintegrants such as starch (preferably corn, potato or tapioca starch), sodium starch glyco
  • Preferred excipients in this regard include starch, a cellulose, milk sugar (lactose) or high molecular weight polyethylene glycols.
  • the agent may be combined with various sweetening or flavoring agents, coloring matter or dyes, with emulsifying and/or suspending agents and with diluents such as water, ethanol, propylene glycol and glycerin, and combinations thereof.
  • the compounds of the present invention are administered parenterally.
  • parenterally examples of such administration include one or more of: intravenously, intraarterially, intraperitoneally, intrathecally, intraventricularly, intraurethrally, intrasternally, intracranially, intramuscularly or subcutaneously administering the compounds; and/or by using infusion techniques.
  • the compounds are best used in the form of a sterile aqueous solution which may contain other substances, for example, enough salts or glucose to make the solution isotonic with blood.
  • the aqueous solutions should be suitably buffered (preferably to a pH of from 3 to 9), if necessary.
  • the preparation of suitable parenteral formulations under sterile conditions is readily accomplished by standard pharmaceutical techniques well known to those skilled in the art.
  • the compounds of the present invention can be administered intranasally or by inhalation and are conveniently delivered in the form of a dry powder inhaler or an aerosol spray presentation from a pressurized container, pump, spray or nebulizer with the use of a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, a hydrofluoroalkane such as 1 ,1 ,1 ,2-tetrafluoroethane (HFA134AT) or 1 , 1 ,1 , 2, 3,3,3- heptafluoropropane (HFA 227EA), carbon dioxide or other suitable gas.
  • a suitable propellant e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, a hydrofluoroalkane such as 1 ,1 ,1 ,2-tetra
  • the dosage unit may be determined by providing a valve to deliver a metered amount.
  • the pressurized container, pump, spray or nebulizer may contain a solution or suspension of the active compound, e. g. using a mixture of ethanol and the propellant as the solvent, which may additionally contain a lubricant, e. g. sorbitan trioleate.
  • a lubricant e. g. sorbitan trioleate.
  • Capsules and cartridges (made, for example, from gelatin) for use in an inhaler or insufflator may be formulated to contain a powder mix of the compound and a suitable powder base such as lactose or starch.
  • the compounds of the present invention can be administered in the form of a suppository or pessary, or it may be applied topically in the form of a gel, hydrogel, lotion, solution, cream, ointment or dusting powder.
  • the compounds of the present invention may also be dermally or transdermally administered, for example, by the use of a skin patch.
  • the compounds may also be administered by the pulmonary or rectal routes. They may also be administered by the ocular route.
  • the compounds can be formulated as micronized suspensions in isotonic, pH was adjusted, sterile saline, or, preferably, as solutions in isotonic, pH was adjusted, sterile saline, optionally in combination with a preservative such as a benzylalkonium chloride. Alternatively, they may be formulated in an ointment such as petrolatum.
  • the compounds of the present invention can be formulated as a suitable ointment containing the active compound suspended or dissolved in, for example, a mixture with one or more of the following: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, emulsifying wax and water.
  • they can be formulated as a suitable lotion or cream, suspended or dissolved in, for example, a mixture of one or more of the following: mineral oil, sorbitan monostearate, a polyethylene glycol, liquid paraffin, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
  • a physician will determine the actual dosage which will be most suitable for an individual subject.
  • the specific dose level and frequency of dosage for any particular individual may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the individual undergoing diagnosis.
  • compositions of the invention can be produced in a manner known per se to the skilled person as described, for example, in Remington's Pharmaceutical Sciences, 15 th Ed., Mack Publishing Co., New Jersey (1975).
  • the compounds of the present invention are useful as an in vitro analytical reference or an in vitro screening tool. They are also useful in in vivo diagnostic methods.
  • the compounds according to the present invention can also be provided in the form of a mixture, a pharmaceutical composition, or a combination, comprising a compound according to the present invention and at least one compound selected from an imaging agent different from the compound according to the invention, a pharmaceutically acceptable excipient, carrier, diluent or adjuvant.
  • the imaging agent different from the compound according to the invention is preferably present in a diagnostically effective amount. More preferably the imaging agent different from the compound according to the invention is an Abeta or Tau imaging agent.
  • the invention provides a method of diagnosing a disease, disorder or abnormality associated with alpha-synuclein aggregates in a subject, the method comprising the steps:
  • said method may further comprise the step of:
  • the invention provides a method of positron emission tomography (PET) imaging of alpha-synuclein aggregates in a tissue of a subject, the method comprising the steps:
  • the invention relates to a method for the detection and optionally quantification (e.g., an in vivo or in vitro method) of alpha-synuclein aggregates in a tissue of a subject, the method comprising the steps:
  • the present invention refers to a method of collecting data for the diagnosis of a disease, disorder or abnormality associated with alpha-synuclein aggregates including the method comprising the steps: (a) Bringing a sample or a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound according to the present invention, or a diagnostic composition which comprises a compound according to the present invention;
  • the amount of the compound bound to the alpha-synuclein aggregates is higher than a normal control value it can be assumed that the patient is suffering from a disease, disorder or abnormality associated with alpha-synuclein aggregates.
  • Yet another embodiment of the present invention refers to a method of collecting data for determining a predisposition to a disease, disorder or abnormality associated with alpha-synuclein aggregates, the method comprising the steps:
  • the present invention relates to a method of collecting data for prognosing a disease, disorder or abnormality associated with alpha-synuclein aggregates, wherein the method comprises the steps:
  • step (e) Optionally repeating steps (a) to (c) and, if present, optional step (d) at least one time.
  • the progression of a disease, disorder or abnormality and/or the prospect (e.g., the probability, duration, and/or extent) of recovery can be estimated by a medical practitioner based on the presence or absence of the compound bound to the alpha-synuclein aggregates, the amount of the compound bound to the alpha-synuclein aggregates or the like. If desired, steps (a) to (c) and, if present, optional step (d) can be repeated over time to monitor the progression of the disease, disorder or abnormality and to thus allow a more reliable estimate.
  • a further aspect is directed to a method of collecting data for monitoring the progression (or evolution) of a disease, disorder or abnormality associated with alpha-synuclein aggregates in a patient, the method comprising the steps:
  • step (e) Optionally repeating steps (a) to (c) and, if present, optional step (d) at least one time.
  • the amount of the compound bound to the alpha- synuclein aggregates can be optionally compared at various points of time during the treatment, for instance, before and after onset of the treatment or at various points of time after the onset of the treatment.
  • the patient is or has been undergoing treatment of the disease, disorder or abnormality associated with alpha-synuclein aggregates or is/has been undergoing treatment of the synucleinopathy.
  • the treatment can involve administration of a medicament which is suitable for treating the disease, disorder or abnormality associated with alpha-synuclein aggregates.
  • the invention relates to a method of collecting data for predicting responsiveness of a patient suffering from a disease, disorder or abnormality associated with alpha- synuclein aggregates to a treatment with a medicament, the method comprising the steps of
  • step (e) Optionally repeating steps (a) to (c) and, if present, optional step (d) at least one time.
  • the method can further comprises steps (i) to (vi) before step (a):
  • step (vi) treating the patient with the medicament.
  • the method can further comprise step (A) after step (d) or step (e):
  • step (A) comparing the amount of the compound bound to the alpha-synuclein aggregates determined in step (iv) to the amount of the compound bound to the alpha-synuclein aggregates determined in step (d).
  • the amount of the compound bound to the alpha- synuclein aggregates can be optionally compared at various points of time during the treatment, for instance, before and after onset of the treatment or at various points of time after the onset of the treatment.
  • a change, especially a decrease, in the amount of the compound bound to the alpha- synuclein aggregates may indicate that the patient has a high potential of being responsive to the respective treatment.
  • the amount of the compound bound to the alpha-synuclein aggregates decreases over time, it can be assumed that the patient is responsive to the treatment. If the amount of the compound bound to the alpha-synuclein aggregates is essentially constant or increases overtime, it can be assumed that the patient is non-responsive to the treatment.
  • the responsiveness can be estimated by determining the amount of the compound bound to the alpha-synuclein aggregates.
  • the amount of the compound bound to the alpha-synuclein 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 the control value of subjects known to be responsive to a certain therapy, or the control value may refer to the control value of subjects known to be non-responsive to a certain therapy.
  • the outcome with respect to responsiveness can either be "responsive" to a certain therapy, "non-responsive" to a certain therapy or “response undetermined” to a certain therapy. Response to the therapy may be different for the respective patients.
  • the diagnostic composition can be used before, during and after, surgical procedures (e.g. deep brain stimulation (DBS)) and non-invasive brain stimulation (such as repetitive transcranial magnetic stimulation (rTMS)), for visualizing alpha-synuclein aggregates before, during and after such procedures.
  • surgical procedures e.g. deep brain stimulation (DBS)
  • non-invasive brain stimulation such as repetitive transcranial magnetic stimulation (rTMS)
  • Surgical techniques including DBS, improve advanced symptoms of PD on top of the best currently used medical therapy.
  • rTMS has been closely examined as a possible treatment for PD (Ying-hui Chou et al. JAMA Neurol. 2015 April 1 ; 72(4): 432-440).
  • the step of optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the alpha- synuclein aggregates in the sample or specific body part or body area; comprises determining the amount of the compound bound to the alpha-synuclein aggregates;
  • the control value can be, e.g., a normal control value, a preclinical control value and/or a clinical control value.
  • a “healthy control subject” or “healthy volunteer (HV) subject” is a person showing no clinical evidence of a disease, disorder or abnormality associated with alpha-synuclein aggregates.
  • the alpha-synuclein aggregates include, but are not limited to, Lewy bodies and/or Lewy neurites.
  • the amount of the compound bound with the alpha- synuclein aggregates is higher than the normal control value, then it can be expected that the patient is suffering from or is likely to from a disease, disorder or abnormality associated with alpha-synuclein aggregates or from a synucleinopathy.
  • a sample or a specific body part or body area suspected to contain alpha-synuclein aggregates is brought into contact with a compound of the present invention.
  • any of the compounds of the present invention can be used in the above summarized methods.
  • Preferably detectably labelled compounds of the present invention are employed in the above summarized methods.
  • the specific body part or body area is preferably of a mammal, more preferably of a human, including the full body or partial body area or body part of the patient suspected to contain alpha-synuclein aggregates.
  • the specific body part or body area can be brain, the central nervous system, eye or a peripheral organ such as the gut, preferably brain.
  • the tissue can be brain tissue, tissue of the central nervous system (CNS), tissue of the eye (such as retinal tissue), tissue of peripheral organs such as the gut or other tissues, or body fluids such as cerebrospinal fluid (CSF) or blood.
  • the tissue is preferably brain tissue.
  • the sample is an in vitro sample from a patient.
  • the compound of the present invention can be brought into contact with the sample or the specific body part or body area suspected to contain the alpha-synuclein aggregates by any suitable method.
  • the compound of the present invention and a liquid sample can be simply mixed.
  • the specific body part or body area can be brought into contact with a compound of the invention by administering an effective amount of a compound of the invention to the patient.
  • the effective amount of a compound of the invention is an amount which is suitable for allowing the presence or absence of alpha-synuclein aggregates in the sample, specific body part or body area to be determined using the chosen analytical technique.
  • the amount is not particularly limited and will depend on the compound of the formula (I), the type of detectable label, the sensitivity of the respective analytical method and the respective device. The amount can be chosen appropriately by a skilled person.
  • the compound is then allowed to bind to the alpha-synuclein aggregates.
  • the step of allowing the compound to bind to the alpha-synuclein aggregates includes allowing sufficient time for the compound of the invention to bind to the alpha-synuclein aggregates.
  • the amount of time required for binding will depend on the type of test (e.g., in vitro or in vivo) and can be determined by a person skilled in the field by routine experiments. In an in vivo method, the amount of time will depend on the time which is required for the compound to reach the specific body part or body area suspected to contain alpha-synuclein aggregates. The amount of time should not be too extended to avoid washout and/or metabolism of the compound of the invention.
  • the compound which has bound to the alpha-synuclein aggregates can be subsequently detected by any appropriate method.
  • the method of detecting the compound bound to the alpha-synuclein aggregates is not particularly limited and depends, among others, on the detectable label, the type of sample, specific body part or body area and whether the method is an in vitro or in vivo method. Examples of possible methods include, but are not limited to, a fluorescence imaging technique or a nuclear imaging technique such as positron emission tomography (PET), single photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), and contrast-enhanced magnetic resonance imaging (MRI). These have been described and enable visualization of alpha- synuclein biomarkers.
  • PET positron emission tomography
  • SPECT single photon emission computed tomography
  • MRI magnetic resonance imaging
  • MRI contrast-enhanced magnetic resonance imaging
  • the fluorescence imaging technique and/or nuclear imaging technique can be employed for monitoring and/or visualizing the distribution of the detectably labelled compound within the sample or a specific body part or body area.
  • the imaging system provides an image of bound detectable label such as radioisotopes, in particular positron emitters or gamma emitters, as present in the tested sample, the tested specific body part or the tested body area.
  • the compound bound to the alpha-synuclein aggregates including is detected by an imaging apparatus such as PET or SPECT scanner, more preferably PET.
  • the amount of the compound bound to the alpha-synuclein aggregates including can be determined by visual or quantitative analysis, for example, using PET scan images.
  • a compound according to the present invention or its precursor can also be incorporated into a test kit for detecting alpha-synuclein protein aggregates.
  • the test kit typically comprises a container holding one or more compounds according to the present invention or its precursor(s) and instructions for using the compound for the purpose of binding to alpha-synuclein aggregates and detecting the formation of the compound bound to the alpha-synuclein aggregates such that presence or absence of the compound bound to the alpha-synuclein aggregates correlates with the presence or absence of the alpha-synuclein aggregates.
  • test kit refers in general to any diagnostic kit known in the art. More specifically, the latter term refers to a diagnostic kit as described in Zrein et al., Clin. Diagn. Lab. Immunol., 1998, 5, 45-49.
  • the dose of the detectably labelled compounds of the present invention preferably compounds of formula (l-F) labelled with 18 F or compounds of formula (l-H*) or (l-H) labelled with 3 H, will vary depending on the exact compound to be administered, the weight of the patient, size and type of the sample, and other variables as would be apparent to a physician skilled in the art. Generally, the dose could preferably lie in the range 0.001 pg/kg to 10 pg/kg, preferably 0.01 pg/kg to 1.0 pg/kg.
  • the radioactive dose can be, e.g., 100 to 600 MBq, more preferably 150 to 450 MBq.
  • the compounds of the present invention may be prepared in accordance with the definition of compound of formula (I) by the routes described in the following Schemes or the Examples. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g. "such as") provided herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed.
  • R 1 , R 2 , ®, LG, and Hal are as previously defined in the above embodiments or limited to designations in the Schemes. Unless otherwise stated, starting materials are either commercially available or are prepared by known methods.
  • the halogen atom of commercially available or custom- made aldehyde derivatives containing an acid sensitive Pg at the triazolo-moiety can be replaced by a heteroaryl boronic acid derivatives via Suzuki coupling to obtain intermediate E after purification.
  • Reductive amination of intermediate E with a suitable amine is employed to obtain intermediate F after purification.
  • the Pg of intermediate F is removed by acid treatment to obtain intermediate D after purification.
  • Intermediate D can be ring cyclized using 1 ,1’-carbonyldiimidazole (CDI) in an appropriate solvent to afford compounds of formula G as a free base.
  • Acid treatment of compounds of formula G is used to afford the salts of compounds of formula (I).
  • intermediate F Reductive amination of intermediate E with a suitable amine is employed to obtain intermediate F after purification.
  • the Pg of intermediate F is removed by acid treatment to obtain intermediate D after purification.
  • Intermediate D can be ring cyclized using 1 ,T- carbonyldiimidazole (CDI) in an appropriate solvent to afford compounds of formula H as a free base.
  • Acid treatment of compounds of formula H can be used to afford the salts of compounds of formula (I).
  • Compounds having the formula (I) which are labelled by 18 F can be prepared by reacting a precursor compound (I I l-F), as described below, with an 18 F-fluorinating agent, so that the LG comprised in the precursor compound is replaced by 18 F.
  • the reagents, solvents and conditions which can be used for the 18 F-fluorination are well-known to a skilled person in the field (L. Cai, S. Lu, V. Pike, Eur. J. Org. Chem 2008, 2853-2873; J.
  • the solvents used in the 18 F-fluorination are DMF, DMSO, acetonitrile, DMA, or mixtures thereof, preferably the solvent is acetonitrile or DMSO.
  • any suitable 18 F-fluorinating agent can be employed. Typical examples include H 18 F, alkali or alkaline earth 18 F-fluorides (e.g., K 18 F, Rb 18 F, Cs 18 F, and Na 18 F).
  • the 18 F-fluorination agent can be used in combination with a chelating agent such as a cryptand (e.g.: 4,7,13,16,21 ,24-hexaoxa-1 ,10- diazabicyclo[8.8.8]-hexacosane - Kryptofix®) or a crown ether (e.g.: 18-crown-6).
  • a cryptand e.g.: 4,7,13,16,21 ,24-hexaoxa-1 ,10- diazabicyclo[8.8.8]-hexacosane - Kryptofix®
  • a crown ether e.g.: 18-crown-6
  • the 18 F-fluorinating agent can be a tetraalkylammonium salt of 18 F or a tetraalkylphosphonium salt of 18 F; e.g., tetra(Ci-6 alkyl)ammonium salt of 18 F or a tetra(Ci-6 alkyl)phosphonium salt of 18 F.
  • the 18 F-fluorination agent is K 18 F, H 18 F, Cs 18 F, Na 18 F, tetra(Ci-e alkyl) ammonium salt of 18 F, Kryptofix[222] 18 F or tetrabutylammonium [ 18 F]fluoride.
  • NMR measurements were performed on a DRX-400 MHz NMR spectrometer, on a Bruker AV-400 MHz NMR spectrometer in deuterated solvents, using or not using TMS as an internal standard. Chemical shifts (6) are reported in ppm downfield from TMS, spectra splitting patterns are designated as singlet (s), doublet (d), triplet (t), quartet (q), quintet (quint), septet (sept), multiplet, unresolved or overlapping signals (m), or broad signal (br).
  • Deuterated solvents are given in parentheses and have chemical shifts of dimethyl sulfoxide (6 2.50 ppm), methanol (6 3.31 ppm), chloroform (6 7.26 ppm), or other solvent as indicated in NMR spectral data. MS were recorded on an Advion CMS mass spectrometer or an UPLC H-Class Plus with Photodiode Array detector and Qda Mass spectrometer from Waters.
  • Flash Column Chromatography System flash purification was conducted with a Biotage Isolera One flash purification system using HP-Sil or KP-NH SNAP cartridges (Biotage) and the solvent gradient indicated in the specific examples.
  • the compound was purchased from the commercial supplier Aurum Pharmatech LLC.
  • Step-A To a stirred solution of 2-fluoroethanol (1.82 mL, 31.2 mmol) and sodium hydride (60% dispersed in mineral oil) (1.25 g, 31.2 mmol) in THF (100 mL) under a nitrogen atmosphere was added 5-bromo-2-fluoropyridine (5.0 g, 28.4 mmol) in portions at 0 °C over a period of 15 min. After the addition was completed, the reaction mixture was stirred at room temperature for 3 h. The reaction mixture was quenched with water (100 mL) and extracted with EtOAc (3 x 100 mL). The combined organic phase was dried over NazSCh and concentrated under vacuum to afford the title compound as a yellow solid (5.0 g, 79%).
  • Step-B In an oven-dried flask was added the title compound from Step A above (5.0 g, 22.7 mmol), bis(pinacalato) diborane (11.5 g, 45.4 mmol), KOAc (6.7 g, 68.9 mmol) and 1 ,4-dioxane (250 mL) under an argon atmosphere. The reaction mixture was degassed with argon for 15 min. Then [1 ,1'- bis(diphenylphosphino)ferrocene]-dichloropalladium(ll), complex with dichloromethane (1.8 g, 22.7 mmol) was added and the mixture was heated to 60 °C for 5 h.
  • the compound was purchased from the commercial supplier SV ChemBioTech, Inc.
  • Step-A 2,5-Dibromopyrazine (5.0 g, 21 mmol), pyrrolidine (1.8 g, 25.2 mmol), CS2CO3 (6.8 g, 21 mmol), and DMSO (50 mL, 10 vol) were heated at 100“C for 16 h under microwave irradiation. After completion, the reaction mixture was quenched with ice cool water (60 mL) and the crude reaction mass was filtered through a buchner funnel. The solid was dried under high vacuum to afford the desired product as a yellow solid (4.3 g, 90%).
  • Step-B To a stirred solution of step-A product (2.0 g, 8.77 mmol) in THF (80 mL, 40 vol.) was added dropwise nBuLi (2.5M in hexane) (4.2 mL, 10.5 mmol) at -78°C under N2 atmosphere and the mixture was stirred for 1 h. Then, 2-isopropoxy-4,4,5,5-tetramethyl-1 ,3,2-dioxaborolane (2.1 mL, 10.5 mmol) was added. The mixture was allowed to stir at -78°C for 2 h and at rt for 3 h. The reaction time was monitored by TLC.
  • nBuLi 2.5M in hexane
  • reaction mixture was cooled to 0°C and quenched with saturated aqueous NH4CI solution.
  • the aqueous layer was extracted with EtOAc (2 x 100 mL) and the combined organic layers were dried over NasSCU and concentrated under vacuum.
  • the crude reaction product was directly used as such in the next step without further purification.
  • Step-B To a solution of the Step A product (3.2 g, 18.48 mmol) in methanol (96 mL, 30 vol.) was added 10% Pd/C (640 mg, 20%w/w) portionwise under N 2 atmosphere. Then, the vessel was filled up with H2 gas and the mixture was stirred at rt for 20 h. The progress of the reaction was monitored by TLC. After completion, the mixture was filtered through celite and the celite was washed with MeOH (3 x 100 mL). The combined filtrates were concentrated in vacuum to yield the desired product as a yellow oil (2.6 g, 93%).
  • Step-A To a solution of 3-nitro-1 H-pyrazole (2.0 g, 17.6 mmol) in DMF (40mL, 20 vol.) was added CS2CO3 (17.3 g, 53.1 mmol) and 3-fluoropropyl 4-methylbenzenesulfonate (8.2 g, 35.4 mmol). The mixture was stirred at rt for 16 h. The progress of the reaction was monitored by TLC. After completion, the solvent was evaporated under vacuum and water (80 mL) was added to the residue. The aqueous layer was extracted with EtOAc (2 x 80 mL) and the combined organic layers were dried over Na2SO4 and concentrated under vacuum. The obtained crude mass was purified by column chromatography over silica gel (100-200 mesh) and eluted in 50% EtOAc in hexane to afford the desired product as a yellow oil (2.3 g, 75%).
  • Step-B To a solution of the Step A product (2.3 g, 13.2 mmol) in methanol (70 mL, 30 vol.) was added 10% Pd/C (230 mg, 10%w/w) portionwise under N2 atmosphere. The reaction vessel was filled up with H 2 gas and the mixture was stirred at rt for 21 h. The progress of the reaction was monitored by TLC. After completion, the mixture was filtered through celite and the celite was washed with MeOH (3 x 100 mL). The combined filtrates were concentrated in vacuum to yield the desired product as a yellow oil (1.7 g, 86%). The compound was pure enough and used as such in the next step without further purification.
  • the reaction mixture was stirred for 4 h and allowed to reach room temperature. Then sodium triacetoxyborohydride (4.0 g, 19.2 mmol) was added and the mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with saturated NaHCO3 (100 mL) and the product was extracted with 5% MeOH in DCM (3 x 250 mL). The combined organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by column chromatography using silica gel (100-200 mesh) employing in 2% MeOH in DCM to afford the title compound as a yellow liquid (3.0 g, 76%).
  • Step B In an oven-dried screw capped vial was added the title compound from Step A above (0.6 g,
  • reaction mixture was quenched with ice-water and extracted in 10% MeOH in DCM (3 x 30 mL).
  • the organic layer was dried over Na2SO4, concentrated and purified by silica gel chromatography (100-200 mesh) using 4% MeOH in DCM to obtain the title compound as a brownish liquid (0.45 g, 68%).
  • Step-C The title compound from Step B above (0.1 g, 0.27 mmol), (R)-3-fiuoropyrrolidine hydrogen chloride (0.051 g, 0.40 mmol), DIPEA (0.14 mL, 0.8 mmol), and NMP (5 mL) were combined in an oven-dried microwave vial under argon atmosphere. The reaction mixture was heated at 160 °C in a microwave oven for 4 h. The reaction mixture was quenched with ice cold water (10 mL) and extracted with 5% MeOH in DCM (3 x 10 mL). The combined organic layers were washed with cold brine solution (2 x 10 mL), dried over Na2SO4, and concentrated under vacuum.
  • Step-D To a stirred solution of the title compound from Step C above (0.050 g, 0.11 mmol) in DCM (2.5 mL) was added a solution of 4 M HCI in 1 ,4-dioxane (0.5 mL) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred for 6 h and allowed to reach room temperature. The solvent was removed under vacuum to afford the title compound as a hydrochloride salt. The yellow solid was washed with hexane (3 x 5 mL) and dried under vacuum.
  • Step-E To an ice cool solution of the title compound from Step D above (0.04 mg, 0.1 mmol) in 1 ,2- DCE (2 mL) was added sodium hydride (60% dispersed in mineral oil) (0.004 g, 0.17 mmol) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 30 min. Then 1 ,1'- carbonyldiimidazole (0.16 g, 1 .0 mmol) was added to the reaction mixture and the mixture was stirred at room temperature for 6 h. The reaction mixture was quenched with ice cold water and the product was extracted with 5% MeOH in DCM. The organic phase was dried over Na2SO4 and concentrated under vacuum. The residue was purified by silica gel chromatography (100-200 mesh) employing 4% MeOH in DCM to yield the title compound as a white solid (0.02 mg, 51%).
  • Step-F To a stirred solution of the title compound from Step E above (0.020 g, 0.05 mmol) in DCM (2 mL) was added a solution of 4 M HCI in 1 ,4-dioxane (0.2 mL) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred for 6 h and allowed to warm to room temperature. The solvent was evaporated, the residue was treated with pentane, and dried under vacuum to afford the title compound as a yellow solid (0-015 g, 71%).
  • Step A To an ice cool solution of ethyl 5-bromo-1/-/-1 ,2,4-triazole-3-carboxylate (2.5 g, 11.3 mmol) in THF (50 mL) was added sodium hydroxide (60% dispersed in mineral oil) (0.59 g, 14.7 mmol) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 30 min.
  • reaction mixture was cooled to 0 °C, 2-(trimethylsilyl)ethoxymethyl chloride (2.4 mL, 13.6 mmol) was dropwise added, and the reaction mixture was stirred for 16 h and allowed to reach room temperature.
  • the reaction mixture was quenched with ice-water and the reaction mixture was extracted with EtOAc (3 x 75 mL).
  • the combined organic phase was washed with brine (2 x 25 mL), dried over NazSCM, and concentrated under vacuum.
  • the residue was purified by silica gel chromatography (100-200 mesh) employing 10% EtOAc in n-hexane to yield the title compound as a yellow oil (0.96 g, 24%).
  • Step B In an oven-dried screw capped vial was added the title compound from Step A above (0.2 g, 0.57 mmol), (6-fluoropyridin-3-yl)boronic acid (0.096 g, 0.68 mmol), CS2CO3 (0.371 g, 1.14 mmol), and 1 ,4-dioxane (8 mL) under an argon atmosphere. The reaction mixture was degassed with argon for 15 min.
  • Step C The title compound from Step B above (3.0 g, 8.2 mmol), (S)-3-fluoropyrrolidine hydrogen chloride (2.0 g, 16.3 mmol), DIPEA (7.13 mL, 41 mmol) and ethanol (30 mL) were combined in an oven-dried microwave vial under argon atmosphere. The reaction mixture was heated at 130 °C in a microwave oven for 3 h. The solvent was removed under vacuum and the obtained crude material was purified by column chromatography using silica gel (100-200 mesh) employing 50% EtOAc in n-hexane to afford the title compound as a white solid (1.8 g, 50%).
  • Step D To an ice cool solution of the title compound from Step C above (1.8 g, 4.13 mmol) in MeOH (90 mL) was added sodium borohydride (1.25 g, 33 mmol) in portions at 0 °C under a nitrogen atmosphere. After the addition was completed, the reaction mixture was stirred at room temperature for 3 h. The reaction mixture was cooled to 0 °C and the reaction was quenched by the addition of sat. ammonium chloride (effervescence!). The solvent was removed under vacuum and the crude material was diluted with water and extracted with DCM (3 x 100 mL). The combined organic layer was washed with cold brine solution (100 mL), dried over NajSCU, and concentrated under vacuum to afford the title compound as a white solid (1 .6 g, 98%).
  • Step E To an ice cool solution of the title compound from Step D above (1.6 g, 4.0 mmol) in DCM (48 mL) was added Dess-Martin periodinane (3.45 g, 8.14 mmol) in portions at 0 °C under a nitrogen atmosphere. After the addition was completed, the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was cooled to 0 °C and the reaction was quenched by the addition of sat. NaHCO3 (50 mL). The reaction mixture was extracted with DCM (3 x 80 mL). The combined organic layer was washed with cold brine solution (80 mL), dried over Na2SO4, and concentrated under vacuum. The crude material was purified by column chromatography using silica gel (100-200 mesh) employing in 50% EtOAc in n-hexane to afford the title compound as a white solid (1.3 g, 82%).
  • Step F To a stirred solution of the title compound from Step E above (0.4 g, 1 .0 mmol) and 1-methyl- 1H-pyrazol-4-amine (0.1 g, 1.0 mmol) in 1 ,2-dichloroethane (16 mL) was added molecular sieves 4A and glacial AcOH (0.8 mL) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 h. Then sodium triacetoxyborohydride (0.43 g, 2.0 mmol) was added and the mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with aqueous sat.
  • Step G To a stirred solution of the title compound from Step F above (0.4mg, 0.85 mmol) in DCM (8.0 mL) was added a solution of 4 M HCI in 1 ,4-dioxane (2.0 mL) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 h, the solvents were evaporated under reduced pressure, the residue was washed with pentane, and dried under vacuum to afford the title compound as an off-white solid (0.28 g, 97%).
  • Step H To an ice cool solution of the title compound from Step G above (0.28 g, 0.82 mmol) in 1 ,2- dichloroethane (17 mL) was added sodium hydride (60% dispersed in mineral oil) (0.033 g, 0.82 mmol) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature and kept for 30 min. Then 1 ,1 '-carbonyldiimidazole (1.33 g, 8.2 mmol) was added to the reaction mixture and the mixture was stirred at room temperature for 24 h. The reaction mixture was quenched with ice cold water and the reaction mixture was extracted with 5% MeOH in DCM (3 x 40 mL).
  • Step I To a stirred solution of the title compound from Step H above (0.006.g, 0.024 mmol) in DCM (1.0 mL) was added a solution of 4M HCI in 1,4-dioxane (0.6 mL,) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 h. The solvents were removed under vacuum to afford the hydrochloride salt. The off-white solid was washed with n-hexane (2 mL) and then dried under vacuum to afford the title compound as a white solid (0.004 g, 40%).
  • Step B To a stirred solution of the title compound from Step A above (0.4 g, 1.3 mmol) and 1 -methyl- 1H-pyrazol-4-amine (0.18 g, 1.9 mmol) in 1 ,2-dichloroethane (16 mL) was added molecular sieves 4 A and glacial AcOH (1.2 mL) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 4 h. Then sodium triacetoxyborohydride (0.55 g, 2.6 mmol) was added and the mixture was stirred at room temperature for 16 h.
  • the reaction mixture was quenched with aqueous saturated NaHCO3 (60 mL) and the product was extracted with 5% MeOH in DCM (3 x 60 mL). The combined organic phase was dried over Na2SO 4 and concentrated under reduced pressure.
  • the crude material was purified by column chromatography using silica gel (100-200 mesh) employing 4% MeOH in DCM to afford the title compound as a brown liquid (0.34 g, 65%).
  • Step C To a stirred solution of the title compound from Step B above (0.26 g, 0.65 mmol) in DCM (13 mL) was added a solution of 4 M HCI in 1 ,4-dioxane (2.6 mL) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 5 h. The solvent was removed under reduced pressure to afford the hydrochloride salt. The yellow solid was washed with n-hexane (3 x 5 mL) and dried under vacuum to afford the title compound (0.2 g, 87%).
  • Step D To an ice cool solution of the title compound from Step C above (0.2 mg, 0.63 mmol) in 1 ,2- dichloroethane (12 mL) was added sodium hydride (60% dispersed in mineral oil) (0.04 g, 0.95 mmol) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 30 min. Then 1 ,T-carbonyldiimidazole (1.02 g, 6.3 mmol) was added and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with ice cold water and the mixture was extracted with 5% MeOH in DCM (3 x 20 mL). The combined organic phase was dried over Na2SC>4 and concentrated under reduced pressure. The residue was purified by silica gel chromatography (100-200 mesh) employing 3% MeOH in DCM to yield the title compound as a white solid (0.09 g, 42%).
  • sodium hydride 60% dispersed in mineral oil
  • Step E To a stirred solution of the title compound from Step D above (0.08 g, 0.23 mmol) in DCM (4 mL) was added a solution of 4 M HCI in 1 ,4-dioxane (0.8 mL) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 5 h and the solvents were evaporated under reduced pressure. The residue was treated with pentane and dried under vacuum to afford the title compound as a white solid (0.08 g, 92%).
  • Example 11 Following the same procedure as described for Example 11, except using the reagents indicated for the specific steps in Table 3 below, the following Example was prepared.
  • Pellets were resuspended in extraction buffer [10 mM Tris-HCI pH 7.4, 10% sucrose, 0.85 M NaCI, 1 % protease inhibitor (Calbiochem 539131 ), 1 mM EGTA, 1% phosphatase inhibitor (Sigma P5726 and P0044)] and centrifuged at 15,000 x g (14,800 RPM, a 70.1 Ti rotor) for 20 minutes at 4°C. Pellets were discarded and sarkosyl (20% stock solution, Sigma L7414) was added to the supernatants to a final concentration of 1% and stirred at room temperature for one hour.
  • PD brain-derived alpha-synuclein aggregates were spotted onto microarray slides.
  • the slides were incubated with [ 3 H]-alpha-synuclein reference at 25nM, 30nM or 40nM and the example compounds (non-radiolabelled) at 1 pM and 100nM.
  • the non-radiolabelled example compounds were further assessed for a range of different concentrations, varying from 0.05nM to 2pM.
  • slides were washed, and scanned by a real-time autoradiography system (BeaQuant, ai4R). Quantification of the signal was performed by using the image analysis software Beamage (ai4R).
  • Non-specific signal was determined with an excess of non-radiolabelled reference alpha- synuclein ligand (2 pM) and specific binding was calculated by subtracting the non-specific signal from the total signal. Competition was calculated as percent, where 0% was defined as the specific binding in the presence of vehicle and 100% as the values obtained in the presence of excess of the non-radiolabelled reference alpha-synuclein ligand.
  • K values were calculated in GraphPad Prism7 by applying a nonlinear regression curve fit using a one site, specific binding model. All measurements were performed with at least two technical replicates. For compounds tested in more than one experiment, the mean of the replicates or Ki values in independent experiments is reported.
  • Example compounds were assessed for their potency to compete with the binding of [ 3 H]- reference alpha-synuclein ligand to PD patient brain-derived alpha-synuclein aggregates.
  • Results of the micro-radiobinding competition assay for the example compounds tested are shown in Table 4 as: % competition at 1 pM and 100 nM.
  • the Table 4 also shows Ki values. Ki measurements for example compounds 1-11 were performed on the same PD brain-derived alpha-synuclein aggregates while Ki measurements for example compounds 13-17, and 25 were performed on PD brain-derived alpha-synuclein aggregates from a different donor. The data shows that the compounds of the invention bind to human brain-derived alpha-synuclein aggregates.
  • Example compounds 4, 5, 11 and 17 exhibited binding with very high affinity (Ki ⁇ 100nM) to human PD patient brain-derived alpha-synuclein aggregates.

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Abstract

The present invention relates to novel compounds of formula (I), or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, that can be employed in the imaging of alpha-synuclein aggregates and determining an amount thereof. Furthermore, the compounds can be used for diagnosing a disease, disorder or abnormality associated with an alpha-synuclein aggregates (such as Parkinson's disease or such as multiple system atrophy (MSA)) determining a predisposition to such a disease, disorder or abnormality, prognosing such a disease, disorder or abnormality, monitoring the evolution of the disease in a patient suffering from such a disease, disorder or abnormality, monitoring the progression of such a disease, disorder or abnormality and predicting responsiveness of a patient suffering from such a disease, disorder or abnormality to a treatment thereof.

Description

NOVEL COMPOUNDS FOR DIAGNOSIS
FIELD OF THE INVENTION
The present invention relates to novel compounds of formula (I), or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, that can be employed in the imaging of alpha-synuclein aggregates and determining an amount thereof. Furthermore, the compounds can be used for diagnosing a disease, disorder or abnormality associated with alpha-synuclein (a-synuclein, A-synuclein, aSynuclein, A-syn, a-syn, aSyn, a-syn) aggregates, such as Parkinson’s disease or multiple system atrophy, determining a predisposition to such a disease, disorder or abnormality, prognosing such a disease, disorder or abnormality, monitoring the evolution of the disease in a patient suffering from such a disease, disorder or abnormality, monitoring the progression of such a disease, disorder or abnormality and predicting responsiveness of a patient suffering from such a disease, disorder or abnormality to a treatment thereof. The present invention also relates to processes for the preparation of the compounds and their precursors, diagnostic compositions comprising the compounds, methods of using the compounds, kits comprising the compounds and their uses thereof.
BACKGROUND OF THE INVENTION
Many diseases of aging are based on or associated with extracellular or intracellular deposits of amyloid or amyloid-like proteins that contribute to the pathogenesis as well as to the progression of the disease. The best characterized amyloid protein that forms extracellular aggregates is amyloid beta (Abeta or Ap).
Amyloid-like proteins that form mainly intracellular aggregates include, but are not limited to, Tau, alpha-synuclein, and huntingtin (HTT). Diseases involving alpha-synuclein aggregates are generally listed as synucleinopathies (or alpha-synucleinopathies) and these include, but are not limited to Parkinson’s disease (PD) or multiple system atrophy (MSA). Synucleinopathies with primarily neuronal aggregates include, but are not limited to, Parkinson's disease (sporadic, familial with SNCA (the gene encoding for the alpha-synuclein protein) mutations or SNCA gene duplication or triplication, familial with mutations in other genes than SNCA, pure autonomic failure and Lewy body dysphagia), SNCA duplication carrier, Lewy Body dementia (LBD), dementia with Lewy bodies (DLB) (“pure” Lewy body dementia), Parkinson’s disease dementia (PDD), diffuse Lewy body disease (DLBD), Alzheimer’s disease, sporadic Alzheimer’s disease, familial Alzheimer's disease with APP mutations, familial Alzheimer's disease with PS-1 , PS-2 or other mutations, familial British dementia, Lewy body variant of Alzheimer’s disease and normal aging in Down syndrome. Synucleinopathies with neuronal and glial aggregates of alpha-synuclein include, but are not limited to, multiple system atrophy (MSA) (Shy-Drager syndrome, striatonigral degeneration and olivopontocerebellar atrophy). Other diseases that may have alpha-synuclein-immunoreactive lesions are, but are not limited to, traumatic brain injury, chronic traumatic encephalopathy, dementia puglistica, tauopathies (Pick's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration and Niemann-Pick type C1 disease, frontotemporal dementia with Parkinsonism linked to chromosome 17), motor neuron disease, Huntington’s disease, amyotrophic lateral sclerosis (sporadic, familial and ALS-dementia complex of Guam), neuroaxonal dystrophy, neurodegeneration with brain iron accumulation type 1 (Hallervorden-Spatz syndrome), prion diseases, Creutzfeldt-Jakob disease, ataxia telangiectatica, Meige’s syndrome, subacute sclerosing panencephalitis, Gerstmann- Straussler-Scheinker disease, inclusion-body myositis, Gaucher disease, Krabbe disease as well as other lysosomal storage disorders (including Kufor-Rakeb syndrome and Sanfilippo syndrome) and rapid eye movement (REM) sleep behavior disorder (Jellinger, Mov. Disord. 2003, 18 Suppl. 6, S2- 12; Galvin et al. JAMA Neurology 2001 , 58 (2), 186-190; Kovari et al., Acta Neuropathol. 2007, 114(3), 295-8; Saito et al., J. Neuropathol. Exp. Neurol. 2004, 63(4), 323-328; McKee et al., Brain, 2013, 136(Pt 1 ), 43-64; Puschmann et al., Parkinsonism Relat. Disord. 2012, 18S1 , S24-S27; Usenovic et al., J. Neurosci. 2012, 32(12), 4240-4246; Winder-Rhodes et al., Mov. Disord. 2012, 27(2), 312-315; Ferman et al., J. Int. Neuropsychol. Soc. 2002, 8(7), 907-914; Smith et al., J. Pathol. 2014; 232:509-521 , Lippa et al., Ann Neurol. 1999 Mar; 45(3):353-7; Schmitz et al., Mol. Neurobiol. 2018 Aug 22; Charles et al., Neurosci. Lett. 2000 Jul 28; 289(1 ):29-32; Wilhelmsen et al., Arch Neurol. 2004 Mar; 61(3):398-406; Yamaguchi et al., J. Neuropathol. Exp. Neurol. 2004, 80th annual meeting, vol. 63; Askanas et al., J. Neuropathol. Exp. Neurol. 2000 Jul; 59(7):592-8).
Alpha-synuclein is a 140 amino acid natively unfolded protein (Iwai et al., Biochemistry 1995, 34(32), 10139-10145). The sequence of alpha-synuclein can be divided into three main domains: 1 ) the N- terminal region comprising of residues 1-60, which contains the 11-mer amphipatic imperfect repeat residues with highly conserved hexamer (KTKEGV). This region has been implicated in regulating alpha-synuclein binding to membranes and its internalization; 2) the hydrophobic Non Amyloid beta Component (NAC) domain spanning residues 61-95; which is essential for alpha-synuclein fibrillization; and 3) the C-terminal region spanning residues 96-140 which is highly acidic and prolinerich and has no distinct structural propensity. Alpha-synuclein has been shown to undergo several posttranslational modifications, including truncations, phosphorylation, ubiquitination, oxidation and/or transglutaminase covalent cross linking (Fujiwara et al., Nat. Cell. Biol. 2002, 4(2); 160-164; Hasegawa et al., J. Biol. Chem. 2002, 277(50), 49071-49076; Li et al., Proc. Natl. Acad. Sci. U S A 2005, 102(6), 2162-2167; Oueslati et al., Prog. Brain Res. 2010, 183, 115-145; Schmid et al., J. Biol. Chem. 2009, 284(19), 13128-13142). Interestingly, the majority of these modifications involve residues within the C-terminal region.
Several phosphorylation sites have been detected in the carboxyl-terminal region on Tyr-125, -133, and -136, and on Ser-129 (Negro et al., FASEB J. 2002, 16(2), 210-212). Tyr-125 residues can be phosphorylated by two Src family protein tyrosine kinases, c-Src and Fyn (Ellis et al., J. Biol. Chem.
2001 , 276(6), 3879-3884; Nakamura et al., Biochem. Biophys. Res. Commun. 2001 , 280(4), 1085- 1092). Phosphorylation by Src family kinases does not suppress or enhance the tendency of alpha- synuclein to polymerize. Alpha-synuclein has proved to be an outstanding substrate for protein tyrosine kinase p72syk (Syk) in vitro-, once it is extensively Tyr-phosphorylated by Syk or tyrosine kinases with similar specificity, it loses the ability to form oligomers, suggesting a putative anti- neurodegenerative role for these tyrosine kinases (Negro et al., FASEB J. 2002, 16(2), 210-212). Alpha-synuclein can be Ser-phosphorylated by protein kinases CKI and CKII (Okochi et al., J. Biol. Chem. 2000, 275(1 ), 390-397). The residue Ser-129 is also phosphorylated by G-protein-coupled receptor protein kinases (Pronin et al., J. Biol. Chem. 2000, 275(34), 26515-26522). Extensive and selective phosphorylation of alpha-synuclein at Ser-129 is evident in synucleinopathy lesions, including Lewy bodies (Fujiwara et al., Nat. Cell. Biol. 2002, 4(2); 160-164). Other post-translational modifications in the carboxyl-terminal, including glycosylation on Ser-129 (McLean et al., Neurosci. Lett. 2002, 323(3), 219-223) and nitration on Tyr-125, -133, and -136 (Takahashi et al., Brain Res.
2002, 938(1-2), 73-80), may affect aggregation of alpha-synuclein. Truncation of the carboxyl- terminal region by proteolysis has been reported to play a role in alpha-synuclein fibrillogenesis in various neurodegenerative diseases (Rochet et al., Biochemistry 2000, 39(35), 10619-10626). Full- length as well as partially truncated and insoluble aggregates of alpha-synuclein have been detected in highly purified Lewy bodies (Crowther et al., FEBS Lett. 1998, 436(3), 309-312).
Abnormal protein aggregation appears to be a common feature in aging brain and in several neurodegenerative diseases (Trojanowski et al., 1998, Cell Death Differ. 1998, 5(10), 832-837, Koo et al., Proc. Natl. Acad. Sci. 1999, 96(18), 9989-9990, Hu et al., Chin. Sci. Bull. 2001 , 46, 1-3); although a clear role in the disease process remains to be defined. In in vitro models, alpha-synuclein (or some of its truncated forms) readily assembles into filaments resembling those isolated from the brain of patients with Lewy Body (LB) dementia and familiar PD (Crowther et al., FEBS Lett. 1998, 436(3), 309-312). Alpha-synuclein and its mutated forms (A53T and A30P) have a random coil conformation and do not form significant secondary structures in aqueous solution at low concentrations; however, at higher concentrations they are prone to self-aggregate, producing amyloid fibrils (Wood et al., J. Biol. Chem. 1999, 274(28), 19509-19512). Several differences in the aggregation behavior of the PD-linked mutants and the wild-type protein have been documented. Monomeric alpha-synuclein aggregates in vitro form stable fibrils via a metastable oligomeric (i.e., protofibril) state (Voiles et al., Biochemistry 2002, 41(14), 4595-4602).
Parkinson’s disease (PD) is the most common neurodegenerative motor disorder. PD is mainly an idiopathic disease, although in at least 5% of the PD patients the pathology is linked to mutations in one or several specific genes. Several point mutations have been described in the alpha-synuclein gene (A30P, E46K, H50Q, G51 D, A53T) which cause familial PD with autosomal dominant inheritance. Furthermore, duplications and triplications of the alpha-synuclein gene have been described in patients that developed PD, underlining the role of alpha-synuclein in PD pathogenesis (Lesage et al., Hum. Mol. Genet., 2009, 18, R48-59). The pathogenesis of PD remains elusive. However, growing evidence suggests a role for the pathogenic folding of the alpha-synuclein protein that leads to the formation of amyloid-like fibrils. Indeed, the hallmarks of PD are the presence of intracellular alpha-synuclein aggregate structures called Lewy Bodies and neurites mainly in the nigral neurons, as well as the death of dopaminergic neurons in the substantia nigra and elsewhere. Alpha-synuclein is a natively unfolded presynaptic protein that can misfold and aggregate into larger oligomeric and fibrillar forms which are linked to the pathogenesis of PD. Recent studies have implicated small soluble oligomeric and protofibrillar forms of alpha-synuclein as the most neurotoxic species (Lashuel et al., J. Mol. Biol., 2002, 322, 1089-102). However, the precise role of alpha- synuclein in the neuronal cell toxicity remains to be clarified (review: Cookson, Annu. Rev. Biochem., 2005, 74, 29-52).
Besides Parkinson's disease, the accumulation of aggregated alpha-synuclein into Lewy bodies is a characteristic of all Lewy body diseases, including Parkinson’s disease with dementia (PDD), and dementia with Lewy bodies (DLB) (Capouch et al., Neurol. Ther. 2018, 7, 249-263). In DLB, Lewy Bodies are diffusely distributed throughout the cortices of the brain and in addition to Lewy bodies and neurites, more threads and dot-like structures (Lewy dots) were found to be immunopositive for alpha-synuclein phosphorylated at Ser-129 (Outeiro et al., Mol. Neurodegener. 2019, 14, 5).
Alpha-synuclein aggregates are also found in multiple system atrophy (MSA). MSA is a rare and sporadic neurodegenerative disorder that manifests with rapidly progressive autonomic and motor dysfunction, as well as variable cognitive decline. Such disorders include Shy-Drager syndrome, striatonigral degeneration and olivopontocerebellar atrophy. The disease can be clinically subclassified in parkinsonian (MSA-P) or cerebellar (MSA-C) variant, depending on the predominant motor phenotype (Fanciulli et al., N. Engl. J. Med. 2015; 372, 249-63). It is characterized by the aggregation of alpha-synuclein in the cytoplasm of oligodendrocytes, forming glial cytoplasmic inclusions (GCIs). GCIs, consisting primarily of fibrillary forms of alpha-synuclein, are the neuropathological hallmark of MSA and are found throughout the neocortex, hippocampus, brainstem, spinal cord and dorsal root ganglia (Galvin et al., Arch Neurol. 2001 , 58,186-90). GCIs are considered a central player in the pathogenesis of MSA. A correlation between the GCI load and the degree of neuronal loss has been reported in both the striatonigral and the olivopontocerebellar regions (Stefanova et al., Neuropathol. Appl. Neurobiol. 2016, 42, 20-32).
Furthermore, a causative link between GCIs and the induction of neuronal loss has been shown in transgenic mice overexpressing human alpha-synuclein in oligodendrocytes under various oligodendroglia-specific promoters. A key event in the pathophysiological cascade is considered to be the permissive templating ('prion-like' propagation) of misfolded alpha-synuclein.
The diagnosis of Parkinson’s disease is largely clinical and depends on the presence of a specific set of symptoms and signs (the initial core feature being bradykinesia, rigidity, rest tremor and postural instability), the absence of atypical features, a slowly progressive course, and the response to a symptomatic drug therapy, mainly limited to a dopamine replacement therapy. The accurate diagnosis requires sophisticated clinical skills and is open to a degree of subjectivity and error, as several other degenerative and non-degenerative diseases can mimic PD symptoms (multiple system atrophy (MSA), progressive supranuclear palsy (PSP), Alzheimer’s disease (AD), essential tremor, dystonic tremor), (Guideline No. 113: Diagnosis and pharmacological management of Parkinson’s disease, January 2010. SIGN). The final confirmation of the pathology can only be made by post-mortem neuropathological analysis.
Computed tomography (CT) and conventional magnetic resonance imaging (MRI) brain scans of people with Parkinson’s disease (PD) usually appear normal. These techniques are nevertheless useful to rule out other diseases that can be secondary causes of parkinsonism, such as basal ganglia tumors, vascular pathology and hydrocephalus. A specific technique of MRI, diffusion MRI, has been reported to be useful at discriminating between typical and atypical parkinsonism, although its exact diagnostic value is still under investigation. Dopaminergic function in the basal ganglia can be measured with different PET and SPECT radiotracers. Examples are ioflupane (123l) (trade name DaTSCAN) and iometopane (Dopascan) for SPECT or fluorodeoxyglucose (18F) (18F-FDG) and dihydrotetrabenazine (11C) (11C-DTBZ) for PET, A pattern of reduced dopaminergic activity in the basal ganglia can aid in diagnosing PD, particularly in the symptomatic stage (Brooks, J. Nucl. Med., 2010, 51 , 596-609; Redmond, Neuroscientist, 2002, 8, 457-88; Wood, Nat. Rev. Neurol., 2014, 10, 305).
Strategies are being developed to apply recent advances in understanding the potential causes of Parkinson’s disease to the development of biochemical biomarkers (Schapira Curr. Opin. Neurol. 2013; 26(4):395-400). Such biomarkers that have been investigated in different body fluids (cerebrospinal fluid (CSF), plasma, saliva) include alpha-synuclein levels but also DJ-1 , Tau and Abeta, as well as neurofilaments proteins, interleukins, osteopontin and hypocrontin (Schapira Curr. Opin. Neurol. 2013; 26(4):395-400), but so far none of these biomarkers alone or in combination can be used as a determinant diagnostic test. To our knowledge, no approved alpha-synuclein diagnostic agent is currently on the market despite a crucial need for Parkinson's disease research and drug development (Eberling et al., J Parkinsons Dis. 2013; 3(4):565-7).
The ability to image alpha-synuclein deposition in the brain would be a huge achievement for alpha- synucleopathies research, including Parkinson’s disease (PD) and MSA research, diagnosis, and drug development. The accumulation of aggregated alpha-synuclein in the brain is considered a key pathological hallmark of PD and MSA and can start many years before the appearance of the symptoms. Therefore, alpha-synuclein is a priority target for drug development given not only its likely contribution to neurodegeneration but also because it can offer the possibility to treat the disease while still in the asymptomatic or prodromal stages. In vivo imaging of alpha-synuclein pathology could be useful as a biomarker to (i) detect the presence of the disease potentially in early stages, (ii) to evaluate disease progression and (iii) to be used as a pharmacodynamics tool for drug development. The development of an alpha-synuclein PET imaging agent is considered nowadays key for an accurate diagnosis of synucleinopathies as well as to support the clinical development of therapeutics targeting alpha-synuclein, starting from the optimal selection of the trial population (Eberling, Dave and Frasier, J. Parkinson's Disease, 3, 565-567 (2013)).
Only recently, the first non-invasive images of pathological alpha-synuclein (a-syn) in human brain were reported and presented positive clinical proof-of-concept data for an a-syn positron emission tomography (PET) tracer, as an imaging agent to identify MSA patients (Capotosti F.; Discovery of a novel and promising PET-tracer for alpha-synuclein; Oral presentation; ADPD 2022 International Conference; Barcelona, Spain; March 18, 2022; Smith R.; Initial scans using a novel PET-tracer for alpha-synuclein; Oral presentation; ADPD 2022 International Conference; Barcelona, Spain; March 18, 2022).
There is a clear need to find molecular probes with high alpha-synuclein selectivity which recognize and bind to the pathological alpha-synuclein. In order to minimize background signal interference resulting from non-specific off-target binding and to reduce dosing requirements, alpha-synuclein imaging compounds should bind with high affinity and selectivity to their target.
For imaging of alpha-synuclein aggregates associated with neurological diseases such as Parkinson's Disease or multiple system atrophy, imaging compounds need to penetrate the blood brain barrier and pass into the relevant regions of the brain. For targeting intracellular amyloid-like inclusions such as alpha-synuclein, cell permeability is a further requirement of imaging compounds. A further prerequisite in order to avoid unnecessary accumulation of the compound which may result in increased risk of unwanted side-effects is a fast compound wash-out from the brain (or other targeting organ).
WO 2011/128455 refers to specific compounds which are suitable for treating disorders associated with amyloid proteins or amyloid-like proteins. US 2012/0302755 relates to certain imaging agents for detecting neurological dysfunction. Further compounds for the diagnosis of neurodegenerative disorders on the olfactory epithelium are discussed in WO 2012/037928.
WO 2010/063701 refers to a certain in vivo imaging agent for use in a method to determine the presence of, or susceptibility to, Parkinson's disease, wherein the in vivo imaging agent comprises an alpha-synuclein binder labelled with an in vivo imaging moiety, and wherein the in vivo imaging agent binds to alpha-synuclein with a binding affinity.
US 2014/0142089 relates to a method for preventing or treating a degenerative brain disease, the method comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising a specific compound, a pharmaceutically acceptable salt, an isomer, a solvate, a hydrate, and a combination thereof.
WO 2009/155017 describes aryl or heteroaryl substituted azabenzoxazole derivatives, which are stated to be useful as tracers in positron emission tomography (PET) imaging to study amyloid deposits in the brain in vivo to allow diagnosis of Alzheimer's disease. WO 2016/033445 refers to a specific compound for imaging huntingtin protein.
WO 2017/153601 , WO 2019/234243 and WO 2021/224489 refer to bicyclic compounds for imaging alpha-synuclein aggregates.
There remains a need for a new class of imaging compounds that bind with reasonably high affinity to alpha-synuclein.
SUMMARY OF THE INVENTION
The present invention provides compounds that can be employed in diagnosing a disease, disorder or abnormality associated with alpha-synuclein aggregates, such as Parkinson's disease or MSA, prognosing such a disease, disorder or abnormality, and monitoring the progression of such a disease, disorder or abnormality. In particular, the compounds should be suitable for determining a predisposition to such a disease, disorder or abnormality, monitoring the progression of the disease, disorder or abnormality, or predicting the responsiveness of a patient who is suffering from such a disease, disorder or abnormality to the treatment with a certain medicament. Furthermore, the compounds should be suitable for positron emission tomography (PET) imaging of a disease, disorder or abnormality associated with alpha-synuclein aggregates and / or detecting and optionally quantifying alpha-synuclein aggregates.
Various embodiments of the invention are described herein.
Within a certain aspect, provided herein is a compound of formula (I):
Figure imgf000009_0001
or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, halo C1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is halo, haloC1-C4alkyl, halo C1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl.
Within a certain aspect, provided herein is a compound of formula (I):
Figure imgf000010_0001
or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
J is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl ;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo; and
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl.
In another aspect, the invention is also directed to a compound having the following subformula (la)
Figure imgf000010_0002
or a detestably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
R1 and R2 are defined above, R3 is halo, or C1-C4alkyl; and q is 0, 1 or 2. In one aspect, the present invention provides a diagnostic composition comprising a compound of formula (I), and optionally at least one pharmaceutically acceptable excipient, carrier, diluent and/or adjuvant.
In one aspect, the present invention provides a compound of formula (I), or a diagnostic composition as defined herein, which can be used in the imaging of alpha-synuclein aggregates.
In another aspect, the compound of formula (I), or the diagnostic composition can be for use in positron emission tomography imaging of alpha-synuclein aggregates.
In another aspect, the compound of formula (I) or the diagnostic composition, as defined herein, can be for use for in vitro imaging, ex vivo imaging, or in vivo imaging, preferably the use is for in vivo imaging, more preferably the use is for brain imaging.
In yet another aspect, the compound of formula (I) or the diagnostic composition, as defined herein, can be for use in diagnosis.
In a further aspect, the present invention refers to a method of diagnosing a disease, disorder or abnormality associated with alpha-synuclein aggregates in a subject, the method comprising the steps:
(a) Administering a compound of formula (I), or a diagnostic composition which comprises a compound of formula (I), as defined herein, to the subject;
(b) Allowing the compound to bind to the alpha-synuclein aggregates; and
(c) Detecting the compound bound to the alpha-synuclein aggregates.
In another aspect, the present invention refers to a method of positron emission tomography (PET) imaging of alpha-synuclein aggregates in a tissue of a subject, the method comprising the steps:
(a) Administering a compound of formula (I), or a diagnostic composition which comprises a compound of formula (I), as defined herein to the subject;
(b) Allowing the compound to bind to the alpha-synuclein aggregates; and
(c) Detecting the compound bound to the alpha-synuclein aggregates by collecting a positron emission tomography (PET) image of the tissue of the subject.
In a further aspect, the present invention is directed to a method for the detection and optionally quantification of alpha-synuclein aggregates in a tissue of a subject, the method comprising the steps: (a) Bringing a sample or a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound of formula (I), or a diagnostic composition which comprises a compound of formula (I), as defined herein;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates; and
(d) Optionally quantifying the amount of the compound bound to the alpha-synuclein aggregates.
The present invention is also directed to a method of collecting data for the diagnosis of a disease, disorder or abnormality associated with alpha-synuclein aggregates wherein the method comprises the steps:
(a) Bringing a sample or a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound of the formula (I), or a diagnostic composition which comprises a compound of formula (I), as defined herein;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates; and
(d) Optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the alpha-synuclein aggregates in the sample or specific body part or body area.
The present invention also refers to a method of collecting data for determining a predisposition to a disease, disorder or abnormality associated with alpha-synuclein aggregates, the method comprising the steps:
(a) Bringing a sample or a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound of the formula (I), or a diagnostic composition which comprises a compound of formula (I), as defined herein;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates; and
(d) Optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the alpha-synuclein aggregates in the sample or specific body part or body area.
In a further aspect, the present invention also relates to a method of collecting data for prognosing a disease, disorder or abnormality associated with alpha-synuclein aggregates, wherein the method comprises the steps: (a) Bringing a sample, a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound of the formula (I), or a diagnostic composition which comprises a compound of formula (I), as defined herein;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates;
(d) Optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the alpha-synuclein in the sample or specific body part or body area; and
(e) Optionally repeating steps (a) to (c) and, if present, optional step (d) at least one time.
In another aspect, the present invention is directed to a method of collecting data for monitoring the progression of a disease, disorder or abnormality associated with alpha-synuclein aggregates in a patient, the method comprising the steps:
(a) Bringing a sample, a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with the compound of the formula (I), or a diagnostic composition which comprises a compound of formula (I), as defined herein;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates;
(d) Optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the alpha-synuclein in the sample or specific body part or body area; and
(e) Optionally repeating steps (a) to (c) and, if present, optional step (d) at least one time.
In a further aspect, the present invention relates to a method of collecting data for predicting responsiveness of a patient suffering from a disease, disorder or abnormality associated with alpha- synuclein aggregates to a medicament, the method comprising the steps:
(a) Bringing a sample, a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound of formula (I), or a diagnostic composition which comprises a compound of formula (I), as defined herein;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates;
(d) Optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the aipha-synuclein aggregates in the sample or specific body part or body area; and
(e) Optionally repeating steps (a) to (c) and, if present, optional step (d) at least one time. In another aspect, the invention is further directed to a compound of formula (lll-F):
Figure imgf000014_0001
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1F is a 4- to 6-membered heterocyclyl, which is optionally substituted with at least one C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably wherein R1F is a 4- to 6-membered heterocyclyl; or
R1F is C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably R1F is C1-C4alkoxy, or C1-C4alkyl; or
R1F is -NH-C3-C6cycloalkyl or Cs-Cacycloalkyl; and
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl ;
R4 and R5 are independently selected from H, and C1-C4alkyl;
LG is a leaving group; and n is at least 1 (e.g., 1 , or 2 or 3), preferably 1.
In a preferred aspect, the invention refers to a compound of formula (lll-F)
Figure imgf000014_0002
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1F is a 4- to 6-membered heterocyclyl; or
R1F is C1-C4alkoxy, or C1-C4alkyl; or
R1F is -NH-C3-C6cycloalkyl or C3-C6cycloalkyl; and
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
LG is a leaving group; and n is at least 1 .
In another preferred aspect, the invention refers to a compound of formula (lll-F’)
Figure imgf000015_0001
(lll-F’) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
(A) is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or CrC4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, CrC4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -Cg-Cgcycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl; and
R2F is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from C1-C4alkoxy, and C1-C4alkyl; and
LG is a leaving group.
A further aspect of the invention relates to a compound of formula (l-F)
Figure imgf000015_0002
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
(A ) is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or CrC4alkyl;
R1F is a 4- to 6-membered heterocyclyl, which is optionally substituted with at least one C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably R1F is a 4- to 6-membered heterocyclyl; or
R1F is C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably R1F is C1-C4alkoxy, or C1-C4alkyl; or
R1F is -NH-C3-C6cycloalkyl or C3-C6cycloalkyl; and R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
R4 and R5 are independently selected from H, and C1-C4alkyl; and n is at least 1 (e.g., 1 , 2 or 3), preferably 1.
In a preferred aspect, the invention is direct to a compound of formula (l-F)
Figure imgf000016_0001
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
6^) is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1F is a 4- to 6-membered heterocyclyi; or
R1F is C1-C4alkoxy, or C1-C4alkyl; or
R1F is -NH-C3-C6cycloalkyl or C3-C6cycloalkyl; and
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and n is at least 1 , preferably 1.
In a further aspect the invention is directed to a compound a compound of formula (l-F’):
Figure imgf000016_0002
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
^-2 is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyi which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl; and R2F is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from C1-C4alkoxy, and C1-C4alkyl.
In another aspect, the invention is further directed to a compound of formula (lll-H)
Figure imgf000017_0001
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
(A) is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably R1 is a 4- to 6- membered heterocyclyl which is optionally substituted with at least one halo; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably halo, haloC1-C4alkyl, haloC1-C4alkoxy, C-i-C4alkoxy, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo; and
R2 is a 5-membered or 6-membered heteroaryl, optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
R4 and R5 are independently selected from H, CrC4alkyl and haloC1-C4alkyl;
X is bromo, chloro or iodo; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; and with the proviso that the compound of formula (lll-H) comprises at least one X.
In a preferred embodiment, a compound of formula (lll-H) is defined
Figure imgf000017_0002
or a stereoisomer, racemic mixture, pharmaceuticaliy acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo; or
R1 is halo, haloC1-C4alkyl, haloCi^alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or
-C3-C6cycloalkyl is optionally substituted with at least one halo; and
R2 is a 5-membered or 6-membered heteroaryl, optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
X is bromo, chloro or iodo; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; and with the proviso that the compound of formula (II l-H) comprises at least one X.
In another aspect, the invention is further directed to a compound of formula (l-H)
Figure imgf000018_0001
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably R1 is a 4- to 6- membered heterocyclyl which is optionally substituted with at least one halo; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo; and
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl; Y is D, CDs, T or CT3; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; with the proviso that the compound of formula (l-H) comprises at least one D, CD3, T or CT3, wherein D is 2H (deuterium) and T is 3H (Tritium). In one embodiment, the compound of formula (l-H) comprises at least one T or CT3. In one embodiment, the compound of formula (l-H) comprises at least one D or CD3.
In a preferred aspect, the invention relates to a compound of formula (l-H)
Figure imgf000019_0001
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof,
CA) is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloCrC4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-Cecycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-Cecycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo; and
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
Y is D, CD3, T or CT3; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; with the proviso that the compound of formula (l-H) comprises at least one D, CD3, T or CT3, wherein D is Deuterium and T is 3H (Tritium).
In another aspect, the invention is further directed to a method of preparing a compound of formula (l-F) by reacting a compound of formula (lll-F) with a 18F-fluorinating agent (e.g., K18F, Rb18F, Cs18F, Na18F, tetra(Ci-6alkyl)ammonium salt of 18F, Kryptofix[222]18F, tetrabutylammonium [18F]fluoride or any other suitable agents), so that the Leaving Group (LG) is replaced by 18F. In another aspect, the invention is further directed to a method of preparing a compound of formula (l-H) by reacting the compound of formula (lll-H) with a 3H radiolabelling agent (e.g., tritium gas or any other suitable agents), so that X is replaced by T or CT3.
In another aspect, the invention is further directed to a method of preparing a compound of formula (l-H) by reacting the compound of formula (lll-H) with a 2H radiolabelling agent comprising D, (e.g., D2O, D4-methanol or any other suitable agents) preferably in the presence of a catalyst like Pd/C, so that X is replaced by D or CD3 (D is deuterium).
In another aspect, the invention is further directed to the use of the compound of formula (I) as an in vitro analytical reference or an in vitro screening tool.
In another aspect, the invention is further directed to a test kit for detection and/or diagnosis of a disease, disorder or abnormality associated with alpha-synuclein aggregates, wherein the test kit comprises at least one compound of formula (I) as defined herein, preferably at least one detectably labelled compound, more preferably at least one compound of formula (l-F) or (l-H).
The invention is further directed to a kit for preparing a radiopharmaceutical preparation, wherein the kit comprises a sealed vial containing at least one compound of formula (lll-F) or (lll-H).
DEFINITIONS
For the purpose of interpreting this specification, the following definitions will apply unless specified otherwise, and when appropriate, terms used in the singular will also include the plural and vice versa. It must also be noted that as used herein and in the appended claims, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "the compound" includes reference to one or more compounds; and so forth.
The term "C1-C4alkyl" refers to a saturated straight or branched hydrocarbon chain consisting solely of carbon and hydrogen atoms, containing no unsaturation, having from one to four carbon atoms, and which is attached to the rest of the molecule by a single bond. Examples of suitable alkyl groups having 1 to 4 carbon atoms include, but are not limited to, methyl, ethyl, propyl, isopropyl, 1 -methylethyl, n-butyl, t-butyl and isobutyl. The term "C1-C4alkoxy" refers to a radical of the formula -ORa where Ra is a C1-C4alkyl radical as generally defined above. Examples of C1-C4alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, and isobutoxy.
The term "halogenC1-C4alkyl" or "haloC1-C4alkyl" refer to a C1-C4alkyl radical as defined above, substituted with one or more (e.g., 1 , 2 or 3, preferably 1 or 2, more preferably 1) halo radicals as defined below. Examples of "haloC1-C4alkyl" include, but are not limited to, trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1 ,3-dibromopropan-2-yl, 3-bromo-2- fluoropropyl and 1 ,4,4-trifluorobutan-2-yl.
The term "halogenC1-C4alkoxy" refers to a C1-C4alkoxy radical as defined above, substituted with one or more (e.g., 1 , 2 or 3, preferably 1 or 2, more preferably 1 ) halo radicals as defined below. Examples of "haloC1-C4alkoxy" include, but are not limited to, trifluoromethoxy, difluoromethoxy, fluoromethoxy, 2,2,2-trifluoroethoxy, 3,3,3-trifluoropropoxy, 4,4,4-trifluorobutoxy, 2,2-difluorobutoxy, and 4-bromobutoxy.
The term "heterocyclyl" refers to a stable 4- to 6-membered non-aromatic monocyclic ring radical which comprises 1 or 2 heteroatoms which are, e.g., selected from N, O or S. The heterocyclyl group can be unsaturated or saturated. The heterocyclyl radical may be bonded via a carbon atom or a heteroatom. Examples include, but are not limited to, azetidinyl, oxetanyl, pyrrolidinyl, pyrrolidyl, tetrahydrofuryl, tetrahydrothienyl, piperidyl, piperazinyl, tetrahydropyranyl, or morpholinyl, preferably azetidinyl, pyrrolidinyl, or piperidyl, more preferably pyrrolidinyl.
The term "heteroaryl" refers to a 5- or 6-membered aromatic monocyclic ring, which comprises 1 , 2, or 3 heteroatoms independently selected from N, O and S. The heteroaryl radical may be bonded via a carbon atom or heteroatom selected from N, O and S. Examples of heteroaryl include, but are not limited to, thiopyranyl, dioxanyl, pyranyl, pyrazinyl, pyridazinyl, pyrimidyl, isothiazolyl, pyrazolyl, thiazolyl or pyridyl, with pyridyl, isothiazolyl, pyrazolyl, and thiazolyl being preferred.
The term "Hal" or "halogen" or "Halo" refers to F, Cl, Br, and I. With respect to diagnostic and pharmaceutical applications, F (e.g., 19F and 18F) is particularly preferred.
The term “leaving group” (LG) as employed herein is any leaving group and means an atom or group of atoms that can be replaced by another atom or group of atoms. Examples are given e.g. in Synthesis (1982), p. 85-125, table 2, Carey and Sundberg, Organische Synthese, (1995), page 279- 281 , table 5.8; or Netscher, Recent Res. Dev. Org. Chem., 2003, 7, 71-83, schemes 1 , 2, 10 and 15 and others). (Coenen, Fluorine-18 Labeling Methods: Features and Possibilities of Basic Reactions, (2006), in: Schubiger P.A., Friebe M., Lehmann L., (eds), PET-Chemistry - The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, pp.15-50, explicitly: scheme 4 pp. 25, scheme 5 pp 28, table 4 pp 30, Figure 7 pp 33). Preferably, the "leaving group" (LG) is selected from halogen, C1-C4 alkylsulfonate and C6-C10arylsulfonate, wherein the Ce-Cwarylsulfonate can be optionally substituted with -CH3 or -NO2.
Unless specified otherwise, the term “compound of the invention” refers to a compound of formula (I), or of subformulae thereof (e.g. (la), (l-F), (l-H*), (l-H)), or a detectably labelled compound, stereoisomer (including diastereomeric mixtures and individual diastereomer, enantiomeric mixture and single enantiomer, mixture of conformers and single conformer), racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof. It is understood that every reference to a compound of formula (I) also covers the subformulae thereof (e.g. (la), (l-F), (l-H*), (l-H)). The compounds of the formulae (II l-F) and (I I l-H) will be referred to as the precursors of the compounds of the present invention.
Compounds of the present invention and their precursors having one or more optically active carbons can exist as racemates and racemic mixtures, stereoisomers (including diastereomeric mixtures and individual diastereomers, enantiomeric mixtures and single enantiomers, mixtures of conformers and single conformers), tautomers, atropoisomers, and rotamers. All isomeric forms are included in the present invention.
"Pharmaceutically acceptable salts" are defined as derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. The pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as, but not limited to, hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as, but not limited to, acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, and the like. The pharmaceutically acceptable salts of the compounds of the present invention and their precursors can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two. Organic solvents include, but are not limited to, nonaqueous media like ethers, ethyl acetate, ethanol, isopropanol, or acetonitrile. Lists of suitable salts can be found in Remington’s Pharmaceutical Sciences, 18th ed., Mack Publishing Company, Easton, PA, 1990, p. 1445, the disclosure of which is hereby incorporated by reference.
"Pharmaceutically acceptable" is defined as those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit/risk ratio.
"Solvates" can be formed from the compound of the present invention and any suitable pharmaceutically acceptable solvent. Examples include C1-4 alcohols (such as methanol or ethanol).
The patients or subjects in the present invention are typically animals, particularly mammals, more particularly humans.
Alpha-synuclein aggregates are multimeric beta-sheet rich assemblies of alpha-synuclein monomers that can form either soluble oligomers or soluble/insoluble protofibrils or mature fibrils which coalesce into intracellular deposits detected as a range of Lewy pathologies in Parkinson’s disease and other synucleinopathies. Alpha-synuclein aggregates that are composing Lewy pathologies can be detected as having the following morphologies: Lewy bodies, Lewy neurites, premature Lewy bodies or pale bodies, perikaryal deposits with diffuse, granular, punctate or pleomorphic patterns. Moreover, alpha-synuclein aggregates are the major component of intracellular fibrillary inclusions detected in oligodendrocytes (also referred to as glial cytoplasmic inclusions) and in neuronal somata, axons and nuclei (referred to as neuronal cytoplasmic inclusions) that are the histological hallmarks of multiple system atrophy. Alpha-synuclein aggregates in Lewy pathologies often display substantial increase in post-translational modifications such as phosphorylation, ubiquitination, nitration, and truncation.
Lewy bodies are abnormal aggregates of protein that develop inside nerve cells in Parkinson’s disease (PD), Lewy body dementia and other synucleinopathies. Lewy bodies appear as spherical masses that displace other cell components. Morphologically, Lewy bodies can be classified as being brainstem or cortical type. Classic brainstem Lewy bodies are eosinophilic cytoplasmic inclusions consisting of a dense core surrounded by a halo of 5-10-nm-wide radiating fibrils, the primary structural component of which is alpha-synuclein; cortical Lewy bodies differ by lacking a halo. The presence of Lewy bodies is a hallmark of Parkinson’s disease.
Lewy neurites are abnormal neuronal processes in diseased neurons, containing granular material, abnormal alpha-synuclein (a-syn) filaments similar to those found in Lewy bodies, dot-like, varicose structures and axonal spheroids. Like Lewy bodies, Lewy neurites are a feature of a- synucleinopathies such as dementia with Lewy bodies and Parkinson's disease.
Glial cytoplasmic inclusions (GCIs or Papp-Lantos bodies) are argyrophilic cytoplasmic aggregates in oligodentroglial cells composed of filamentous alpha-synuclein. Morphologically appear as triangles, half-moon or sickle shapes. In MSA, besides GCIs, inclusions composed of alpha-synuclein filaments are detected in neurons in the cytoplasm or beneath the nuclear membrane termed neuronal cytoplasmic inclusions and neuronal nuclear inclusions respectively. GCIs are recognized as the defining morphological feature of MSA; their widespread distribution is a criterion for the definite post-mortem neuropathological diagnosis of MSA.
The terms "disease", "disorder" or "abnormality" are used interchangeably herein.
The compounds of formula (I) can bind to alpha-synuclein aggregates. The type of bonding with the compounds of formula (I) has not been elucidated and any type of bonding is covered by the present invention. The wording "compound bound to the alpha-synuclein aggregates" and the like are used interchangeably herein and are not considered to be limited to any specific type of bonding.
The preferred definitions given in the "Definition'-section apply to all of the embodiments described below unless stated otherwise. Various embodiments of the invention are described herein, it will be recognized that features specified in each embodiment may be combined with other specified features to provide further embodiments of the present invention. DETAILED DESCRIPTION OF THE INVENTION
Compounds of the invention
The compounds of the present invention and their precursors are described in the following. It is to be understood that all possible combinations of the following definitions are also envisaged. It is also understood that all of the embodiments and preferred embodiments which are given with respect to the formula (I) apply analogously to the formulae (la), (lll-F), (l-F), (II l-H), (l-H) and (l-H*), etc. and vice versa. It is also understood that the preferred embodiments of which are given with respect to the formula (lll-F) apply analogously to the formula (l-F) and vice versa. It is also understood that the preferred embodiments of which are given with respect to the formula (I I l-H) apply analogously to the formula (l-H) and (l-H*), respectively, and vice versa.
The present invention relates to a compound of formula (I):
Figure imgf000025_0001
or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl, if
Figure imgf000025_0002
substituted, <-^is substituted with 1 , 2 or 3, preferably 1 or 2, more preferably with 1 halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloCr C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo; and
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and R4 and R5 are independently selected from H, CrC4alkyl and haloC1-C4alkyl. Throughout the present application, R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl, preferably at least one of R4 and R5 is not H.
In one embodiment,
Figure imgf000026_0001
is selected from the group consisting of
Figure imgf000026_0002
which can be attached at any available position to the triazolo heterocycle;
( A-j— and wherein can be optionally substituted by one or more substituents selected from halo; haloCi-TXialkyl, haloC1-C4alkoxy, C1-C4alkoxy, or (XUalkyl.
( AT—
In a preferred embodiment, is selected from the group consisting of
Figure imgf000026_0003
which can be attached at any available position to the triazolo heterocycle; and wherein
Figure imgf000026_0004
can be optionally substituted by one or more substituents selected from halo; haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl.
The optional substituent of
Figure imgf000026_0005
is preferably halo or C1-C4alkyl.
In a preferred embodiment,
Figure imgf000026_0006
is a 6-membered heteroaryl comprising at least one N, preferably
® (A) is pyridyl which can be substituted at any available position by R1, more preferably R1-V> is
Figure imgf000026_0007
being as defined above.
In one embodiment, R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl (e.g., 1 to 3, preferably 1 or 2, more preferably 1 ). In a preferred embodiment, R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo (e.g., 1 to 3, preferably 1 or 2, more preferably 1 ). In a preferred embodiment, R1 is a 4- to 6-membered heterocyclyl which is substituted with at least one halo. Preferably, the heterocyclyl is substituted with at least one halo, more preferably with one or two halo, even more preferably with one halo. In another preferred embodiment, R1 is a 4- to 6- membered heterocyclyl which is unsubstituted.
In an embodiment, R1 is selected from the following:
Figure imgf000027_0001
wherein R1’ is independently halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; and s = 0, 1 , 2 or 3.
In one embodiment, R1 is a 4- to 6-membered heterocyclyl selected from the following:
Figure imgf000027_0002
wherein R1a is F or H, preferably F. In another preferred embodiment R1a is H.
Preferably, R1 is a 4- or 5-membered heterocyclyl selected from the following:
Figure imgf000027_0003
wherein R1a is F or H, preferably F. In another preferred embodiment R1a is H.
In yet another embodiment R1 is a 5-membered heterocyclyl which is:
Figure imgf000027_0004
In yet another embodiment R1 is a 5-membered heterocyclyl which is: o .
In another embodiment, R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl. In an embodiment R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl. Preferably, R1 is haloC1-C4alkyl or halo, more preferably R1 is -O-(CH2)X-Hal (x = 1 to 4, preferably 1 to 3, more preferably 1 or 2, even more preferably 2), even more preferably -O-CH2-CH2-F. In another preferred embodiment, R1 is NR4R5, preferably wherein R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl.
In yet a further embodiment, R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo.
In each of the embodiments, F is preferably 19F or 18F.
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl, preferably haloC1-C4alkyl, or C1-C4alkyl.
In preferred embodiment, R2 is a 5-membered or 6-membered heteroaryl selected from the following:
Figure imgf000028_0001
wherein
R2a is independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
R2b is selected from H, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and s is 0, 1 or 2 (preferably 0 or 1 , more preferably 0). In another preferred embodiment s is 1 .
More preferably, R2 is a 5-membered or 6-membered heteroaryl selected from the following:
Figure imgf000028_0002
wherein
R2b is selected from haloC1-C4alkyl, haloC1-C4alkoxy, alkoxy, H or C1-C4alkyl, preferably H, haloC1-C4alkyl (preferably wherein halo is F) or C1-C4alkyl, preferably H or C1-C4alkyl. In another preferred embodiment, the invention provides a compound having the formula (la):
Figure imgf000029_0001
or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein R1 and R2 are as defined above, and R3 is halo, or C1-C4alkyl; and q is 0, 1 or 2.
In one embodiment, the present invention provides a compound of formula (I), wherein the compound is selected from
Figure imgf000029_0002
Figure imgf000030_0001
Figure imgf000031_0001
Figure imgf000032_0002
or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof. In one embodiment, the present invention provides a compound of formula (I), wherein the compound is selected from
Figure imgf000032_0001
Figure imgf000033_0001
Figure imgf000034_0001
Figure imgf000035_0003
or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof.
In one embodiment, the present invention provides a compound of formula (I) which is a detectably labelled compound. The detectable label can be a radioisotope. In one embodiment, the compound of formula (I) comprises at least one radioisotope. Preferably, the detectable label is a radioisotope selected from 18F, 2H and 3H. Most preferably, the radioisotope is selected from 18F and 3H.
In one embodiment, the present invention provides a compound of formula (I) wherein R1 is
Figure imgf000035_0001
In another embodiment, the present invention provides a compound of formula (I) wherein R1 is -O-CH2-CH2-18F.
In another embodiment, the present invention provides a compound of formula (I) wherein Rz is
Figure imgf000035_0002
, wherein R2b is -CH2-CH2-18F or -CH2-CH2-CH2-18F.
In another embodiment, the present invention provides a compound of formula (I) wherein R2 is
Figure imgf000036_0001
In one embodiment the present invention provides a compound of formula (I), wherein the compound is a detectably labelled compound of formula (l-F):
Figure imgf000036_0002
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl; which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1F is a 4- to 6-membered heterocyclyl, which is optionally substituted with at least one C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably a 4- to 6-membered heterocyclyl; or
R1F is C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably C1-C4alkoxy, or C1-C4alkyl; or
R1F is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, and
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
R4 and R5 are independently selected from H, and C1-C4alkyl; and n is at least 1 (e.g., 1, 2 or 3), preferably 1.
In one embodiment, R1F is -NH-C3-C6cycloalkyl, C3-C6cycloalkyl, C1-C4alkoxy, or C1-C4alkyl or heterocyclyl. Preferably -R1F-(18F)n is selected from the following:
Figure imgf000036_0003
wherein R1’ is 18F; and s = 1 , 2 or 3, preferably s = 1.
In a preferred embodiment, -R1F-(18F)n is selected from the following:
Figure imgf000036_0004
More preferably, -R1F-(18F)n is selected from the following:
Figure imgf000037_0001
Even more preferably, -R1F-(18F)n is:
Figure imgf000037_0002
In another embodiment, -R1F-(18F)n is -O-(CH2)X-18F (x = 1 to 4, preferably 1 to 3, more preferably 1 or 2, even more preferably 2), even more preferably -O-CH2-CH2-18F.
In another embodiment the present invention provides a compound of formula (I), wherein the compound is a detectably labelled compound of formula (l-F’):
Figure imgf000037_0003
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloCr
C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl; and
R2F is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from C1-C4alkoxy.
In a preferred embodiment, -R2F-(18F)n is selected from the following
Figure imgf000037_0004
Figure imgf000038_0001
The detectably labelled compound of formula (l-F) or (l-F’) comprises at least one 18F. Preferably, the detectably labelled compound of formula (l-F) or (l-F’) comprises one 18F.
In one embodiment, the present invention provides a compound of formula (I), wherein the compound is a detectably labelled compound of formula (l-H*)
Figure imgf000038_0002
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof,
(A'
7) is a 6-membered heteroaryl which is optionally substituted with at least one halo; haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or (XUalkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or CrC4alkyl, preferably halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl; with the proviso that the compound of formula (l-H*) comprises at least one 2H (deuterium “D”) or 3H (Tritium “T”), preferably 1 , 2, or 3 D or T, even more preferably 2 or 3 D or T. Preferably, the compound of formula (l-H*) comprises at least one 3H (Tritium “T”), preferably 1 , 2, or 3 T, even more preferably 2 or 3 T. The 3H can be present as T or as -CT3. The 2H can be present as D or as -CD3.
In a preferred embodiment, the compound is a detectably labelled compound of formula (l-H)
Figure imgf000039_0001
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloCrC4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, CrC4alkoxy, NR4R5, or C1-C4alkyl, preferably a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or CrC4alkyl, preferably halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo; and
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and CrC4alkyl;
R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl;
Y is D, CD3, T or CT3; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; with the proviso that the compound of formula (l-H) comprises at least one D, CD3, T or CT3, wherein D is 2H (Deuterium) and T is 3H (Tritium). In one embodiment, the compound of formula (l-H) comprises at least one D or CD3. In another embodiment, the compound of formula (l-H) comprises at least one T or CT3.
It is understood that the deuterium or tritium can present at any available position at which a hydrogen is present. For instance, in the group R2 deuterium or tritium can be present either directly bound to the 5-membered or 6-membered heteroaryl (such as in the form of D or T) or can be present in the haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl (such as in the form of CD3 or CT3). In the 4- to 6-membered heterocyclyl of R1, deuterium or tritium can be, e.g., directly bound to the 4- to 6-membered heterocyclyl. (A)
In one preferred embodiment, is a 6-membered heteroaryl optionally substituted with one halo or C1-C4alkyl and m is 1 , 2 or 3, preferably 1 .
In one embodiment, R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl, and p is 1 , 2 or 3, preferably 1.
Preferably, R2 is a 5-membered or 6-membered heteroaryl selected from the following:
Figure imgf000040_0001
wherein
R2a is independently selected from T, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; R2b is selected from H, T, haloC1-C4alkyl and C1-C4alkyl, wherein haloC1-C4alkyl and C1-C4alkyl optionally comprise one or more T (preferably R2b is selected from T or CT3); and s is 0, 1 or 2 (preferably 0 or 1 , more preferably 0).
Preferably, R2a is -T, -OCH3, -CH3 or -H; and R2b is selected from -H, -T or -CT3.
Preferably, R2 is a 5-membered or 6-membered heteroaryl selected from the following:
Figure imgf000040_0002
wherein
R2b is selected from T, H or C1-C4alkyl (e.g., CT3), preferably T, H or C1-C4alkyl (e.g., CT3).
In a preferred embodiment, the detectably labelled compound of formula (l-H*) or (l-H) comprises one, two or three T. Preferably, the detectably labelled compound of formula (l-H*) or (l-H) comprises one T. In another embodiment, the detectably labelled compound of formula (l-H*) or (l-H) comprises two T. In another embodiment, the detectably labelled compound of formula (l-H*) or (l-H) comprises three T such as -CT3.
In another embodiment R2 is a 5-membered or 6-membered heteroaryl selected from the following:
Figure imgf000041_0001
wherein
R2a is independently selected from D, haloC1-C4alkyl, haloC1-C4alkoxy, CrC4alkoxy, and C1-C4alkyl; R2b is selected from H, D, haloC1-C4alkyl and C1-C4alkyl, wherein haloC1-C4alkyl and C1-C4alkyl optionally comprise one or more D (preferably R2b is selected from D or CD3); and s is 0, 1 or 2 (preferably 0).
Preferably, R2a is -D, -OCH3, -CHsor -H; and R2b is selected from -H, -D or -CD3.
Preferably, R2 is a 5-membered or 6-membered heteroaryl selected from the following:
Figure imgf000041_0002
wherein
R2b is selected from D, H or C1-C4alkyl (e.g., CD3).
In one embodiment, the detectably labelled compound of formula (l-H*) or (l-H) comprises one, two or three D. Preferably, the detectably labelled compound of formula (l-H*) or (l-H) comprises one D. In another embodiment, the detectably labelled compound of formula (l-H*) or (l-H) comprises two D. In another embodiment, the detectably labelled compound of formula (l-H*) or (l-H) comprises three D such as -CD3.
In another embodiment, the invention provides a detectably labelled compound of formula (l-H*) or (l-H) wherein 3H Tritium (“T”) can be replaced by 2H Deuterium (“D”). The deuterated compound can be prepared by reacting a compound of formula (lll-H) with a 2H radiolabelling agent. The compounds of the present invention and their precursors can be detectably labelled. The type of the label is not specifically limited and will depend on the detection method chosen. Examples of possible labels include isotopes such as radionuclides, positron emitters, and gamma emitters, preferably the detectable label is a radioisotope. With respect to the detectably labelled compounds of the present invention and their precursors which include a radioisotope, a positron emitter, or a gamma emitter, it is to be understood that the radioisotope, positron emitter, or gamma emitter is to be present in an amount which is not identical to the natural amount of the respective radioisotope, positron emitter, or gamma emitter. Furthermore, the employed amount should allow detection thereof by the chosen detection method. Examples of suitable isotopes such as radionuclides, positron emitters and gamma emitters include 2H, 3H, 11C, 13N, 15O, and 18F, preferably 2H, 3H and 18F.
18F-labelled compounds are particularly suitable for imaging applications such as PET. The corresponding compounds which include fluorine having a natural 19F isotope are also of particular interest as they can be used as analytical standards and references during manufacturing, quality control, release, and clinical use of their 18F-analogs.
Further, substitution with isotopes such as deuterium, i.e. 2H or D, may afford certain diagnostic and therapeutic advantages resulting from greater metabolic stability by reducing for example defluorination, increased in vivo half-life or reduced dosage requirements, while keeping or improving the original compound efficacy.
Isotopic variations of the compounds of the invention and their precursors can generally be prepared by conventional procedures such as by the illustrative methods or by the preparations described in the Examples and Preparative Examples hereafter using appropriate isotopic variations of suitable reagents, which are commercially available or prepared by known synthetic techniques.
Radionuclides, positron emitters and gamma emitters can be included into the compounds of the present invention and their precursors by methods which are usual in the field of organic synthesis. Typically, they will be introduced by using a correspondingly labelled starting material when the desired compound of the present invention and its precursor is prepared. Illustrative methods of introducing detectable labels are described, for instance, in US 2012/0302755.
The position at which the detectable label is to be attached to the compounds of the present invention and their precursors is not particularly limited. The radionuclides, positron emitters and gamma emitters, for example, can be attached at any position where the corresponding non-emitting atom can also be attached. For instance, 18F can be attached at any position which is suitable for attaching F. The same applies to the other radionuclides, positron emitters and gamma emitters. Due to the ease of synthesis, preferably R1 is substituted with 18F. 3H can be attached at any available position at which H is present. If 2H is employed as a detectable label it can be attached at any available position at which H is present.
In another embodiment, the present invention relates further to a compound of formula (lll-F) that is a precursor of the compound of formula (l-F)
Figure imgf000043_0001
(lll-F) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein fA') is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1F is a 4- to 6-membered heterocyclyl, which is optionally substituted with at least one C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably R1F is a 4- to 6-membered heterocyclyl; or
R1F is C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably R1F is C1-C4alkoxy, or C1-C4alkyl; or
R1F is -NH-C3-C6cycloalkyl or C3-C6cycloalkyl, and
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
R4 and R5 are independently selected from H, and C1-C4alkyl;
LG is a leaving group; and n is at least 1 (e.g., 1, 2 or 3; preferably 1 ).
In another preferred embodiment, (LG)n-R1F is selected from the following:
Figure imgf000043_0002
wherein x is at least 1 (e.g., 1 to 3), preferably 1 or 2, even more preferably 2.
More preferably, (LG)n-R1F is selected from the following:
Figure imgf000043_0003
wherein x is 1 or 2, more preferably 2.
Even more preferably, (LG)n-R1F is :
Figure imgf000044_0001
In another embodiment, the present invention relates further to a compound of formula (lll-F') that is a precursor of the compound of formula ( l-F’):
Figure imgf000044_0002
(lll-F’) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or CrC4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl ; and
R2F is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from C1-C4alkoxy, and C1-C4alkyl; and
LG is a leaving group.
In a preferred embodiment, -R2F-(LG) is selected from the following: wherein R2b is -CH2-CH2-LG or -CH2-CH2-CH2-LG; or
Figure imgf000044_0003
Preferably, the Leaving Group (LG) in (lll-F) or (lll-F’) is halogen, C1-C4 alkylsulfonate, C1-C4alkyl ammonium, or Ce-Cwarylsulfonate, wherein the Ce-C-ioarylsulfonate can be optionally substituted with -CH3 or -NO2. More preferably, the Leaving Group (LG) is bromo, chloro, iodo, C6-C4alkylsulfonate, or Ce-Cwarylsulfonate, wherein the Ce-Cwarylsulfonate can be optionally substituted with -CH3 or -NO2. Even more preferably, the Leaving Group (LG) is mesylate, tosylate or nosylate. Even more preferably, the Leaving Group (LG) is mesylate, or nosylate. More preferably the Leaving Group (LG) is mesylate.
In another embodiment, the present invention relates to a compound of formula (lll-H), a precursor of the compound of formula (l-H):
Figure imgf000045_0001
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
( A ) is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is' optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl, preferably optionally substituted with at least one halo; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or CrC4alkyl, preferably halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
R2 is a 5-membered or 6-membered heteroaryl, optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and
R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; and
X is bromo, chloro or iodo; with the proviso that the compound of formula (lll-H) comprises at least one X (e.g., 1 , 2 or 3 X, preferably 1 or 2 X).
In the formula (lll-H), halo is preferably F, 18F or 19F. X is attached to the 6-membered heteroaryl of and or the 5-membered or 6-membered heteroaryl of R2. If halo is present as a substituent of
Figure imgf000046_0001
and X is present, the X can be present in addition of halo.
In a preferred embodiment, R2 is selected from the following:
Figure imgf000046_0002
wherein
R2a is independently selected from X, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; wherein haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkyl, or C1-C4alkoxy are optionally substituted with one or more X; s is 0, 1 or 2 (preferably 0 or 1 ); and
R2b is selected from H, X, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl.
Preferably, R2 is selected from the following:
Figure imgf000046_0003
wherein
R2a is X;
R2b is selected from H, X, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl, preferably H, X, haloC1-C4alkyl, and C1-C4alkyl, more preferably X. In a further preferred embodiment, R2b is selected from haloC1-C4alkyl, and haloC1-C4alkoxy; s is 0, 1 or 2 (preferably 0); and wherein C1-C4alkyl is optionally substituted by one or more X. In a preferred embodiment, the detectably labelled compound of formula (lll-H) comprises one, two or three X. In a preferred embodiment, the detectably labelled compound of formula (lll-H) comprises one or two X. In a more preferred embodiment, the detectably labelled compound of formula (lll-H) comprises two X, X is selected from bromo, chloro and iodo. In a preferred embodiment X is bromine.
Methods of synthesis of detectably labelled compounds
The present invention relates further to a method for preparing a compound of formula (I), or of subformulae thereof (e.g. (I-F), (l-F’), (l-H*), (l-H)), and in particular a compound of formula (lll-F), (lll-F’), or (lll-H).
In one embodiment, the present invention relates to a method for preparing a compound of formula (l-F) by reacting a compound of formula (lll-F) with a 18F-fluorinating agent, so that LG is replaced by 18F.
Figure imgf000047_0001
wherein , R1F, R2, n, and LG are as defined herein above.
In another embodiment, the present invention relates to a method for preparing a compound of formula (l-F’) by reacting a compound of formula (lll-F’) with a 18F-fluorinating agent, so that LG is replaced by 18F.
Figure imgf000047_0002
wherein , R1, R2F and LG are as defined herein above.
Suitable solvents for the 18F-fluorination comprise DMF, DMSO, acetonitrile, DMA, or mixtures thereof, preferably acetonitrile or DMSO. Suitable agents for the 18F-fluorination are selected from K18F, Rb18F, Cs18F, Na18F, tetra(Ci-6alkyl)ammonium salt of 18F, Kryptofix[222]18F and tetrabutylammonium [18F]fluoride. In one embodiment, the present invention relates to a method of preparing a compound of formula (l-H) by reacting a compound of formula (lll-H) with a 3H radiolabelling agent.
Figure imgf000048_0001
wherein R1, R2, X, Y, m, and p are as defined herein above.
In one embodiment, the present invention relates to a method of preparing a compound of formula (l-H*), by reacting a compound of formula (lll-H*) with a 3H or D radiolabelling agent.
Figure imgf000048_0002
wherein , R1, R2, X, m, and p are as defined herein above and wherein Y is D, CDs, T or CT3.
The 3H radiolabelling agent can be tritium gas. The method can be conducted in the presence of a catalyst such as palladium on carbon (Pd/C), a solvent such as dimethylformamide (DMF) and a base such as N,N-diisopropylethylamine (DIEA).
In a further embodiment, the compound of formula (lll-H*) can be reacted with a 2H radiolabelling agent comprising D (e.g., D2O, D4-methanol or any other suitable agents), preferably in the presence of a catalyst like Pd/C, so that X is replaced by D (D is deuterium, 2H).
Alternatively, in another embodiment, the present invention relates to a method for preparing a compound of formula (l-H*) by radiolabelling a compound of formula (lll-H*) (reacting the compound of formula (lll-H*)) with a 3H radiolabelling agent like CT3 radiolabelling agent, wherein T is 3H, so that X is replaced by CT3. The CT3 radiolabelling agent can be ICT3 (derivative of iodomethane with 3H). The method can be conducted in the presence of a solvent such as dimethylformamide (DMF) and a base such as caesium carbonate or sodium hydride. Kits
The precursor compounds of the present invention can also be employed in kits for the preparation of radiopharmaceutical preparations. Due to the radioactive decay, the radiopharmaceuticals are usually prepared immediately before use. The kit typically comprises a precursor of the compound of the present invention, and an agent which reacts with the precursor to introduce a radioactive label into the compound of the present invention. The precursor of the compound of the present invention, can, for example, be a compound having the formula (lll-F), (lll-H*) or (lll-H). The agent can be an agent which introduces a radioactive label such as 18F, 3H, or D.
In one embodiment, the kit of parts is a test kit for the detection and/or diagnosis of a disease, disorder or abnormality associated with alpha-synuclein aggregates, wherein the test kit comprises at least one precursor of the compound of the present invention (e.g. a compound having the formula (lll-F), (lll-H*) or (lll-H)).
In another embodiment, the kit of parts is a kit for preparing a radiopharmaceutical preparation, wherein the kit comprises a sealed vial containing at least one precursor of the compound of the present invention (e.g. a compound having the formula (lll-F), (lll-H*) or (lll-H)).
In one preferred embodiment, the kit is for use in the imaging of alpha-synuclein aggregates, wherein the imaging is preferably conducted by positron emission tomography, or is for use for in vitro imaging, ex vivo imaging, or in vivo imaging, preferably the use is for in vivo imaging. More preferably, the use is for brain imaging.
Diagnostic Compositions
The compounds of the present invention are particularly suitable for imaging of alpha-synuclein aggregates including. With respect to alpha-synuclein protein, the compounds are particularly suitable for binding to various types of alpha-synuclein aggregates including, but not limited to, Lewy bodies and/or Lewy neurites. The imaging can be conducted in mammals, preferably in humans. The imaging is preferably in vitro imaging, ex vivo imaging, or in vivo imaging. More preferably the imaging is in vivo imaging: Even more preferably, the imaging is preferably brain imaging. The imaging can also be eye/retinal imaging. The compounds of the present invention are particularly suitable for use in diagnostics. The diagnostics can be conducted for mammals, preferably for humans. The tissue of interest on which the diagnostic is conducted can be brain, tissue of the central nervous system, tissue of the eye (such as retinal tissue), tissue of peripheral organs such as the gut or other tissues, or body fluids such as cerebrospinal fluid (CSF) or blood. The tissue is preferably brain tissue.
In one embodiment, the present invention provides a diagnostic composition comprising a compound of the invention, and optionally at least one pharmaceutically acceptable excipient, carrier, diluent and/or adjuvant.
Due to their design and to the binding characteristics, the compounds of the present invention are suitable for use in the diagnosis of diseases, disorders and abnormalities associated with alpha- synuclein aggregates. In another embodiment, the diagnostic composition which comprises a compound of the present invention is also suitable for use in the diagnosis of diseases, disorders and abnormalities associated with alpha-synuclein aggregates.
In yet another embodiment, the compound of the present invention, or the diagnostic composition comprising a compound of the invention, is suitable for use in imaging, such as in vitro imaging, ex vivo imaging, or in vivo imaging, preferably the use is for in vivo imaging, more preferably the use is for brain imaging. In particular, the use is in humans.
In another embodiment, the compounds of the present invention or the diagnostic composition are particularly suitable for use in positron emission tomography imaging of alpha-synuclein aggregates.
Diseases involving alpha-synuclein aggregates are generally listed as synucleinopathies (or a- synucleinopathies). The compounds of the present invention are suitable for use in the diagnosis of diseases, disorders or abnormalities associated with alpha-synuclein aggregates including, but not limited to, Lewy bodies and/or Lewy neurites, or a predisposition therefor, wherein the diseases, disorders or abnormalities are selected from (including, but not limited to) Parkinson's disease (sporadic, familial with alpha-synuclein mutations, familial with mutations other than alpha-synuclein, pure autonomic failure and Lewy body dysphagia), SNCA duplication carrier, dementia with Lewy bodies (“pure” Lewy body dementia), Alzheimer’s disease, sporadic Alzheimer’s disease, familial Alzheimer's disease with APP mutations, familial Alzheimer's disease with PS-1 , PS-2 or other mutations, familial British dementia, Lewy body variant of Alzheimer’s disease and normal aging in Down syndrome). The compounds of the present invention are suitable for use in the diagnosis of diseases, disorders or abnormalities associated with alpha-synuclein aggregates including, but not limited to, neuronal and glial aggregates of alpha synuclein, includingmultiple system atrophy (MSA) (Shy-Drager syndrome, striatonigral degeneration and olivopontocerebellar atrophy). Other diseases that may have alpha-synuclein-immunoreactive lesions include traumatic brain injury, chronic traumatic encephalopathy, tauopathies (Pick's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration and Niemann-Pick type C1 disease), motor neuron disease, amyotrophic lateral sclerosis (sporadic, familial and ALS-dementia complex of Guam), neuroaxonal dystrophy, neurodegeneration with brain iron accumulation type 1 (Hallervorden-Spatz syndrome), prion diseases, ataxia telangiectatica, Meige’s syndrome, subacute sclerosing panencephalitis, Gaucher disease as well as other lysosomal storage disorders (including Kufor- Rakeb syndrome and Sanfilippo syndrome) and rapid eye movement (REM) sleep behavior disorder (Jellinger, Mov Disord 2003, 18 Suppl. 6, S2-12; Galvin et al. JAMA Neurology 2001 , 58 (2), 186- 190; Kovari et al., Acta Neuropathol. 2007, 114(3), 295-8; Saito et al., J Neuropathol Exp Neurol. 2004, 63(4), 323-328; McKee et al., Brain, 2013, 136(Pt 1), 43-64; Puschmann et al., Parkinsonism Relat Disord 2012, 18S1 , S24-S27; Usenovic et al., J Neurosci. 2012, 32(12), 4240-4246; Winder- Rhodes et al., Mov Disord. 2012, 27(2), 312-315; Ferman et al., J Int Neuropsychol Soc. 2002, 8(7), 907-914). Preferably, the compounds of the present invention are suitable for use in the diagnosis of Parkinson's disease, multiple system atrophy, dementia with Lewy bodies, Parkinson’s disease dementia, SNCA duplication carrier, or Alzheimer’s disease, more preferably Parkinson’s disease (PD) or multiple system atrophy (MSA).
In the methods of diagnosing a disease, disorder or abnormality associated with alpha-synuclein aggregates, e.g. Parkinson's disease or MSA, or a predisposition therefor in a subject, the method comprises the steps of:
(a) administering to the subject a diagnostically effective amount of a compound of the present invention, or a diagnostic composition which comprises a compound of the present invention;
(b) allowing the compound of the present invention to distribute into the tissue of interest (such as brain tissue, tissue of the central nervous system (CNS), tissue of the eye, tissue of peripheral organs or other tissues), or body fluid (such as cerebrospinal fluid (CSF) or blood); and
(c) imaging the tissue of interest or body fluid.
If the amount of the compound bound to the alpha-synuclein aggregates is increased compared to a normal control level the subject is suffering from or is at risk of developing a disease, disorder or abnormality associated with alpha-synuclein aggregates. The compounds of the present invention can be used for imaging of alpha-synuclein aggregates in any sample or a specific body part or body area of a patient which is suspected to contain alpha- synuclein aggregates. The compounds are able to pass the blood-brain barrier. Consequently, they are particularly suitable for imaging of alpha-synuclein aggregates in the brain, tissue of the central nervous system (CNS), tissue of the eye (such as retinal tissue), tissue of peripheral organs such as the gut or other tissues, or body fluids such as cerebrospinal fluid (CSF) or blood.
In diagnostic applications, the compounds of the present invention are preferably administered in the form of a diagnostic composition comprising the compound of the invention. A "diagnostic composition" is defined in the present invention 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 which is suitable for use in the diagnosis of the specific disease, disorder or abnormality at issue. Preferably a diagnostic composition further comprises a pharmaceutically acceptable excipient, carrier, diluent or adjuvant. Administration is preferably carried out as defined below. More preferably by injection of the composition as an aqueous solution. Such a composition may optionally contain further ingredients such as buffers; pharmaceutically acceptable solubilizers (e.g., cyclodextrins or surfactants such as Pluronic, Tween or phospholipids); and pharmaceutically acceptable stabilisers or antioxidants (such as ascorbic acid, gentisic acid or para-aminobenzoic acid). The dose of the compound of the present invention will vary depending on the exact compound to be administered, the weight of the patient, and other variables as would be apparent to a physician skilled in the art.
While it is possible for the compounds of the present invention to be administered alone, it is preferable to formulate them into a diagnostic composition in accordance with standard pharmaceutical practice. Thus, the invention also provides a diagnostic composition which comprises a diagnostically effective amount of a compound of the present invention in admixture with, optionally, at least one pharmaceutically acceptable excipient, carrier, diluent or adjuvant.
Pharmaceutically acceptable excipients are well known in the pharmaceutical art, and are described, for example, in Remington's Pharmaceutical Sciences, 15th Ed., Mack Publishing Co., New Jersey (1975). The pharmaceutical excipient can be selected with regard to the intended route of administration and standard pharmaceutical practice. The excipient must be acceptable in the sense of being not deleterious to the recipient thereof. Pharmaceutically useful excipients, carriers, adjuvants and diluents that may be used in the formulation of the diagnostic composition of the present invention may comprise, for example, solvents such as monohydric alcohols such as ethanol, isopropanol and polyhydric alcohols such as glycols and edible oils such as soybean oil, coconut oil, olive oil, safflower oil, cottonseed oil, oily esters such as ethyl oleate, isopropyl myristate, binders, adjuvants, solubilizers, thickening agents, stabilizers, disintegrants, glidants, lubricating agents, buffering agents, emulsifiers, wetting agents, suspending agents, sweetening agents, colorants, flavors, coating agents, preservatives, antioxidants, processing agents, drug delivery modifiers and enhancers such as calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose, sodium carboxymethyl cellulose, dextrose, hydroxypropyl-B-cyclodextrin, polyvinylpyrrolidone, low melting waxes, and ion exchange resins.
The routes for administration (delivery) of the compounds of the invention include, but are not limited to, one or more of: intravenous, gastrointestinal, intraspinal, intraperitoneal, intramuscular, oral (e. g. as a tablet, capsule, or as an ingestible solution), topical, mucosal (e. g. as a nasal spray or aerosol for inhalation), nasal, parenteral (e. g. by an injectable form), intrauterine, intraocular, intradermal, intracranial, intratracheal, intravaginal, intracerebroventricular, intracerebral, subcutaneous, ophthalmic (including intravitreal or intracameral), transdermal, rectal, buccal, epidural and sublingual. Preferably, the route of administration (delivery) of the compounds of the invention is intravenous.
For example, the compounds can be administered orally in the form of tablets, capsules, ovules, elixirs, solutions or suspensions, which may contain flavoring or coloring agents, for immediate-, delayed-, modified-, sustained-, pulsed- or controlled-release applications.
The tablets may contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate and glycine, disintegrants such as starch (preferably corn, potato or tapioca starch), sodium starch glycolate, croscarmellose sodium and certain complex silicates, and granulation binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin and acacia. Additionally, lubricating agents such as magnesium stearate, stearic acid, glyceryl behenate and talc may be included. Solid compositions of a similar type may also be employed as fillers in gelatin capsules. Preferred excipients in this regard include starch, a cellulose, milk sugar (lactose) or high molecular weight polyethylene glycols. For aqueous suspensions and/or elixirs, the agent may be combined with various sweetening or flavoring agents, coloring matter or dyes, with emulsifying and/or suspending agents and with diluents such as water, ethanol, propylene glycol and glycerin, and combinations thereof.
Preferably, in diagnostic applications, the compounds of the present invention are administered parenterally. If the compounds of the present invention are administered parenterally, then examples of such administration include one or more of: intravenously, intraarterially, intraperitoneally, intrathecally, intraventricularly, intraurethrally, intrasternally, intracranially, intramuscularly or subcutaneously administering the compounds; and/or by using infusion techniques. For parenteral administration, the compounds are best used in the form of a sterile aqueous solution which may contain other substances, for example, enough salts or glucose to make the solution isotonic with blood. The aqueous solutions should be suitably buffered (preferably to a pH of from 3 to 9), if necessary. The preparation of suitable parenteral formulations under sterile conditions is readily accomplished by standard pharmaceutical techniques well known to those skilled in the art.
As indicated, the compounds of the present invention can be administered intranasally or by inhalation and are conveniently delivered in the form of a dry powder inhaler or an aerosol spray presentation from a pressurized container, pump, spray or nebulizer with the use of a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, a hydrofluoroalkane such as 1 ,1 ,1 ,2-tetrafluoroethane (HFA134AT) or 1 , 1 ,1 , 2, 3,3,3- heptafluoropropane (HFA 227EA), carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. The pressurized container, pump, spray or nebulizer may contain a solution or suspension of the active compound, e. g. using a mixture of ethanol and the propellant as the solvent, which may additionally contain a lubricant, e. g. sorbitan trioleate. Capsules and cartridges (made, for example, from gelatin) for use in an inhaler or insufflator may be formulated to contain a powder mix of the compound and a suitable powder base such as lactose or starch.
Alternatively, the compounds of the present invention can be administered in the form of a suppository or pessary, or it may be applied topically in the form of a gel, hydrogel, lotion, solution, cream, ointment or dusting powder. The compounds of the present invention may also be dermally or transdermally administered, for example, by the use of a skin patch.
They may also be administered by the pulmonary or rectal routes. They may also be administered by the ocular route. For ophthalmic use, the compounds can be formulated as micronized suspensions in isotonic, pH was adjusted, sterile saline, or, preferably, as solutions in isotonic, pH was adjusted, sterile saline, optionally in combination with a preservative such as a benzylalkonium chloride. Alternatively, they may be formulated in an ointment such as petrolatum.
For application topically to the skin, the compounds of the present invention can be formulated as a suitable ointment containing the active compound suspended or dissolved in, for example, a mixture with one or more of the following: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, emulsifying wax and water. Alternatively, they can be formulated as a suitable lotion or cream, suspended or dissolved in, for example, a mixture of one or more of the following: mineral oil, sorbitan monostearate, a polyethylene glycol, liquid paraffin, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
Typically, a physician will determine the actual dosage which will be most suitable for an individual subject. The specific dose level and frequency of dosage for any particular individual may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the individual undergoing diagnosis.
The diagnostic compositions of the invention can be produced in a manner known per se to the skilled person as described, for example, in Remington's Pharmaceutical Sciences, 15th Ed., Mack Publishing Co., New Jersey (1975).
The compounds of the present invention are useful as an in vitro analytical reference or an in vitro screening tool. They are also useful in in vivo diagnostic methods.
The compounds according to the present invention can also be provided in the form of a mixture, a pharmaceutical composition, or a combination, comprising a compound according to the present invention and at least one compound selected from an imaging agent different from the compound according to the invention, a pharmaceutically acceptable excipient, carrier, diluent or adjuvant. The imaging agent different from the compound according to the invention is preferably present in a diagnostically effective amount. More preferably the imaging agent different from the compound according to the invention is an Abeta or Tau imaging agent. Methods of using the invention
In one embodiment, the invention provides a method of diagnosing a disease, disorder or abnormality associated with alpha-synuclein aggregates in a subject, the method comprising the steps:
(a) Administering a compound of the invention, or a diagnostic composition which comprises a compound of the invention to the subject;
(b) Allowing said compound to bind to the alpha-synuclein aggregates; and
(c) Detecting the compound bound to the alpha-synuclein aggregates.
Optionally, said method may further comprise the step of:
(d) Generating an image representative of the location and/or amount of the compound bound to the alpha-synuclein aggregates including.
In another embodiment, the invention provides a method of positron emission tomography (PET) imaging of alpha-synuclein aggregates in a tissue of a subject, the method comprising the steps:
(a) Administering a compound of the invention, or a diagnostic composition which comprises a compound of the invention to the subject;
(b) Allowing the compound to bind to the alpha-synuclein aggregates; and
(c) Detecting the compound bound to the alpha-synuclein aggregates by collecting a positron emission tomography (PET) image of the tissue of the subject;
In another embodiment, the invention relates to a method for the detection and optionally quantification (e.g., an in vivo or in vitro method) of alpha-synuclein aggregates in a tissue of a subject, the method comprising the steps:
(a) Bringing a sample or a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound of the invention, or a diagnostic composition which comprises a compound of the invention;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates; and
(d) Optionally quantifying the amount of the compound bound to the alpha-synuclein aggregates.
In an embodiment, the present invention refers to a method of collecting data for the diagnosis of a disease, disorder or abnormality associated with alpha-synuclein aggregates including the method comprising the steps: (a) Bringing a sample or a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound according to the present invention, or a diagnostic composition which comprises a compound according to the present invention;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates; and
(d) Optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the alpha-synuclein aggregates in the sample or specific body part or body area.
If the amount of the compound bound to the alpha-synuclein aggregates is higher than a normal control value it can be assumed that the patient is suffering from a disease, disorder or abnormality associated with alpha-synuclein aggregates.
Yet another embodiment of the present invention refers to a method of collecting data for determining a predisposition to a disease, disorder or abnormality associated with alpha-synuclein aggregates, the method comprising the steps:
(a) Bringing a sample or a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound according to the present invention, or a diagnostic composition which comprises a compound according to the present invention;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates; and
(d) Optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the alpha-synuclein aggregates in the sample or specific body part or body area.
If the amount of the compound bound to the alpha-synuclein aggregates is higher than a normal control value of a healthy/reference subject this indicates that the patient is suffering from or is at risk of developing a disease, disorder or abnormality associated with alpha-synuclein aggregates. In particular, if the amount of the compound bound to the alpha-synuclein aggregates is higher than what expected in a person showing no clinical evidence of a disease, disorder or abnormality associated with alpha-synuclein aggregates, it can be assumed that the patient has a disposition to a disease, disorder or abnormality associated with alpha-synuclein aggregates. In a further aspect, the present invention relates to a method of collecting data for prognosing a disease, disorder or abnormality associated with alpha-synuclein aggregates, wherein the method comprises the steps:
(a) Bringing a sample, a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound according to the present invention, or a diagnostic composition which comprises a compound according to the present invention;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates;
(d) Optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the alpha-synuclein aggregates in the sample or specific body part or body area; and
(e) Optionally repeating steps (a) to (c) and, if present, optional step (d) at least one time.
The progression of a disease, disorder or abnormality and/or the prospect (e.g., the probability, duration, and/or extent) of recovery can be estimated by a medical practitioner based on the presence or absence of the compound bound to the alpha-synuclein aggregates, the amount of the compound bound to the alpha-synuclein aggregates or the like. If desired, steps (a) to (c) and, if present, optional step (d) can be repeated over time to monitor the progression of the disease, disorder or abnormality and to thus allow a more reliable estimate.
A further aspect is directed to a method of collecting data for monitoring the progression (or evolution) of a disease, disorder or abnormality associated with alpha-synuclein aggregates in a patient, the method comprising the steps:
(a) Bringing a sample, a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with the compound according to the present invention, or a diagnostic composition which comprises a compound according to the present invention;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates;
(d) Optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the alpha-synuclein aggregates in the sample or specific body part or body area; and
(e) Optionally repeating steps (a) to (c) and, if present, optional step (d) at least one time.
In the method for monitoring the progression the amount of the compound bound to the alpha- synuclein aggregates can be optionally compared at various points of time during the treatment, for instance, before and after onset of the treatment or at various points of time after the onset of the treatment.
Typically, the patient is or has been undergoing treatment of the disease, disorder or abnormality associated with alpha-synuclein aggregates or is/has been undergoing treatment of the synucleinopathy. In particular, the treatment can involve administration of a medicament which is suitable for treating the disease, disorder or abnormality associated with alpha-synuclein aggregates.
In another embodiment, the invention relates to a method of collecting data for predicting responsiveness of a patient suffering from a disease, disorder or abnormality associated with alpha- synuclein aggregates to a treatment with a medicament, the method comprising the steps of
(a) Bringing a sample, a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound of the invention, or a diagnostic composition which comprises a compound of the invention;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates;
(d) Optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the alpha-synuclein aggregates in the sample or specific body part or body area; and
(e) Optionally repeating steps (a) to (c) and, if present, optional step (d) at least one time.
In the method for predicting the responsiveness, the method can further comprises steps (i) to (vi) before step (a):
(i) bringing a sample or specific body part or body area suspected to contain alpha-synuclein aggregates into contact with the compound of the present invention, which compound specifically binds to the alpha-synuclein aggregates;
(ii) allowing the compound to bind to the alpha-synuclein aggregates;
(iii) detecting the formation of the compound bound to the alpha-synuclein aggregates;
(iv) optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of alpha-synuclein aggregates in the sample or specific body part or body area;
(v) optionally comparing the amount of the compound bound to the alpha-synuclein aggregates to a normal control value; and
(vi) treating the patient with the medicament. Optionally the method can further comprise step (A) after step (d) or step (e):
(A) comparing the amount of the compound bound to the alpha-synuclein aggregates determined in step (iv) to the amount of the compound bound to the alpha-synuclein aggregates determined in step (d).
In the method for predicting responsiveness the amount of the compound bound to the alpha- synuclein aggregates can be optionally compared at various points of time during the treatment, for instance, before and after onset of the treatment or at various points of time after the onset of the treatment. A change, especially a decrease, in the amount of the compound bound to the alpha- synuclein aggregates may indicate that the patient has a high potential of being responsive to the respective treatment.
If the amount of the compound bound to the alpha-synuclein aggregates decreases over time, it can be assumed that the patient is responsive to the treatment. If the amount of the compound bound to the alpha-synuclein aggregates is essentially constant or increases overtime, it can be assumed that the patient is non-responsive to the treatment.
Alternatively, the responsiveness can be estimated by determining the amount of the compound bound to the alpha-synuclein aggregates. The amount of the compound bound to the alpha-synuclein aggregates can be compared to a control value such as a normal control value, a preclinical control value or a clinical control value. Alternatively, the control value may refer to the control value of subjects known to be responsive to a certain therapy, or the control value may refer to the control value of subjects known to be non-responsive to a certain therapy. The outcome with respect to responsiveness can either be "responsive" to a certain therapy, "non-responsive" to a certain therapy or “response undetermined” to a certain therapy. Response to the therapy may be different for the respective patients.
Optionally, the diagnostic composition can be used before, during and after, surgical procedures (e.g. deep brain stimulation (DBS)) and non-invasive brain stimulation (such as repetitive transcranial magnetic stimulation (rTMS)), for visualizing alpha-synuclein aggregates before, during and after such procedures. Surgical techniques, including DBS, improve advanced symptoms of PD on top of the best currently used medical therapy. During the past 2 decades, rTMS has been closely examined as a possible treatment for PD (Ying-hui Chou et al. JAMA Neurol. 2015 April 1 ; 72(4): 432-440). In any of the above methods, the step of optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the alpha- synuclein aggregates in the sample or specific body part or body area; comprises determining the amount of the compound bound to the alpha-synuclein aggregates;
- correlating the amount of the compound bound to the alpha-synuclein aggregates with the amount of the alpha-synuclein aggregates in the sample or specific body part or body area; and optionally comparing the amount of the compound bound with the alpha-synuclein aggregates in the sample or specific body part or body area to a normal control value in a healthy control subject.
The control value can be, e.g., a normal control value, a preclinical control value and/or a clinical control value.
A “healthy control subject” or “healthy volunteer (HV) subject” is a person showing no clinical evidence of a disease, disorder or abnormality associated with alpha-synuclein aggregates.
In an embodiment of any of the above methods the alpha-synuclein aggregates include, but are not limited to, Lewy bodies and/or Lewy neurites.
If in any of the above summarized methods the amount of the compound bound with the alpha- synuclein aggregates is higher than the normal control value, then it can be expected that the patient is suffering from or is likely to from a disease, disorder or abnormality associated with alpha-synuclein aggregates or from a synucleinopathy.
A sample or a specific body part or body area suspected to contain alpha-synuclein aggregates, is brought into contact with a compound of the present invention.
Any of the compounds of the present invention can be used in the above summarized methods. Preferably detectably labelled compounds of the present invention are employed in the above summarized methods.
The specific body part or body area is preferably of a mammal, more preferably of a human, including the full body or partial body area or body part of the patient suspected to contain alpha-synuclein aggregates. The specific body part or body area can be brain, the central nervous system, eye or a peripheral organ such as the gut, preferably brain.
The tissue can be brain tissue, tissue of the central nervous system (CNS), tissue of the eye (such as retinal tissue), tissue of peripheral organs such as the gut or other tissues, or body fluids such as cerebrospinal fluid (CSF) or blood. The tissue is preferably brain tissue. Preferably, the sample is an in vitro sample from a patient.
In the above methods, the compound of the present invention can be brought into contact with the sample or the specific body part or body area suspected to contain the alpha-synuclein aggregates by any suitable method.
In in vitro methods the compound of the present invention and a liquid sample can be simply mixed.
In an in vivo method, the specific body part or body area can be brought into contact with a compound of the invention by administering an effective amount of a compound of the invention to the patient.
The effective amount of a compound of the invention is an amount which is suitable for allowing the presence or absence of alpha-synuclein aggregates in the sample, specific body part or body area to be determined using the chosen analytical technique. The amount is not particularly limited and will depend on the compound of the formula (I), the type of detectable label, the sensitivity of the respective analytical method and the respective device. The amount can be chosen appropriately by a skilled person.
The compound is then allowed to bind to the alpha-synuclein aggregates. The step of allowing the compound to bind to the alpha-synuclein aggregates includes allowing sufficient time for the compound of the invention to bind to the alpha-synuclein aggregates. The amount of time required for binding will depend on the type of test (e.g., in vitro or in vivo) and can be determined by a person skilled in the field by routine experiments. In an in vivo method, the amount of time will depend on the time which is required for the compound to reach the specific body part or body area suspected to contain alpha-synuclein aggregates. The amount of time should not be too extended to avoid washout and/or metabolism of the compound of the invention.
The compound which has bound to the alpha-synuclein aggregates can be subsequently detected by any appropriate method. The method of detecting the compound bound to the alpha-synuclein aggregates is not particularly limited and depends, among others, on the detectable label, the type of sample, specific body part or body area and whether the method is an in vitro or in vivo method. Examples of possible methods include, but are not limited to, a fluorescence imaging technique or a nuclear imaging technique such as positron emission tomography (PET), single photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), and contrast-enhanced magnetic resonance imaging (MRI). These have been described and enable visualization of alpha- synuclein biomarkers. The fluorescence imaging technique and/or nuclear imaging technique can be employed for monitoring and/or visualizing the distribution of the detectably labelled compound within the sample or a specific body part or body area. The imaging system provides an image of bound detectable label such as radioisotopes, in particular positron emitters or gamma emitters, as present in the tested sample, the tested specific body part or the tested body area. Preferably, the compound bound to the alpha-synuclein aggregates including is detected by an imaging apparatus such as PET or SPECT scanner, more preferably PET.
The amount of the compound bound to the alpha-synuclein aggregates including can be determined by visual or quantitative analysis, for example, using PET scan images.
A compound according to the present invention or its precursor can also be incorporated into a test kit for detecting alpha-synuclein protein aggregates. The test kit typically comprises a container holding one or more compounds according to the present invention or its precursor(s) and instructions for using the compound for the purpose of binding to alpha-synuclein aggregates and detecting the formation of the compound bound to the alpha-synuclein aggregates such that presence or absence of the compound bound to the alpha-synuclein aggregates correlates with the presence or absence of the alpha-synuclein aggregates.
The term "test kit" refers in general to any diagnostic kit known in the art. More specifically, the latter term refers to a diagnostic kit as described in Zrein et al., Clin. Diagn. Lab. Immunol., 1998, 5, 45-49.
The dose of the detectably labelled compounds of the present invention, preferably compounds of formula (l-F) labelled with 18F or compounds of formula (l-H*) or (l-H) labelled with 3H, will vary depending on the exact compound to be administered, the weight of the patient, size and type of the sample, and other variables as would be apparent to a physician skilled in the art. Generally, the dose could preferably lie in the range 0.001 pg/kg to 10 pg/kg, preferably 0.01 pg/kg to 1.0 pg/kg. The radioactive dose can be, e.g., 100 to 600 MBq, more preferably 150 to 450 MBq. Methods of synthesizing the compounds of the invention
The compounds of the present invention may be prepared in accordance with the definition of compound of formula (I) by the routes described in the following Schemes or the Examples. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g. "such as") provided herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. In the following general methods, R1, R2, ®, LG, and Hal are as previously defined in the above embodiments or limited to designations in the Schemes. Unless otherwise stated, starting materials are either commercially available or are prepared by known methods.
General synthetic schemes for the preparation of compounds of this invention:
Scheme 1
Figure imgf000064_0001
Commercially available or custom-made aldehyde derivatives containing an acid sensitive protecting group (Pg) at the triazolo-moiety can be reacted with a suitable amine via reductive amination to obtain intermediate A after purification. The halogen atom is then replaced by a heteroaryl boronic acid derivative containing a leaving group (LG) via Suzuki coupling to obtain intermediate B after purification. The LG is then substituted with a suitable amine derivative via aromatic nucleophilic substitution to obtain intermediate C after purification. The Pg is removed by acid treatment to obtain intermediate D after purification. Alternatively, the halogen atom of commercially available or custom- made aldehyde derivatives containing an acid sensitive Pg at the triazolo-moiety can be replaced by a heteroaryl boronic acid derivatives via Suzuki coupling to obtain intermediate E after purification. Reductive amination of intermediate E with a suitable amine is employed to obtain intermediate F after purification. The Pg of intermediate F is removed by acid treatment to obtain intermediate D after purification. Intermediate D can be ring cyclized using 1 ,1’-carbonyldiimidazole (CDI) in an appropriate solvent to afford compounds of formula G as a free base. Acid treatment of compounds of formula G is used to afford the salts of compounds of formula (I).
Scheme 2
Figure imgf000065_0001
Commercially available or custom-made triazolo derivatives containing an ester moiety are treated with a suitable reagent to introduce an acid sensitive Pg after purification. The halogen atom is then replaced by a heteroaryl boronic acid derivative containing a leaving group (LG) via Suzuki coupling to obtain intermediate B after purification. The LG is then substituted with a suitable amine derivative via aromatic nucleophilic substitution to obtain intermediate C after purification. Reduction of the ester moiety to the corresponding alcohol is used to obtain intermediate D after purification. Mild oxidation of the alcohol moiety with reagents like Dess-Martin periodinane (DMP) is employed to obtain the aldehyde containing intermediate E. Reductive amination of intermediate E with a suitable amine is employed to obtain intermediate F after purification. The Pg of intermediate F is removed by acid treatment to obtain intermediate D after purification. Intermediate D can be ring cyclized using 1 ,T- carbonyldiimidazole (CDI) in an appropriate solvent to afford compounds of formula H as a free base. Acid treatment of compounds of formula H can be used to afford the salts of compounds of formula (I).
General synthesis of 18F-labelled compounds of the present invention:
Compounds having the formula (I) which are labelled by 18F can be prepared by reacting a precursor compound (I I l-F), as described below, with an 18F-fluorinating agent, so that the LG comprised in the precursor compound is replaced by 18F. The reagents, solvents and conditions which can be used for the 18F-fluorination are well-known to a skilled person in the field (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 P.A., Friebe M., Lehmann L, (eds), PET-Chemistry - The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, pp.15-50). Preferably, the solvents used in the 18F-fluorination are DMF, DMSO, acetonitrile, DMA, or mixtures thereof, preferably the solvent is acetonitrile or DMSO.
Any suitable 18F-fluorinating agent can be employed. Typical examples include H18F, alkali or alkaline earth 18F-fluorides (e.g., K18F, Rb18F, Cs18F, and Na18F). Optionally, the 18F-fluorination agent can be used in combination with a chelating agent such as a cryptand (e.g.: 4,7,13,16,21 ,24-hexaoxa-1 ,10- diazabicyclo[8.8.8]-hexacosane - Kryptofix®) or a crown ether (e.g.: 18-crown-6). Alternatively, the 18F-fluorinating agent can be a tetraalkylammonium salt of 18F or a tetraalkylphosphonium salt of 18F; e.g., tetra(Ci-6 alkyl)ammonium salt of 18F or a tetra(Ci-6 alkyl)phosphonium salt of 18F. Preferably, the 18F-fluorination agent is K18F, H18F, Cs18F, Na18F, tetra(Ci-e alkyl) ammonium salt of 18F, Kryptofix[222]18F or tetrabutylammonium [18F]fluoride.
Although the reaction is shown above with respect to 18F as a radioactive label, other radioactive labels can be introduced following similar procedures.
The invention is illustrated by the following examples which, however, should not be construed as limiting.
EXAMPLES
Compounds of the present disclosure may be prepared by methods known in the art of organic synthesis. In all of the methods it is understood that protecting groups for sensitive or reactive groups may be employed where necessary in accordance with general principles of chemistry. Protecting groups are manipulated according to standard methods of organic synthesis (T. W. Green and P. G. M. Wuts (2014) Protective Groups in Organic Synthesis, 5th edition, John Wiley & Sons). These groups are removed at a convenient stage of the compound synthesis using methods that are readily apparent to those skilled in the art.
Unless otherwise noted, all reagents and solvents were obtained from commercial sources and used without further purification. The chemical names were generated using ChemBioDraw Ultra v20 from CambridgeSoft.
Temperatures are given in degrees Celsius. If not mentioned otherwise, all evaporations are performed under reduced pressure, typically between about 15 mm Hg and 100 mm Hg (= 20 - 133 mbar). The structure of final products, intermediates and starting materials was confirmed by standard analytical methods, e.g., microanalysis and spectroscopic characteristics, e.g., MS, IR, NMR. Abbreviations:
Abbreviations used are those conventional in the art.
Figure imgf000067_0001
Figure imgf000068_0001
Analytical details, preparative and analytical methods
NMR measurements were performed on a DRX-400 MHz NMR spectrometer, on a Bruker AV-400 MHz NMR spectrometer in deuterated solvents, using or not using TMS as an internal standard. Chemical shifts (6) are reported in ppm downfield from TMS, spectra splitting patterns are designated as singlet (s), doublet (d), triplet (t), quartet (q), quintet (quint), septet (sept), multiplet, unresolved or overlapping signals (m), or broad signal (br). Deuterated solvents are given in parentheses and have chemical shifts of dimethyl sulfoxide (6 2.50 ppm), methanol (6 3.31 ppm), chloroform (6 7.26 ppm), or other solvent as indicated in NMR spectral data. MS were recorded on an Advion CMS mass spectrometer or an UPLC H-Class Plus with Photodiode Array detector and Qda Mass spectrometer from Waters.
Column 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 Column Chromatography System: flash purification was conducted with a Biotage Isolera One flash purification system using HP-Sil or KP-NH SNAP cartridges (Biotage) and the solvent gradient indicated in the specific examples.
TLC was carried out on silica gel plates with UV detection. Preparative examples
Figure imgf000069_0001
The compound was purchased from the commercial supplier Aurum Pharmatech LLC.
Preparative Example 2
Figure imgf000069_0002
Figure imgf000069_0003
,
Step A
Step B
Step-A: To a stirred solution of 2-fluoroethanol (1.82 mL, 31.2 mmol) and sodium hydride (60% dispersed in mineral oil) (1.25 g, 31.2 mmol) in THF (100 mL) under a nitrogen atmosphere was added 5-bromo-2-fluoropyridine (5.0 g, 28.4 mmol) in portions at 0 °C over a period of 15 min. After the addition was completed, the reaction mixture was stirred at room temperature for 3 h. The reaction mixture was quenched with water (100 mL) and extracted with EtOAc (3 x 100 mL). The combined organic phase was dried over NazSCh and concentrated under vacuum to afford the title compound as a yellow solid (5.0 g, 79%).
1H NMR (CDCI3) δ 8.17 (dd, 1 H), 7.66 (dd, 1H), 6.73 (dd, 1 H), 4.80 (m, 1 H), 6 4.68 (m, 1 H), 4.57 (m, 1 H), 4.50 (m, 1 H).
Step-B: In an oven-dried flask was added the title compound from Step A above (5.0 g, 22.7 mmol), bis(pinacalato) diborane (11.5 g, 45.4 mmol), KOAc (6.7 g, 68.9 mmol) and 1 ,4-dioxane (250 mL) under an argon atmosphere. The reaction mixture was degassed with argon for 15 min. Then [1 ,1'- bis(diphenylphosphino)ferrocene]-dichloropalladium(ll), complex with dichloromethane (1.8 g, 22.7 mmol) was added and the mixture was heated to 60 °C for 5 h. The solvent was removed under reduced pressure, the crude material was treated with 40% EtOAc in n-hexane (200 mL) and filtered through a celite pad. The filtrate was dried over NazSO^ concentrated and the residue was purified by silica gel chromatography (100-200 mesh) employing 10% EtOAc in n-hexane to afford the title compound as a pale-yellow solid (3.5 g, 57%). 1H NMR (CDCh) δ 8.51 (q, 1 H), 7.94 (dd, 1 H), 6.78 (dd, 1 H), 6 4.81 (m, 1 H), 4.69 (m, 1 H), 4.64 (m, 1 H), 4.57 (m, 1 H), 6 1.35 (d, 1 H), 1 .34 (s, 12H)
Preparative Example 3
Figure imgf000070_0001
The compound was purchased from the commercial supplier SV ChemBioTech, Inc.
Preparative Example 4
Figure imgf000070_0002
(2-Chloropyrimidin-5-yl)boronic acid (1.0g, 6.32 mmol), pyrrolidine (0.7 mL, 8.2 mmol), DIPEA (1.6 mL, 9.4 mmol), and NMP (10 mL, 10 vol) were heated at 160°C for 1 h under microwave irradiation. After completion, the reaction mixture was quenched with ice cool water (20 mL) and the crude reaction mass was filtered through a buchner funnel. The crude solid was washed with pentane (5 mL x 3), and dried under high vacuum to afford the desired intermediate as a yellow solid (80 mg, 70%).
1H NMR (DMSO-d6) δ 8.59 (s, 2H), 7.97 (s, 2H), 3.48 (t, 4H), 1.91 (m, 4H).
MS (ESI): 193.73 [M+H],
Preparative Example 5
Figure imgf000071_0001
Step-A: 2,5-Dibromopyrazine (5.0 g, 21 mmol), pyrrolidine (1.8 g, 25.2 mmol), CS2CO3 (6.8 g, 21 mmol), and DMSO (50 mL, 10 vol) were heated at 100“C for 16 h under microwave irradiation. After completion, the reaction mixture was quenched with ice cool water (60 mL) and the crude reaction mass was filtered through a buchner funnel. The solid was dried under high vacuum to afford the desired product as a yellow solid (4.3 g, 90%).
1H NMR (CDC13) δ 8.10 (d, 1 H), 7.61 (d, 1 H), 3.44 (t, 4H), 2.06-2.02 (m, 4H).
MS (ESI): 229.9 [M+H],
Step-B: To a stirred solution of step-A product (2.0 g, 8.77 mmol) in THF (80 mL, 40 vol.) was added dropwise nBuLi (2.5M in hexane) (4.2 mL, 10.5 mmol) at -78°C under N2 atmosphere and the mixture was stirred for 1 h. Then, 2-isopropoxy-4,4,5,5-tetramethyl-1 ,3,2-dioxaborolane (2.1 mL, 10.5 mmol) was added. The mixture was allowed to stir at -78°C for 2 h and at rt for 3 h. The reaction time was monitored by TLC. After completion, the reaction mixture was cooled to 0°C and quenched with saturated aqueous NH4CI solution. The aqueous layer was extracted with EtOAc (2 x 100 mL) and the combined organic layers were dried over NasSCU and concentrated under vacuum. The crude reaction product was directly used as such in the next step without further purification.
MS (ESI): 276.3 [M+H],
Preparative Example 6
Figure imgf000071_0002
Step-A: To a solution of 4-nitro-1 H-pyrazole (2.5 g, 21.9 mmol) in DMF (50 mL, 20 vol.) was added CS2CO3 (21.5 g, 65.9 mmol) and 3-fluoropropyl 4-methylbenzenesulfonate (10.2 g, 43.9 mmol). The mixture was stirred at rt for 16 h. The progress of the reaction was monitored by TLC. After completion, the solvent was evaporated under vacuum and water (100 mL) was added to the residue. The aqueous layer was extracted with EtOAc (2 x 100 mL) and the combined organic layers were dried over NazSO4 and concentrated under vacuum. The crude product was purified by column chromatography over silica gel (100-200 mesh) and eluted in 50% EtOAc in hexane to afford the desired product as a yellow oil (3.0 g, 78%).
1H NMR (CDC13) δ 8.18 (s, 1 H), 8.10 (s, 1 H), 4.52 (t, 1 H), 4.41 (t, 1 H), 4.33 (t, 2H), 2.37-2.25 (m, 2H). MS (ESI) 174.05 [M+H],
Step-B: To a solution of the Step A product (3.2 g, 18.48 mmol) in methanol (96 mL, 30 vol.) was added 10% Pd/C (640 mg, 20%w/w) portionwise under N2 atmosphere. Then, the vessel was filled up with H2 gas and the mixture was stirred at rt for 20 h. The progress of the reaction was monitored by TLC. After completion, the mixture was filtered through celite and the celite was washed with MeOH (3 x 100 mL). The combined filtrates were concentrated in vacuum to yield the desired product as a yellow oil (2.6 g, 93%).
1H NMR (CDC13) δ 7.04 (s, 1 H), 6.91 (s, 1 H), 4.43 (t, 1 H), 4.33 (t, 1 H), 4.01 (t, 2H), 3.89 (s, 2H), 2.06 (tt, 2H).
MS (ESI) 144.0 [M+H].
Preparative Example 7
Figure imgf000072_0001
Step-A: To a solution of 3-nitro-1 H-pyrazole (2.0 g, 17.6 mmol) in DMF (40mL, 20 vol.) was added CS2CO3 (17.3 g, 53.1 mmol) and 3-fluoropropyl 4-methylbenzenesulfonate (8.2 g, 35.4 mmol). The mixture was stirred at rt for 16 h. The progress of the reaction was monitored by TLC. After completion, the solvent was evaporated under vacuum and water (80 mL) was added to the residue. The aqueous layer was extracted with EtOAc (2 x 80 mL) and the combined organic layers were dried over Na2SO4 and concentrated under vacuum. The obtained crude mass was purified by column chromatography over silica gel (100-200 mesh) and eluted in 50% EtOAc in hexane to afford the desired product as a yellow oil (2.3 g, 75%).
1H NMR (CDCI3) 3 7.50 (d, 1 H), 6.91 (d, 1 H), 4.51 (t, 1 H), 4.40-4.36 (m, 3H), 2.45-2.27 (m, 2H).
MS (ESI) 174.11 [M+H],
Step-B: To a solution of the Step A product (2.3 g, 13.2 mmol) in methanol (70 mL, 30 vol.) was added 10% Pd/C (230 mg, 10%w/w) portionwise under N2 atmosphere. The reaction vessel was filled up with H2 gas and the mixture was stirred at rt for 21 h. The progress of the reaction was monitored by TLC. After completion, the mixture was filtered through celite and the celite was washed with MeOH (3 x 100 mL). The combined filtrates were concentrated in vacuum to yield the desired product as a yellow oil (1.7 g, 86%). The compound was pure enough and used as such in the next step without further purification.
1H NMR (DMSO-D6) δ 7.32 (d, 1 H), 5.40-5.39 (m, 1 H), 4.87 (s, 2H), 4.45 (t, 1 H), 4.33 (t, 1H), 3.93 (t, 2H), 2.11-2.08 (m, 1 H), 2.06 -1.98 (m, 1 H).
MS (ESI) 144.01 [M+H],
Examples according to the invention
Example 1 (R)-2-(6-(3-fluoropyrrolidin-1-yl)-2-methylpyridin-3-yl)-6-(pyridin-3-yl)-6,7-dihvdro-5/-/- imidazofl ,5-£>l[1 ,2,4]triazol-5-one hydrochloride
Figure imgf000073_0001
Step A: To a stirred solution of (3-bromo-1-(tetrahydro-2/-/-pyran-2-yl)-1H-1 ,2,4-triazole-5- carbaldehyde (3.0 g, 9.6 mmol) and pyridin-3-amine (1.17 g, 12.5 mmol) in 1 ,2 dichloroethane (120 mLI) was added molecular sieves 4 A and glacial AcOH (7.5 mL) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred for 4 h and allowed to reach room temperature. Then sodium triacetoxyborohydride (4.0 g, 19.2 mmol) was added and the mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with saturated NaHCO3 (100 mL) and the product was extracted with 5% MeOH in DCM (3 x 250 mL). The combined organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by column chromatography using silica gel (100-200 mesh) employing in 2% MeOH in DCM to afford the title compound as a yellow liquid (3.0 g, 76%).
1H NMR (400 MHz, DMSO- d6) δ 8.03 (d, 1 H), 7.81 (dd, 1 H), 7.09 (dd, 1H), 6.99 (dq, 1 H), 6.49 (t, 1H), 5.70 (dd, 1H), 4.53 (m, 2H), 3.88 (m, 1 H), 3.69 (m, 1 H), 2.09 (m, 1H), 1.93 (m, 2H), 1.60 (m, 3H) MS: 338.24 [M+H]+
Step B: In an oven-dried screw capped vial was added the title compound from Step A above (0.6 g,
1.78 mmol), (6-fluoro-2-methylpyridin-3-yl)boronic acid (0.55 g, 3.5 mmol), K2CO3 (0.49 g, 3.5 mmol), and a mixture of 1 ,4-dioxane and water (30 mL, 4/1) under an argon atmosphere. The reaction mixture was degassed with argon for 15 min. Then [1 ,1 '-bis(diphenylphosphino)ferrocene]- dichloropalladium(ll), complex with dichloromethane (0.145 g, 0.178 mmol) was added and the mixture was heated to 90 °C for 4 h. The reaction mixture was quenched with ice-water and extracted in 10% MeOH in DCM (3 x 30 mL). The organic layer was dried over Na2SO4, concentrated and purified by silica gel chromatography (100-200 mesh) using 4% MeOH in DCM to obtain the title compound as a brownish liquid (0.45 g, 68%).
1H NMR (400 MHz, DMSO-d6) δ 8.37 (t, 1 H), 8.09 (d, 1 H), 7.81 (q, 1 H), 7.09 (m, 1 H), 6.51 (t, 1 H),
5.78 (m, 1 H), 4.60 (ddd, 2H), 3.91 (d, 1 H), 3.73 (m, 1 H), 2.70 (s, 3H), 2.24 (m, 1 H), 2.01 (m, 3H), 1.69 (t, 1 H), 1.57 (q, 2H), 1.34 (s, 1 H), 1.23 (d, 4H), 1.17 (q, 1 H)
MS: 369.15 [M+H]+
Step-C: The title compound from Step B above (0.1 g, 0.27 mmol), (R)-3-fiuoropyrrolidine hydrogen chloride (0.051 g, 0.40 mmol), DIPEA (0.14 mL, 0.8 mmol), and NMP (5 mL) were combined in an oven-dried microwave vial under argon atmosphere. The reaction mixture was heated at 160 °C in a microwave oven for 4 h. The reaction mixture was quenched with ice cold water (10 mL) and extracted with 5% MeOH in DCM (3 x 10 mL). The combined organic layers were washed with cold brine solution (2 x 10 mL), dried over Na2SO4, and concentrated under vacuum. The crude material was purified by column chromatography using silica gel (100-200 mesh) employing 3% MeOH in DCM to afford the title compound as a white solid (0.050 g, 42%). 1H NMR (400 MHz, DMSO-d6) δ 8.09 (d, 1H), 7.99 (d, 1 H), 7.80 (q, 1 H), 7.08 (dd, 2H), 6.44 (m, 2H), 5.71 (dd, 1 H), 5.44 (m, 1 H), 4.54 (ddd, 2H), 3.91 (d, 1 H), 3.67 (m, 4H), 3.43 (td, 1 H), 2.63 (s, 3H), 2.20 (m, 3H), 1 .98 (m, 2H), 1 .63 (m, 3H)
Step-D: To a stirred solution of the title compound from Step C above (0.050 g, 0.11 mmol) in DCM (2.5 mL) was added a solution of 4 M HCI in 1 ,4-dioxane (0.5 mL) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred for 6 h and allowed to reach room temperature. The solvent was removed under vacuum to afford the title compound as a hydrochloride salt. The yellow solid was washed with hexane (3 x 5 mL) and dried under vacuum.
MS: 354.25 [M+H]+
Step-E: To an ice cool solution of the title compound from Step D above (0.04 mg, 0.1 mmol) in 1 ,2- DCE (2 mL) was added sodium hydride (60% dispersed in mineral oil) (0.004 g, 0.17 mmol) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 30 min. Then 1 ,1'- carbonyldiimidazole (0.16 g, 1 .0 mmol) was added to the reaction mixture and the mixture was stirred at room temperature for 6 h. The reaction mixture was quenched with ice cold water and the product was extracted with 5% MeOH in DCM. The organic phase was dried over Na2SO4 and concentrated under vacuum. The residue was purified by silica gel chromatography (100-200 mesh) employing 4% MeOH in DCM to yield the title compound as a white solid (0.02 mg, 51%).
1H NMR (400 MHz, DMSO-d6) δ 8.98 (d, 1 H), 8.46 (m, 1 H), 8.19 (d, 1 H), 8.16 (d, 1 H), 7.54 (d, 1 H), 6.51 (d, 1 H), 5.46 (d, 1 H), 5.26 (s, 2H), 3.72 (m, 3H), 3.48 (m, 1 H), 2.73 (s, 3H), 2.22 (m, 2H) MS: 379.9 [M+H]+
Step-F: To a stirred solution of the title compound from Step E above (0.020 g, 0.05 mmol) in DCM (2 mL) was added a solution of 4 M HCI in 1 ,4-dioxane (0.2 mL) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred for 6 h and allowed to warm to room temperature. The solvent was evaporated, the residue was treated with pentane, and dried under vacuum to afford the title compound as a yellow solid (0-015 g, 71%).
1H NMR (400 MHz, DMSO-d6) δ 9.09 (d, 1 H), 8.57 (m, 1 H), 8.40 (d, 1 H), 7.74 (q, 1 H), 6.94 (s, 1 H), 5.55 (d, 1 H), 5.34 (s, 2H), 3.77 (m, 4H), 3.57 (s, 1 H), 2.91 (s, 3H), 2.30 (m, 2H) MS: 379.9 [M+H]+
Examples 2 to 5
Following the same procedure as described for Example 1 , except using the reagents indicated for the specific steps in Table 1 below, the following Examples were prepared. Table 1:
Figure imgf000076_0001
Figure imgf000077_0001
Figure imgf000078_0001
Figure imgf000079_0001
Figure imgf000080_0001
Figure imgf000081_0002
Example 7 (S)-2-(6-(3-fluoropyrrolidin-1-yl)pyridin-3-yl)-6-(1-methyl-1 H-pyrazol-4-yl)-6,7-dihydro-
5/7-imidazo[1 ,5-b1[1 ,2,41triazol-5-one hydrochloride
Figure imgf000081_0001
Step A: To an ice cool solution of ethyl 5-bromo-1/-/-1 ,2,4-triazole-3-carboxylate (2.5 g, 11.3 mmol) in THF (50 mL) was added sodium hydroxide (60% dispersed in mineral oil) (0.59 g, 14.7 mmol) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 30 min. The reaction mixture was cooled to 0 °C, 2-(trimethylsilyl)ethoxymethyl chloride (2.4 mL, 13.6 mmol) was dropwise added, and the reaction mixture was stirred for 16 h and allowed to reach room temperature. The reaction mixture was quenched with ice-water and the reaction mixture was extracted with EtOAc (3 x 75 mL). The combined organic phase was washed with brine (2 x 25 mL), dried over NazSCM, and concentrated under vacuum. The residue was purified by silica gel chromatography (100-200 mesh) employing 10% EtOAc in n-hexane to yield the title compound as a yellow oil (0.96 g, 24%). 1H NMR (400 MHz, CDCI3) δ 5.86 (s, 2H), 4.48 (q, 2H), 3.67 (m, 2H), 1.45 (d, 3H), 0.92 (m, 2H), -17.45 (s, 9H)
MS: 352.79 [M+2H]+
Step B: In an oven-dried screw capped vial was added the title compound from Step A above (0.2 g, 0.57 mmol), (6-fluoropyridin-3-yl)boronic acid (0.096 g, 0.68 mmol), CS2CO3 (0.371 g, 1.14 mmol), and 1 ,4-dioxane (8 mL) under an argon atmosphere. The reaction mixture was degassed with argon for 15 min. Then [1 ,T-bis(diphenylphosphino)ferrocene]dichloropalladium(ll), complex with dichloromethane (0.048 g, 0.06 mmol) and 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (0.023 g, 0.06 mmol) were added and the mixture was heated to 80 °C for 3 h. The reaction was quenched with ice-water and the reaction mixture was extracted with DCM (3 x 30 mL). The combined organic phase was washed with brine (30 mL), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (100-200 mesh) employing 20% EtOAc in n-hexane to obtain the title compound as a yellow liquid (0.170 g, 81%).
1H NMR (400 MHz, CDCI3) δ 9.01 (d, 1 H), 8.54 (td, 1 H), 7.02 (dd, 1H), 5 5.95 (s, 2H), 4.54 (q, 2H), 3.71 (t, 2H), 1.48 (t, 3H), 0.95 (t, 2H), -22.7 (s, 9H)
MS: 367.25 [M+H]+
Step C: The title compound from Step B above (3.0 g, 8.2 mmol), (S)-3-fluoropyrrolidine hydrogen chloride (2.0 g, 16.3 mmol), DIPEA (7.13 mL, 41 mmol) and ethanol (30 mL) were combined in an oven-dried microwave vial under argon atmosphere. The reaction mixture was heated at 130 °C in a microwave oven for 3 h. The solvent was removed under vacuum and the obtained crude material was purified by column chromatography using silica gel (100-200 mesh) employing 50% EtOAc in n-hexane to afford the title compound as a white solid (1.8 g, 50%).
1H NMR (400 MHz, CDC13) δ 8.94 (d, 1 H), 8.20 (dd, 1 H), 6.44 (d, 1 H), 5.92 (s, 2H), 5.40 (dt, 1H), 4.52 (q, 2H), 3.91 (q, 1 H), 3.69 (m, 4H), 3.64 (m, 2H), 2.41 (m, 1H), 2.17 (m, 1H), 1.48 (t, 3H), -23.4 (s, 9H)
MS: 436.58 [M+H]+ Step D: To an ice cool solution of the title compound from Step C above (1.8 g, 4.13 mmol) in MeOH (90 mL) was added sodium borohydride (1.25 g, 33 mmol) in portions at 0 °C under a nitrogen atmosphere. After the addition was completed, the reaction mixture was stirred at room temperature for 3 h. The reaction mixture was cooled to 0 °C and the reaction was quenched by the addition of sat. ammonium chloride (effervescence!). The solvent was removed under vacuum and the crude material was diluted with water and extracted with DCM (3 x 100 mL). The combined organic layer was washed with cold brine solution (100 mL), dried over NajSCU, and concentrated under vacuum to afford the title compound as a white solid (1 .6 g, 98%).
1H NMR (400 MHz, CDC13) δ 8.90 (d, 1 H), 8.11 (dd, 1 H), 6.44 (d, 1 H), 5.55 (s, 2H), 5.39 (dt, 1H), 4.88 (s, 2H), 3.90 (q, 1 H), 3.80 (s, 1 H), 3.68 (m, 6H), 2.41 (m, 1 H), 2.17 (m, 1 H), 0.93 (t, 2H), 0.85 (m, 1 H), -16.2 (s, 9H) MS: 394.62 [M+H]+
Step E: To an ice cool solution of the title compound from Step D above (1.6 g, 4.0 mmol) in DCM (48 mL) was added Dess-Martin periodinane (3.45 g, 8.14 mmol) in portions at 0 °C under a nitrogen atmosphere. After the addition was completed, the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was cooled to 0 °C and the reaction was quenched by the addition of sat. NaHCO3 (50 mL). The reaction mixture was extracted with DCM (3 x 80 mL). The combined organic layer was washed with cold brine solution (80 mL), dried over Na2SO4, and concentrated under vacuum. The crude material was purified by column chromatography using silica gel (100-200 mesh) employing in 50% EtOAc in n-hexane to afford the title compound as a white solid (1.3 g, 82%).
1H NMR (400 MHz, CDCI3) δ 10.06 (s, 1 H), 8.95 (d,1 H), 8.18 (dd, 1 H), 6.47 (d, 1 H), 5.86 (s, 2H), 5.41 (dt, 1 H), 3.93 (q, 1 H), 3.71 (m, 6H), 2.43 (m, 1 H), 2.18 (m, 1H), 0.94 (t, 2H), -22.03 (s, 9H) MS: 392.26 [M+H]+
Step F: To a stirred solution of the title compound from Step E above (0.4 g, 1 .0 mmol) and 1-methyl- 1H-pyrazol-4-amine (0.1 g, 1.0 mmol) in 1 ,2-dichloroethane (16 mL) was added molecular sieves 4A and glacial AcOH (0.8 mL) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 h. Then sodium triacetoxyborohydride (0.43 g, 2.0 mmol) was added and the mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with aqueous sat. NaHCO3 (20 mL) and the reaction mixture was extracted with 5% MeOH in DCM (3 x 50 mL). The combined organic layer was dried over Na2SO4 and concentrated under vacuum. The crude material was purified by column chromatography using silica gel (100-200 mesh) employing 3% MeOH in DCM to afford the title compound as a yellow liquid (0.4 g, 83%).
MS: 473.59 [M+H]+ Step G: To a stirred solution of the title compound from Step F above (0.4mg, 0.85 mmol) in DCM (8.0 mL) was added a solution of 4 M HCI in 1 ,4-dioxane (2.0 mL) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 h, the solvents were evaporated under reduced pressure, the residue was washed with pentane, and dried under vacuum to afford the title compound as an off-white solid (0.28 g, 97%).
MS: 341.13 [M+H]+
Step H: To an ice cool solution of the title compound from Step G above (0.28 g, 0.82 mmol) in 1 ,2- dichloroethane (17 mL) was added sodium hydride (60% dispersed in mineral oil) (0.033 g, 0.82 mmol) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature and kept for 30 min. Then 1 ,1 '-carbonyldiimidazole (1.33 g, 8.2 mmol) was added to the reaction mixture and the mixture was stirred at room temperature for 24 h. The reaction mixture was quenched with ice cold water and the reaction mixture was extracted with 5% MeOH in DCM (3 x 40 mL). The combined organic phase was dried over Na2SO 4 and concentrated under vacuum. The residue was purified by preparative TLC employing 5% MeOH in DCM as a mobile phase. The solvent was removed under vacuum to afford the title compound as a yellow white solid (0.009 g, 3%).
1H NMR (400 MHz, DMSO- d6) δ 8.10 (dd, 1 H), 8.01 (s, 1H), 7.64 (s, 1 H), 6.61 (d, 1H), 5.44 (d, 1 H), 4.99 (s, 2H), 3.83 (m, 3H), 3.68 (m, 3H), 3.45 (m, 1 H), 2.19 (m, 2H)
MS: 368.95 [M+H]+
Step I: To a stirred solution of the title compound from Step H above (0.006.g, 0.024 mmol) in DCM (1.0 mL) was added a solution of 4M HCI in 1,4-dioxane (0.6 mL,) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 h. The solvents were removed under vacuum to afford the hydrochloride salt. The off-white solid was washed with n-hexane (2 mL) and then dried under vacuum to afford the title compound as a white solid (0.004 g, 40%).
1H NMR (400 MHz, DMSO- d6) δ 9.08 (d, 1 H), 8.70 (d, 1 H), 8.56 (d, 1 H), 8.38 (dd, 2H), 7.72 (q, 1 H), 7.00 (d, 1 H), 5.54 (d, 1 H), 5.32 (s, 2H), 3.85 (m, 3H), 3.61 (td, 1H), 2.26 (m, 2H)
MS: 368.90 [M+H]+
Examples 8 to 10
Following the same procedure as described for Example 7, except using the reagents indicated for the specific steps in Table 2 below, the following Examples were prepared. Table 2:
Figure imgf000085_0001
Example 11 (2-(6-(2-fluoroethoxy)pyridin-3-yl)-6-(1-methyl-1/7-pyrazol-4-yl)-6,7-dihvdro-5/-/- imidazo[1 ,5-b][1 ,2,4]triazol-5-one
Figure imgf000086_0001
In an oven-dried screw capped vial was added 3-bromo-1-(tetrahydro-2/7-pyran-2-yl)-1/7-
1 ,2,4-triazole-5-carbaldehyde (1.5 g, 5.7 mmol), the title compound from Preparative Example 2 (3.1 g, 11.5 mmol), K2CO3 (1.6 g, 11.5 mmol) and a mixture of 1 ,4-dioxane and water (5 mL, 4/1 ) under an argon atmosphere. The reaction mixture was degassed with argon for 15 min. Then [1 ,1 - bis(diphenylphosphino)ferrocene]dichloropalladium(ll), complex with dichloromethane (0.47 g, 0.57 mmol) was added and the mixture was heated to 60 °C for 5 h. The reaction mixture was quenched with ice-water and extracted with EtOAc (3 x 60 mL). The combined organic phase was dried over NazSCX concentrated and the residue was purified by silica gel chromatography (100-
200 mesh) employing 25% EtOAc in n-hexane to afford the title compound as a brownish solid (1.0 g, 55%).
1H NMR (400 MHz, CDCI3) δ 10.7 (s, 1 H), 8.92 (d, 1H), 8.33 (dd, 1H), 6.89 (d, 1 H), 6.25 (dd, 1 H), 4.83 (t, 1 H), 4.74 (t, 1 H), 4.68 (t, 1 H), 4.62 (t, 1 H), 4.12 (m, 1 H), 3.78 (td, 1 H), 2.42 (m, 1 H), 2.16 (m, 1 H), 2.02 (m, 1 H), 1.77 (m, 2H), 1.66 (m, 1 H)
MS: 321.35 [M+H]+
Step B: To a stirred solution of the title compound from Step A above (0.4 g, 1.3 mmol) and 1 -methyl- 1H-pyrazol-4-amine (0.18 g, 1.9 mmol) in 1 ,2-dichloroethane (16 mL) was added molecular sieves 4 A and glacial AcOH (1.2 mL) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 4 h. Then sodium triacetoxyborohydride (0.55 g, 2.6 mmol) was added and the mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with aqueous saturated NaHCO3 (60 mL) and the product was extracted with 5% MeOH in DCM (3 x 60 mL). The combined organic phase was dried over Na2SO 4 and concentrated under reduced pressure. The crude material was purified by column chromatography using silica gel (100-200 mesh) employing 4% MeOH in DCM to afford the title compound as a brown liquid (0.34 g, 65%). 1H NMR (400 MHz, DMSO-c/6) δ 8.73 (d, 1 H), 8.23 (dd, 1 H), 7.16 (s, 1 H), 7.03 (s, 1 H), 6.97 (d, 1 H), 5.72 (dd, 1H), 5.02 (s, 1 H), 4.81 (t, 1 H), 4.72 (t, 1 H), 4.56 (dt, 2H), 4.28 (dd, 2H), 3.94 (d, 1 H), 3.70 (m, 1H), 3.68 (s, 3H), 2.22 (ddd, 1 H), 2.01 (m, 1H), 1.89 (td, 1H), 1.65 (m, 1H), 1.58 (m, 2H) MS: 402.43 [M+H]+
Step C: To a stirred solution of the title compound from Step B above (0.26 g, 0.65 mmol) in DCM (13 mL) was added a solution of 4 M HCI in 1 ,4-dioxane (2.6 mL) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 5 h. The solvent was removed under reduced pressure to afford the hydrochloride salt. The yellow solid was washed with n-hexane (3 x 5 mL) and dried under vacuum to afford the title compound (0.2 g, 87%).
1H NMR (400 MHz, DMSO-b6) δ 8.81 (d, 1 H), 8.29 (dd, 1 H), 7.87 (s, 1 H), 7.54(s, 1 H), 7.07 (d, 1H), 4.82 (t, 1H), 4.73 (dd, 1 H), 4.61 (t, 1 H), 4.54 (s, 4H), 3.82 (s, 3H) MS: 318.35 [M+H]+
Step D: To an ice cool solution of the title compound from Step C above (0.2 mg, 0.63 mmol) in 1 ,2- dichloroethane (12 mL) was added sodium hydride (60% dispersed in mineral oil) (0.04 g, 0.95 mmol) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 30 min. Then 1 ,T-carbonyldiimidazole (1.02 g, 6.3 mmol) was added and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was quenched with ice cold water and the mixture was extracted with 5% MeOH in DCM (3 x 20 mL). The combined organic phase was dried over Na2SC>4 and concentrated under reduced pressure. The residue was purified by silica gel chromatography (100-200 mesh) employing 3% MeOH in DCM to yield the title compound as a white solid (0.09 g, 42%).
1H NMR (400 MHz, DMSO- d6) δ 8.88 (d, 1 H), 8.36 (dd, 1 H), 8.07 (s, 1 H), 7.69 (s, 1 H), 7.05 (d, 1 H), 5.07 (s, 2H), 4.79 (m, 2H), 4.60 (m, 2H), 3.88 (s, 3H) MS: 344.15 [M+Hf
Step E: To a stirred solution of the title compound from Step D above (0.08 g, 0.23 mmol) in DCM (4 mL) was added a solution of 4 M HCI in 1 ,4-dioxane (0.8 mL) at 0 °C under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 5 h and the solvents were evaporated under reduced pressure. The residue was treated with pentane and dried under vacuum to afford the title compound as a white solid (0.08 g, 92%).
1H NMR (400 MHz, DMSO-d6) δ 8.88 (d, 1 H), 8.36 (dd, 1 H), 8.07 (s, 1 H), 7.69 (s, 1 H), 7.05 (d, 1 H), 5.07 (s, 2H), 4.83 (t, 1 H), 4.74 (t, 1 H), 4.63 (t, 1 H), 4.57 (t, 1 H), 3.88 (s, 3H) MS: 344.20 [M+H]+ Example 12
Following the same procedure as described for Example 11, except using the reagents indicated for the specific steps in Table 3 below, the following Example was prepared.
Table 3:
Figure imgf000088_0001
BIOLOGICAL ASSAY DESCRIPTION AND CORRESPONDING RESULTS
1 . Preparation of human Parkinson’s disease (PD) brain-derived alpha-synuclein (a-syn) aggregates
The procedure was adapted from the protocol described in Spillantini et al., 1998. Frozen tissue blocks from PD donors were thawed on ice and homogenized using a glass dounce homogenizer. The homogenate was then centrifuged at 11 ,000 x g (12,700 RPM) in an ultracentrifuge (Beckman, XL100K) for 20 minutes at 4°C using a pre-cooled 70.1 rotor (Beckman, 342184). Pellets were resuspended in extraction buffer [10 mM Tris-HCI pH 7.4, 10% sucrose, 0.85 M NaCI, 1 % protease inhibitor (Calbiochem 539131 ), 1 mM EGTA, 1% phosphatase inhibitor (Sigma P5726 and P0044)] and centrifuged at 15,000 x g (14,800 RPM, a 70.1 Ti rotor) for 20 minutes at 4°C. Pellets were discarded and sarkosyl (20% stock solution, Sigma L7414) was added to the supernatants to a final concentration of 1% and stirred at room temperature for one hour. This solution was then centrifuged at 100,000 x g (38,000 RPM, 70.1 Ti rotor) for one hour at 4°C. Pellets containing enriched alpha- synuclein aggregates were resuspended in PBS and stored at -80°C until use. 2. Micro-radiobindinq competition assay for the determination of binding affinity in PD brain-derived alpha-syn aggregates
PD brain-derived alpha-synuclein aggregates were spotted onto microarray slides. The slides were incubated with [3H]-alpha-synuclein reference at 25nM, 30nM or 40nM and the example compounds (non-radiolabelled) at 1 pM and 100nM. In some cases, the non-radiolabelled example compounds were further assessed for a range of different concentrations, varying from 0.05nM to 2pM. After incubation, slides were washed, and scanned by a real-time autoradiography system (BeaQuant, ai4R). Quantification of the signal was performed by using the image analysis software Beamage (ai4R). Non-specific signal was determined with an excess of non-radiolabelled reference alpha- synuclein ligand (2 pM) and specific binding was calculated by subtracting the non-specific signal from the total signal. Competition was calculated as percent, where 0% was defined as the specific binding in the presence of vehicle and 100% as the values obtained in the presence of excess of the non-radiolabelled reference alpha-synuclein ligand. K, values were calculated in GraphPad Prism7 by applying a nonlinear regression curve fit using a one site, specific binding model. All measurements were performed with at least two technical replicates. For compounds tested in more than one experiment, the mean of the replicates or Ki values in independent experiments is reported.
Results: Example compounds were assessed for their potency to compete with the binding of [3H]- reference alpha-synuclein ligand to PD patient brain-derived alpha-synuclein aggregates. Results of the micro-radiobinding competition assay for the example compounds tested are shown in Table 4 as: % competition at 1 pM and 100 nM. The Table 4 also shows Ki values. Ki measurements for example compounds 1-11 were performed on the same PD brain-derived alpha-synuclein aggregates while Ki measurements for example compounds 13-17, and 25 were performed on PD brain-derived alpha-synuclein aggregates from a different donor. The data shows that the compounds of the invention bind to human brain-derived alpha-synuclein aggregates. Example compounds 4, 5, 11 and 17 exhibited binding with very high affinity (Ki<100nM) to human PD patient brain-derived alpha-synuclein aggregates.
Table 4
Figure imgf000090_0001
Table 4: Assessment of binding affinity by micro-radiobinding competition assay on human PD brain- derived alpha-synuclein aggregates. Percent (%) competition over the tritiated [3H]-reference alpha- synuclein ligand in the presence of 1 pM and 100 nM of example compounds 1 -29 and Ki values are also shown for selected example compounds in PD brain homogenates, (n.d. = not determined;* = two independent experiments were performed)

Claims

A compound of formula (I):
Figure imgf000091_0001
or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-Cg-Cecycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and
R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl.
2. The compound according to claim 1 , having a formula (la):
Figure imgf000091_0002
or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
R1 and R2 are defined in claim 1 ,
R3 is halo or C1-C4alkyl; and q is 0, 1 or 2.
3. The compound according to claim 1 or 2, wherein R1 is a 4- to 6-membered heterocyclyl selected from the following:
Figure imgf000092_0003
wherein R1a is F or H; or R1 is haloCi-C4alkoxy-.
4. The compound according to claim 3, wherein R1 is a 5-membered heterocyclyl which is:
Figure imgf000092_0004
, preferably F is 19F or 18F, more preferably 18F; or R1 is -OCH2-CH2-F.
5. The compound according to claim 1 or 2, wherein R2 a 5-membered or 6-membered heteroaryl selected from the following:
Figure imgf000092_0005
Figure imgf000092_0006
wherein
R2a is selected from haloCi-C4alkyl, haloCi-C4alkoxy, Ci-C4alkoxy, and Ci-C4alkyl;
R2b is selected from H, haloCi-C4alkyl, haloCi-C4alkoxy, Ci-C4alkoxy, and Ci-C4alkyl; and s is 0, 1 or 2.
6. The compound according to claim 5, wherein R2 a 5-membered or 6-membered heteroaryl selected from the following:
Figure imgf000092_0001
wherein
R2b is selected from haloCi-C4alkyl, haloCi-C4alkoxy, alkoxy, H, or Ci-C4alkyl.
7. The compound according to claim 1 , wherein the compound is selected from:
Figure imgf000092_0002
Figure imgf000093_0001
Figure imgf000094_0003
Figure imgf000094_0002
or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof.
Figure imgf000094_0001
Figure imgf000095_0001
Figure imgf000095_0002
or a detectably labelled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof.
9. The compound according to any one of the preceding claims, wherein the compound is a detectably labelled compound.
10. The compound according to claim 9, wherein the detectably labelled compound comprises a radioisotope selected from 18F, ZH and 3H. r L~N J
11. The compound according to claim 9 or 10, wherein R1 is 5
12. A diagnostic composition comprising a compound according to any one of claims 9 to 11 , and optionally at least one pharmaceutically acceptable excipient, carrier, diluent and/or adjuvant.
13. The compound according to any one of claims 9 to 11 , or the diagnostic composition according to claim 12, for use in the imaging of alpha-synuclein aggregates.
14. The compound according to any one of claims 9 to 11 , or the diagnostic composition according to claim 12, for use in positron emission tomography imaging of alpha-synuclein aggregates.
15. The compound for use or the diagnostic composition for use according to claim 13 or 14, wherein the use is for in vitro imaging, ex vivo imaging, or in vivo imaging, preferably the use is for in vivo imaging, more preferably the use is for brain imaging.
16. The compound according to any one of claims 9 to 11 , or the diagnostic composition according to claim 12, for use in diagnostics.
17. The compound for use or the diagnostic composition for use according to claim 16, wherein the diagnosis is the diagnosis of a disease, disorder or abnormality associated with alpha-synuclein aggregates or a predisposition therefor, wherein the disease, disorder or abnormality is optionally selected from Parkinson's disease (including sporadic, familial with alpha-synuclein mutations, familial with mutations other than alpha-synuclein, pure autonomic failure or Lewy body dysphagia), SNCA duplication carrier, Lewy Body dementia (LBD), dementia with Lewy bodies (DLB) (including “pure” Lewy body dementia), Parkinson’s disease dementia (PDD), diffuse Lewy body disease (DLBD), Alzheimer’s disease, sporadic Alzheimer’s disease, familial Alzheimer's disease with APP mutations, familial Alzheimer's disease with PS-1 , PS-2 or other mutations, familial British dementia, Lewy body variant of Alzheimer’s disease, Down syndrome, multiple system atrophy (MSA) (including Shy-Drager syndrome, striatonigral degeneration or olivopontocerebellar atrophy), traumatic brain injury, chronic traumatic encephalopathy, dementia puglistica, tauopathies (including Pick's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration, Niemann-Pick type C1 disease, frontotemporal dementia with Parkinsonism linked to chromosome 17), Creutzfeldt- Jakob disease, Huntington's disease, motor neuron disease, amyotrophic lateral sclerosis (including sporadic, familial or ALS-dementia complex of Guam), neuroaxonal dystrophy, neurodegeneration with brain iron accumulation type 1 (including Hallervorden-Spatz syndrome), prion diseases, ataxia telangiectatica, Meige’s syndrome, subacute sclerosing panencephalitis, Gerstmann-Straussler-Scheinker disease, inclusion-body myositis, Gaucher disease, Krabbe disease as well as other lysosomal storage disorders (including Kufor-Rakeb syndrome and Sanfilippo syndrome) and rapid eye movement (REM) sleep behavior disorder.
18. The compound for use or the diagnostic composition for use according to claim 17, wherein the disease is Parkinson's disease.
19. The compound for use or the diagnostic composition for use according to claim 17, wherein the disease is multiple system atrophy.
20. The compound for use or the diagnostic composition for use according to claim 17, wherein the disease is dementia with Lewy bodies.
21 . The compound for use or the diagnostic composition for use according to claim 17, wherein the disease is Parkinson’s disease dementia.
22. The compound for use or the diagnostic composition for use according to claim 16, wherein the disease is SNCA duplication carrier.
23. The compound for use or the diagnostic composition for use according to claim 17, wherein the disease is Alzheimer’s disease.
24. The compound for use or the diagnostic composition for use according to any one of claims 13 to 23, wherein the use is in a human.
25. A method of diagnosing a disease, disorder or abnormality associated with alpha-synuclein aggregates, in a subject, the method comprising the steps:
(a) Administering a compound according to any one of claims 1 to 11 , or a diagnostic composition according to claim 12 which comprises a compound according to any one of claims 1 to 11 , to the subject;
(b) Allowing the compound to bind to the alpha-synuclein aggregates; and
(c) Detecting the compound bound to the alpha-synuclein aggregates.
26. The method of diagnosing according to claim 24, the method further comprising the step of:
(d) Generating an image representative of the location and/or amount of the compound bound to the alpha-synuclein aggregates.
27. A method of positron emission tomography (PET) imaging of alpha-synuclein aggregates in a tissue of a subject, the method comprising the steps:
(a) Administering a compound according to any one of claims 1 to 11 , or a diagnostic composition according to claim 11 which comprises a compound according to any one of claims 1 to 11 , to the subject;
(b) Allowing the compound to bind to the alpha-synuclein aggregates; and
(c) Detecting the compound bound to the alpha-synuclein aggregates by collecting a positron emission tomography (PET) image of the tissue of the subject. The method of positron emission tomography imaging according to claim 27, wherein the tissue is a tissue of the central nervous system (CNS), an eye tissue, tissue of a peripheral organ, or a brain tissue, preferably wherein the tissue is brain tissue. A method for the detection and optionally quantification of alpha-synuclein aggregates in a tissue of a subject, the method comprising the steps:
(a) Bringing a sample or a specific body part or body area suspected to contain an alpha- synuclein aggregates into contact with a compound according to any one of claims 1 to 11 , or a diagnostic composition according to claim 12 which comprises a compound according to any one of claims 1 to 11 ;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates using positron emission tomography; and
(d) Optionally quantifying the amount of the compound bound to the alpha-synuclein aggregates. A method of collecting data for the diagnosis of or for determining a predisposition to a disease, disorder or abnormality associated with alpha-synuclein aggregates, the method comprising the steps:
(a) Bringing a sample or a specific body part or body area suspected to contain alpha- synuclein aggregates into contact with a compound according to any one of claims 1 to 11 , or a diagnostic composition according to claim 12 which comprises a compound according to any one of claims 1 to 11 ;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates; and
(d) Optionally correlating the presence or absence of the compound bound to the alpha- synuclein aggregates with the presence or absence of the alpha-synuclein in the sample or specific body part or body area. A method of collecting data for prognosing a disease, disorder or abnormality associated with alpha-synuclein aggregates, for monitoring the progression of a disease, disorder or abnormality associated with alpha- synuclein aggregates in a patient; or for predicting responsiveness of a patient suffering from a disease, disorder or abnormality associated with alpha-synuclein aggregates to a treatment of the disease, disorder or abnormality associated with alpha-synuclein aggregates, wherein the method comprises the steps:
(a) Bringing a sample, a specific body part or body area suspected to contain alpha-synuclein aggregates into contact with a compound according to any one of claims 1 to 11 , or a diagnostic composition according to claim 12 which comprises a compound according to any one of claims 1 to 11 ;
(b) Allowing the compound to bind to the alpha-synuclein aggregates;
(c) Detecting the compound bound to the alpha-synuclein aggregates
(d) Optionally correlating the presence or absence of the compound bound to the alpha- synuclein aggregates with the presence or absence of the alpha-synuclein aggregates in the sample or specific body part or body area; and
(e) Optionally repeating steps (a) to (c) and, if present, optional step (d) at least one time.
32. The method of claims 30 or 31 , wherein the step of optionally correlating the presence or absence of the compound bound to the alpha-synuclein aggregates with the presence or absence of the alpha-synuclein aggregates in the sample or specific body part or body area; comprises the steps of
- determining the amount of the compound bound to the alpha-synuclein aggregates;
- correlating the amount of the compound bound to the alpha-synuclein aggregates with the amount of the alpha-synuclein aggregates in the sample or specific body part or body area; and
- optionally comparing the amount of the compound bound with the alpha-synuclein aggregates in the sample or specific body part or body area to a normal control value in a healthy control subject.
33. A compound of formula (lll-F)
Figure imgf000099_0001
(lll-F) or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
(A) is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1F is a 4- to 6-membered heterocyclyl, which is optionally substituted with at least one C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1F is C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1F is -NH-C3-C6cycloalkyl or C3-C6cycloalkyl; and
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
R4 and R5 are independently selected from H, and CrC4alkyl; LG is a leaving group; and n is at least 1 . 34.. A compound of formula (I I l-F)
Figure imgf000100_0001
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl ; and
R2F is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from C1-C4alkoxy, and C1-C4alkyl; and
LG is a leaving group. , The compound of formula (lll-F) according to claim 33, or the compound of formula (II l-F’) according to claim 34, wherein LG is selected from bromo, chloro, iodo, C1-C4alkylsulfonate and Ce-Cioarylsulfonate, wherein the Ce-Cwarylsulfonate can be optionally substituted with -CH3 or -NO2. . A compound of formula (l-F)
Figure imgf000100_0002
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein
(A) is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl; R1F is a 4- to 6-membered heterocyclyl, which is optionally substituted with at least one C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1F is C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1F is -NH-C3-C6cycloalkyl or C3-C6cycloalkyl;
R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
R4 and R5 are independently selected from H, and C1-C4alkyl; and n is at least 1 , preferably 1 .
37. A compound of formula (l-F’)
Figure imgf000101_0001
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl ; and
R2F is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from C1-C4alkoxy, and C1-C4alkyl.
38. A compound of formula (fll-H)
Figure imgf000101_0002
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, wherein is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-C6cycloalkyl is optionally substituted with at least one halo;
R2 is a 5-membered or 6-membered heteroaryl, optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl; and
R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl;
X is bromo, chloro or iodo; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; and with the proviso that the compound of formula (I I l-H) comprises at least one X. A compound of formula (l-H)
Figure imgf000102_0001
or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, is a 6-membered heteroaryl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, or C1-C4alkyl;
R1 is a 4- to 6-membered heterocyclyl which is optionally substituted with at least one halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is halo, haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, NR4R5, or C1-C4alkyl; or
R1 is -NH-C3-C6cycloalkyl or -C3-Cecycloalkyl, wherein the C3-C6cycloalkyl of -NH-C3-C6cycloalkyl or -C3-Cecycloalkyl is optionally substituted with at least one halo; and R2 is a 5-membered or 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from haloC1-C4alkyl, haloC1-C4alkoxy, C1-C4alkoxy, and C1-C4alkyl;
R4 and R5 are independently selected from H, C1-C4alkyl and haloC1-C4alkyl;
Y is D, CD3, T or CT3; m is 0, 1 , 2 or 3; p is 0, 1 , 2 or 3; with the proviso that the compound of formula (l-H) comprises at least one D, CDs, T or CT3, wherein D is Deuterium and T is 3H (Tritium).
5 40. A method of preparing the compound of formula (l-F) according to claim 36, the method comprising reacting the compound of formula (lll-F) according to claim 33 or 34 with a bpfluorinating agent, so that LG is replaced by 18F.
41. A method of preparing the compound of formula (l-F’) according to claim 37, the method comprising reacting the compound of formula (lll-F’) according to claim 35 with a 18F-
10 fluorinating agent, so that LG is replaced by 18F.
42. The method according to claim 40 or 41 , wherein the 18F-fluorinating agent is selected from K18F, Rb18F, CS18F, Na18F, Kryptofix[222]18F, tetra(Ci.6alkyl)ammonium salt of 18F, and tetrabutylammonium [18F]fluoride.
43. A method of preparing the compound of formula (l-H) according to claim 39, the method
15 comprising reacting the compound of formula (lll-H) according to claim 38 with a 2H radiolabelling agent, so that X is replaced by D or CDs.
44. A method of preparing the compound of formula (l-H) according to claim 39, comprising reacting the compound of formula (lll-H) according to claim 38 with a 3H radiolabelling agent like CT3, so that X is replaced by T or CT3.
20 45. The compound according to any one of claims 1 to 11 , for use as an in vitro analytical reference or an in vitro screening tool.
46. A test kit for the detection and/or diagnosis of a disease, disorder or abnormality associated with alpha-synuclein aggregates, wherein the test kit comprises at least one compound as defined in any one of claims 9 to 11.
25 47. A kit for preparing a radiopharmaceutical preparation, wherein the kit comprises a sealed vial containing at least one compound as defined in any one of claims 33, 34, 35 or 38.
48. The kit according to claim 46, wherein the radiopharmaceutical preparation is for use in the imaging of alpha-synuclein aggregates, wherein the imaging is preferably conducted by positron emission tomography.
3049. The kit according to claim 46, wherein the radiopharmaceutical preparation is for use for in vitro imaging, ex vivo imaging, or in vivo imaging, preferably the use is for in vivo imaging.
50. The kit according to claim 46, wherein the radiopharmaceutical preparation is for use in brain imaging.
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