Methods for purifying compounds

Abstract

The present invention is 1~4 The present invention relates to a method for purifying a compound of formula (IV-F) containing an alkyl sulfonate as a leaving group, or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof. The compound of formula (IV-F) is useful for imaging and determining the amount of alpha-synuclein aggregates. 18 It can be used as a precursor to an F-labeled agent. 18 F-labeled agents can be used to diagnose diseases, disorders, or conditions associated with alpha-synuclein aggregates (e.g., multiple system atrophy (MSA)), determine a predisposition to such diseases, disorders, or conditions, prognose such diseases, disorders, or conditions, monitor the progression of disease in patients with such diseases, disorders, or conditions, monitor the progression of such diseases, disorders, or conditions, and predict the responsiveness of patients with such diseases, disorders, or conditions to treatment. JPEG2025539063000038.jpg20170

Description

[Technical Field]

[0001] The present invention relates to a method for purifying a compound of formula (IV-F). The present invention also relates to a kit for preparing a radiopharmaceutical preparation and a method for preparing a detectably labeled compound of formula (III-F). The compound of formula (III-F) can be used to image and determine the amount of alpha-synuclein aggregates. Furthermore, the compound of formula (III-F) can be used to diagnose diseases, disorders, or disorders associated with alpha-synuclein (α-synuclein, A-synuclein, a-synuclein, A-syn, α-syn, aSyn, a-syn) aggregates (e.g., multiple system atrophy (MSA)), determine predisposition to such diseases, disorders, or disorders, prognose such diseases, disorders, or disorders, monitor the progression of such diseases, disorders, or disorders in patients affected by such diseases, disorders, or disorders, monitor the progression of such diseases, disorders, or disorders, and predict the responsiveness of patients affected by such diseases, disorders, or disorders to treatment. [Background technology]

[0002] Many age-related diseases are based on or associated with the extracellular or intracellular deposition of amyloid or amyloid-like proteins, which contribute to the pathogenesis and progression of the disease. The best-characterized amyloid protein that forms extracellular aggregates is amyloid beta (Abeta or Aβ).

[0003] Amyloid-like proteins that primarily form intracellular aggregates include, but are not limited to, tau, alpha-synuclein, and huntingtin (HTT). Diseases associated with alpha-synuclein aggregates are generally classified as synucleinopathies (or α-synucleinopathies), including, but not limited to, Parkinson's disease (PD) and multiple system atrophy (MSA). Synucleinopathies involving primarily neuronal aggregates include, but are not limited to, Parkinson's disease (sporadic, familial with SNCA (the gene encoding the alpha-synuclein protein) mutations or SNCA gene duplication or triplication, familial with mutations in other genes other than SNCA, pure autonomic failure, and Lewy body dysphagia), SNCA duplication carriers, dementia with Lewy bodies (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's syndrome. Synucleinopathies involving 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 include traumatic brain injury, chronic traumatic encephalopathy, dementia pugilistica, tauopathies (Pick's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration and Niemann-Pick disease type C1, frontotemporal dementia with chromosome 17-linked parkinsonism), motor neuron diseases, Huntington's disease, amyotrophic lateral sclerosis (sporadic, familial and Guam ALS-dementia complex), neuroaxonal dystrophies, neurodegeneration type 1 with cerebral iron accumulation (Hallervorden-Spatz syndrome), prion diseases, Creutzfeldt-Jakob disease, ataxia-telangiectasia, Meige syndrome, subacute sclerosing panencephalitis, Gerstmann-Sträussler-Scheinker disease, inclusion body myositis, Gaucher disease, Krabbe disease,and other lysosomal storage disorders (including Kufor-Rakev syndrome and Sanfilippo syndrome) and rapid eye movement (REM) sleep behavior disorder, but are not limited to these disorders (Jellinger, Mov Disord 2003, 18 Suppl. 6, S2-12; Galvin et al. JAMA Neurology 2001, 58 (2), pp. 186-190; Kovari et al., Acta Neuropathol. 2007, 114 (3), pp. 295-8; Saito et al., J Neuropathol Exp Neurol. 2004, 63 (4), pp. 323-328; McKee et al., Brain 2013, 136 (Pt 1), pp. 43-64; Puschmann et al., Parkinsonism Relat Disord 2012, 18S1, S24-S27; Usenovic et al., J Neurosci. 2012, 32(12), pp. 4240-4246; Winder-Rhodes et al., Mov Disord. 2012, 27(2), pp. 312-315; Ferman et al., J Int Neuropsychol Soc. 2002, 8(7), pp. 907-914; Smith et al., J Pathol. 2014; pp. 232:509-521; Lippa et al., Ann Neurol. 1999 March; 45(3):353-7; Schmitz et al., Mol Neurobiol. 2018 August 22; Charles et al., Neurosci Lett. 2000 July 28; pp. 289(1):29-32; Wilhelmsen et al., Arch Neurol. 2004 March;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 July;59(7):592-8).

[0004] Alpha-synuclein is a naturally occurring unfolded protein of 140 amino acids (Iwai et al., Biochemistry 1995, 34(32), pp. 10139-10145). The alpha-synuclein sequence can be divided into three major domains: 1) the N-terminal region encompassing residues 1-60, which contains an 11-mer amphipathic imperfect repeat with a 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 proline-rich and lacks unique structural trends. Parkinson's disease (PD) is the most common neurodegenerative movement disorder. The etiology of PD remains unknown. However, increasing evidence suggests a role for pathogenic misfolding of the alpha-synuclein protein, which leads to the formation of amyloid-like fibrils. Indeed, a hallmark of PD is the presence of intracellular alpha-synuclein aggregate structures called Lewy bodies and neurites, primarily in substantia nigra neurons, and the death of dopaminergic neurons in the substantia nigra and elsewhere. Alpha-synuclein is a naturally unfolded presynaptic protein that can misfold and aggregate into large oligomeric and fibrillar forms, which have been linked to the pathogenesis of PD. Recent studies have implicated small soluble oligomeric and prefibrillar forms of alpha-synuclein as the most neurotoxic species (Lashuel et al., J. Mol. Biol., 2002, 322, 1089-102).

[0005] In addition to Parkinson's disease, the accumulation of aggregated alpha-synuclein into Lewy bodies is a characteristic of all Lewy body diseases, including Parkinson's disease dementia (PDD) and dementia with Lewy bodies (DLB) (Capouch et al., Neurol Ther. 2018, 7, 249-263). In DLB, Lewy bodies are widely distributed throughout the brain cortex, and in addition to Lewy bodies and neurites, many thread-like and punctate structures (Lewy puncta) have been found to be immunopositive for alpha-syn, phosphorylated at Ser-129 (Outeiro et al., Mol. Neurodegener. 2019, 14, 5).

[0006] Alpha-synuclein aggregates are also found in multiple system atrophy (MSA). MSA is a rare, sporadic neurodegenerative disorder manifested by rapidly progressive autonomic and motor dysfunction and heterogeneous cognitive decline. Examples of such disorders include Shy-Drager syndrome, striatonigral degeneration, and olivopontocerebellar atrophy. This disease can be clinically subclassified into parkinsonian (MSA-P) or cerebellar (MSA-C) variants depending on the predominant motor phenotype (Fanciulli et al., N Engl J Med 2015;372, 249-63). It is characterized by aggregation of alpha-synuclein in the cytoplasm of oligodendrocytes, forming glial cytoplasmic inclusions (GCIs). GCIs, consisting primarily of fibrillar forms of α-synuclein, are a 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 thought to play a central role in the pathogenesis of MSA. Correlations between GCI volume and the degree of neuronal loss have been reported in both the striatonigral and olivopontocerebellar regions (Stefanova et al., Neuropathol Appl Neurobiol. 2016, 42, 20-32).

[0007] The ability to image alpha-synuclein deposits in the brain would be a major advancement for alpha-synucleopathy research, including Parkinson's disease 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 begin years before the onset of symptoms. Therefore, alpha-synuclein is a priority target for drug development, not only considering its potential contribution to neurodegeneration but also potentially offering treatment options for those still in asymptomatic or presymptomatic stages of the disease. In vivo imaging of alpha-synuclein pathology could be useful as a biomarker (i) to potentially detect the presence of disease early, (ii) to assess disease progression, and (iii) as a pharmacodynamic tool for drug development. The development of alpha-synuclein PET imaging agents is now considered important for the accurate diagnosis of synucleinopathies and to support the clinical development of alpha-synuclein-targeting therapeutics, starting with the optimal selection of study populations (Eberling, Dave and Frasier, J. Parkinson's Disease, 3, 565-567 (2013)).

[0008] Only recently have the first noninvasive imaging of pathological alpha-synuclein (a-syn) in the human brain been reported, and positive clinical proof-of-concept data have been presented for the a-syn positron emission tomography (PET) tracer ACI-12589 as an imaging agent to identify patients with MSA (Capotosti F.; Discovery of [18F]ACI-12589, 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 [18F]ACI-12589, a novel PET-tracer for alpha-synuclein; Oral presentation; ADPD 2022 International Conference; Barcelona, ​​Spain; March 18, 2022).

[0009] Therefore, there is a clear need to find molecular probes with high alpha-synuclein selectivity that recognize and bind to pathological alpha-synuclein. To reduce background signal interference resulting from nonspecific off-target binding and to ease administration requirements, alpha-synuclein imaging compounds should bind to their targets with high affinity and selectivity. To image alpha-synuclein aggregates associated with neurological diseases, such as multiple system atrophy (MSA), imaging compounds must cross the blood-brain barrier and enter relevant brain regions. To target intracellular amyloid-like inclusions, such as alpha-synuclein, cell permeability is a further requirement for imaging compounds. Rapid washout of the compound from the brain (or other target organs) is another prerequisite for avoiding unnecessary compound accumulation, which may increase the risk of undesired side effects.

[0010] WO2021 / 224489 discloses a new class of compounds of formula (I) that are capable of binding to alpha-synuclein. The compounds are therefore capable of binding to suitable radioisotopes, in particular 18 When radiolabeled with F, it is a suitable PET tracer for imaging pathological alpha-syn aggregates in PD and other alpha-synucleinopathies. The precursor of the PET tracer was purified by flash chromatography. This method exhibits low yields and is therefore not suitable for industrial-scale production. [Prior art documents] [Patent documents]

[0011] [Patent Document 1] WO2021 / 224489 [Non-patent literature]

[0012] [Non-Patent Document 1] Jellinger, Mov Disord 2003, 18 Suppl. 6, S2~12 [Non-patent document 2] Galvin et al. JAMA Neurology 2001, 58 (2), pp. 186-190 [Non-patent document 3] Kovari et al., Acta Neuropathol. 2007, 114(3), pp. 295-8. [Non-patent document 4] Saito et al., J Neuropathol Exp Neurol. 2004, 63(4), pp. 323-328 [Non-Patent Document 5] McKee et al., Brain, 2013, 136(Pt 1), pp. 43-64 [Non-patent document 6] Puschmann et al., Parkinsonism Relat Disord 2012, 18S1, S24-S27 [Non-Patent Document 7] Usenovic, J Neurosci. 2012, 32(12), pp. 4240-4246 [Non-licensed Document 8] Winder-Rhodes, Mov Disord. 2012, 27(2), pp. 312-315 [Non-licensed Document 9] Ferman, J Int Neuropsychol Soc. 2002, 8(7), pp. 907-914 [Non-licensed Document 10] Smith, J. Pathol. 2014; 232: 509-521. [Non-licensed Document 11] Lippa, Ann Neurol. March 1999; 45(3): 353-7 pages [Non-licensed Document 12] Schmitz, Mol Neurobiol. August 22, 2018 [Non-licensed Document 13] Charles, Neurosci Lett. July 28, 2000; 289(1): 29-32 [Non-licensed Document 14] Wilhelmsen, Arch Neurol. March 2004; 61(3): 398-406 [Non-licensed Document 15] Yamaguchi, J Neuropathol Exp. Neurol. 2004, 80th Annual Meeting, Volume 63 [Non-licensed Document 16] Askanas, J Neuropathol Exp Neurol. July 2000; 59(7): 592-8 pages [Non-licensed Document 17] Iwai, Biochemistry 1995, 34(32), pages 10139~10145 [Non-licensed Document 18] Lashuel, J. Mol. Biol., 2002, 322, pp. 1089-102. [Non-licensed Document 19] Capouchら, Neurol. Ther. 2018, 7, pages 249~263 [Non-licensed Document 20] Outeiro, Mol. Neurodegener. 2019, pp. 14, 5 [Non-licensed Document 21] Fanciulli, N Engl J Med, 2015; pp. 372, 249-63 [Non-licensed Document 22] Galvin, Arch Neurol. 2001, 58, pp. 186-90. [Non-licensed Document 23] Stefanova, Neuropathol. Appl Neurobiol. 2016, 42, 20~32 pages [Non-licensed Document 24] Eberling, Dave and Frasier, J. Parkinson's Disease, 3, pages 565~567 (2013) [Non-licensed Document 25] Capotosti F.; Discovery of [18F] ACI-12589, a novel and promising PET-tracer for alpha-synuclein; Oral presentation; ADPD 2022 International Conference; Barcelona,​​ Spain; March 18, 2022 [Non-licensed Document 26] Smith R.; Initial scans using [18F] ACI-12589, a novel PET-tracer for alpha-synuclein; Oral presentation; ADPD 2022 International Conference; Barcelona,​​ Spain; March 18, 2022 [Non-licensed Document 27] Synthesis (1982), pp. 85-125, Table 2, Carey and Sundberg [Non-licensed Document 28] Organische Synthese (1995), pp. 279-281, table 5.8 [Non-licensed Document 29] Netscher, Recent Res. Dev. Org. Chem., 2003, 7, 71~83 pages, スキーム1, 2, 10 and 15, etc. [Non-licensed Document 30] Coenen, Fluorine-18 Labeling Methods: Features and Possibilities of Basic Reactions (2006), Schubiger PA, Friebe M., Lehmann L. (eds.), PET-Chemistry - The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, pp. 15-50, page 25 Skim 4, page 28 Skim 5, page 30 Table 4, page 33 Illustration 7 [Non-licensed Document 31] Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Company, Easton, PA, 1990, 1445 pages [Non-licensed Document 32] Ph. Eur. 2902 "Chemical Precursors for Radiopharmaceutical Preparations" [Non-licensed Document 33] L. Cai, S. Lu, V. Pike, Eur. J. Org. Chem 2008, pp. 2853-2873 [Non-licensed Document 34] J. Fluorine Chem., 27 (1985): 177-191 [Non-licensed Document 35] Remington's Pharmaceutical Sciences, 15th edition, Mack Publishing Co., New Jersey (1975) [Non-licensed Document 36] Ying-hui Chou, JAMA Neurol. April 1, 2015; 72(4): 432-440 [Non-licensed Document 37] Zrein et al., Clin. Diagn. Lab. Immunol., 1998, 5, 45-49 Summary of the Invention [Problem to be solved by the invention]

[0013] 18 Because the half-life of F is short (approximately 110 minutes), radiolabeled compounds, e.g., radiopharmaceuticals, are typically sterilized immediately prior to use. 18 The precursor is provided to the user in the form of a precursor that reacts with an F-fluorinating agent. The precursor has been observed to be unstable under normal storage conditions. An object of the present invention is therefore to provide a method for preparing the precursor compound with improved stability. A further object of the present invention is to provide a method that produces only small amounts of impurities and is suitable for the production of precursors for diagnostic applications on an industrial scale. [Means for solving the problem]

[0014] In a first aspect, the present invention provides a compound of formula (IV-F) as defined herein

[0015] [ka]

[0016] or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, comprising the steps of: (i) dissolving the crude compound of formula (IV-F) in DMSO and ethanol to obtain a solution; and (ii) carrying out crystallization from the solution to obtain the purified compound of formula (IV-F) The present invention covers a method including:

[0017] In a second aspect, the present invention relates to a compound obtainable by a method according to the first aspect.

[0018] A third aspect of the present invention refers to a kit for preparing a radiopharmaceutical preparation, the kit comprising a sealed vial containing at least one compound obtainable by the method according to the first aspect.

[0019] A fourth aspect of the present invention relates to a detectably labeled compound of formula (III-F) as defined herein.

[0020] [ka]

[0021] or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, comprising reacting a compound obtainable by the method according to the first aspect with a compound of formula (I) 18 so that it can be replaced by F 18 The present invention relates to a method comprising the step of reacting a compound with an F-fluorinating agent.

[0022] definition For purposes of interpreting this specification, the following definitions will apply unless otherwise specified and where appropriate, and terms used in the singular will include the plural and vice versa.

[0023] "Alkyl" refers to a saturated, straight-chain or branched organic moiety consisting of carbon and hydrogen atoms. Alkyl groups typically do not contain any saturation and are usually attached to the remainder of the molecule by a single bond. Examples of suitable alkyl groups have 1 to 6 carbon atoms, preferably 1 to 4 carbon atoms. The term "C1-C4 alkyl" should be construed accordingly. Examples of "C1-C4 alkyl" include, but are not limited to, methyl, ethyl, propyl, isopropyl, 1-methylethyl, n-butyl, t-butyl, and isobutyl, such as methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, and isobutyl.

[0024] "C1-C4 alkoxy" refers to a radical of the formula -ORa, where Ra is a C1-C4 alkyl radical as generally defined above. Examples of C1-C4 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, and isobutoxy.

[0025] "Halogen C1-C4 alkyl" or "halo C1-C4 alkyl" refers to a C1-C4 alkyl group as defined above that is substituted by one or more halo groups as defined below. Examples of "halo C1-C4 alkyl" 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.

[0026] "C3-C6 cycloalkyl" refers to a stable monocyclic saturated hydrocarbon group consisting solely of carbon and hydrogen atoms, having from 3 to 6 carbon atoms. Examples of C3-C6 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0027] "Heterocyclyl" refers to a stable 4- to 6-membered non-aromatic monocyclic ring group containing one or two heteroatoms selected from, for example, N, O, or S. Heterocyclyl groups can be unsaturated or saturated. Heterocyclyl groups can be attached via a carbon atom or a heteroatom. Examples include, but are not limited to, azetidinyl, oxetanyl, pyrrolinyl, pyrrolidyl, tetrahydrofuryl, tetrahydrothienyl, piperidyl, piperazinyl, tetrahydropyranyl, morpholinyl, or perhydroazepinyl. Examples of preferred heterocyclyl groups include, but are not limited to, azetidinyl, morpholinyl, piperazinyl, pyrrolidinyl, or piperidinyl.

[0028] "Aryl" refers to a homocyclic aromatic organic moiety (e.g., containing one or two rings) consisting of carbon and hydrogen atoms, preferably having from 5 to 12 carbon atoms, preferably from 6 to 12 carbon atoms, more preferably from 6 to 10 carbon atoms, even more preferably from 5 to 10 carbon atoms, and even more preferably 5 or 6 carbon atoms. Examples include, but are not limited to, phenyl, biphenyl, and naphthyl.

[0029] "Heteroaryl" refers to an aryl group, as defined above, in which at least one of the carbon atoms is replaced by a heteroatom selected from, for example, N, O, or S, or a moiety containing a heteroatom (e.g., N, O, and / or S). Typically, the heteroaryl is a 5- to 8-membered ring system, preferably a 5- to 6-membered ring system, in which at least one of the carbon atoms is replaced by a heteroatom selected from, for example, N, O, or S. Examples of possible heteroaryl groups include, but are not limited to, furyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazinyl, pyridazinyl, pyrimidyl, or pyridyl. Preferred examples thereof include pyridine, pyrazole, etc., more preferably pyridine.

[0030] "Hal" or "halogen" or "halo" refers to F, Cl, Br, and I. For diagnostic and pharmaceutical applications, F (e.g., 19 F and 18 F) is particularly preferred.

[0031] Unless otherwise defined, the term "leaving group" (LG) as used herein refers to any leaving group, an atom or group of atoms that can be displaced by another atom or group of atoms. Examples include, for example, Synthesis (1982), pp. 85-125, table 2; Carey and Sundberg, Organische Synthese (1995), pp. 279-281, table 5.8; or Netscher, Recent Res. Dev. Org. Chem., 2003, pp. 7, 71-83, Schemes 1, 2, 10, and 15). (Coenen, Fluorine-18 Labeling Methods: Features and Possibilities of Basic Reactions (2006), Schubiger PA, Friebe M., Lehmann L. (eds.), PET-Chemistry - The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, pp. 15-50, Scheme 4 on page 25, Scheme 5 on page 28, Table 4 on page 30, and Figure 7 on page 33.) Preferably, the "leaving group" (LG) is a halogen, C 1~4 Alkyl sulfonates and C 6~10 aryl sulfonates, C 6~10 The aryl may be optionally substituted with -CH or -NO. 3 In the moiety, the leaving group LG is C 1~4 It is an alkyl sulfonate.

[0032] The compounds of formula (III-F) and their precursors, such as compounds of formula (IV-F), having one or more optically active carbons can exist as racemates and racemic mixtures, stereoisomers (including diastereomeric mixtures and individual diastereoisomers, enantiomeric mixtures and single enantiomers, conformer mixtures and single conformers), tautomers, atropisomers, and rotamers. All isomeric forms are included in the present invention. Compounds described herein containing olefinic double bonds include E and Z geometric isomers.

[0033] All salt forms, polymorphs, hydrates and solvates (eg, ethanolates) are included in the present invention.

[0034] "Pharmaceutically acceptable salts" are defined as derivatives of the disclosed compounds in which the unchanged form has been modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues, such as amines; alkali or organic salts of acidic residues, such as carboxylic acids; and the like. Pharmaceutically acceptable salts include the conventional non-toxic salts or quaternary ammonium salts of the unchanged form, formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like; and organic acids such as, but not limited to, acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, and the like. Pharmaceutically acceptable salts of the compounds of formula (III-F) and their precursors can be synthesized from the unchanged form containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of an appropriate base or acid in water or an organic solvent, or a mixture of the two. Organic solvents include, but are not limited to, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Company, Easton, PA, 1990, p. 1445, the disclosure of which is hereby incorporated by reference.

[0035] "Pharmaceutically acceptable" is defined as those compounds, materials, compositions and / or dosage forms that are suitable for use in contact with the tissues of human beings and animals within the bounds of good medical practice and without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0036] The compounds of formula (III-F) may also be provided in the form of a prodrug, ie, a compound that is metabolized in vivo to an active metabolite.

[0037] The patient or subject in the present invention is typically an animal, particularly a mammal, and more particularly a human.

[0038] Alpha-synuclein aggregates are multimeric beta-sheet-rich aggregates of alpha-synuclein monomers that can form soluble oligomers or soluble / insoluble prefibrils or mature fibrils, associated with intracellular deposits detected in Parkinson's disease, multiple system atrophy (MSA), and other synucleinopathies. Alpha-synuclein aggregates constituting Lewy pathology can be detected as having the following morphologies: Lewy bodies, Lewy neurites, immature Lewy bodies or pale bodies, and perikarya deposits with diffuse, granular, punctate, or polymorphic patterns. Furthermore, alpha-synuclein aggregates are the major component of intracellular fibrillar inclusions (also called glial cytoplasmic inclusions) detected in oligodendrocytes, as well as intracellular fibrillar inclusions (called neuronal cytoplasmic inclusions) in neuronal cell bodies, axons, and nuclei, which are histological hallmarks of multiple system atrophy. Alpha-synuclein aggregates in Lewy pathology and glial cytoplasmic inclusions often display a substantial increase in post-translational modifications, such as phosphorylation, ubiquitination, nitration, and truncation.

[0039] Lewy bodies are abnormal aggregates of proteins that develop inside nerve cells in Parkinson's disease (PD), dementia with Lewy bodies, and other synucleinopathies. Lewy bodies appear as spherical masses that replace other cellular components. Morphologically, Lewy bodies can be classified as brainstem or cortical. Typical brainstem Lewy bodies are eosinophilic cytoplasmic inclusions consisting of a dense core surrounded by a halo of 5-10 nm-wide radial fibers, the main structural component of which is alpha-synuclein. Cortical Lewy bodies differ in the absence of the halo. The presence of Lewy bodies is a hallmark of Parkinson's disease.

[0040] Lewy neurites are abnormal neuronal processes in pathological neurons that contain granules, abnormal alpha-synuclein (a-syn) filaments similar to those found in Lewy bodies, punctate varicose structures, and axonal spheroids. Lewy neurites are characteristic of α-synucleinopathies, such as dementia with Lewy bodies, Parkinson's disease, and multiple system atrophy (MSA).

[0041] Glial cytoplasmic inclusions (GCIs or Papp-Lantos bodies) are argyrophilic cytoplasmic aggregates in oligodendrocytes composed of filamentous alpha-synuclein. They appear morphologically triangular, crescentic, or sickle-shaped. In MSA, in addition to GCIs, inclusions composed of alpha-synuclein filaments are detected in neurons in the cytoplasm or beneath the nuclear membrane; these are called neuronal cytoplasmic inclusions and neuronal nuclear inclusions, respectively. GCIs are recognized as the defining morphological feature of MSA, and their widespread distribution is the criterion for a definite postmortem neuropathological diagnosis of MSA.

[0042] The terms "disease," "disorder," or "condition" are used interchangeably herein.

[0043] The compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, may bind to alpha-synuclein aggregates. The type of bond between the compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, is not specified, and any type of bond is covered by the present invention. The terms "compound that binds to alpha-synuclein aggregates," "compound / (alpha-synuclein aggregate) complex," "compound / alpha-synuclein aggregate complex," "compound / protein aggregate complex," and the like are used interchangeably herein and are not intended to be limited to any particular type of bond.

[0044] The preferred definitions provided in the "Definitions" section apply to all of the embodiments described below unless otherwise specified. Various embodiments of the invention are described herein, and it is recognized that the features specified in each embodiment may be combined with other specified features to obtain further embodiments of the invention. [Brief explanation of the drawings]

[0045] [Figure 1] FIG. 1 shows the purity profile over time of the crude compound of Preparative Example 1 and the purified compound of Example 1. DETAILED DESCRIPTION OF THE INVENTION

[0046] The present invention relates to a compound of formula (IV-F)

[0047] [ka] or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, comprising the steps of: (i) dissolving the crude compound of formula (IV-F) in dimethyl sulfoxide (DMSO) and ethanol to obtain a solution; and (ii) carrying out crystallization from the solution to obtain the purified compound of formula (IV-F) The present invention relates to a method comprising:

[0048] The presently claimed method for recrystallizing compounds of formula (IV-F) is particularly suitable for industrial scale.

[0049] Compounds of formula (IV-F), as well as methods for synthesizing them, are disclosed in WO2021 / 224489.

[0050] R 3 teeth,

[0051] [ka]

[0052] Preferably, R 3 teeth,

[0053] [ka]

[0054] More preferably, R 3 teeth,

[0055] [ka]

[0056] is.

[0057] R 3 The leaving group LG in C 1~4 Preferably, the leaving group (LG) is:

[0058] [ka]

[0059] , i.e., mesylate.

[0060] R 4 is aryl or 5- or 6-membered heteroaryl, and R 4 teeth,

[0061] [ka]

[0062] is selected from R 2a , R 2a' is independently selected from H or F; R 2b are independently selected from F, —OH, C1-C4 alkyl, haloC1-C4 alkyl, —NH2, —CN, or C1-C4 alkoxy; R 2c , R 2c' are independently selected from H, F, OH, OCH3 or CH3; R 2d is selected from H, F or —OH; R 2e is selected from H, OH, CH3 or F, Z is independently N, NH, N(C1-C4 alkyl), N(haloC1-C4 alkyl), O, or S; Z 1 are independently N, NH, O or S, p is 0, 1 or 2; m is 0 or 1; As allowed by valence,

[0063] [ka]

[0064] is a combination of a single bond and a double bond, * is the position of the bond.

[0065] Preferably R 4 teeth,

[0066] [ka]

[0067] Selected from R 2a , R 2a' , R 2b , R 2c , R 2c' , R 2d , R 2e and p are as defined herein above, and R z is selected from H, C1-C4 alkyl or haloC1-C4 alkyl.

[0068] More preferably, R 4 is the following:

[0069] [ka]

[0070] is selected from R 2a , R 2a' , R 2b , R 2c , R 2c' , R 2d , R 2e , R z and p is as defined herein above.

[0071] More preferably, R 4 is the following:

[0072] [ka]

[0073] Selected from R 2a , R 2a' , R 2b , R 2c , R 2c' , R 2e , R zand p is as defined herein above.

[0074] More preferably, R 4 teeth,

[0075] [ka]

[0076] is.

[0077] In a further preferred embodiment, R 4 teeth,

[0078] [ka]

[0079] is.

[0080] Even more preferably, R 4 teeth,

[0081] [ka]

[0082] is.

[0083] Preferred compounds are

[0084] [ka]

[0085] or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0086] In one embodiment, preferred compounds of formula (I) are the following stereoisomers:

[0087] [ka]

[0088] or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0089] More preferred compounds are

[0090] [ka]

[0091] or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0092] In one embodiment, more preferred compounds of formula (I) are

[0093] [ka]

[0094] or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0095] The crude compound of formula (IV-F) used in the method of the present invention can be prepared by the method set forth in WO2021 / 224489. In one embodiment, the crude compound of formula (IV-F) can be prepared as shown in the following scheme:

[0096] Scheme 1

[0097] [ka]

[0098] Pg is a protecting group and LG is a leaving group.

[0099] Commercially available hydrazines can be condensed with an appropriate ketone to give the corresponding hydrazones. The crude hydrazones can be cyclized using DMF / DMA to give intermediate A. SNAr can be carried out with a suitable nucleophile in a suitable solvent to give intermediate D.

[0100] Scheme 1A

[0101] [ka]

[0102] An alternative approach (Scheme 1A) involves deprotection of intermediate A followed by an SNAr reaction with a suitable nucleophile, preferably carried out in the presence of CsF in DMSO. Intermediate D can be further functionalized, preferably using copper(I) in the presence of a base and a solvent (Ullmann reaction), to give intermediate E. Finally, LG can be introduced into intermediate E to give the crude compound of formula (IV-F).

[0103] In the first reaction step, 2-LG1-5-hydrazinylpyridine can be reacted with a suitably protected 2,4-dioxopyrrolidine (e.g., tert-butyl 2,4-dioxopyrrolidine-1-carboxylate). The leaving group LG1 is not specifically limited and can be any leaving group that can be displaced in a subsequent SNAr reaction. Examples include Hal (e.g., Cl, Br, I, F). The reaction can be carried out, for example, in a suitable solvent, e.g., C 1~4 This reaction may be carried out in an alcohol (preferably ethanol) at elevated temperature. Ring closure of the resulting hydrazone may be carried out, if appropriate, using, for example, DMF-DMA, to give intermediate A.

[0104] The protecting group Pg can be removed using suitable conditions depending on the protecting group chosen, for example acid cleavage.

[0105] Desirable R 3 Precursor of the group R *The group can be introduced via an SNAr reaction (nucleophilic aromatic substitution reaction), which gives intermediate D. This type of reaction is well known in the chemical arts and can be carried out under any suitable conditions. A suitable catalyst, such as cesium fluoride, can be used if necessary.

[0106] R * teeth,

[0107] [ka]

[0108] or the corresponding moiety in which OH is protected by a protecting group.

[0109] R 4 The group can be, for example, intermediates D and R 4 -Hal or R 4 -triflate coupling to give compound E. Coupling reagents include Cu-, Pd- and Ni-containing compounds, such as CuI.

[0110] In the final step, the crude compound of formula (IV-F) is obtained by converting the OH group of intermediate E to C 1~4 This reaction can be carried out, for example, by converting intermediate E to C 1~4 This can be carried out by reacting with an alkylsulfonate-Hal (Hal is for example Cl, eg mesyl chloride).

[0111] The above reaction scheme is advantageous because it allows the preparation of compound of formula (IV-F) in gram quantities rather than milligram quantities. Furthermore, the crude compound of formula (IV-F) obtained by this method typically has an initial purity of more than 90%. Therefore, it is an efficient starting material for GMP-grade purification processes.

[0112] Scheme 1B

[0113] [ka]

[0114] A further general approach is depicted in Scheme 1B, following the same preferred conditions described in general Scheme 1 or 1A.

[0115] In Scheme 1B, Lg is a leaving group and Pg is a protecting group;

[0116] [ka]

[0117] teeth

[0118] [ka]

[0119] and R 1 is R 3 and R 2 is R 4 and R 0 is H.

[0120] Or, 18 The F-precursor can be obtained by treating intermediate A with hydroxypyrrolidine in a suitable solvent under heating. 4 The group can be introduced by palladium-catalyzed amidation or Ullmann reaction. Finally, the alcohol intermediate E can be modified to a leaving group using standard conditions to give crude compounds of formula (IV-F).

[0121] Process (i) In step (i), the crude compound of formula (IV-F) is dissolved in a solvent containing (preferably consisting of) DMSO and ethanol. The ratio of DMSO to ethanol (v:v) is not particularly limited. The ratio of DMSO to ethanol is typically within the range of about 5:1 to about 0.1:1, preferably about 4:1 to about 0.5:1, more preferably about 3:1 to about 0.5:1, even more preferably about 2:1 to about 0.75:1, and most preferably about 1:1. In this context, the ratio (v:v) refers to the ratio of a unit volume of DMSO to a unit volume of ethanol, for example, ml:ml.

[0122] The ratio of the solvent to the crude compound of formula (IV-F) is not particularly limited, as long as the amount of solvent is sufficient to dissolve the crude compound of formula (IV-F). If the amount of solvent is too large, the cost will increase and the crystallization time will be long. Therefore, the ratio of the solvent to the crude compound of formula (IV-F) (v:w) is preferably within the range of about 5:1 to about 500:1, preferably about 25:1 to about 300:1, more preferably about 40:1 to about 250:1. It is understood that this ratio may vary depending on, for example, the conditions used during the dissolution step, such as the temperature selected. In this context, the ratio (v:w) refers to the ratio of the unit volume of the total solvent to the unit mass of the crude compound of formula (IV-F), for example, ml:g.

[0123] The crude compound of formula (IV-F) can be dissolved in a solvent by any suitable means.

[0124] In one embodiment, ethanol and DMSO can be mixed, and then the crude compound of formula (IV-F) can be added. In a further embodiment, the crude compound of formula (IV-F) can be mixed with ethanol, and then DMSO can be added. In a further preferred embodiment, the crude compound of formula (IV-F) can be mixed with DMSO, and then ethanol can be added.

[0125] If necessary, dissolution can be facilitated by using elevated temperatures, for example, temperatures within the range of about 50 to about 90° C., preferably about 60 to about 90° C., and more preferably about 65 to about 85° C. These temperatures are given relative to ambient pressure (approximately 1 atm). If elevated or reduced pressure is applied, the temperature ranges may be adapted accordingly.

[0126] A major amount of crude compound of formula (IV-F) should be dissolved during dissolution step (i), typically at least about 50 wt.%, preferably at least about 60 wt.%, even more preferably at least about 70 wt.%, even more preferably at least about 80 wt.%, even more preferably at least about 90 wt.%, and most preferably 100%.

[0127] Conventional techniques, such as stirring, may be used during dissolution step (i) to enhance or accelerate dissolution.

[0128] Process (ii) After the dissolution step (i), the compound of formula (IV-F) is crystallized from the solution to obtain the purified compound of formula (IV-F).

[0129] Crystallization can be carried out by various techniques, such as cooling the solution, removing the solvent, etc. The solvent can be removed, for example, by reducing the pressure or by heating the solution so as to evaporate the solvent.

[0130] In a preferred embodiment, the solution is cooled to crystallize the compound of formula (IV-F). The temperature can be appropriately selected, for example, within the range of about 0°C to about 50°C, preferably about 0°C to about 40°C, and more preferably about 0°C to about 30°C. If necessary, the temperature can be varied within this range during the crystallization process. For example, crystallization can be initially carried out at about 10°C to about 50°C (preferably about 0°C to about 40°C, more preferably about 0°C to about 30°C), and then the temperature can be lowered to about 0°C to about 10°C to improve the yield.

[0131] Optional step (iii) Finally, at least a portion (typically all) of the crystallization solution may be removed to provide the purified compound of formula (IV-F). Filtration, centrifugation, etc. may be used.

[0132] If necessary, the obtained crystals may be washed with a solvent, such as ethanol, in step (iii) and may be dried.

[0133] Further steps Step (i), step (ii) and optionally step (iii) may be carried out one or more times (e.g., at least once, preferably at least twice, more preferably at least three times, even more preferably at least four times, and even more preferably at least five times) by using the purified compound of formula (IV-F) in step (i) as a crude compound of formula (IV-F) to further increase the purity to a desired level.

[0134] In a preferred embodiment, steps (i), (ii) and optionally step (iii) are carried out at least twice, preferably at least three times.

[0135] In another preferred embodiment, steps (i), (ii) and optionally step (iii) are performed at least four times. In a further preferred embodiment, steps (i), (ii) and optionally step (iii) are performed at least five times.

[0136] In one embodiment, steps (i), (ii), and optionally step (iii) are performed twice. In another embodiment, steps (i), (ii), and optionally step (iii) are performed three times. In a preferred embodiment, steps (i), (ii), and optionally step (iii) are performed four times. In another preferred embodiment, steps (i), (ii), and optionally step (iii) are performed five times.

[0137] In other embodiments, step (i), step (ii) and optional step (iii) may be performed at least 6 times, at least 7 times, at least 8 times, at least 9 times or at least 10 times.

[0138] The maximum number of repetitions is not particularly limited. The method of the present invention may be repeated as many times as desired until the required purity is achieved. However, since the overall yield decreases when the method of the present invention is repeated, step (i), step (ii) and optional step (iii) are typically carried out at most 30 times, preferably at most 25 times, more preferably at most 20 times, and even more preferably at most 15 times.

[0139] Optional Purification Step The process of the present invention may, if desired, be combined with other purification steps conventional in the art. Examples of further purification steps include recrystallization using other solvent systems or chromatography.

[0140] In one embodiment, a solvent system using DMSO and 2-butanone or ethyl acetate can be used to dissolve the compound of formula (IV-F). The compound of formula (IV-F) can then be crystallized, and then at least a portion of the solution can be removed, optionally followed by drying and / or washing.

[0141] Purification using DMSO and 2-butanone or ethyl acetate can be performed at any time, but is typically performed before or after the method of the invention, more typically after the method of the invention. It can be performed after the complete sequence of steps (i), (ii) and optional step (iii), including any optional repetitions thereof, has been performed, or it can be performed after the sequence of one or more of steps (i), (ii) and optional step (iii), but before any repetition of steps (i), (ii) and optional step (iii) occurs. In a preferred embodiment, purification using DMSO and 2-butanone or ethyl acetate is used after the complete sequence of steps (i), (ii) and optional step (iii), including any repetitions thereof, has been performed (e.g., after steps (i), (ii) and optional step (iii) have been performed at least two times, preferably at least three times), as this sequence of steps improves purity compared to performing purification using DMSO and 2-butanone or ethyl acetate at an earlier stage.

[0142] The ratio of DMSO to 2-butanone (v:v) is not particularly limited. The ratio of DMSO to 2-butanone is typically within the range of about 5:1 to about 0.1:1, preferably about 4:1 to about 0.5:1, more preferably about 3:1 to about 0.5:1, even more preferably about 2:1 to about 0.75:1, and most preferably about 1:1. In this context, the ratio (v:v) refers to the ratio of a unit volume of DMSO to a unit volume of 2-butanone, for example, ml:ml.

[0143] The ratio of DMSO to ethyl acetate (v:v) is not particularly limited. The ratio of DMSO to ethyl acetate is typically within the range of about 5:1 to about 0.1:1, preferably about 4:1 to about 0.5:1, more preferably about 3:1 to about 0.5:1, even more preferably about 2:1 to about 0.75:1, and most preferably about 1:1. In this context, the ratio (v:v) refers to the ratio of a unit volume of DMSO to a unit volume of ethyl acetate, for example, ml:ml.

[0144] The ratio of the solvent to the crude compound of formula (IV-F) is not particularly limited, as long as the amount of solvent is sufficient to dissolve the crude compound of formula (IV-F). If the amount of solvent is too large, the cost will increase and the crystallization time will be long. Therefore, the ratio of the solvent to the crude compound of formula (IV-F) (v:w) is preferably within the range of about 5:1 to about 500:1, preferably about 25:1 to about 300:1, more preferably about 40:1 to about 250:1. It is understood that this ratio may vary depending on, for example, the conditions used during the dissolution step, such as the temperature selected. In this context, the ratio (v:w) refers to the ratio of the unit volume of the total solvent to the unit mass of the crude compound of formula (IV-F), for example, ml:g.

[0145] The crude compound of formula (IV-F) may be dissolved in a solvent by any suitable means.

[0146] In one embodiment, 2-butanone or ethyl acetate and DMSO can be mixed, and then the crude compound of formula (IV-F) can be added. In a further embodiment, the crude compound of formula (IV-F) can be suspended in 2-butanone or ethyl acetate, and then DMSO can be added. In a further preferred embodiment, the crude compound of formula (IV-F) can be suspended in DMSO, and then 2-butanone or ethyl acetate can be added.

[0147] If necessary, dissolution can be facilitated by using elevated temperatures, for example, temperatures within the range of about 50 to about 90° C., preferably about 60 to about 90° C., and more preferably about 65 to about 85° C. These temperatures are given relative to ambient pressure (approximately 1 atm). If elevated or reduced pressure is applied, the temperature ranges may be adapted accordingly.

[0148] A major amount of crude compound of formula (IV-F) should be dissolved during dissolution step (i), typically at least about 50 wt.%, preferably at least about 60 wt.%, even more preferably at least about 70 wt.%, even more preferably at least about 80 wt.%, even more preferably at least about 90 wt.%, and most preferably 100%.

[0149] Conventional techniques, such as stirring, may be used during the dissolution process to enhance or accelerate dissolution.

[0150] After the dissolution step, the compound of formula (IV-F) is crystallized from the solution to obtain the purified compound of formula (IV-F).

[0151] Crystallization can be carried out by various techniques, such as cooling the solution, removing the solvent, etc. The solvent can be removed, for example, by reducing the pressure or by heating the solution so as to evaporate the solvent.

[0152] In a preferred embodiment, the solution is cooled to crystallize the purified compound of formula (IV-F). The temperature can be appropriately selected, for example, within the range of about 0° C. to about 50° C., preferably about 0° C. to about 40° C., and more preferably about 0° C. to about 30° C. For example, crystallization can be initially carried out at about 10° C. to about 50° C. (preferably about 0° C. to about 40° C., more preferably about 0° C. to about 30° C.), and then the temperature can be lowered to about 0° C. to about 10° C. to improve the yield.

[0153] Typically, the purified compound of formula (IV-F) is isolated from the crystallization solution. Filtration, centrifugation, etc. may be used.

[0154] If necessary, the obtained crystals may be washed with a solvent such as 2-butanone or ethyl acetate. The crystals may be dried if necessary.

[0155] The desired final purity depends on the intended use. For diagnostic applications, a purity of at least 97% is typically required (Ph. Eur. 2902 "Chemical Precursors for Radiopharmaceutical Preparations"). The conditions in steps (i), (ii) and optional step (iii) may be the same or different during the repetitions.

[0156] It has been unexpectedly discovered that the purified compound of formula (IV-F) prepared by the method of the present invention not only has higher purity than the crude compound of formula (IV-F), but also has improved stability. For example, the purified compound of formula (IV-F) that has been subjected to three recrystallizations is stable under accelerated storage conditions (40°C, 4 weeks). In contrast, the crude compound of formula (IV-F) decomposed to 15.2 wt.% under these conditions.

[0157] The advantage of the present invention is that LG 1~4 This is observed with compounds that are alkyl sulfonates, especially mesylates.

[0158] Methods for synthesizing detectably labeled compounds The compounds obtainable by the methods of the present invention are suitable as precursors to detectably labeled compounds that can be used in imaging applications.

[0159] In one embodiment, the present invention provides a compound of formula (IV-F) containing a radioisotope. 18 The present invention relates to a process for preparing a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, by radiolabeling with F.

[0160] 18Reagents, solvents and conditions that can be used for F-fluorination are well known to those skilled in the art (L. Cai, S. Lu, V. Pike, Eur. J. Org. Chem 2008, pp. 2853-2873; J. Fluorine Chem., 27 (1985): pp. 177-191; Coenen, Fluorine-18 Labeling Methods: Features and Possibilities of Basic Reactions, (2006), Schubiger PA, Friebe M., Lehmann L. (eds.), PET-Chemistry - The Driving Force in Molecular Imaging. Springer, Berlin Heidelberg, pp. 15-50). Preferably, 18 The solvent used for F-fluorination is DMF, DMSO, acetonitrile, DMA, or a mixture thereof, preferably the solvent is acetonitrile or DMSO.

[0161] Any suitable 18 F-fluorinating agents may be used. Typical examples are H 18 F, alkali or alkaline earth 18 F-fluoride (e.g., K 18 F, Rb 18 F, Cs 18 F and Na 18 F). Optionally, 18 The F-fluorinating agent may 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). 18 F-fluorinating agent is 18 a tetraalkylammonium salt of F or 18 Tetraalkylphosphonium salts of F; for example, 18 F Tetra (C 1~6 alkyl)ammonium salts or 18 F Tetra (C 1~6 The alkyl phosphonium salt may be:18 F-fluorinating agent is K 18 F, H 18 F, Cs 18 F, Na 18 F, 18 F Tetra (C 1~6 (Alkyl)ammonium salt, kryptofix

[0222] 18 F or tetrabutylammonium [ 18 F] fluoride.

[0162] Diagnostic Compositions The compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, is particularly suitable for imaging alpha-synuclein aggregates. With respect to alpha-synuclein protein, the compounds are particularly suitable for binding to various types of alpha-synuclein aggregates. Imaging can be performed in mammals, preferably humans. Imaging is preferably in vitro imaging, ex vivo imaging, or in vivo imaging. More preferably, imaging is in vivo imaging; even more preferably, imaging is preferably brain imaging. Imaging can also be ocular / retinal imaging. The compounds of formula (III-F), or stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates thereof, are particularly suitable for use in diagnostic methods.

[0163] The diagnostic method can be performed on a mammal, preferably a human. The tissue of interest on which the diagnostic method is performed can be the brain, tissue of the central nervous system, tissue of the eye (e.g., retinal tissue) or other tissue, a peripheral organ, such as, but not limited to, the intestine, or a body fluid, such as cerebrospinal fluid (CSF). The tissue is preferably brain tissue.

[0164] Due to their design and binding properties, compounds of formula (III-F), or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, are suitable for use in the diagnosis of diseases, disorders, and disorders associated with alpha-synuclein aggregates. Compounds of formula (III-F), or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, are particularly suitable for positron emission tomography imaging of alpha-synuclein aggregates. Diseases associated with alpha-synuclein aggregates are generally listed as synucleinopathies (or α-synucleinopathies). The compounds of Formula (III-F), or stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates thereof, are suitable for use in the diagnosis of diseases, disorders, or abnormalities including, but not limited to, Parkinson's disease (sporadic, familial with alpha-synuclein mutations, familial with non-alpha-synuclein mutations, pure autonomic failure, and Lewy body dysphagia), SNCA duplication carriers, 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's syndrome. Synucleinopathies associated with neuronal and glial aggregates of alpha-synuclein include multiple 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 disease type C1), motor neuron diseases, amyotrophic lateral sclerosis (sporadic, familial, and Guam ALS-dementia complex), neuroaxonal dystrophies, neurodegeneration type 1 with cerebral iron accumulation (Hallervorden-Spatz syndrome), prion diseases, ataxia-telangiectasia, Meige syndrome, subacute sclerosing panencephalitis, Gaucher disease, and other lysosomal storage diseases (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), pp. 186-190; Kovari et al., Acta Neuropathol. 2007, 114(3), pp. 295-298; Saito et al., J Neuropathol Exp Neurol. 2004, 63(4), pp. 323-328; McKee et al., Brain, 2013, 136(Pt1), pp. 43-64; Puschmann et al., Parkinsonism Relat Disord 2012, 18S1, S24-S27; Usenovic et al., J Neurosci. 2012, 32(12), pp. 4240-4246; Winder-Rhodes et al., Mov Disord. 2012, 27(2), pp. 312-315; Ferman et al., J Int Neuropsychol Soc. 2002, 8(7), pp. 907-914). Preferably, the compound of formula (III-F) is suitable for use in the diagnosis of Parkinson's disease, multiple system atrophy, dementia with Lewy bodies, Parkinson's disease dementia, SNCA duplication carriers or Alzheimer's disease, more preferably multiple system atrophy (MSA).

[0165] In a method for diagnosing an alpha-synuclein aggregate-associated disease, disorder, or condition, e.g., multiple system atrophy (MSA), or a predisposition thereto, in a subject, the method comprises: a) administering to a subject a diagnostically effective amount of a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; b) distributing the compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, into a tissue of interest (e.g., the brain, tissue of the central nervous system (CNS), ocular tissue, or other tissue, a peripheral organ, such as, but not limited to, the intestine, or a body fluid, such as cerebrospinal fluid (CSF)); and c) imaging the tissue of interest, wherein increased binding of the compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, to the tissue of interest compared to normal control levels of binding indicates that the subject is suffering from or at risk of developing a disease, disorder, or condition associated with alpha-synuclein aggregates. Includes.

[0166] The compounds of formula (III-F), or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, can be used to image alpha-synuclein aggregates in any patient sample or specific body part or region suspected of containing alpha-synuclein aggregates. The compounds can cross the blood-brain barrier. As a result, they are particularly suitable for imaging alpha-synuclein aggregates in the brain, tissues of the central nervous system (CNS), or ocular tissue, or in peripheral organs, such as, but not limited to, the intestine, as well as in body fluids, such as cerebrospinal fluid (CSF).

[0167] For diagnostic applications, the compounds of formula (III-F), or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, are preferably administered in the form of a diagnostic composition comprising the compounds of formula (III-F), or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates. A "diagnostic composition" is defined herein as a composition comprising one or more compounds of formula (III-F), or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates or solvates, in a form suitable for administration to a patient, e.g., a mammal, e.g., a human, and suitable for use in diagnosing a specific disease, disorder or abnormality in a tissue. Preferably, the diagnostic composition further comprises a physiologically acceptable excipient, carrier, diluent, or adjuvant. Administration is preferably carried out by injecting the composition as an aqueous solution, as defined below, more preferably. Such compositions may optionally contain additional ingredients, such as a buffer; a pharmaceutically acceptable solubilizer (e.g., a cyclodextrin or a surfactant, such as Pluronic, Tween, or a phospholipid); and a pharmaceutically acceptable stabilizer or antioxidant (e.g., ascorbic acid, gentisic acid, or para-aminobenzoic acid). The dosage of the compound of formula (III-F), or its stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate, will vary depending on the exact compound administered, the patient's weight, and other variables apparent to a practitioner skilled in the art.

[0168] While it is possible for the compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, to be administered alone, it is preferable to formulate it into a diagnostic composition according to standard pharmaceutical practice. Accordingly, the present invention also provides a diagnostic composition comprising a diagnostically effective amount of a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, optionally in admixture with at least one pharmaceutically acceptable excipient, carrier, diluent, or adjuvant.

[0169] Pharmaceutically acceptable excipients are well known in the pharmaceutical industry and are described, for example, in Remington's Pharmaceutical Sciences, 15th ed., Mack Publishing Co., New Jersey (1975). Pharmaceutical excipients can be selected with reference to the intended route of administration and standard pharmaceutical practice. An excipient must be acceptable in the sense of not being harmful to the recipient thereof.

[0170] Pharmaceutically useful excipients, carriers, adjuvants, and diluents that can be used to formulate the diagnostic compositions of the present invention may include, for example, solvents, such as monohydric alcohols, e.g., ethanol, isopropanol, and polyhydric alcohols, e.g., glycols, and edible oils, e.g., soybean oil, coconut oil, olive oil, safflower oil, cottonseed oil, oily esters, e.g., ethyl oleate, isopropyl myristate, binders, adjuvants, solubilizers, thickeners, stabilizers, disintegrants, glidants, lubricants, buffers, emulsifiers, wetting agents, suspending agents, sweeteners, colorants, flavorings, coating agents, preservatives, antioxidants, processing agents, drug delivery modifiers and enhancers, e.g., calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, methylcellulose, sodium carboxymethylcellulose, dextrose, hydroxypropyl-β-cyclodextrin, polyvinylpyrrolidone, low melting point waxes, and ion exchange resins.

[0171] The route for administering (delivering) the compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, is not limited, but is preferably via infusion, more preferably via intravenous infusion.

[0172] Preferably, for diagnostic applications, the compound of formula (III-F), or its stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate, is administered parenterally. When the compound of formula (III-F), or its stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate, is administered parenterally, examples of such administration include one or more of intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, ​​intrasternal, intracranial, intramuscular, or subcutaneous administration of the compound, and / or administration by using injection techniques. For parenteral administration, the compound is best used in the form of a sterile aqueous solution, which may contain other substances, for example, sufficient salts or glucose to make the solution isotonic with blood. The aqueous solution should be suitably buffered (preferably to a pH of 3 to 9), if necessary. The preparation of suitable parenteral formulations under sterile conditions is readily accomplished by standard pharmaceutical techniques well known to those skilled in the art.

[0173] Typically, a physician will determine the actual dosage that will be most suitable for an individual subject. The specific dose level and frequency of administration for any particular individual may vary and will depend on a variety of factors, including the activity of the particular compound used, the metabolic stability and length of action of that compound, age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the precise severity of the condition, and the individual diagnosis.

[0174] The diagnostic compositions of the present invention can be produced by means known per se to those skilled in the art, for example as described in Remington's Pharmaceutical Sciences, 15th Edition, Mack Publishing Co., New Jersey (1975).

[0175] The compounds of formula (III-F), or their stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates, are useful as in vitro analytical standards or in vitro screening tools. They are also useful in in vivo diagnostic methods.

[0176] The compound of formula (III-F), or its stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate, may also be provided in the form of a mixture containing the compound of formula (III-F), or its stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate, and at least one compound selected from an imaging agent other than the compound of formula (III-F), a pharmaceutically acceptable excipient, carrier, diluent or adjuvant. The imaging agent other than the compound of formula (III-F) is preferably present in a diagnostically effective amount. More preferably, the imaging agent other than the compound of formula (III-F) is an amyloid beta or tau imaging agent.

[0177] Diagnosis of an alpha-synuclein aggregate-associated disease, disorder or disorder, or a predisposition to an alpha-synuclein aggregate-associated disease, disorder or disorder in a patient can be achieved by detecting specific binding of a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, to alpha-synuclein aggregates in a sample or in a specific body part or body region, which can be achieved by: (a) contacting a sample or a specific body part or region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, that binds to alpha-synuclein aggregates; (b) binding a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, to alpha-synuclein aggregates to form a compound / (alpha-synuclein aggregate) complex (hereinafter, "compound / (alpha-synuclein aggregate) complex" is abbreviated as "compound / protein aggregate complex"); (c) detecting the formation of a compound / protein aggregate complex. (d) optionally correlating the presence or absence of the compound / protein aggregate complex with the presence or absence of alpha-synuclein aggregates in the sample or in a particular body part or region; and (e) optionally, comparing the amount of the compound / protein aggregate complex to a normal control value, wherein an increase in the amount of the compound / protein aggregate complex compared to the normal control value may indicate that the patient is suffering from or at risk of developing a disease, disorder, or condition associated with alpha-synuclein aggregates. Includes.

[0178] The compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, can be contacted with a sample or specific body part or region suspected of containing alpha-synuclein aggregates by a suitable method. In in vitro methods, the compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, and a liquid sample can simply be mixed. For in vivo testing, the compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, is typically administered to a patient by any suitable means, such as parenteral administration, with intravenous infusion being preferred.

[0179] After contacting the sample or specific body part or region with a compound of Formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, the compound is allowed to bind to alpha-synuclein aggregates. The amount of time required for binding depends on the type of test (e.g., in vitro or in vivo) and can be determined by one of ordinary skill in the art through routine experimentation.

[0180] Compounds bound to alpha-synuclein aggregates can then be detected by any suitable method. The particular method selected will depend on the detectable label selected. Examples of possible methods include, but are not limited to, fluorescent or nuclear imaging techniques, such as positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), and contrast-enhanced magnetic resonance imaging (MRI). Fluorescent and / or nuclear imaging techniques can be used to monitor and / or visualize the distribution of detectably labeled compounds within a sample or a specific body part or region.

[0181] The presence or absence of the compound / protein aggregate complex is then optionally correlated with the presence or absence of alpha-synuclein aggregates in the sample or in a particular body part or region. Finally, the amount of the compound / protein aggregate complex may be compared to a normal control value determined in the sample or in a particular body part or region of healthy subjects, and an increased amount of the compound / protein aggregate complex compared to the normal control value may indicate that the patient is suffering from or at risk of developing a disease, disorder, or condition associated with alpha-synuclein aggregates.

[0182] The present invention also relates to a method for determining the amount of alpha-synuclein aggregates in tissues and / or body fluids, the method comprising: (a) preparing samples representative of the tissue and / or body fluid under investigation; (b) testing the sample for the presence of alpha-synuclein aggregates with a compound of formula (III-F); (c) determining the amount of the compound that binds to the alpha-synuclein aggregates; and (d) calculating the amount of alpha-synuclein aggregates in the tissue and / or body fluid Includes.

[0183] A sample can be tested for the presence of alpha-synuclein aggregates using a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, by contacting the sample with a compound of formula (III-F), allowing the compound of formula (III-F) to bind to alpha-synuclein aggregates to form a compound / protein aggregate complex, and detecting the formation of the compound / protein aggregate complex as described above.

[0184] Monitoring minimal residual disease, disorder, or disorder in patients suffering from a disease, disorder, or disorder associated with alpha-synuclein aggregates who are being treated with a medicament having a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof, comprises: (a) contacting a sample or a specific body part or region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to the alpha-synuclein aggregates to form a compound / protein aggregate complex; (c) detecting the formation of a compound / protein aggregate complex. (d) optionally correlating the presence or absence of the compound / protein aggregate complex with the presence or absence of alpha-synuclein aggregates in the sample or in a particular body part or region; and (e) Optionally, this may be accomplished by comparing the amount of compound / protein aggregate complex to a normal control value, where an increase in the amount of aggregate compared to the normal control value may indicate that the patient is also likely to be suffering from minimal residual disease, disorder or abnormality.

[0185] How steps (a) to (e) can be carried out has already been explained above.

[0186] In a method for monitoring minimal residual disease, disorder or abnormality, the method may comprise, prior to step (a), steps (i) to (vi): (i) contacting a sample or a specific body part or region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound specifically binds to alpha-synuclein aggregates; (ii) binding the compound to the alpha-synuclein aggregates to form a compound / (alpha-synuclein aggregate) complex; (iii) detecting the formation of a compound / (alpha-synuclein aggregate) complex; (iv) correlating the presence or absence of the compound / (alpha-synuclein aggregate) complex with the presence or absence of alpha-synuclein aggregates in the sample or in a particular body part or region; (v) optionally, comparing the amount of compound / (alpha-synuclein aggregate) complex with a normal control value; and (vi) treating the patient with a medicine It may further include:

[0187] Optionally, the method further comprises, after step (d) or step (e), step (A): (A) comparing the amount of compound / (alpha-synuclein aggregate) complex determined in step (iv) with the amount of compound / (alpha-synuclein aggregate) complex determined in step (d). It may further include:

[0188] Steps (a) through (c), and optionally steps (d) and (e) of the method for monitoring for minimal residual disease, disorder or abnormality, may be repeated one or more times over an extended period of time to monitor for minimal residual disease, disorder or abnormality.

[0189] In methods for monitoring minimal residual disease, disorder, or abnormality, the amount of compound / protein aggregate complex can optionally be compared at various time points during treatment, for example, before and after treatment begins, or at various time points after treatment begins. A change, particularly a decrease, in the amount of compound / protein aggregate complex can indicate a reduction in residual disease, disorder, or abnormality.

[0190] Predicting the response of, and treating with pharmaceuticals, patients suffering from diseases, disorders or conditions associated with alpha-synuclein aggregates (a) contacting a sample or a specific body part or region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to the alpha-synuclein aggregates to form a compound / protein aggregate complex; (c) detecting the formation of a compound / protein aggregate complex; (d) optionally correlating the presence or absence of the compound / protein aggregate complex with the presence or absence of alpha-synuclein aggregates in the sample or in a particular body part or region; and (e) Optionally, this can be achieved by comparing the amount of compound / protein aggregate complex with a normal control value.

[0191] How steps (a) to (e) can be carried out has already been explained above.

[0192] In a method for predicting reactivity, the method may include, prior to step (a), steps (i) to (vi): (i) contacting a sample or a specific body part or region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the compound specifically binds to alpha-synuclein aggregates; (ii) binding the compound to the alpha-synuclein aggregates to form a compound / (alpha-synuclein aggregate) complex; (iii) detecting the formation of a compound / (alpha-synuclein aggregate) complex; (iv) correlating the presence or absence of the compound / (alpha-synuclein aggregate) complex with the presence or absence of alpha-synuclein aggregates in the sample or in a particular body part or region; (v) optionally, comparing the amount of compound / (alpha-synuclein aggregate) complex with a normal control value; and (vi) treating the patient with a medicine It may further include:

[0193] Optionally, the method further comprises, after step (d) or step (e), step (A): (A) comparing the amount of compound / (alpha-synuclein aggregate) complex determined in step (iv) with the amount of compound / (alpha-synuclein aggregate) complex determined in step (d). It may further include:

[0194] To determine reactivity over an extended period of time, steps (a) through (c), and optionally steps (d) and (e) of the method for predicting reactivity, may be repeated one or more times.

[0195] In methods for predicting responsiveness, the amount of compound / protein aggregate complex can optionally be compared at various time points during treatment, for example, before and after initiation of treatment, or at various time points after initiation of treatment. A change, particularly a decrease, in the amount of compound / protein aggregate complex can indicate that the patient is likely to be responsive to the respective treatment.

[0196] Optionally, the diagnostic compositions can be used to visualize alpha-synuclein aggregates before, during, and after surgical procedures (e.g., deep brain stimulation (DBS)) and non-invasive brain stimulation (e.g., repetitive transcranial magnetic stimulation (rTMS)). Surgical techniques, including DBS, in addition to currently available optimal medical treatments improve advanced symptoms of PD. Over the past 20 years, rTMS has been closely examined as a possible treatment for PD (Ying-hui Chou et al., JAMA Neurol. 2015 Apr 1;72(4):432-440).

[0197] In a further embodiment of the invention, the diagnostic composition may be used in a method of collecting data for monitoring residual disease, disorder or abnormality in a patient suffering from a disease, disorder or abnormality associated with. It will be understood that the term "monitoring minimal residual disease" referred to herein relates to monitoring disease progression, for example, monitoring disease progression, disorder or abnormality in a patient suffering from a disease, disorder or abnormality associated with alpha-synuclein aggregates.

[0198] The compounds of formula (III-F) or (IV-F), or stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates thereof, may also be incorporated into test kits for detecting alpha-synuclein protein aggregates. The test kits typically include a container holding one or more compounds of formula (III-F) or one or more compounds of formula (IV-F), or stereoisomers, racemic mixtures, pharmaceutically acceptable salts, hydrates, or solvates thereof, and instructions for use of the compounds to bind to alpha-synuclein aggregates to form compound / protein aggregate complexes and to detect the formation of the compound / protein aggregate complexes, thereby correlating the presence or absence of the compound / protein aggregate complexes with the presence or absence of alpha-synuclein aggregates.

[0199] The term "test kit" generally refers to any diagnostic kit known in the art. More specifically, the term refers to any diagnostic kit described in Zrein et al., Clin. Diagn. Lab. Immunol., 1998, 5, 45-49.

[0200] The dose of the detectably labeled compound of formula (III-F), or a racemic mixture, its pharmaceutically acceptable salt, hydrate, or solvate, will vary depending on the exact compound to be administered, the patient's weight, the size and type of sample, and other variables known to a practitioner in the art. Generally, the dose can be in the range of 0.001 μg / kg to 10 μg / kg, preferably 0.01 μg / kg to 1.0 μg / kg. The radioactive dose can be, for example, 100 to 600 MBq, more preferably 150 to 450 MBq.

[0201] In another embodiment, the present invention provides a method for imaging a disease, disorder or abnormality associated with alpha-synuclein aggregates in a sample or in a specific body part or region, particularly in a sample obtained from the brain or the brain of a patient, comprising: (a) contacting a sample, specific body part, or body region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to alpha-synuclein aggregates; and (c) imaging the sample, specific body part, or body region with an imaging system. The present invention provides a method comprising:

[0202] In another embodiment, the present invention provides a method for determining the amount of alpha-synuclein aggregates in a sample or in a particular body part or body region, comprising: (a) contacting a sample, specific body part, or body region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to alpha-synuclein aggregates; (c) detecting a compound that binds to alpha-synuclein aggregates (d) determining the amount of the compound that binds to the alpha-synuclein aggregates; and (e) optionally calculating the amount of alpha-synuclein aggregates in the sample, a particular body part, or a body region; The present invention provides a method comprising:

[0203] In another embodiment, the present invention provides a method of imaging an alpha-synuclein aggregate-associated disease, disorder, or condition, comprising: (a) contacting a sample, specific body part, or body region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to alpha-synuclein aggregates; (c) detecting a compound that binds to alpha-synuclein aggregates; and (d) correlating the presence or absence of a compound that binds to alpha-synuclein aggregates with a disease, disorder, or abnormality associated with alpha-synuclein aggregates. The present invention provides a method comprising:

[0204] In another embodiment, the present invention provides a method of collecting data for diagnosing a disease, disorder, or condition associated with alpha-synuclein aggregates, comprising: (a) contacting a sample or a specific body part or region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to alpha-synuclein aggregates; (c) detecting a compound that binds to alpha-synuclein aggregates; and (d) optionally correlating the presence or absence of the compound that binds to alpha-synuclein aggregates with the presence or absence of alpha-synuclein aggregates in the sample or in a particular body part or body region. The present invention provides a method comprising:

[0205] In another embodiment, the present invention provides a method of collecting data for determining a predisposition to a disease, disorder, or condition associated with alpha-synuclein aggregates, comprising: (a) contacting a sample or a specific body part or region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to alpha-synuclein aggregates; (c) detecting a compound that binds to alpha-synuclein aggregates; and (d) optionally correlating the presence or absence of the compound that binds to alpha-synuclein aggregates with the presence or absence of alpha-synuclein aggregates in the sample or in a particular body part or body region. The present invention provides a method comprising:

[0206] If the amount of compound that binds to alpha-synuclein aggregates is higher than the normal control value for healthy / reference subjects, this indicates that the patient is suffering from or at risk of developing a disease, disorder or abnormality associated with alpha-synuclein aggregates. In particular, if the amount of compound that binds to alpha-synuclein aggregates is higher than that expected in someone who does not show clinical evidence of a neurodegenerative disease, the patient can be presumed to have a predisposition to a disease, disorder or abnormality associated with alpha-synuclein aggregates or a synucleinopathy.

[0207] In another embodiment, the present invention provides a method of collecting data for prognosing an alpha-synuclein aggregate-associated disease, disorder, or condition, comprising: (a) contacting a sample, specific body part, or body region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to alpha-synuclein aggregates; (c) detecting a compound that binds to alpha-synuclein aggregates; (d) optionally correlating the presence or absence of the compound that binds to alpha-synuclein aggregates with the presence or absence of alpha-synuclein aggregates in the sample or in a particular body part or region; and (e) optionally repeating steps (a) through (c), and, if present, optional step (d), at least once; The present invention provides a method comprising:

[0208] The likelihood (e.g., likelihood, duration, and / or extent) of progression and / or recovery from the disease, disorder, or disorder can be estimated by a medical professional based on the presence or absence of a compound that binds to alpha-synuclein aggregates, the amount of compound that binds to alpha-synuclein aggregates, etc. If necessary, steps (a) through (c), and, if present, optional step (d), can be repeated over time to monitor the progression of the disease, disorder, or disorder, and thus to make the estimation more reliable.

[0209] In another embodiment, the present invention provides a method of collecting data to monitor disease progression in a patient suffering from a disease, disorder, or condition associated with alpha-synuclein aggregates, comprising: (a) contacting a sample, specific body part, or body region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a detectably labeled compound, stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to alpha-synuclein aggregates; (c) detecting a compound that binds to alpha-synuclein aggregates; (d) optionally correlating the presence or absence of the compound that binds to alpha-synuclein aggregates with the presence or absence of alpha-synuclein aggregates in the sample or in a particular body part or region; and (e) optionally repeating steps (a) through (c), and, if present, optional step (d), at least once; The present invention provides a method comprising:

[0210] Typically, the patient is undergoing or has undergone treatment for a disease, disorder or disorder associated with alpha-synuclein aggregates, or is undergoing / has undergone treatment for a synucleinopathy. In particular, treatment may involve the administration of a medicament suitable for the treatment of a disease, disorder or disorder associated with alpha-synuclein aggregates.

[0211] In another embodiment, the present invention provides a method of collecting data to monitor the progression of an alpha-synuclein aggregate-associated disease, disorder, or condition in a patient, comprising: (a) contacting a sample, specific body part, or body region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to alpha-synuclein aggregates; (c) detecting a compound that binds to alpha-synuclein aggregates; (d) optionally correlating the presence or absence of the compound that binds to alpha-synuclein aggregates with the presence or absence of alpha-synuclein aggregates in the sample or in a particular body part or region; and (e) optionally repeating steps (a) through (c), and, if present, optional step (d), at least once; The present invention provides a method comprising:

[0212] Typically, the patient is undergoing or has undergone treatment for a disease, disorder or condition associated with alpha-synuclein aggregates, or is undergoing or has undergone treatment for a synucleinopathy. In particular, treatment may involve the administration of a medicament suitable for treating a disease, disorder or condition associated with alpha-synuclein aggregates.

[0213] In another embodiment, the present invention provides a method of collecting data for predicting the responsiveness of a patient suffering from an alpha-synuclein aggregate associated disease, disorder or disorder to treatment of the alpha-synuclein aggregate associated disease, disorder or disorder, comprising: (a) contacting a sample, specific body part, or body region suspected of containing alpha-synuclein aggregates with a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to alpha-synuclein aggregates; (c) detecting a compound that binds to alpha-synuclein aggregates; (d) optionally correlating the presence or absence of the compound that binds to alpha-synuclein aggregates with the presence or absence of alpha-synuclein aggregates in the sample or in a particular body part or region; and (e) optionally repeating steps (a) through (c), and, if present, optional step (d), at least once; The present invention provides a method comprising:

[0214] Typically, the patient is undergoing or has undergone treatment for a disease, disorder or condition associated with alpha-synuclein aggregates, or is undergoing or has undergone treatment for a synucleinopathy. In particular, treatment may involve the administration of a medicament suitable for treating a disease, disorder or condition associated with alpha-synuclein aggregates.

[0215] If the amount of compound that binds to alpha-synuclein aggregates decreases over time, the patient can be presumed to be responsive to treatment. If the amount of compound that binds to alpha-synuclein aggregates remains essentially constant or increases over time, the patient can be presumed to be non-responsive to treatment.

[0216] Alternatively, responsiveness can be estimated by determining the amount of compound that binds to alpha-synuclein aggregates. The amount of compound that binds to 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 can refer to a control value of a subject known to be responsive to a treatment, or the control value can refer to a control value of a subject known to be non-responsive to a treatment. The outcome for responsiveness can be "responsive" to a treatment, "non-responsive" to a treatment, or "unknown response" to a treatment. Response to treatment can vary for each patient.

[0217] In yet another embodiment, the present invention relates to a method for detecting alpha-synuclein aggregates, the method comprising: optionally correlating the presence or absence of a compound that binds to alpha-synuclein aggregates with the presence or absence of alpha-synuclein aggregates in a sample or a particular body part or body region; - determining the amount of the compound that binds to the alpha-synuclein aggregates - correlating the amount of compound that binds to alpha-synuclein aggregates with the amount of alpha-synuclein aggregates in the sample or in a particular body part or body region; and - optionally comparing the amount of alpha-synuclein aggregates in the sample or in a particular body part or body region with a normal control value in a healthy control subject. The method as defined herein comprises:

[0218] The control value can be, for example, a normal control value, a preclinical control value and / or a clinical control value.

[0219] A "healthy control subject" or "healthy volunteer (HV) subject" is one who does not show clinical evidence of a neurodegenerative disease.

[0220] If the amount of compound that binds to alpha-synuclein aggregates in any of the methods summarized above is greater than the normal control value, the patient can be predicted to be suffering from or susceptible to developing a disease, disorder or condition associated with alpha-synuclein aggregates or a synucleinopathy.

[0221] Any compound of formula (III-F) can be used in the methods summarized above.

[0222] The particular body part or body region is preferably that of a mammal, more preferably a human, including the whole body or a partial body region or body part of a patient suspected of containing alpha-synuclein aggregates.

[0223] The sample may be selected from tissues or body fluids suspected of containing alpha-synuclein aggregates, and the sample is obtained from a patient. Preferably, the tissue is selected from brain tissue, central nervous system tissue, and eye tissue (e.g., retinal tissue), more preferably brain tissue. Examples of body fluids include cerebrospinal fluid (CSF) or blood. The sample may be from a peripheral organ, such as, but not limited to, the intestine. The sample is obtained from a mammal, more preferably a human. Preferably, the sample is an in vitro sample from a patient.

[0224] In in vivo methods, a particular body part or region can be contacted with a compound of Formula (III-F) by administering to a patient an effective amount of the compound of Formula (III-F), the effective amount being an amount suitable to allow the presence or absence of alpha-synuclein aggregates to be determined in the particular body part or region using a selected analytical technique.

[0225] The step of binding the compound to alpha-synuclein aggregates includes allowing a sufficient time for the compound of Formula (III-F) to bind to alpha-synuclein aggregates. The amount of time required for binding depends on the type of test (e.g., in vitro or in vivo) and can be determined by one skilled in the art through routine experimentation. For in vivo methods, the amount of time depends on the time required for the compound to reach a particular body part or region suspected of containing alpha-synuclein aggregates. The amount of time should not be excessively prolonged to avoid washout and / or metabolism of the compound of Formula (III-F).

[0226] The method for detecting a compound that binds to alpha-synuclein aggregates is not particularly limited and depends, inter alia, on the type of sample, the specific body part or region, and whether the method is an in vitro or in vivo method. Possible detection methods include, but are not limited to, fluorescent or nuclear imaging techniques, such as positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), and contrast-enhanced magnetic resonance imaging (MRI). Fluorescent and / or nuclear imaging techniques can be used to monitor and / or visualize the distribution of the compound of formula (III-F) in a sample or in the body. The imaging system obtains images of the bound detectable label, such as a radioisotope, particularly a positron emitter or gamma emitter, present in the tested sample, the specific body part, or the tested body region. Preferably, the compound that binds to alpha-synuclein aggregates is detected by an imaging device, such as a PET or SPECT scanner.

[0227] The amount of compound that binds to alpha-synuclein aggregates can also be determined by visual or quantitative analysis, for example, using PET scan images.

[0228] In any of the above methods, steps (a) to (c), and, if present, optional step (d), can be repeated at least once. Repeating the steps is particularly useful for methods of collecting data for prognosis, monitoring disease progression, progression, and predicting responsiveness. In these methods, it may be advantageous to monitor the patient over a long period of time and repeat the above steps after a certain period of time. The time intervals at which the above-mentioned steps are repeated can be determined by a physician depending on the severity of the disease, disorder, or abnormality associated with alpha-synuclein aggregates or synucleinopathy.

[0229] In a further aspect, the present invention provides a method of imaging an alpha-synuclein aggregate-associated disease, disorder, or abnormality in a subject, comprising: (a) administering to a subject a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to alpha-synuclein aggregates; and (c) detecting a compound that binds to alpha-synuclein aggregates This refers to a method including:

[0230] In a further aspect, the present invention provides a method of imaging an alpha-synuclein aggregate-associated disease, disorder, or abnormality in a subject, comprising: (a) administering to a subject a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; and (b) imaging the subject's brain; The present invention covers a method including:

[0231] The subject's brain should be imaged if the compound binds to the alpha-synuclein aggregates. Compounds that bind to the alpha-synuclein aggregates can then be imaged in the subject's brain.

[0232] In a further aspect, the present invention provides a method for positron emission tomography (PET) imaging of alpha-synuclein aggregates in tissue of a subject, comprising: (a) administering to a subject a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) allowing the compound to pass into the tissue of the subject; and (c) acquiring a positron emission tomography (PET) image of the tissue of interest. wherein the tissue is central nervous system (CNS) tissue, ocular tissue, or brain tissue, preferably the tissue is brain tissue.

[0233] PET imaging should be performed as the compound passes into the tissue and as the compound binds to alpha-synuclein aggregates.

[0234] In a further aspect, the present invention provides a method for detecting a neurological disease, disorder, or abnormality associated with alpha-synuclein aggregates in a subject, comprising: (a) administering to a subject a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to alpha-synuclein aggregates; and (c) measuring the radioactive signal of the compound that binds to the alpha-synuclein aggregates. The present invention covers a method including:

[0235] The radioactive signal referred to herein is observed when a detectably labeled compound of formula (III-F) comprising at least one radioactively labeled atom binds to alpha-synuclein aggregates.

[0236] In a further aspect, the present invention provides a method (e.g., an in vivo or in vitro method) for detecting and / or quantitating alpha-synuclein aggregates in tissue of a subject, comprising: (a) contacting a tissue in a subject with a compound of Formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; (b) binding the compound to alpha-synuclein aggregates; and (c) detecting and / or quantifying compounds that bind to alpha-synuclein aggregates using positron emission tomography. The present invention covers a method including:

[0237] In yet another aspect, the present invention provides a method of imaging the brain of a subject, comprising: (a) administering to a subject a compound of formula (III-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate, or solvate thereof; and (b) obtaining an image of the subject's brain using positron emission tomography; This refers to a method including:

[0238] In the methods of the present invention, the compound of formula (III-F), or its stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate, is typically administered in a detectable amount, i.e., an amount that can be detected by the device used to detect the compound in each method. The amount is not particularly limited and depends on the compound of formula (III-F), the type of detectable label, the sensitivity of each analytical method, and each device. The amount can be appropriately selected by those skilled in the art.

[0239] Radiopharmaceutical Preparations The compound of formula (IV-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, may also be used in a kit for preparing a radiopharmaceutical preparation. Due to radioactive decay, the radiopharmaceutical is usually prepared immediately before use. The kit typically contains a compound of formula (IV-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, and a compound of formula (IV-F) reacted with the compound to produce a radiolabel ( 18F) into a compound of formula (IV-F), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof.

[0240] This invention is illustrated by the following examples which should not be construed as limiting. [Example]

[0241] All reagents and solvents were obtained from commercial sources and used without further purification.

[0242] HPLC-UV Purity: A 1 mg / mL solution of each compound was prepared in 2% TFA in water, and 5 μL was injected onto an Ascentis Express 160Å ES-C18, 5 μm, 150 × 4.6 mm column. Each peak was eluted with 0.1% TFA (mobile phase A) and 0.1% TFA in MeCN (mobile phase B), with a linear gradient from 5% to 100% B within 10 min at a flow rate of 1.5 mL / min. Detection was performed by UV at a wavelength of 254 nm.

[0243] HPLC-UV chiral purity: A solution of approximately 0.5 mg / mL of each compound is prepared in MeCN / IPA / 5 mM NHOAc = 3 / 4 / 1 (samples are filtered if necessary), and 10 μL is injected onto a Daicel Chiralpak OX-RH, 5 μm, 150 × 4.6 mm column. The precursor S-enantiomer peak (room temperature, approximately 7.9 min) is eluted isocratically at 40 °C with a mixture of MeCN:water containing 5 mM NHOAc in a 75:25 by volume ratio, using a flow rate of 1 mL / min. Detection is by UV at a wavelength of 254 nm.

[0244] Identity: The identity of each compound was obtained using a Bruker AV500 spectrometer. 1 H and 13C nuclear magnetic resonance (NMR) spectra were confirmed, while ESI-MS molecular ion confirmation was obtained using a Thermo Fisher Scientific Surveyor MSQ Plus mass detector in positive ionization mode. LC analysis was performed on an XSelect Peptide CSH C18 column, 130 Å, 3.5 μm, 150 × 2.1 mm. For sample preparation, 0.5 mg of each compound was dissolved in 1 mL MeCN / HO (1:1), and the suspension was filtered. After injection, each compound was eluted with 0.1% HCOH (mobile phase B) and 0.1% HCOH in MeCN (mobile phase A) at a flow rate of 0.4 mL / min, with a linear gradient from 5% to 100% A within 15 min.

[0245] (Preparation Example 1)

[0246] [ka]

[0247] Process A: A suspension of 2-bromo-5-hydrazinylpyridine (3.21 g, 17.07 mmol) and tert-butyl 2,4-dioxopyrrolidine-1-carboxylate (3.40 g, 17.07 mmol) in ethanol (150 mL) was refluxed for 3 hours and monitored by TLC. The crude product was concentrated under reduced pressure and diluted with dichloromethane and water. The layers were separated, and the aqueous layer was extracted twice with dichloromethane. The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by flash chromatography (silica, ethyl acetate:hexane=4:1) to give (E)-tert-butyl 4-(2-(6-bromopyridin-3-yl)hydrazono)-2-oxopyrrolidine-1-carboxylate as a brown solid (4.97 g, 79%).

[0248] Process B: The compound from Step A (3.9 g, 10.56 mmol) was stirred in 1,1-dimethoxy-N,N-dimethylmethanamine (80 mL) at 50° C. for 3 hours. The reaction was concentrated to approximately 10 mL, and ethanol was added. The solid was filtered and washed with a small amount of ethanol to give tert-butyl 2-(6-bromopyridin-3-yl)-4-oxo-4,6-dihydropyrrolo[3,4-c]pyrazole-5(2H)-carboxylate as a light brown powder (2.30 g, 57.4%).

[0249] Process C: The compound from Step B (1000 mg, 2.64 mmol) was stirred in 4 M HCl in dioxane (37 mL) at room temperature (RT) for 4 hours. The solvent was evaporated under reduced pressure, and the solid was suspended in dichloromethane. A solution of saturated NaHCO was added, and the aqueous phase was extracted twice. The combined organic layers were filtered to give 2-(6-bromopyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazol-4(2H)-one as a beige solid (682 mg, 93%).

[0250] Process N: In a round-bottom flask under argon, 2-(6-bromopyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazol-4(2H)-one (7.03 g, 25.17 mmol), (S)-pyrrolidinol (3.29 g, 37.76 mmol), and cesium fluoride (6.69 g, 50.35 mmol) were mixed in dry DMSO (70 mL). The resulting mixture was flushed with argon and stirred at 120 °C for 3 h. The reaction mixture was cooled and poured into cold water pre-cooled in an ice bath. The resulting suspension was filtered, and the solid was rinsed with ice-cold water. The solid was directly triturated in the frit using a 3 mL volume of isopropanol to give the product as a beige solid (6.75 g, 7.23 mmol, 94%).

[0251] Process O: In a flask under argon, the compound from Step N (6.26 g, 21.94 mmol), 3-bromopyridine (4.65 mL, 48.3 mmol), potassium carbonate (5.27 g, 87.75 mmol), and copper(I) iodide (1.67 g, 8.78 mmol) were mixed, and the system was flushed with argon. Dioxane (772 mL) and N1,N2-dimethylethane-1,2-diamine (1.87 mL, 17.55 mmol) were added, and the mixture was stirred at 110 °C for 22 h. The crude product was concentrated under reduced pressure and suspended in water. Ammonia was added until the solution was basic (pH 13). The aqueous layer was extracted 20 times with a solution of DCM / MeOH (9:1). The combined organic layers were dried over Na2SO4, filtered, and concentrated to dryness to give the product as a beige solid (6.36 g, 17.56 mmol, 80%).

[0252] Process P: In a vial, under argon and cooled to 0° C., (S)-2-(6-(3-hydroxypyrrolidin-1-yl)pyridin-3-yl)-5-(pyridin-3-yl)-5,6-dihydropyrrolo[3,4-c]pyrazol-4(2H)-one (100 mg, 0.276 mmol) and 4-dimethylaminopyridine (337 mg, 2.76 mmol) were mixed in pyridine (4 mL). Mesyl chloride (0.108 mL, 1.380 mmol) was added, and the mixture was flushed with argon. Pyridine (13 mL) was added, and the reaction mixture was transferred to a flask. The reaction mixture was heated to room temperature and stirred for 2 hours, after which time 4-dimethylaminopyridine (169 mg, 1.380 mmol) and mesyl chloride (0.054 mL, 0.690 mmol) were added at 0° C. The reaction was complete after 40 minutes. Then 0.1 N NaOH in water (5 mL) was added to the mixture to make it basic. If it was not basic enough, additional 1 N NaOH in water (15 mL) was added. The reaction mixture turned brown with a pH of 14. The solution was poured into cold water and filtered. The solid residue was washed with water until the pH of the water was 7. The resulting solid was dried under high vacuum for 30 minutes to give the compound as an orange solid (86 mg, 0.195 mmol, 70.8%).

[0253] It was observed that the product was not stable and was sensitive to temperature, oxygen and light.

[0254] Example 1 First crystallization: The compound from Preparation 1 (2.97 g, 6.7 mmol, purity: 93.7%) was suspended in ethanol (148 mL) and heated to reflux in an oil bath. DMSO (233 mL) was added at 80° C. until a solution was obtained. The oil bath was then removed, and the solution was cooled to room temperature. The flask was sealed and stored at 2 to 8° C. overnight. The precipitate was isolated by suction filtration and washed three times with ethanol (1.8 g, 4.1 mmol, 61% yield, 95.5% HPLC purity).

[0255] Second crystallization: The crystallized mesylate precursor from cycle 1 (1.8 g, 4.1 mmol) was suspended in DMSO (60 mL) at room temperature. The suspension was heated to 80 °C (a solution formed at approximately 70 °C). At 80 °C, ethanol (60 mL) was added, and the resulting slightly cloudy solution was stirred at room temperature for 10 min. The oil bath was removed, and the solution was allowed to cool to room temperature. The flask was sealed and stored at 2 to 8 °C overnight. The precipitate was isolated by suction filtration and washed three times with ethanol (1.6 g, 3.6 mmol, 89% yield, 96.4% HPLC purity).

[0256] Third crystallization: The crystallized mesylate precursor from Cycle 2 (1.6 g, 3.6 mmol) was further crystallized (1.5 g, 3.3 mmol, 93% yield, 96.9% HPLC purity) by repeating the conditions of Cycle 2.

[0257] Example 2 Stability Test The compound of Preparative Example 1 was transferred to a glass vial and flushed with argon. These vials were stored frozen (-20°C ± 5°C), refrigerated (2-8°C), room temperature (20-25°C), or at 40°C, both protected and unprotected from light. The material was retested by HPLC-UV weekly for the first month and monthly thereafter. Decomposition of the compound of Preparative Example 1 was noted under all conditions tested; however, this was significantly minimized when the material was stored refrigerated at -20°C and protected from light. The first major degradation product formed was detected by HPLC-UV at RT 1.6 min and identified by HPLC-ESI-MS as a pyrrole degradation product generated by elimination of the mesylate group and subsequent aromatization. The pyrrole degradation product likely has the following structure:

[0258] [ka]

[0259] The stability of the purified compound of Example 1 was also evaluated under the same stress test study conditions at 40° C. As can be seen from Figure 1, no degradation of the purified compound of Example 1 was observed upon storage at 40° C. (over 1 month), demonstrating that purification by the method of the present invention significantly improved stability.

[0260] Example 3 First crystallization: 1.21 g of the compound of Preparation 1 (HPLC purity: 93.2%) was suspended in 40 ml DMSO and heated to 80° C. The compound was completely dissolved at 80° C. 40 ml ethanol was added at 80° C. and the mixture was stirred at 80° C. for 10 minutes. The stirrer was switched off and the mixture was cooled to room temperature. The flask was stored at 2 to 8° C. overnight to complete the crystallization. The product was isolated by suction filtration of the still cold suspension and washed three times with ethanol. Yield: 963 mg, 80%, HPLC purity: 93.6%

[0261] Second crystallization: 963 mg was crystallized using the method described above to yield 880 mg, 92%, HPLC purity: 96.1%.

[0262] Third crystallization: 880 mg was crystallized using the method described above to yield 810 mg, 92%, HPLC purity: 96.7%.

[0263] Fourth crystallization: 250 mg of the material isolated from the third crystallization above was suspended in DMSO (8.9 mL) at room temperature. The suspension was heated to 80°C (a solution formed at approximately 70°C). At 80°C, ethyl acetate (8.9 mL) was added, and the resulting solution was stirred at 80°C for 10 minutes. The oil bath was removed, and the solution was allowed to cool to room temperature. The flask was sealed and stored at 2 to 8°C overnight. The precipitate was isolated by suction filtration and washed three times with ethyl acetate (200 mg, 0.45 mmol, 80% yield, 97.6% HPLC purity).

[0264] Example 4 First crystallization: 1.21 g of the compound of Preparation 1 (HPLC purity: 93.2%) was suspended in 40 ml DMSO and heated to 80° C. The compound was completely dissolved at 80° C. 40 ml ethanol was added at 80° C. and the mixture was stirred at 80° C. for 10 minutes. The stirrer was switched off and the mixture was cooled to room temperature. The flask was stored at 2 to 8° C. overnight to complete the crystallization. The product was isolated by suction filtration of the still cold suspension and washed three times with ethanol. Yield: 963 mg, 80%, HPLC purity: 93.6%

[0265] Second crystallization: 963 mg was crystallized using the method described above to yield 880 mg, 92%, HPLC purity: 96.1%.

[0266] Third crystallization: 880 mg was crystallized using the method described above to yield 810 mg, 92%, HPLC purity: 96.7%.

[0267] Fourth crystallization: 250 mg of the material isolated from the third crystallization above was suspended in DMSO (8.9 mL) at room temperature. The suspension was heated to 80°C (a solution formed at approximately 70°C). At 80°C, 2-butanone (8.9 mL) was added, and the resulting solution was stirred at 80°C for 10 minutes. The oil bath was removed, and the solution was allowed to cool to room temperature. The flask was sealed and stored at 2 to 8°C overnight. The precipitate was isolated by suction filtration and washed three times with ethyl acetate (175 mg, 0.40 mmol, 70% yield, 97.9% HPLC purity).

[0268] Example 5 First crystallization: 14.7 g of the compound of Preparation 1 (HPLC purity: 92.2%) was suspended in 484 ml DMSO and heated to 80° C. The compound was completely dissolved at 80° C. 484 ml ethanol was added at 80° C. and the mixture was stirred at 80° C. for 10 minutes. The stirrer was switched off and the mixture was cooled to room temperature. The flask was stored at 2 to 8° C. overnight to complete the crystallization. The product was isolated by suction filtration of the still cold suspension and washed three times with 118 ml ethanol. The product was transferred to a flask and dried under high vacuum for 3 hours, yield: 12.8 g, 29.1 mmol, 87.1%, HPLC purity: 96.2%.

[0269] Second crystallization: 12.8 g was crystallized using the method described above in a yield of 11.9 g, 27.0 mmol, 93%, HPLC purity: 96.9%.

[0270] Third crystallization: 11.9 g was crystallized using the method described above in a yield of 11.1 g, 25.2 mmol, 93.3%, HPLC purity: 97.1%.

[0271] Fourth crystallization: 11.1 g was crystallized using the method described above in a yield of 10.4 g, 23.6 mmol, 93.7%, HPLC purity: 97.3%.

[0272] Fifth crystallization: 10.4 g was crystallized using the method described above in a yield of 9.9 g, 22.5 mmol, 95.2%, HPLC purity: 98.1%.

[0273] The overall yield was 67.4% after the crystallization cycle.

[0274] Stability Test The stability of the purified compound prepared according to the purification process of Example 5 was evaluated under storage at -20°C ± 5°C and 5°C ± 3°C for 9 months, with impurities measured by HPLC every 3 months, and under short-term stress testing at 40°C ± 2°C and 75% ± 5% relative humidity for 2 weeks, with impurities measured by HPLC on days 0, 3, 7, and 14. The purified compound was observed to be stable over 9 months of storage at -20°C ± 5°C and 5°C ± 3°C. No significant changes in the detected impurities were observed over the long term at both temperatures. The purified compound was also found to be stable over 14 days of storage at 40°C ± 2°C and 75% ± 5% relative humidity. These results demonstrate the good stability of the purified compound prepared according to the method of the present invention.

[0275] (Example 6 (Comparative)) Comparative compounds of formula (IV-F) in which LG is nosylate or tosylate instead of mesylate were synthesized according to the method of Preparation Example 1. Tosylate compounds were not obtained by this method.

[0276] Five mg of the mesylate compound from Preparation 1 and 5 mg of the corresponding nosylate compound were radiolabeled in the presence of 10 mg Kryptofix and 1.24 mg KCO in 0.7 ml DMSO at 140° C. for 10 minutes. The labeling yield for the mesylate compound was 60%, while the labeling yield for the comparative nosylate compound was only 7%.

[0277] (Example 7 (Comparative)) The compound of Preparation 1 was recrystallized twice using ethyl acetate:DMSO as the solvent system.

[0278] First crystallization: The compound of Preparation 1 (325 mg, 0.74 mmol, purity: 93.6%) was suspended in DMSO (11.6 mL) at room temperature. The suspension was heated to 80° C. (a solution formed at approximately 70° C.). At 80° C., ethyl acetate (11.6 mL) was added, and the resulting solution was stirred at 80° C. for 10 minutes. The oil bath was removed, and the solution was cooled to room temperature. The flask was sealed and stored at 2 to 8° C. overnight. The precipitate was isolated by suction filtration and washed three times with ethyl acetate (156 mg, 0.35 mmol, 47% yield, 94.7% HPLC purity).

[0279] Second crystallization: The crystallized mesylate precursor from cycle 1 (156 mg, 0.35 mmol) was further crystallized by repeating the conditions of cycle 1 (50 mg, 0.11 mmol, 32% yield, 94.1% HPLC purity).

[0280] As can be seen, the solvent mixture of Example 7 resulted in much lower purity and yield compared to the solvent mixture used in the process of the present invention.

[0281] (Example 8 (Comparative)) The compound of Preparation 1 was recrystallized twice using 2-butanone:DMSO as the solvent system.

[0282] First crystallization: The compound of Preparation 1 (325 mg, 0.74 mmol, purity: 93.6%) was suspended in DMSO (11.6 mL) at room temperature. The suspension was heated to 80° C. (a solution formed at approximately 70° C.). At 80° C., 2-butanone (11.6 mL) was added, and the resulting solution was stirred at 80° C. for 10 minutes. The oil bath was removed, and the solution was cooled to room temperature. The flask was sealed and stored at 2 to 8° C. overnight. The precipitate was isolated by suction filtration and washed three times with 2-butanone (157 mg, 0.36 mmol, 48% yield, 94.2% HPLC purity).

[0283] Second crystallization: The crystallized mesylate precursor from cycle 1 (157 mg, 0.36 mmol) was further crystallized by repeating the conditions of cycle 1 (50 mg, 0.11 mmol, 32% yield, 94.4% HPLC purity).

[0284] As can be seen, the solvent mixture of Example 8 resulted in lower purity and much lower yield compared to the solvent mixture used in the process of the present invention.

Claims

1. Formula (IV-F) 【Chemistry 1】 (In the formula, R 3 teeth, 【Chemistry 2】 LG is selected from C 1～4 alkyl sulfonates, R 4 is aryl or 5- or 6-membered heteroaryl, and R 4 but, 【Transformation 3】 is selected from R 2a , R 2a' is independently selected from H or F; R 2b are F, -OH, C 1 ~C 4 Alkyl, HaloC 1 ~C 4 Alkyl, -NH 2 , -CN or C 1 ~C 4 independently selected from alkoxy; R 2c , R 2c' are H, F, OH, OCH 3 or CH 3 are independently selected from R 2d is selected from H, F or —OH; R 2e are H, OH, and CH 3 or F, Z is independently N, NH, N(C 1 ~C 4 alkyl), N(haloC 1 ~C 4 alkyl), O or S; Z 1 are independently N, NH, O or S, p is 0, 1 or 2; m is 0 or 1; As allowed by valence, 【Chemistry 4】 is a combination of a single bond and a double bond, * is the position of the bond), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, comprising: (i) dissolving the crude compound of formula (IV-F) in DMSO and ethanol to obtain a solution; and (ii) carrying out crystallization from the solution to obtain the purified compound of formula (IV-F) A method comprising:

2. The compound of formula (IV-F) 【Transformation 5】 or a pharmaceutically acceptable salt, hydrate or solvate thereof.

3. 3. The method of claim 1 or 2, wherein LG is mesylate.

4. (iii) isolating the purified compound of formula (IV-F) from the solution; and Optionally washing and drying the isolated purified compound of formula (IV-F).

4. The method of claim 1, further comprising:

5. 5. The method of any one of claims 1 to 4, wherein steps (i), (ii), and optionally step (iii) are carried out at least two times, preferably at least three times, more preferably at least four times, and even more preferably at least five times.

6. Step (i) is mixing the crude compound of formula (IV-F) with DMSO, and then adding ethanol to the resulting mixture.

6. The method of any one of claims 1 to 5, comprising:

7. 7. The method of any one of claims 1 to 6, wherein the ratio of DMSO to ethanol (v:v) is within the range of about 5:1 to about 0.1:1, preferably about 4:1 to about 0.5:1, more preferably about 3:1 to about 0.5:1, and even more preferably about 2:1 to about 0.75:

1.

8. 8. The method according to any one of claims 1 to 7, wherein the crude compound of formula (IV-F) is dissolved at a temperature ranging from about 50 to about 90°C, preferably from about 60 to about 90°C, more preferably from about 65 to about 85°C.

9. 9. The method according to any one of claims 1 to 8, wherein the crystallization is carried out at a temperature ranging from about 0°C to about 40°C, preferably from about 0°C to about 40°C, more preferably from about 0°C to about 30°C.

10. After step (ii) or, if present, optional step (iii), (iv) dissolving the compound of formula (IV-F) in DMSO and 2-butanone or ethyl acetate to obtain a solution; and (v) carrying out crystallization from the solution to obtain the purified compound of formula (IV-F) 10. The method of claim 1, further comprising:

11. 11. The method of any one of claims 1 to 10, wherein the purified compound of formula (IV-F) has a purity of at least 97%.

12. A compound obtainable by the method according to any one of claims 1 to 11.

13. 13. A kit for preparing a radiopharmaceutical preparation, comprising a sealed vial containing at least one compound of claim 12.

14. The radiopharmaceutical preparation is for use in a method for the diagnosis of, or a predisposition to, a disease, disorder or abnormality associated with alpha-synuclein aggregates, wherein the disease, disorder or abnormality is optionally selected from the group consisting of Parkinson's disease (including sporadic, familial with alpha-synuclein mutations, familial with non-alpha-synuclein mutations, pure autonomic failure or Lewy body dysphagia), SNCA duplicate carriers, dementia with Lewy bodies (LBD), dementia with Lewy bodies (DLB) ("pure" Lewy body dementia), (including APP mutations), 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's syndrome, multiple system atrophy (MSA) (including Shy-Drager syndrome, striatonigral degeneration or olivopontocerebellar atrophy), traumatic brain injury, chronic traumatic encephalopathy, dementia pugilistica, tauopathies (including P. Schwartz's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration, Niemann-Pick disease type C1, frontotemporal dementia with chromosome 17-linked parkinsonism), Creutzfeldt-Jakob disease, Huntington's disease, motor neuron disease, amyotrophic lateral sclerosis (including sporadic, familial or Guam ALS-dementia complex), neuroaxonal dystrophies, neurodegeneration type 1 with cerebral iron accumulation (including Hallervorden-Spatz syndrome), prion diseases, ataxia-telangiectasia, Meige syndrome, subacute myelopathy, 14. The kit of claim 13, wherein the disease is selected from sclerosing panencephalitis, Gerstmann-Sträussler-Scheinker disease, inclusion body myositis, Gaucher disease, Krabbe disease, and other lysosomal storage diseases (including Kufor-Rakeb syndrome and Sanfilippo syndrome) and rapid eye movement (REM) sleep behavior disorder, and the disease is preferably selected from Parkinson's disease, multiple system atrophy, dementia with Lewy bodies, Parkinson's disease dementia, SNCA duplication carriers, and Alzheimer's disease, more preferably multiple system atrophy.

15. 14. The kit of claim 13, wherein the radiopharmaceutical preparation is for use in imaging alpha-synuclein aggregates, the imaging preferably being performed by positron emission tomography.

16. 14. The kit of claim 13, 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, more preferably the use is for brain imaging.

17. Detectably labeled compound of formula (III-F) 【Transformation 6】 (In the formula, R 3 but, 【Transformation 7】 is selected from R 4 is aryl or 5- or 6-membered heteroaryl, and R 4 but, 【Transformation 8】 is selected from R 2a , R 2a' is independently selected from H or F; R 2b But F, -OH, C 1 ~C 4 Alkyl, HaloC 1 ~C 4 Alkyl, -NH 2 , -CN or C 1 ~C 4 independently selected from alkoxy; R 2c , R 2c' But, H, F, OH, OCH 3 or CH 3 are independently selected from R 2d is selected from H, F or —OH; R 2e But H, OH, CH 3 or F, Z is independently N, NH, N(C 1 ~C 4 alkyl), N(haloC 1 ~C 4 alkyl), O or S; Z 1 are independently N, NH, O or S; p is 0, 1 or 2; m is 0 or 1, As allowed by valence, 【Chemistry 9】 is a combination of a single bond and a double bond, * is the position of attachment), or a stereoisomer, racemic mixture, pharmaceutically acceptable salt, hydrate or solvate thereof, comprising: The compound of claim 12 or the compound prepared by the method of any one of claims 1 to 11, 18 To be replaced by F, 18 A method comprising the step of reacting with an F-fluorinating agent.

18. 18 F-fluorinating agent is K 18 F, Rb 18 F, Cs 18 F, Na 18 F, Rb 18 F, Kryptofix[222]K 18 F, 18 F Tetra (C 1～6 alkyl) ammonium salts, and tetrabutylammonium [ 18 18. The method of claim 17, wherein the compound is selected from the group consisting of [F] fluorides.

19. The detectably labeled compound of formula (III-F) is for use in a method for diagnosing a disease, disorder or abnormality associated with, or a predisposition to, alpha-synuclein aggregates, wherein the disease, disorder or abnormality is optionally Parkinson's disease (including sporadic, familial with alpha-synuclein mutations, familial with non-alpha-synuclein mutations, pure autonomic failure or Lewy body dysphagia), SNCA duplication carriers, dementia with Lewy bodies (LBD), dementia with Lewy bodies (DLB) ("pure"). Lewy body dementia (including dementia with Lewy bodies), 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's syndrome, multiple system atrophy (MSA) (including Shy-Drager syndrome, striatonigral degeneration or olivopontocerebellar atrophy), traumatic brain injury, chronic traumatic encephalopathy, dementia pugilistica, tau Pathologies (including Pick's disease, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration, Niemann-Pick disease type C1, and frontotemporal dementia with chromosome 17-linked parkinsonism), Creutzfeldt-Jakob disease, Huntington's disease, motor neuron disease, amyotrophic lateral sclerosis (including sporadic, familial, or Guam ALS-dementia complex), neuroaxonal dystrophies, neurodegeneration type 1 with cerebral iron accumulation (including Hallervorden-Spatz syndrome), prion diseases, ataxia-telangiectasia, Meige syndrome, subacute myelopathy, and encephalopathy.

19. The method of claim 17 or 18, wherein the disease is selected from chronic sclerosing panencephalitis, Gerstmann-Sträussler-Scheinker disease, inclusion body myositis, Gaucher disease, Krabbe disease, and other lysosomal storage diseases (including Kufor-Rakeb syndrome and Sanfilippo syndrome) and rapid eye movement (REM) sleep behavior disorder, and the disease is preferably selected from Parkinson's disease, multiple system atrophy, dementia with Lewy bodies, Parkinson's disease dementia, SNCA duplication carriers, and Alzheimer's disease, more preferably the disease is multiple system atrophy.

20. 19. The method of claim 17 or 18, wherein the detectably labeled compound of formula (III-F) is for use in imaging alpha-synuclein aggregates, and the imaging is preferably performed by positron emission tomography.

21. The method of claim 17 or 18, wherein the detectably labeled compound of formula (III-F) 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.

Images