Compounds for the diagnosis, treatment and prevention of diseases associated with aggregation of alpha-synuclein

Abstract

The present invention relates to compounds represented by general formula (Ia), general formula (Ib), general formula (IIa), or general formula (IIb). These compounds are suitable for imaging α-synuclein and for diagnosing diseases associated with the aggregation of α-synuclein. The compounds can also be used to treat and prevent diseases associated with the aggregation of α-synuclein. #imgabs0#

Description

SUMMARY OF THE INVENTION

[0002] The present invention relates to novel compounds suitable for imaging alpha-synuclein and for diagnosing diseases associated with alpha-synuclein aggregation. The compounds can also be used to treat and prevent diseases associated with alpha-synuclein aggregation. Background of the Invention

[0004] A large number of neurological diseases and neurodegenerative diseases are known, many of which are currently incurable and difficult to diagnose. All common neurodegenerative diseases are characterized by misfolding, aggregation and deposition of specific proteins in the brain. These diseases include medical conditions such as Parkinson's disease (PD), dementia with Lewy bodies (DLB), multiple system atrophy (MSA), Alzheimer's disease, progressive supranuclear palsy, corticobasal degeneration, frontotemporal dementia, Creutzfeldt-Jakob disease and many other diseases.

[0005] Synucleinopathy (synucleinopathy) is a group of diseases characterized by the accumulation and deposition of the alpha-synuclein (αSYN) of aggregation and misfolding.Synucleinopathy includes Parkinson's disease (PD), dementia with Lewy bodies (DLB) and multiple system atrophy (MSA).These diseases are different in the distribution of the accumulation and deposition of the alpha-synuclein of aggregation and misfolding in the central nervous system and the peripheral nervous system.In neuropathology, they can be distinguished from other neurodegenerative diseases according to the disease-specific accumulation and deposition of the alpha-synuclein of aggregation and misfolding, and other neurodegenerative diseases are characterized by the accumulation and deposition of other aggregation and misfolded proteins.For example, Alzheimer's disease is characterized by the accumulation and misfolding Abeta (A β) protein and tau protein, progressive supranuclear palsy and corticobasal degeneration are characterized by the accumulation and misfolding tau protein, and in addition, some cases of frontotemporal dementia are characterized by the accumulation and misfolding tau protein. Creutzfeldt-Jakob disease is characterized by aggregated and misfolded prion proteins.

[0006] Disease-specific accumulation and deposits of aggregated and misfolded proteins provide targets for both therapeutic and diagnostic approaches through compounds that bind to these aggregated and misfolded protein deposits.

[0007] An option for the diagnosis detection of the disease-specific accumulation and deposition of aggregated and misfolded proteins is to use a detectably labeled compound, which shows the specificity and selective high affinity of the deposits combined with the aggregated protein. This can be achieved, for example, by using a compound labeled with a suitable radioisotope and by using PET imaging for detection. Although the compound can be used for the clinical use of the PET imaging of A β and τ, there is no invention to date of the compound (Kotzbauer, PT, Tu, Z. and Mach, RH, Current status of the development of PET radiotracers for imaging alpha synuclein aggregates in Lewy bodies and Lewy neurites. Clin. Transl. Imaging, 2017.5: pages 3-14) of the diagnosis PET imaging of alpha synuclein deposits in synuclein diseases. To date, compounds developed and tested by other groups lack the appropriate combination of properties, including high binding affinity to aggregated and misfolded α-synuclein; sufficient selectivity in binding affinity compared to aggregated and misfolded other proteins, particularly Aβ and τ (which is required for differential diagnosis of different diseases characterized by disease-specific accumulation and deposits of aggregated and misfolded proteins, and for the ability to accurately detect aggregated and misfolded α-synuclein also in patients with more than one disease, namely Lewy body dementia and Alzheimer's disease). In addition, desirable properties include the ability to cross the blood-brain barrier and bind to intracellular deposits, low nonspecific binding to brain tissue, and rapid clearance of unbound compounds from the brain.

[0008] WO 2009 / 146343 relates to certain pyrazoles, 1,2,4-oxadiazoles and 1,3,4-oxadiazoles, which are tracers in positron emission tomography (PET) imaging to study amyloid deposits in the brain in vivo to allow diagnosis of Alzheimer's disease. Alzheimer's disease is characterized by the aggregation of Aβ proteins and tau proteins, and the PET tracers mentioned bind to aggregates of these proteins. In contrast, the present invention targets diseases characterized by the aggregation of α-synuclein. For selective diagnostic imaging of aggregated α-synuclein, selective binding to α-synuclein is required, as well as no to low binding to aggregated Aβ and tau.

[0009] WO 00 / 66578 describes specific NPY antagonists that are useful for treating NPY-mediated diseases / conditions, such as obesity. It is mentioned that, in addition to the "direct" effects of the compounds of WO0066578 on the NPY5 isoform, there are also diseases / conditions that would benefit from weight loss, such as insulin resistance, impaired glucose tolerance, type II diabetes, hypertension, hyperlipidemia, cardiovascular disease, gallstones, certain cancers, sleep apnea, etc.

[0010] WO 2010 / 000372 relates to a specific compound that can be used to treat or prevent diseases and / or neurodegenerative diseases associated with protein aggregation. The compound of WO 2010 / 000372 is particularly suitable for treating diseases associated with protein aggregation, including Parkinson's disease. It has been shown that they bind to proteins of several different aggregations including A β and τ. Therefore, they can be used to diagnose the disorder associated with protein aggregation, but it is impossible to reliably distinguish between the disorder associated with α-synuclein aggregation and other disorders associated with amyloid aggregation, such as tauopathy (tauopathy) or Alzheimer's disease. However, this will be important because the clinical manifestations of the disorder associated with protein aggregation are very similar, and distinguishing, for example, multiple disorders will be very desirable to adjust treatment accordingly. In addition, the molecule described in WO 2010 / 000372 has high nonspecific binding to lipids and hydrophobic proteins, which results in strong nonspecific binding in brain tissue and other tissues. Therefore, they do not achieve the desired specific binding to α-synuclein and signal-to-noise ratio required for PET tracers.

[0011] In view of the foregoing, there is a need for compounds with improved properties as diagnostic agents. In particular, the specificity of binding to α-synuclein should be improved. Nonspecific binding in brain and other tissues should be reduced, binding affinity should be increased, and the physiological half-life should be reduced.

[0012] US 2005 / 0075375 discloses specific heterocyclic compounds for the treatment of hepatitis C virus. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 : Autoradiography of human brain tissue from a patient with Lewy body dementia using [3H]-Compound 1. Detailed Description of the Invention

[0016] The present invention relates to compounds represented by general formula Ia, general formula Ib, general formula IIa or general formula IIb

[0017]

[0018] X 1 、X 2 and X 3 Independently selected from CR 2 , N and NR 1 , the condition is X 1 、X 2 and X 3 At least two of them are N or NR 1 It should be understood that N and NR 1 According to the existence allowed by the valence, N can only exist in the position =X 1 -、=X 2 - or =X 3 - and NR 1 Can only exist at position -X 1 -or-X 2 -place.

[0019] Examples include

[0020] Examples include

[0021] In a preferred embodiment,

[0022] yes (More preferably ),or yes (More preferably ).

[0023] Y 1 、Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 Independently selected from CR 3 and N, conditional on Y 1 、Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 At least one of them is N.

[0024] In preferred embodiments of Formula Ia or Formula Ib, Y 1、Y 2 、Y 3 、Y 4 、Y 5 and Y 6 Independently selected from CR 3 and N, conditional on Y 1 、Y 2 、Y 3 、Y 4 、Y 5 and Y 6 In a more preferred embodiment, at least one of Y 1 、Y 2 、Y 3 、Y 4 、Y 5 and Y 6 One, two or three of are N, and even more preferably Y 1 、Y 2 、Y 3 、Y 4 、Y 5 and Y 6 One or both are N, still more preferably Y 1 、Y 2 、Y 3 、Y 4 、Y 5 and Y 6 One of is N. In a preferred embodiment, Y 1 It's N.

[0025] In preferred embodiments of Formula Ia or Formula Ib, Y 1 、Y 3 、Y 4 and Y 6 At least one of is N. In another preferred embodiment of Formula Ia or Formula Ib, Y 1 is N and Y 3 、Y 4 and Y 6 At least one of them is N.

[0026] In preferred embodiments of Formula Ia or Formula Ib, Y 1 、Y 3 、Y 4 and Y 6 At least one of them is N and Y 1 、Y 2 、Y 3 、Y 4 、Y 5 and Y 6 The others are CR 3 (such as CH). In another preferred embodiment of Formula Ia or Formula Ib, Y1 It's N, Y 3 、Y 4 and Y 6 At least one of them is N and Y 2 、Y 3 、Y 4 、Y 5 and Y 6 The others are CR 3 (such as CH).

[0027] In a preferred embodiment of Formula Ia or Formula Ib, Y 1 is N and the others are Y 2 、Y 3 、Y 4 、Y 5 and Y 6 It's CR 3 , more preferably Y 1 is N and Y 2 、Y 3 、Y 4 、Y 5 and Y 6 It is CH.

[0028] In a preferred embodiment of Formula IIa or Formula IIb, Y 1 、Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 Independently selected from CR 3 and N, conditional on Y 1 、Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 In a more preferred embodiment, at least one of Y 1 、Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 One, two or three of are N, and even more preferably Y 1 、Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y8 One or both are N, still more preferably Y 1 、Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 One of is N. In a preferred embodiment, Y 1 It's N.

[0029] In a preferred embodiment of Formula IIa or Formula IIb, Y 1 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 At least one of is N. In a preferred embodiment of Formula IIa or Formula IIb, Y 1 、Y 3 、Y 4 、Y 5 and Y 7 At least one of is N. In another preferred embodiment of Formula IIa or Formula IIb, Y 1 is N and Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 At least one of them is N.

[0030] In a preferred embodiment of Formula IIa or Formula IIb, Y 1 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 At least one of them is N and Y 1 、Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 The others are CR 3 (such as CH). In preferred embodiments of Formula IIa or IIb, Y 1 、Y 3 、Y 4 、Y 5 and Y 7 At least one of them is N and Y 1、Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 The others are CR 3 (such as CH). In another preferred embodiment of Formula IIa or Formula IIb, Y 1 It's N, Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 At least one of them is N and Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 The others are CR 3 (such as CH). In preferred embodiments of Formula IIa or IIb, Y 1 It's N, Y 3 、Y 4 、Y 5 and Y 7 At least one of them is N and Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 The others are CR 3 (such as CH).

[0031] In a preferred embodiment of Formula IIa or Formula IIb, Y 1 is N and the others are Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 It's CR 3 , more preferably Y 1 is N and Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 It is CH.

[0032] It has surprisingly been found that the introduction of one or more nitrogen atoms as Y 1 、Y 2 、Y 3 、Y 4 、Y 5 、Y 6 、Y 7 and Y 8 The selectivity of binding to α-synuclein is strongly increased, allowing corresponding disorders associated with α-synuclein aggregation, such as Parkinson's disease, Lewy body dementia, and multiple system atrophy, to be distinguished from disorders with aggregation of other proteins, such as Alzheimer's disease, in which Aβ and tau aggregate predominantly. In addition, the signal-to-noise ratio is improved.

[0033] R 1 Selected from hydrogen, C 1-4 Alkyl and -(CH2)-OP(=O)(OR)(OR), where C 1-4 The alkyl group may be optionally substituted with one or more halogens. In a preferred embodiment, R 1 is hydrogen, methyl or 2-fluoroethyl. If present, preferably, -(CH2)-OP(=O)(OR)(OR) is attached to X 1 or X 2 .

[0034] R is hydrogen or a cation. The cation can be any pharmaceutically acceptable cation. Preferably, the cation is a monovalent cation. Examples are sodium, lithium, potassium, ammonium, and the protonated forms of ethanolamine, choline, lysine, meglumine, piperazine, and tromethamine. Preferably, the cation is sodium. In the compounds of the present invention, both R can be hydrogen, both R can be cations (the same or different cations), or one R can be hydrogen and the other R can be a cation. Preferably, both R are sodium. A divalent cation such as Ca 2+ Mg 2+ and Zn 2+ or trivalent cations such as Al 3+ is possible, but not preferred, because the resulting salt is poorly water soluble. 1 Compounds that are -(CH2)-OP(=O)(OR)(OR) and their synthesis are described in WO 2017 / 102893, which is incorporated herein by reference.

[0035] R 2 independently selected from hydrogen, halogen and C 1-4 Alkyl, where C 1-4 The alkyl group may be optionally substituted with one or more halogens. In a preferred embodiment, R 2 It's hydrogen.

[0036] R 3 It is hydrogen, halogen, C 1-4 Alkyl, OH and C 1-4 Alkoxy, where C 1-4 Alkyl and C 1-4 Alkoxy may be optionally substituted with one or more halogens. In a preferred embodiment, R 3 is hydrogen or fluorine. Even more preferably, R 3 It's hydrogen.

[0037] R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 The alkyl group may be optionally substituted with one or more halogens, or wherein R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a 4- to 6-membered saturated heterocyclic ring, wherein the 4- to 6-membered saturated heterocyclic ring has R 4 and R 5 In addition to the nitrogen atom bound to it, it optionally contains one or more heteroatoms selected from O and N, wherein the 4- to 6-membered saturated heterocyclic ring may be optionally replaced by one or more R 6 replace.

[0038] In one embodiment, R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 The alkyl group may be optionally substituted with one or more halogens. 4 and R 5 Independently selected from H and C 1-4 In a preferred embodiment, R 4 and R 5 At least one of them is C 1-4 Alkyl, where C 1-4 The alkyl group may be optionally substituted with one or more halogens. In a more preferred embodiment, R 4 is hydrogen and R 5 It is C 1-4 Alkyl, where C 1-4 The alkyl group may be optionally substituted with one or more halogens. In a more preferred embodiment, R 4 and R 5 At least one of them is C 1-4 In a more preferred embodiment, R 4 is hydrogen and R 5 It is C 1-4 alkyl.

[0039] In another embodiment, R4 and R 5 Together with the nitrogen atom to which they are bound, they form a 4- to 6-membered saturated heterocyclic ring, wherein the 4- to 6-membered saturated heterocyclic ring has R 4 and R 5 In addition to the nitrogen atom bonded to it, it optionally contains one or more (eg, one) heteroatoms selected from O and N, wherein the 4- to 6-membered saturated heterocyclic ring may be optionally replaced by one or more R 6 In a preferred embodiment, the 4- to 6-membered saturated heterocycle is selected from azetidinyl, pyrrolidinyl, piperidinyl, morpholinyl and piperazinyl, wherein azetidinyl, pyrrolidinyl, piperidinyl, morpholinyl and piperazinyl may be optionally replaced by one or more R 6 More preferably, the 4- to 6-membered saturated heterocyclic ring is selected from azetidinyl, pyrrolidinyl, piperidinyl, morpholinyl and piperazinyl, wherein the N atom of the piperazinyl group may be optionally replaced by R 6 replace.

[0040] R 6 independently selected from halogen, C 1-4 Alkyl, OH and C 1-4 Alkoxy, where C 1-4 Alkyl and C 1-4 The alkoxy group may be optionally substituted by one or more halogens, preferably, R 6 It is C 1-4 Alkyl or fluorine.

[0041] Hal is a halogen such as Br, Cl and F. Preferably, Hal is Br or F.

[0042] m is the number of cells that contain Y 1 、Y 2 、Y 3 and Y 4 R other than hydrogen in the ring 3 The number of groups. m is 0 to m 最大 An integer, where m 最大 To include Y 1 、Y 2 、Y 3 and Y 4 Preferably, m is 0.

[0043] n is the number of cells that contain Y 5 、Y 6 、Y 7 and Y 8 In the ring (Formula IIa and Formula IIb), R other than hydrogen 3 The number of groups. n is 0 to n 最大 An integer, where n 最大 To include Y 5 、Y6 、Y 7 and Y 8 Preferably, n is 0.

[0044] p is the number of 5 and Y 6 In the ring (Formula Ia and Formula Ib), R other than hydrogen 3 The number of groups. p is 0 to p 最大 integer, where p 最大 To include Y 5 and Y 6 Preferably, p is 0.

[0045] The compounds of the present invention may also exist in the form of prodrugs, solvates or salts thereof.

[0046] The compounds of the present invention form salts, which are also within the scope of the present invention. Unless otherwise indicated, reference to the compounds of the present invention herein should be understood to include reference to their salts. For example, during preparation or in an isolation step or purification step that can be used in an in vitro method, pharmaceutically acceptable (i.e., nontoxic, physiologically acceptable) salts are preferred, although other salts are also useful. The salts of the compounds of the present invention can be, for example, reacted with a compound and an amount of acid (such as an equivalent amount of acid) in a medium (such as a medium in which the salt is precipitated) or in an aqueous medium, followed by lyophilization to form.

[0047] Compounds comprising a basic moiety can form salts with a variety of organic and inorganic acids. Exemplary acid addition salts include acetates (such as acetates formed with acetic acid or trihaloacetic acids, e.g., trifluoroacetic acid), adipates, alginate, ascorbate, aspartate, benzoate, benzenesulfonate, bisulfate, borate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, hydroxyethanesulfonate (e.g., 2-hydroxyethanesulfonate), lactate, maleate, methanesulfonate, methyl ...

[0014] Examples of the present invention include succinates, naphthalenesulfonates (e.g., 2-naphthalenesulfonate), nicotinates, nitrates, oxalates, pectinates, persulfates, phenylpropionates (e.g., 3-phenylpropionate), phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates (e.g., sulfates formed with sulfuric acid), sulfonates (e.g., the sulfonates mentioned herein), tartrates, thiocyanates, toluenesulfonates such as tosylate, undecanoates, and similar acid addition salts.

[0048] Prodrugs and solvates of the compounds of the invention are also contemplated herein. The term "prodrug" as used herein refers to a compound that, upon administration to a subject, undergoes chemical transformation by metabolic or chemical processes to produce a compound of the invention or a salt and / or solvate thereof.

[0049] Solvates of the compounds of the present invention include, for example, hydrates.

[0050] All stereoisomers of the compounds herein (e.g., those that may exist due to asymmetric carbons on various substituents), including enantiomeric and diastereomeric forms, are contemplated within the scope of the present invention. Individual stereoisomers of the compounds of the present invention may, for example, be substantially free of other isomers (e.g., as pure or substantially pure optical isomers having a particular activity), or may be admixed, for example, as racemates or with all other stereoisomers or other selected stereoisomers. The chiral centers of the compounds of the present invention may have the S-configuration or the R-configuration as defined by the IUPAC 1974 Recommendations.

[0051] Racemic forms can be resolved by physical methods such as fractional crystallization, separation or crystallization of diastereomeric derivatives or separation by chiral column chromatography. Individual optical isomers can be obtained from the racemate by any suitable method, including but not limited to salt formation with an optically active acid, followed by crystallization.

[0052] All configurational isomers of the compounds of the present invention are contemplated, either in admixture or in pure or substantially pure form. The definition of the compounds of the present invention includes both cis (Z) olefin isomers and trans (E) olefin isomers, as well as cis and trans isomers of cyclic hydrocarbons or heterocycles.

[0053] Deuterated forms of the claimed compounds may also be provided. The position of deuteration is not particularly limited, but may be, for example, at R 4 and R 5 C 1-4 In the alkyl group.

[0054] Throughout the specification, groups and substituents thereof may be chosen to provide stable moieties and compounds.

[0055] The compounds of the present invention may be provided in the form of diagnostic or pharmaceutical compositions, optionally comprising a pharmaceutically acceptable carrier or excipient.

[0056] According to the present invention, the term "diagnostic composition" relates to a composition for determining the presence of aggregated alpha-synuclein in diseases associated with alpha-synuclein aggregation.

[0057] In preferred embodiments, the compounds of the present invention are detectable or detectably labeled. According to the present invention, it is understood that a compound is detectable or detectably labeled if its presence can be monitored by conventional techniques such as NMR spectroscopy, single photon emission computed tomography (SPECT), optical detection, positron emission tomography (PET), electron microscopy, magnetic resonance imaging (MRI), spectrometry, chromatography, ELISA assays, detection of radioactive emissions, preferably by PET, scintillation counting or gamma counting, more preferably by PET.

[0058] When the compounds of the invention are used as probes for imaging aggregated α-synuclein, they should be labeled. The specific nature of the label will depend on the method to be used for imaging. Typically, radiolabels that emit positrons (PET) and have a short half-life, such as 18 F. 11 C. 125 I. 123 I. 131 I. 77 Br and 76 Br, especially 18 F and 11 C, will be useful. Due to their short half-life, the labeled compounds of the present invention should be prepared shortly before they are used for testing. Therefore, the diagnostic composition of the present invention can also be provided in the form of a kit, which is composed of at least two precursors of the compound of the present invention, which react to form the desired compound of the present invention.

[0059] The skilled artisan will be able to devise methods by which detectable labels can be attached to the compounds of the invention.The following schemes may be used as illustrative examples.

[0060] use 18 F Mark :

[0061] 2-[ 18 F] Fluoroethyl tosylate is used for the incorporation of 18Useful precursors of F, such as described in WO2010 / 000372, page 32, Scheme A, for the preparation of compounds 21 and 22 starting from compound 23. Compounds A and B can be prepared in the same manner. Another method includes direct nucleophilic substitution of a suitable leaving group, as shown for the conversion of C to D in Scheme 1 (see J. Med. Chem. 2013, 56, 4568-4579, Scheme 2) or the conversion of compound 24 to compound 12.

[0062] Solution 1

[0063]

[0064] use 11 C Mark:

[0065] Marked with 11 The compound of the present invention can be prepared by using [ 11 C]MeI can be prepared by direct nucleophilic alkylation of a suitable precursor, as described in WO 2010 / 000372, page 32, Scheme B, for the preparation of compounds 27 and 28 starting from compound 23. Compound 1 of the present invention can be synthesized similarly starting from compound 2. In the same manner, compounds 18 and 19 can be synthesized starting from compound 9, as shown in Scheme 2. Alternatively, for the preparation of compounds 27 and 28 by direct nucleophilic alkylation of a suitable precursor with [C]MeI under mild conditions without loss of specific activity, 11 C] methyl iodide is converted to [ 11 C] formaldehyde to produce [ 11 A simple and available method for C] formaldehyde (JM Hooker et al., Angew. Chem. Int. Ed. 2008, 47, 5989-5992) can be used to convert compound 2 to compound 1 via reductive amination.

[0066] Option 2:

[0067]

[0068] The present invention provides a method for imaging deposits of aggregated α-synuclein, the method comprising the steps of:

[0069] (i) introducing into a subject a detectable amount of a composition comprising a detectably labeled compound of the invention;

[0070] (ii) allowing sufficient time for the compound to associate with aggregated α-synuclein; and

[0071] (iii) Detecting compounds that associate with aggregated α-synuclein.

[0072] Compositions comprising detectably labeled compounds can be introduced into a subject by any route of administration described below, such as, for example, oral or parenteral. The labeled compound can be introduced into the patient, and after a time span sufficient for the compound to become associated with aggregated α-synuclein, the labeled compound can be non-invasively detected within the patient. Alternatively, the labeled compound can be introduced into the patient, allowing sufficient time for the compound to become associated with aggregated α-synuclein, and then extracting a sample from the patient's tissue, and detecting the labeled compound in the tissue leaving the patient. Tissue samples can also be removed from the patient before the labeled compound is introduced into the tissue sample. After allowing a sufficient amount of time for the compound to become bound to aggregated α-synuclein, the compound can be detected.

[0073] Imaging of aggregated α-synuclein can also be performed quantitatively, allowing the amount of aggregated α-synuclein to be determined.

[0074] The present invention also relates to the compounds of the present invention and their prodrugs, solvates or salts for use in the treatment or prevention of diseases associated with the aggregation of α-synuclein.

[0075] Another embodiment is the use of a compound of the present invention for the preparation of a pharmaceutical composition for treating or preventing a disease associated with the aggregation of alpha-synuclein, and a method for treating or preventing a disease associated with the aggregation of alpha-synuclein, the method comprising administering a therapeutically effective amount of a compound of the present invention to a patient in need thereof. This includes administering the compound as a "pharmaceutical composition," as described below.

[0076] According to the present invention, the term "aggregation" refers to the formation of oligomeric or multimeric complexes of α-synuclein, which aggregation may be accompanied by the incorporation of additional biomolecules such as carbohydrates, nucleic acids, lipids and / or metal ions into the complexes.

[0077] As used herein, the term "disease associated with aggregation of α-synuclein" refers to those diseases characterized by the presence of aggregated α-synuclein. Such aggregated α-synuclein may form deposits in specific tissues, more preferably in neural tissue or brain tissue. The extent of aggregation depends on the specific disease.

[0078] According to the present invention, the term "pharmaceutical composition" refers to a composition for administration to a patient, preferably a mammal, more preferably a human patient. The pharmaceutical composition of the present invention comprises the compound described above and optionally another molecule capable of changing the properties of the compound of the present invention, such as stabilizing, regulating and / or activating its function. The composition can be in solid form, liquid form or gaseous form, and can especially be in the form of a powder, tablet, solution or aerosol. The pharmaceutical composition of the present invention can optionally and additionally comprise a pharmaceutically acceptable carrier or excipient. Examples of suitable pharmaceutical carriers and excipients are well known in the art and include phosphate-buffered saline solutions, water, emulsions such as oil / water emulsions, various types of wetting agents, sterile solutions, organic solvents including DMSO, etc. Compositions comprising such carriers can be formulated by well-known conventional methods.

[0079] The pharmaceutical composition will be formulated and administered in a manner consistent with good medical practice, taking into account the clinical condition of the individual patient, the site of delivery of the pharmaceutical composition, the method of administration, the schedule of administration, and other factors known to practitioners. Thus, the "effective amount" of the pharmaceutical composition for the purposes herein is determined by such considerations. The skilled artisan will appreciate that the effective amount of the pharmaceutical composition administered to an individual will depend, among other things, on the nature of the compound.

[0080] The pharmaceutical compositions of the present invention can be administered orally, rectally, parenterally, intracisternal, intravaginal, intraperitoneal, topically (e.g., by powders, ointments, drops, or transdermal patches), buccally, or as an oral or nasal spray. Preferably, when used as PET tracers for diagnosis, they will be administered intravenously, and when used for treatment or prevention of disease, they will be administered orally.

[0081] "Pharmaceutically acceptable carrier" means a non-toxic solid, semisolid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.

[0082] The term "parenteral" as used herein refers to modes of administration that include intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous, and intraarticular injection and infusion.

[0083] For therapeutic purposes, pharmaceutical composition can also be suitably used by sustained release system.Sustained release composition suitable example includes the semi-permeable polymer matrix in the form of formed articles such as film or microcapsule.Sustained release matrix includes polylactide (USP the 3rd, 773, No. 919, EP 58 481), L-glutamic acid and γ-ethyl-L-glutamate copolymer (Sidman, U. et al., Biopolymers 22:547-556 (1983)), poly (2-hydroxyethyl methacrylate) (R.Langer et al., J.Biomed.Mater.Res.15:167-277 (1981) and R.Langer, Chem.Tech.12:98-105 (1982)), ethylene vinyl acetate (R.Langer et al., ditto) or poly-D-(-)-3-hydroxybutyric acid (EP 133988). Sustained-release pharmaceutical compositions may also comprise liposomally entrapped compounds. Liposomes containing pharmaceutical compositions are prepared by methods known per se: DE 32 18 121; Epstein et al., Proc. Natl. Acad. Sci. (USA) 82: 3688-3692 (1985); Hwang et al., Proc. Natl. Acad. Sci. (USA) 77: 4030-4034 (1980); EP 52 322; EP 36 676; EP 88 046; EP 143 949; EP 142 641; Japanese Patent Application No. 83-118008; U.S. Pat. Nos. 4,485,045 and 4,544,545; and EP 102 324. Typically, liposomes are of the small (about 200-800 angstroms) unilamellar type, in which the lipid content is greater than about 30 mol% cholesterol, the selected ratio being adjusted for optimal therapy.

[0084] For parenteral administration, the pharmaceutical composition is generally formulated by mixing it at the desired degree of purity in a unit dose injectable form (solution, suspension or emulsion) with a pharmaceutically acceptable carrier, i.e., a carrier that is non-toxic to the recipient at the dosages and concentrations employed and is compatible with the other ingredients of the formulation.

[0085] Typically, the preparation is prepared by contacting the components of the pharmaceutical composition uniformly and closely with a liquid carrier or a subdivided solid carrier or both. Then, if necessary, the product is shaped into the desired preparation. Preferably, the carrier is a parenteral carrier, more preferably a solution isotonic with the recipient's blood. The example of such a carrier vehicle includes water, saline, Ringer's solution and dextrose solution. Non-aqueous vehicles such as fixed oils and ethyl oleate and liposomes are also useful herein. The carrier suitably includes a small amount of additives, such as substances that enhance isotonicity and chemical stability. Such materials are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, succinate, acetic acid, and other organic acids or salts thereof; antioxidants such as ascorbic acid; low molecular weight (less than about 10 residues) (poly)peptides, for example, polyarginine or tripeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamic acid, aspartic acid, or arginine; monosaccharides, disaccharides, and other carbohydrates, including cellulose or its derivatives, glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; counterions such as sodium; and / or nonionic surfactants such as polysorbates, poloxamers, or PEG.

[0086] The components of the pharmaceutical composition to be used for therapeutic administration must be sterile. Sterility is readily achieved by filtration through a sterile filtration membrane (e.g., a 0.2 μm membrane). The therapeutic components of the pharmaceutical composition are typically placed into a container with a sterile access port, e.g., an intravenous solution bag or vial with a stopper pierceable by a hypodermic injection needle.

[0087] The components of the pharmaceutical composition will typically be stored as an aqueous solution or as a lyophilized formulation for reconstitution in unit dose containers or multidose containers, such as sealed ampoules or vials. As an example of a lyophilized formulation, a 10 ml vial is filled with 5 ml of a sterile filtered 1% (w / v) aqueous solution, and the resulting mixture is lyophilized. The infusion solution is prepared by reconstituting the lyophilized compound with bacteriostatic water for injection.

[0088] The present invention also relates to a method for treating or preventing a disease associated with α-synuclein aggregation, which comprises administering a therapeutically effective amount of the compound of the present invention to a patient in need thereof.

[0089] As used herein, the term "therapeutically effective amount" refers to an amount sufficient to elicit a desired biological response. In the present invention, the desired biological response is to inhibit α-synuclein aggregation and / or reduce the amount of aggregated α-synuclein present in tissues.

[0090] The present invention also relates to the use of a compound as defined above for inhibiting α-synuclein aggregation in vitro or ex vivo.

[0091] The disease associated with α-synuclein aggregation is not particularly limited, and is usually selected from Parkinson's disease, Lewy body dementia, and multiple system atrophy.

[0092] The following examples are intended to illustrate the present invention. However, they should not be construed as limiting. Example

[0093] Example 1: Synthesis and testing

[0094] Chemical synthesis procedures

[0095] The following methods are presented in detail with respect to the preparation and illustrative examples of the compounds of the present invention.The compounds of the present invention can be prepared by one skilled in the art of organic synthesis from known or commercially available starting materials and reagents.

[0096] All starting materials and solvents were of commercial grade and used as received unless otherwise stated.

[0097] Thin layer chromatography (TLC) was performed using Macherey-Nagel pre-coated sheets, 0.25 mm The detection was performed on SILG / UV254 plates with UV detection and / or by carbonization with 10 wt% ethanolic phosphomolybdic acid reagent followed by heating at 200°C.

[0098] Flash column chromatography was performed using Merck silica gel 60 (0.063 mm-0.100 mm).

[0099] Analytical high performance liquid chromatography (HPLC) was performed using a Waters HPLC system with a Waters 996 photodiode array detector. All separations involved a mobile phase of 0.1% trifluoroacetic acid (TFA) (v / v) in water and 0.1% TFA in acetonitrile. HPLC was performed using a reverse phase (RP) column, Eurospher RP 18, 5μm, 250×4.6mm, 1mL·min -1 flow rate to carry out.

[0100] Electrospray ionization mass spectrometry (ESI-MS) and liquid chromatography / mass spectrometry (LC / MS) analyses were obtained using a Waters Micromass ZQ 4000 mass spectrometer coupled to the Waters HPLC instrument described above.

[0101] NMR spectra were recorded using a 400 MHz Bruker Avance spectrometer equipped with a TXI HCN z-gradient probe (Bruker AG, Rheinstetten, Germany). All spectra were processed using TOPSPIN 3.1 (Bruker AG, Karlsruhe, Germany). 1 H NMR chemical shifts (δ) are reported in parts per million (ppm) relative to CHCl 3 , DMSO-d 5 , and TFA as internal standards. Data are reported as follows: chemical shift, multiplicity (s = singlet, d = doublet, t = triplet, q = quartet, qi = quintet, dd = doublet of doublet, dt = doublet of triplet, b = broadened, m = multiplet), coupling constant (J, given in Hz), integration. 13 C NMR chemical shifts (δ) are reported in parts per million (ppm) relative to CDCl3, DMSO-d6, and TFA-d1 as internal standards. The following protocol was used to record the resonances of the compounds: 1 H-1D, 13 C-1D NMR spectroscopy and 13 C-APT (attached proton test with a single J evolution time of 1 / 145 s, the spectrum was processed so that the quaternary ammonium groups and methylene groups have positive signs, and the methyl groups and methine groups have negative signs). 1 Resonance overlap and recovery of undetectable resonances were resolved in H and APT spectra, and 2D-[ 13 C, 1 H]-HSQC (heteronuclear single quantum coherence), 2D-[ 13 C, 1 H]-HMBC (heteronuclear multiple bond correlation) and 2D-NOESY.

[0102] Selected compounds (Compound 1, Compound 2, Compound 12, Compound 18) were tritiated for binding assays using H / T exchange labeling with 99% tritium gas / Kerr's catalyst by RC TRITEC AG, Teufen, Switzerland. Compounds were delivered as ethanol solutions with a packing of 185 MBq, a concentration of 37 MBq / mL, and specific activities ranging from 1.1 GBq / mmol to 2.3 GBq / mmol.

[0103] Method A: Synthesis of 1H-pyrazoles

[0104] Those skilled in the art will recognize that compound 11A and compound 11B depicted below are two tautomeric forms of the same compound. All such tautomeric forms are considered part of the present invention. As an illustration, all tautomeric forms of the pyrazole moiety, as depicted, for example, for compound 11 below, are included within the present invention. In a related manner, those skilled in the art will recognize that the compound names contained herein are based on a naming convention in which tautomeric configurations are depicted relative to compound 11A below. Thus, the 1,3-benzodioxole substituent is located at the three-position. An alternative naming convention would be based on the tautomer compound 11B below, and in this convention, the 1,3-benzodioxole substituent is located at the five-position.

[0105]

[0106] Illustrative Example: 2-[3-(1,3-Benzodioxol-5-yl)-1H-pyrazol-5-yl]-6-fluoropyridine Compound 11

[0107]

[0108] Sodium hydride (FW 24.00, 60% in oil, 3.9 mmol, 156 mg) was added to a solution of 1-(1,3-benzodioxol-5-yl)ethanone (FW 164.16, 492 mg, 3.00 mmol) and methyl 6-fluoropyridine-2-carboxylate (FW 155.13, 605 mg, 3.9 mmol) in DMSO (7.5 mL) and THF (1.9 mL), and the reaction mixture was stirred at 20 ° C for 15 h. The reaction mixture was poured into 60 mL of ice and water containing AcOH (450 μL). The mixture was stirred for 1 h. The resulting precipitate was filtered out, washed with water (10 mL), hexane: EtOH=5: 1 (10 mL), hexane (10 mL), and dried in air to obtain the crude intermediate 1- (1,3-benzodioxole-5-yl) -3- (6-fluoropyridine-2-yl) propane -1,3- dione (594 mg) as a yellow solid. To a suspension of the crude intermediate in EtOH (20 mL) was added hydrazine hydrate (146 μL, 150 mg, 3 mmol). The reaction mixture was stirred at 70 ° C for 5 h, cooled and concentrated in vacuo. The residue was suspended in MeOH (10 mL), boiled under stirring for 5 min, cooled, filtered out, washed with MeOH (10 mL), and dried at 20 ° C in a high vacuum for 15 h to provide the pure product compound 11 (471 mg, 1.66 mmol, 55% over two steps) as a white solid.

[0109] Method B: Synthesis of 1H-pyrazoles

[0110] Illustrative Example: 4-[3-(1,3-Benzodioxol-5-yl)-1H-pyrazol-5-yl]-2-bromopyridine Compound 9

[0111]

[0112] Under nitrogen, a solution of potassium tert-butoxide (FW 112.21, 281 mg, 2.5 mmol) in dry THF (5 mL) was added to a solution of 1-(1,3-benzodioxol-5-yl)ethanone (FW 164.16, 328 mg, 2 mmol) and 2-bromopyridine-4-formic acid methyl ester (FW 216.03, 518 mg, 2.4 mmol) in dry THF (5 mL). The reaction mixture was stirred at 20 ° C for 15 h. 20 μ L aliquots were sampled, quenched with 1 M phosphate buffer pH 7, extracted with EtOAc, and analyzed by TLC. Complete conversion of ketone was observed. The mixture was poured into 1 M phosphate buffer pH 7 (15 mL) and ice water (15 mL), and stirred at 0 ° C for 30 min. The obtained yellow solid was filtered out, washed with water (5×10 mL) and air-dried to give 656 mg (1.88 mmol, 94%) of crude intermediate 1-(1,3-benzodioxol-5-yl)-3-(2-bromopyridin-4-yl)propane-1.3-dione, which was used in the next step without purification. A mixture of this intermediate and hydrazine monohydrate (FW 50.06, d 1.03; 274 μL, 282 mg, 5.64 mmol) in THF (10 mL) was stirred at 50° C. for 15 h. The cooled mixture was poured into water (40 mL) and stirred at 0° C. for 30 min. The obtained precipitate was filtered out, washed with water and air-dried. The crude product was crystallized from n-BuOH (10 mL) and DMF (1 mL) to give the pure product Compound 9 (458 mg, 1.33 mmol, 67% over 2 steps) as a white powder.

[0113] Method C: Removal of Boc protecting group

[0114] Illustrative Example: 4-[5-(2-bromopyridin-4-yl)-1H-pyrazol-3-yl]aniline Compound 7

[0115]

[0116] Trifluoroacetic acid (2mL, 2.96g, 26mmol) is added to a suspension of crude { 4- [5- (2-bromopyridin-4-yl) -1H- pyrazol-3-yl] phenyl} tert-butyl carbamate (FW 415.28, 865mg, 2.08mmol) prepared according to method B from N- (4- acetylphenyl) carbamate and 2- bromopyridine -4- methyl formate. The mixture is stirred at room temperature for 15h and concentrated in vacuo. 1M phosphate buffer pH 7 (20mL) is added, the resulting precipitate is filtered out, washed with water (2 × 10mL) and air-dried to give 543mg (1.72mmol, 69% over 3 steps) of the desired product as a yellow-orange solid.

[0117] Method D: Synthesis of 1H-pyrazoles

[0118] Illustrative Example: 4-[3-(1,3-Benzodioxol-5-yl)-1H-pyrazol-5-yl]-2-fluoropyridine Compound 12

[0119]

[0120] iPrNEt (1.79mL, 1.33g, 10.26mmol) is added to 1-(1,3-benzodioxole-5-yl) ethyl ketone (561mg, 3.42mmol) and MgBrEtO (1.56g, 6.04mmol) in CHCl(35mL) stirring mixture. Obtained suspension is stirred for 5min, and then dropwise added in CHCl2 (7mL) 2-fluoropyridine-4-formic acid pentafluorophenyl ester (1.37g, 4.45mmol). Reaction mixture is stirred for 48h. Then 1N aqueous HCl (20mL) is added, and stirring is continued for 5min. With aqueous layer CHCl(20mL) extraction, and the organic extract dried (MgSO4) combined and concentrated in a vacuum. With residue EtO (10mL) grind and filter. By solid Et o washing and air drying, to give crude product (1.14g, orange solid). The crude product is recrystallized from EtOAc (8mL) and hexane (4mL) to provide the pure intermediate 1- (1,3-benzodioxole -5- bases) -3- (2- fluoropyridine -4- bases) propane -1,3- dione (900mg, 3.13mmol, 92%) as an orange solid. The intermediate is suspended in THF (20mL), and hydrazine monohydrate (292 μ L, 300mg, 6mmol) is added. Reactant mixture is stirred for 5h at 70 ° C, cooled and concentrated in a vacuum. The residue was suspended in Et2O (10 mL), boiled with stirring for 5 min, cooled, filtered off, washed with Et2O (2×10 mL), and dried at 20°C under high vacuum for 15 h to afford the product compound 12 (722 mg, 2.55 mmol, 76% over 2 steps) as a white solid.

[0121] Method E: Synthesis of pentafluorophenyl esters

[0122] Illustrative Example: 2-Fluoropyridine-4-carboxylic acid pentafluorophenyl ester Compound 59

[0123]

[0124] DCC (FW 206.33, 2.27 g, 11 mmol) was added to a stirred suspension of 2-fluoropyridine-4-carboxylic acid (1.41 g, 10 mmol) and pentafluorophenol (1.84 g, 10 mmol) in 1,4-dioxane (40 mL). Stirring was continued for 15 h, at which time a colorless precipitate had formed. The mixture was passed through Filtration and evaporation gave a semisolid.Chromatography on silica gel gave ester compound 59 (2.21 g, 7.2 mmol, 72%) as a pure colorless oil.

[0125] Method F: Synthesis of 1H-pyrazoles

[0126] Illustrative Example: 6-[3-(3-bromophenyl)-1H-pyrazol-5-yl]-1,3-dioxolo[4,5-c]pyridine Compound 20

[0127]

[0128] Step 1

[0129] To a suspension of 1,3-dioxol [4,5-c] pyridine-6-carbaldehyde (WO / 2019 / 208509) (35 mg, 0.23 mmol) and 1-(3-bromophenyl) ethanone (46 mg, 0.23 mmol) in methanol (0.7 mL) was added Ba(OH)2*8H2O (5 mg) and NaOH (0.5 mg), and the resulting mixture was stirred at room temperature (RT) overnight. After evaporation of methanol in vacuo, the residue was ground in water (5 mL), the solid was collected by filtration, washed with cold methanol (0.5 mL) and dried to produce the intermediate compound 1-(3-bromophenyl)-3-([1,3]dioxol [4,5-c] pyridin-6-yl) prop-2-ene-1-one (67 mg, 88%) as a white solid.

[0130] Step 2

[0131] To a vigorously stirred suspension of 1-(3-bromophenyl)-3-([1,3]dioxolo[4,5-c]pyridin-6-yl)prop-2-en-1-one (67 mg, 0.2 mmol) in DMSO (0.8 mL) was added an aqueous solution of H O (30%, 45 mg, 0.4 mmol), followed by dropwise addition of aqueous NaOH (10%, 16 μL, 0.04 mmol). The yellow mixture was stirred at RT for 1.5 h and poured into cold phosphate buffer (20 mL, 0.1 M, pH 7). The oily precipitate was extracted with ethyl acetate (2*15 mL), the combined organic fractions were dried over Na SO, concentrated in vacuo, and the residue was resuspended in toluene (0.8 mL). The suspension in toluene is treated with hydrazine hydrate (35mg, 0.7mmol) and PTSA hydrate (5mg), and the mixture is stirred under reflux for 1.5h. After cooling, phosphate buffer (0.3M, 20mL) is added, and the product is extracted with ethyl acetate (2*20mL). The combined organic fractions are washed with salt water (5mL), through Na2SO4 and concentrated in vacuo. The crude residue is purified by column chromatography (15g, silica gel 63-100, CHCl3 / MeOH=100 / 1) to provide a yellow solid, which is washed with Et2O (1mL) to provide compound 20 (25mg, 36%) as a white solid.

[0132] Method G: Synthesis of 1H-pyrazoles

[0133] Illustrative Example: 4-[3-(1,3-Benzodioxin-5-yl)-1H-pyrazol-5-yl]-3,6-dichloropyridazine Compound 48

[0134]

[0135] A mixture of 1-(1,3-benzodioxol-5-yl)ethanone (138 mg, 0.84 mmol), MgBr2·Et2O (542 mg, 2.1 mmol) in DCM (5 mL) was treated with DIPEA (323 mg, 426 μL, 2.5 mmol) and stirred at RT for 10 minutes. Next, a crude mixture of 1H-benzotriazol-1-yl(3,6-dichloropyridazin-4-yl)methanone, prepared separately by stirring 3,6-dichloropyridazine-4-carboxylic acid (203 mg, 1.05 mmol), benzotriazole (125 mg, 1.05 mmol) and DCC (216 mg, 1.05 mmol) in dry DCM (5 mL) at 25°C was added dropwise over 5 minutes. The resulting mixture was stirred at 25 ° C for 12h, then treated with 0.5M aqueous HCl (2mL), and stirred at 25 ° C for another 10 minutes. After adding water (20mL), the mixture was extracted with DCM (2*15mL). The combined organic fractions were washed with salt water, dried over Na2SO4 and concentrated in vacuo. The crude product was purified by column chromatography to provide an intermediate 1- (1,3-benzodioxole-5-yl) -3- (3,6-dichloropyridazine-4-yl) propane -1,3- dione (190mg, 67%) as an orange solid, which was used for the next step without further purification. The intermediate was suspended in THF (4mL), and hydrazine monohydrate (40mg, 0.8mmol) was added. The reaction mixture was stirred overnight at 40 ° C, cooled and concentrated in vacuo. The crude product was purified by column chromatography (silica gel 63-100, 20 g, chloroform / methanol = 100 / 1) to provide compound 48 (100 mg, 36% over two steps) as a pale yellow solid.

[0136] Method H: Synthesis of 1H-imidazoles

[0137] Illustrative Example: 4-[4-(1,3-Benzodioxol-5-yl)-1H-imidazol-2-yl]-2-bromopyridine Compound 33

[0138]

[0139] Step 1

[0140] A suspension of 1-(1,3-benzodioxol-5-yl)-2-bromoethanone (729 mg, 3 mmol), 2-bromopyridine-4-carboxylic acid (606 mg, 3 mmol) and KCO (414 mg, 3 mmol) in DMF (6 mL) was stirred at 55 to 60° C. for 6 h. After cooling, the mixture was poured into water (60 mL), stirred for 10 minutes, and the resulting precipitate was collected by filtration, washed with water (20 mL) on the filter, and dried to provide the intermediate compound 2-(1,3-benzodioxol-5-yl)-2-oxoethyl 2-bromopyridine-4-carboxylate (899 mg, 87%), which was used in the next step without further purification.

[0141] Step 2

[0142] By intermediate compound 2- (1,3-benzodioxole -5- bases) -2- oxoethyl 2- bromopyridine -4- formate (364mg, 1mmol) and AcONH4 (924mg, 12mmol) suspension in toluene (7mL) at 100 DEG C under strong stirring and heating for 4h.After cooling, the reaction mixture is treated with phosphate buffer (50mL, 0.25M, pH 7) and extracted with ethyl acetate (2*50mL).The organic fractions merged are washed with salt water, through Na2SO4 dry and concentrated in vacuo.The residue is purified by column chromatography (silica gel 63-100,30g, CHCl3 / MeOH=100 / 1→100 / 3).The purified product is recrystallized from aqueous ethanol (90%) to provide compound 33 (140mg, 41%) as off-white solid.

[0143] Method I: Synthesis of 1H-1,2,4-triazoles

[0144] Illustrative Example 4-[3-(1,3-Benzodioxol-5-yl)-1H-1,2,4-triazol-5-yl]-2-bromopyridine Compound 30

[0145]

[0146] To a solution of t-BuOK (84mg, 0.75mmol) in n-BuOH (2mL) was added benzo [1,3] dioxole -5- formamidine hydrochloride (100mg, 0.5mmol) at 0°C, and the mixture was stirred at RT for 20 minutes. After adding 2- bromoisonicotinic acid hydrazide (108mg, 0.5mmol), yellow suspension was stirred at 85°C for 3h. After cooling to room temperature, ethanol (4mL) was added and CO was bubbled into the suspension for 5 minutes. After removing the solvent in a vacuum, the residue was ground in water (10mL), and the resulting precipitate was collected by filtration, washed with water (5mL) and dried to provide compound 30 (85mg, 49%) as a grey solid.

[0147] Method J: Synthesis of 1H-1,2,4-triazoles

[0148] Illustrative Example 4-[3-(2-bromopyridin-4-yl)-1H-1,2,4-triazol-5-yl]-N,N-dimethylaniline Compound 44

[0149]

[0150] A mixture of 2-bromoisonicotinic acid hydrazide (151 mg, 0.7 mmol), 4-(dimethylamino)benzonitrile (307 mg, 2.1 mmol) and K CO (48 mg, 0.5 mmol) in n-BuOH (2 mL) was stirred at 145° C. for 8 h. The mixture was concentrated in vacuo and the crude product was purified by column chromatography twice (silica gel 63-100, 20 g, CHCl / MeOH = 100 / 1) and (silica gel 63-100, 20 g, acetone / hexane = 1 / 5) to provide compound 44 (20 mg, 8%) as a beige solid.

[0151] Method K: Synthesis of 4-chloro-1H-pyrazoles

[0152] 4-[3-(1,3-Benzodioxol-5-yl)-4-chloro-1H-pyrazol-5-yl]-2-bromopyridine, Compound 28

[0153]

[0154] A suspension of compound 9 (100 mg, 0.29 mmol) and NCS (80 mg, 0.6 mmol) in water (3 mL) was stirred at 70 ° C for 16 h, TLC showed the consumption of the starting material. After cooling to RT, the precipitate was collected by filtration, washed with water (2 * 5 mL) and recrystallized from ethanol (8 mL) to provide compound 28 (52 mg, 47%) as a gray solid.

[0155] Method L: Synthesis of 1-[4-(4-R-piperazin-1-yl)phenyl]ethanones

[0156] Illustrative Example 1-{4-[4-(2-methoxyethyl)piperazin-1-yl]phenyl}ene ketone

[0157]

[0158] A mixture of 1-[4-(piperazin-1-yl)phenyl]ethanone (408 mg, 2 mmol), 1-bromo-2-methoxyethane (417 mg, 3 mmol) and CsCO (1304 mg, 4 mmol) in DMF (5 mL) was stirred at 25° C. overnight. After evaporation of DMF in vacuo, the residue was partitioned between water (25 mL) and ethyl acetate (35 mL), the aqueous phase was extracted with ethyl acetate (25 mL), and the combined organic fractions were washed with brine, dried over NaSO and concentrated in vacuo. The crude product was purified by column chromatography (30 g silica gel 63-100, chloroform → chloroform / MeOH = 100 / 2) to provide 1-{4-[4-(2-methoxyethyl)piperazin-1-yl]phenyl}vinyl ketone (525 mg, 99%) as a pale yellow solid.

[0159] 2-Methoxyethyl 2,6-dibromopicolinate and 2-methoxyethyl 2,6-dichloropicolinate were prepared according to published protocols (Journal of Medicinal Chemistry (2012), 55, 10564-10571).

[0160] tert-Butyl (4-acetylphenyl)methylcarbamate, (4-acetylphenyl)-N-( 2 H3) tert-Butyl methylcarbamate, tert-butyl (4-acetylphenyl)ethylcarbamate and tert-butyl (4-acetylphenyl)(2-fluoroethyl)carbamate were prepared according to published protocols (Organic Letters (2020), 22, 5522-5527).

[0161] 1-[4-(4-Fluoropiperidin-1-yl)phenyl]ethanone and 1-[4-(3-fluoroazetidin-1-yl)phenyl]ethanone were prepared according to published protocols (WO 2011071570 A1).

[0162] Pentafluorophenyl 2-fluoropyridine-4-carboxylate and pentafluorophenyl 2-chloropyridine-4-carboxylate were prepared according to Method E. 1-{4-[4-(2-methoxyethyl)piperazin-1-yl]phenyl}eneketone and 1-{4-[4-(2-fluoroethyl)piperazin-1-yl]phenyl}eneketone were prepared according to Method L.

[0163] Exemplary compounds of the present invention are shown in Tables 1 and 2. Table 1 shows the name, structure, IUPAC name, starting materials used for preparation, method of synthesis, and chemical yield of specific exemplary compounds. Table 2 shows the high performance liquid chromatography (HPLC) retention time, molecular weight found using low resolution mass spectrometry coupled with HPLC, and proton nuclear magnetic resonance ( 1 H-NMR).

[0164] Table 1:

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

[0171]

[0172] Table 2:

[0173]

[0174]

[0175]

[0176]

[0177]

[0178]

[0179] a Gradient 50% CH3CN / 50% H2O→100% CH3CN in 30 min

[0180] b Gradient 5% CH3CN / 100% H2O→100% CH3CN in 30 min

[0181] Example 2: Binding studies on aggregated α-synuclein, τ, and Aβ

[0182] To analyze the binding affinity and target selectivity of the compounds, two types of in vitro fibril binding assays were used: a saturation assay and a competition assay.

[0183] 1) Preparation of fibrils

[0184] α-Synuclein and τ46 were purified from Escherichia coli according to established protocols (Nuscher B, et al., J. Biol. Chem., 2004; 279(21): 21966-75). 1-42 Obtained from rPeptide, Watkinsville, GA, USA. Protofibrils were prepared by aggregating the recombinant protein with constant stirring. Specifically, 70 μM α-synuclein mixed with 100 mM NaCl in 50 mM Tris, pH 7.0 + 0.02% NaN3 was incubated at 1,400 rpm, 37°C for 96 h. 10 μM τ46 was incubated with 0.03 mg / ml heparin in 50 mM Tris, pH = 7.0 at 1,000 rpm, 37°C for 72 h. Aβ 1-42 Dissolved in 20 mM NaP i , pH = 8.0 + 0.2 mM EDTA + 0.02% NaN3 (Deeg AA, et al., Biochim. Biophys. Acta, 2015; 1850(9): 1884-90; Goedert M, et al. Nature, 1996; 383(6600): 550-3).

[0185] 2a) Saturation binding assay

[0186] Sonicated αSYN (0.04 μM) or τ46 (0.4 μM) or Aβ diluted in PBS were added to the 1-42 (6μM) fibrils with decreasing concentrations of [ 3 [H]-labeled compounds (48 nM or 24 nM-23 pM) were incubated in 50 mM Tris-base, 10% ethanol, 0.05% Tween 20, pH 7.4. To determine nonspecific binding, the corresponding unlabeled compounds (400 nM) were added to duplicate sets of binding reactions.

[0187] The assay plate was incubated at 37°C under agitation for 2h and covered with plastic foil (resealable tape, PerkinElmer, Waltham, MA, USA). Before collection, a filter (Printed filtermat B, PerkinElmer, Waltham, MA, USA) was incubated with 5mg / mL polyethyleneimine (PEI, poly (ethyleneimine) solution, Sigma Aldrich Chemie GmbH, Taufkirchen, Germany) at 4°C for 30min. After incubation, a harvester (harvester) (Filtermate harvester, PerkinElmer, Waltham, MA, USA) was used to separate the bound ligand and the free ligand by vacuum filtration. The filter was washed three times with a buffer solution cooled to 4°C and then dried in a microwave oven at medium power for 2min. A melt scintillator sheet (Melt-on scintillator sheet) (MeltiLex TM B / HS, PerkinElmer, Waltham, MA, USA) was melted into the filter using a hot plate set at 120° C. After solidification at room temperature, the filter was sealed in a plastic bag ( The accumulation of tritium was immediately detected in a liquid scintillation counter (Wallac The radioactivity was counted in a TriLux, 1450 LSC&Luminescence Counter, PerkinElmer, Waltham, MA, USA. Radioactivity was plotted against increasing concentrations of tritiated or cold compounds. Data points were fitted using nonlinear regression analysis in GraphPad Prism (GraphPad Software, Inc., Version 7.03, La Jolla, CA, USA).

[0188] 2b) Modified saturation binding assay

[0189] For saturation binding assays, fixed concentrations of sonicated human recombinant αSYN (15 nM / well) or τ46 (250 nM / well) or Aβ diluted in phosphate-buffered saline (PBS) were added to the PBS. 1-42 (1 μM / well) fibrils were incubated with increasing concentrations (from 0.05 nM to 12 nM / 24 nM) of [ 3 H]-compound 1 or [ 3[H]-Compound 2 was incubated in 30 mM Tris HCl, 10% ethanol, 0.05% Tween 20, pH 7.4 (hereinafter referred to as incubation buffer) in a total volume of 200 μL / well. Nonspecific binding of the radiotracer was determined by co-incubation with 400 nM unlabeled Compound 1 or Compound 2. A concentration determination assay was used to determine the optimal fibril concentration.

[0190] The plate, covered with removable sealing tape (PerkinElmer), was placed on a shaker (MaxQ TM 6000, track diameter 1.9 cm, Thermo Fisher Scientific Inc., Marietta, OH, USA) at 45 rpm at 37 ° C for two hours. After incubation, the bound radioligand and free radioligand were separated by vacuum filtration through a glass fiber filtermat B (PerkinElmer) using a filtermat collector (PerkinElmer). In order to collect the radioligand containing αSYN and Aβ 1-42 The filtermat was incubated with 5 mg / mL polyethyleneimine at 4°C for 30 minutes before collection. The filtermat was washed three times with 100 mL (approximately 1 mL / well) of ice-cold incubation buffer and then dried in a microwave oven at medium power for 2.5 minutes. Melt scintillator sheets (MeltiLex TM B / HS, PerkinElmer) was melted into the filter using a hot plate set at 120°C. After hardening at room temperature, the filter was sealed in a plastic sample bag (PerkinElmer). Tritium accumulation was immediately detected by Wallac The radioactivity was counted in a TriLux liquid scintillation counter (PerkinElmer). 3 The H-labeled compound concentrations were plotted. Data points were fitted using nonlinear regression analysis in GraphPad Prism (GraphPad Software, Inc., version 7.03, La Jolla, CA, USA).

[0191] 3a) Competition binding assay

[0192] Fixed concentrations of sonicated human recombinant αSYN (200 nM / well) or τ46 (208 nM / well) fibrils diluted in phosphate-buffered saline (PBS) were plated in low-binding plates (96-well micro-assay plates, Ratiolab GmbH, Dreieich, Germany) together with 1 nM [ 3H]-Compound 1 and a 1:4 serial dilution of the cold compound of interest starting at 1 μM were incubated in 50 mM Tris-base, 10% EtOH, 0.05% Tween 20, pH 7.4. To calculate K i The K value of compound 1 against sonicated human recombinant αSYN fibrils D The value was set to 3 nM.

[0193] The assay plate was incubated at 37°C for 2h under agitation and covered with plastic foil (resealable tape, PerkinElmer, Waltham, MA, USA). Before collection, a filter (Printed filtermat B, PerkinElmer, Waltham, MA, USA) was incubated with 5mg / mL polyethyleneimine (PEI, poly (ethyleneimine) solution, Sigma Aldrich Chemie GmbH, Taufkirchen, Germany) at 4°C for 30min. After incubation, a collector (Filtermate collector, PerkinElmer, Waltham, MA, USA) was used to separate the bound ligand and free ligand by vacuum filtration. The filter was washed three times with a buffer solution cooled to 4°C and subsequently dried in a microwave oven at medium power for 2min. The filter was sealed in a plastic bag ( ) together with an added scintillator (BETAPLATE SCINT, PerkinElmer, Waltham, MA, USA). The accumulation of tritium was immediately detected in a liquid scintillation counter (Wallac The radioactivity was counted in a TriLux, 1450 LSC&Luminescence Counter, PerkinElmer, Waltham, MA, USA. Radioactivity was plotted against increasing concentrations of tritiated or cold compounds. Data points were fitted using nonlinear regression analysis in GraphPad Prism (GraphPad Software, Inc., Version 7.03, La Jolla, CA, USA).

[0194] 3b) Modified competition binding assay

[0195] For competition binding experiments, a fixed concentration of recombinant αSYN fibrils (sonicated, 15 nM / well) was mixed with 1 nM [ 3 H]-compound 1 or [ 3[H]-Compound 2 and serial dilutions of decreasing concentrations of unlabeled competitor (serial dilutions 1:4 or 1:3.5 or 1:3, providing concentration ranges of 1 μM-1 pM, 1 μM-4 nM, or 1 μM-17 pM of unlabeled competitor, respectively) were incubated in 30 mM Tris-HCl, 10% ethanol, 0.05% Tween 20, pH 7.4, in a total volume of 200 μL / well. The plate was incubated at RT for 4.5 h and covered with removable sealing tape (PerkinElmer, Waltham, MA, USA).

[0196] Filtering and readout were performed as described for the saturation binding assay. 3 H]-labeled ligand binding [in CMP] was plotted against increasing competitor concentrations, and the data points were fitted using nonlinear regression analysis to calculate the IC 50 and K i The values ​​were calculated using GraphPad Software, Inc., version 7.03, La Jolla (CA), USA. In a competition binding assay using sonicated recombinant αSYN fibrils, K i The values ​​were calculated based on the K values ​​of 0.6 nM and 0.2 nM for compound 1 and compound 2, respectively. D value to complete.

[0197] Fibril binding saturation assay provides K D This assay uses directly radiolabeled compounds (e.g. 3 H mark) to carry out. K D The values ​​are shown in Table 3a and Table 3b. 1-42 The fibril aggregates produced by τ46 and τ46 were used as target structures for saturation binding assays.

[0198] The results showed high affinity for α-synuclein fibrils. Compound 1, Compound 2 and Compound 12 showed very high affinity, with K D The values ​​were below 10 nM. Compound 1 and Compound 2 were also tested for binding to Aβ and τ46 fibrils and showed good to excellent selectivity, demonstrating the suitability of these compounds for the diagnostic detection of aggregated α-synuclein.

[0199] Table 3a: Saturation determination according to protocol 2a

[0200]

[0201] Table 3b: Saturation determination according to protocol 2b

[0202] Compound number <![CDATA[K for aggregated α-synuclein D [nM]]]> 1 0.6 2 0.2

[0203] Fibril competition assays provide K i Value. K i is a quantitative measure of the concentration of non-radioactive test compound (competitor ligand) required to displace 50% of the binding of a reference compound to the target structure (1 nM), in this case [ 3 H]-compound 1 (1 nM) or [ 3 H]-compound 2 (1 nM), in our case the target structures were recombinant α-synuclein and τ46 fibrils. This assay is suitable for the screening of non-radioactive ligands. The K values ​​obtained for the tested compounds were i The values ​​are shown in Table 4a, Table 4b and Table 5.

[0204] Table 4a: Competition assay according to protocol 3a (reference ligand [ 3 H]-compound 1 (1 nM)

[0205]

[0206] *repeat

[0207] Table 4b: Competition assay according to protocol 3b (reference ligand [ 3 H]-compound 1 (1 nM)

[0208]

[0209] Table 5: Competition assay according to protocol 3b (reference ligand [ 3 H]-compound 2 (1 nM)

[0210]

[0211]

[0212]

[0213] The results of various competition assays indicated high binding affinity of α-synuclein fibrils for the tested compounds (low K i =A value indicates a higher binding affinity). In the case of competition assays relative to compound 1 (Table 4a), similar K values ​​for α-synuclein and τ46 fibrils were observed. i The values ​​indicate similar selectivity to compound 1, with K values ​​for those τ46 fibrils i The K value of α-synuclein fibrils i For compounds with higher values, the data indicate further improved selectivity.

[0214] In addition, compounds 63, 64, and 65 were tested in direct comparison with their counterparts having a nitrogen atom in one of the phenyl rings. However, a K of 0.4 nM was obtained for compound 1 in Table 6. i The K value of compound 65 (which lacks a nitrogen atom in the benzene ring) i The value was much higher (1.9 nM), indicating a significantly improved binding affinity for compound 1. i Analysis of the binding properties of the N-containing compounds revealed significantly improved binding affinities. 3 H]-K obtained for the tested compounds in the competition binding assay of compound 1 (1 nM) i The values ​​are shown in Table 6.

[0215] Table 6: Competition assay according to protocol 3b (reference ligand [ 3 H]-compound 1 (1 nM)

[0216]

[0217] *Sonicated α-synuclein fibrils

[0218] Example 3: Therapeutic effects in cell culture

[0219] To determine potential therapeutically useful effects, compounds were tested in two cell models of aggregated α-synuclein-dependent toxicity in H4 cells. These cell models allow for the induction of either α-synuclein hemi-Venus fusion constructs (V1S+SV2) or full-length α-synuclein (described in detail in: Bartels M, Weckbecker D, Kuhn PH, Ryazanov S, Leonov A, Griesinger C, Lichtenthaler SF, K, Giese A: Iron-mediated aggregation and toxicity in a novel neuronal cell culture model with inducible alpha-synuclein expression, Sci.Rep. 2019 Jun 24; 9(1): 9100) of inducible overexpression. In addition, the addition of DMSO and FeCl3 promoted the aggregation of alpha-synuclein, which was associated with increased cytotoxicity. Cell culture assays used these effects by determining changes in cell number and changes in the percentage of condensed nuclei indicating apoptosis to obtain information about which compounds showed the most promising effects and were therefore potentially useful therapeutically.

[0220] Data are summarized in table 7 hereinafter.In these experiments, in the presence of 10 μM anle138c, compound 1 to compound 19 (10 μM) as positive controls or as the DMSO of negative controls, cells and 100 μM FeCl 3 and 0.75% DMSO were incubated. After 48h, cell OPERA high-throughput imaging system was used to image the cells, and Acapella software (Perkin Elmer) was used to analyze the cells. The table shows the change in the cell number relative to the DMSO control and the change in the score of the cells (i.e., apoptotic cells) with condensed nuclei. The reduction (in the absence of the strong minimization of cell number) of the score of condensed nuclei indicates a useful therapeutic effect.

[0221] Table 7: Effects of therapeutic compounds in cell models

[0222]

[0223] Example 4: 11 In vivo biodistribution of [C]-labeled Compounds 1 and 2

[0224] [ 11 C]-labeled Compound 1 and Compound 2 were injected intravenously into the tail vein of mice. The mice were then imaged in a small animal PET instrument. Good blood-brain barrier penetration was observed for both compounds, with SUV values ​​>1.5. In addition, rapid clearance from the brain was observed. 11 The clearance half-lives of [C]-labeled Compound 1 and Compound 2 were 12 minutes and 9 minutes, respectively.

[0225] Example 5: Autoradiography using human brain tissue

[0226] Autoradiography was performed on frozen brain tissue and histological sections of brain tissue from the cingulate gyrus of a patient with Lewy body dementia treated with tritiated Compound 1. 3 When [H]-compound 1 was incubated with tissue followed by a wash step, preferential binding to gray matter was observed, a pattern consistent with the known distribution of aggregated α-synuclein ( Figure 1 , left panel). When specific binding was blocked by excess non-tritiated (i.e., "cold") Compound 1, no to very low nonspecific binding of Compound 1 to brain tissue was observed ( Figure 1 , right).

[0227] These findings indicate that Compound 1 binds specifically and with high affinity to pathologically aggregated α-synuclein present in human synucleinopathy patients and allows for the diagnostic detection of aggregated α-synuclein.

Claims

1. A compound represented by Formula Ia, Formula Ib, Formula IIa or Formula IIb or a salt thereof, in In Formula Ia: yes R 1 It's H, R 2 is H or halogen; Y 5 and Y 6 It's CH, Y 1 It's N, Y 2 、Y 3 and Y 4 It is CH; or yes R 1 and R 2 It is H; Y 5 It's CH, Y 6 is N; and Y 1 、Y 2 、Y 3 and Y 4 It is CH; In Formula Ib: yes R 1 It's H, R 2 is H or halogen; Y 5 and Y 6 It's CH, Y 1 It's N, Y 2 、Y 3 and Y 4 It is CH; or yes R 1 and R 2 It is H; Y 5 It's CH, Y 6 is N; and Y 1 、Y 2 、Y 3 and Y 4 It is CH; or yes R 1 and R 2 It is H; Y 5 and Y 6 It's CH, Y 1 is N, and Y 2 、Y 3 and Y 4 It is CH; In the general formula IIa: yes R 1 and R 2 It is H; R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 Alkyl can be optionally substituted with one or more halogens; Y 5 、Y 6 、Y 7 and Y 8 It's CH, Y 1 It's N, Y 2 and Y 4 is CH, and Y 3 is CH or N; or yes R 1 and R 2 It is H; R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 Alkyl can be optionally substituted with one or more halogens; Y 5 It's N, Y 6 、Y 7 and Y 8 It's CH, Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH; or yes R 1 It is H; R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 Alkyl can be optionally substituted with one or more halogens; Y 5 、Y 6 、Y 7 and Y 8 It is CH; Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH; or yes R 1 and R 2 It is H; R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a 4- to 6-membered saturated heterocyclic ring, said 4- to 6-membered saturated heterocyclic ring containing no additional heteroatoms, and said 4- to 6-membered saturated heterocyclic ring is optionally substituted with halogen; or R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a morpholinyl or piperazinyl group, wherein the nitrogen atom of the piperazinyl group is a C 1-4 Alkyl substituted; Y 5 、Y 6 、Y 7 and Y 8 It is CH; Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH; or yes R 1 and R 2 It is H; R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a 4- to 6-membered saturated heterocyclic ring, said 4- to 6-membered saturated heterocyclic ring containing no additional heteroatoms, and said 4- to 6-membered saturated heterocyclic ring is optionally substituted with halogen; or R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a morpholinyl or piperazinyl group, wherein the nitrogen atom of the piperazinyl group is a C 1-4 Alkyl substituted; Y 5 It is N; Y 6 、Y 7 and Y 8 It is CH; Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH; In the general formula IIb: yes R 1 and R 2 It is H; R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 Alkyl can be optionally substituted with one or more halogens; Y 5 、Y 6 、Y 7 and Y 8 It's CH, Y 1 It's N, Y 2 and Y 4 is CH, and Y 3 is CH or N; or yes R 1 and R 2 It is H; R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 Alkyl can be optionally substituted with one or more halogens; Y 5 It's N, Y 6 、Y 7 and Y 8 It's CH, Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH; or yes R 1 It is H; R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 Alkyl can be optionally substituted with one or more halogens; Y 5 、Y 6 、Y 7 and Y 8 It is CH; Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH; or yes R 1 and R 2 It is H; R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a 4- to 6-membered saturated heterocyclic ring, said 4- to 6-membered saturated heterocyclic ring containing no additional heteroatoms, and said 4- to 6-membered saturated heterocyclic ring is optionally substituted with halogen; or R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a morpholinyl or piperazinyl group, wherein the nitrogen atom of the piperazinyl group is a C 1-4 Alkyl substituted; Y 5 、Y 6 、Y 7 and Y 8 It is CH; Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH; or yes R 1 and R 2 It is H; R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a 4- to 6-membered saturated heterocyclic ring, said 4- to 6-membered saturated heterocyclic ring containing no additional heteroatoms, and said 4- to 6-membered saturated heterocyclic ring is optionally substituted with halogen; or R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a morpholinyl or piperazinyl group, wherein the nitrogen atom of the piperazinyl group is a C 1-4 Alkyl substituted; Y 5 It is N; Y 6 、Y 7 and Y 8 It is CH; Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH; or yes R 1 and R 2 It is H; R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 Alkyl can be optionally substituted with one or more halogens; Y 5 、Y 6 、Y 7 and Y 8 It is CH; Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH; Hal in Formula Ia, Formula Ib, Formula IIa or Formula IIb is halogen; or wherein the compound is:

2. The compound or salt thereof according to claim 1, wherein: In the general formula Ia yes or in Formula Ib yes R 1 It's H, R 2 is H or halogen; Y 5 and Y 6 It's CH, Y 1 It's N, Y 2 、Y 3 and Y 4 It is CH.

3. The compound or salt thereof according to claim 1, wherein: In the general formula Ia yes or in Formula Ib yes R 1 and R 2 It is H; Y 5 It's CH, Y 6 is N; and Y 1 、Y 2 、Y 3 and Y 4 It is CH.

4. The compound or salt thereof according to claim 1, wherein in the general formula Ib: yes R 1 and R 2 It is H; Y 5 and Y 6 It's CH, Y 1 is N, and Y 2 、Y 3 and Y 4 It is CH.

5. The compound or salt thereof according to claim 1, wherein: In the general formula IIa yes or in Formula IIb yes R 1 and R 2 It is H; R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 Alkyl can be optionally substituted with one or more halogens; Y 5 、Y 6 、Y 7 and Y 8 It's CH, Y 1 It's N, Y 2 and Y 4 is CH, and Y 3 It is CH or N.

6. The compound or salt thereof according to claim 1, wherein: In the general formula IIa yes or in Formula IIb yes R 1 and R 2 It is H; R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 Alkyl can be optionally substituted with one or more halogens; Y 5 It's N, Y 6 、Y 7 and Y 8 It's CH, Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH.

7. The compound or salt thereof according to claim 1, wherein: In the general formula IIa yes or in Formula IIb yes R 1 It is H; R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 Alkyl can be optionally substituted with one or more halogens; Y 5 、Y 6 、Y 7 and Y 8 It is CH; Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH.

8. The compound or salt thereof according to claim 1, wherein: In the general formula IIa yes or in Formula IIb yes R 1 and R 2 It is H; R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a 4- to 6-membered saturated heterocyclic ring, said 4- to 6-membered saturated heterocyclic ring containing no additional heteroatoms, and said 4- to 6-membered saturated heterocyclic ring is optionally substituted with halogen; or R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a morpholinyl or piperazinyl group, wherein the nitrogen atom of the piperazinyl group is a C 1-4 Alkyl substituted; Y 5 、Y 6 、Y 7 and Y 8 It is CH; Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH.

9. The compound or salt thereof according to claim 1, wherein: In the general formula IIa yes or in Formula IIb yes R 1 and R 2 It is H; R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a 4- to 6-membered saturated heterocyclic ring, said 4- to 6-membered saturated heterocyclic ring containing no additional heteroatoms, and said 4- to 6-membered saturated heterocyclic ring is optionally substituted with halogen; or R 4 and R 5 Together with the nitrogen atom to which they are bound, they form a morpholinyl or piperazinyl group, wherein the nitrogen atom of the piperazinyl group is a C 1-4 Alkyl substituted; Y 5 It is N; Y 6 、Y 7 and Y 8 It is CH; Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH.

10. The compound or salt thereof according to claim 1, wherein in the general formula IIb: yes R 1 and R 2 It is H; R 4 and R 5 Independently selected from H and C 1-4 Alkyl, where C 1-4 Alkyl can be optionally substituted with one or more halogens; Y 5 、Y 6 、Y 7 and Y 8 It is CH; Y 1 is N; and Y 2 、Y 3 and Y 4 It is CH.

11. A compound selected from the group consisting of:

12. The compound or salt thereof according to any one of claims 1 to 11, wherein the compound is detectably labeled.

13. The compound or salt thereof according to claim 12, wherein the compound is 18 F. 11 C. 125 I. 123 I. 131 I. 77 Br or 76 Br is detectably labeled.

14. The compound or salt thereof according to claim 13, wherein the compound is 18 F or 11 C is detectably labeled.

15. A diagnostic composition comprising a compound as defined in any one of claims 1 to 14 or a salt thereof, and optionally a pharmaceutically acceptable carrier.

16. A pharmaceutical composition comprising a compound as defined in any one of claims 1 to 11 or a salt thereof, and optionally a pharmaceutically acceptable carrier.

17. Use of a compound according to any one of claims 1 to 14 or a salt thereof for non-diagnostic or non-therapeutic inhibition of α-synuclein aggregation in vitro or ex vivo.

18. A kit for preparing a detectably labeled compound as defined in any one of claims 12 to 14, or a salt thereof, wherein the kit comprises at least two precursor compounds which, upon reaction, form the compound as defined in any one of claims 12 to 14, or a salt thereof.

19. Use of the compound or salt thereof according to any one of claims 1 to 14 in the preparation of a diagnostic composition for diagnosing a disease associated with α-synuclein aggregation.

20. Use of the compound or salt thereof according to any one of claims 1 to 11 in the preparation of a pharmaceutical composition for treating diseases associated with α-synuclein aggregation.

21. The use according to claim 19 or 20, wherein the disease associated with α-synuclein aggregation is selected from Parkinson's disease, Lewy body dementia and multiple system atrophy.

22. Use of a compound or salt thereof according to any one of claims 1 to 14 for non-diagnostic or non-therapeutic imaging of α-synuclein aggregates.

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