Tau protein inhibitors

Abstract

The present invention relates to compounds and pharmaceutical compositions thereof for use in the prevention or treatment of tauopathies.

Description

[Technical field]

[0001] TECHNICAL FIELD OF THEINVENTION The present invention relates to compounds and pharmaceutical compositions thereof for use in the prevention or treatment of tauopathies. [Background technology]

[0002] Tau is a microtubule-associated protein found primarily in the axons and dendrites of neurons, where it functions in the assembly and regulation of microtubules. The accumulation of insoluble tau aggregates characterizes several neurodegenerative diseases known as tauopathies, including, for example, Alzheimer's disease (AD), Pick's disease (PiD), Huntington's disease (HD), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), chronic traumatic encephalopathy (CTE), argyrophilic grain disease (AGD), and frontotemporal dementia with parkinsonism (FTDP-17). 1 The neuropathological hallmarks of AD are extracellular amyloid plaques composed of amyloid-β (Aβ) and intracellular inclusions of aggregated tau called neurofibrillary tangles (NFTs). 2 .

[0003] The extent of aggregated tau pathology is associated with the degree of clinical dementia in AD 3、4 Tau pathology is thought to spread throughout the brain in a classic manner, with tau aggregates first forming in the locus coeruleus and then spreading through the entorhinal cortex next to the hippocampus and the neocortex. 5 Therefore, potential therapeutic strategies for AD include targeting tau aggregation and spreading. Moreover, tau aggregation can be observed at least 20 years before the clinical manifestation of AD, suggesting that inhibiting this process may be a preventative measure for AD. 6、7 .

[0004] Pharmacological agents developed to target AD amyloid plaques or tau aggregates are classified as "disease-modifying therapies (DMTs)" for AD, according to the nomenclature proposed by the Common Alzheimer's and Related Dementias Research Ontology (CADRO). 8 Over the past 5 years, most DMTs in the AD drug development pipeline have been Aβ-directed drugs, with tau-directed drugs lagging behind. 9 Notable examples of Aβ-targeting DMTs are aducanumab and lecanemab, which were approved by the US Federal Drug Authority (FDA) in 2021 and 2023, respectively. However, some AD patients treated with these two antibodies have experienced cerebral hemorrhages, suggesting a low risk-to-benefit ratio. Many clinical trials for other Aβ-directed drugs have failed, mostly due to lack of efficacy. 6 In the case of tau-based DMTs, progress has been made in identifying small molecules that disrupt tau aggregation, but to date, no tau-based DMTs have been approved by the FDA.

[0005] There is therefore a need for drugs that can prevent or treat tauopathies. Summary of the Invention

[0006] The present invention relates to a compound of formula (I) for use in the prevention or treatment of a tauopathy. The present invention relates to a compound of formula (I) or a pharma- ceutically acceptable salt thereof, During the ceremony, G is TIFF2025515141000002.tif25128, preferably TIFF2025515141000003.tif24128; X is N or CH, preferably N; Y is N or CH, preferably N; L is TIFF2025515141000004.tif12128, preferably TIFF2025515141000005.tif12128; A is N or CR; preferably CR, more preferably CH; E is N or CR 1 ;Preferably CR 1 , more preferably CH; R is hydrogen, fluoro, chloro, bromo, -(C1-C6)alkyl, cyano, or -O(C1-C6)alkyl, preferably hydrogen; R 1 is hydrogen, fluoro, chloro, bromo, -(C1-C6)alkyl, cyano, or -O(C1-C6)alkyl, preferably hydrogen; R 2 is hydrogen, fluoro, chloro, bromo, -(C1-C6)alkyl, cyano, -O(C1-C6)alkyl, or difluoromethoxy, preferably -O(C1-C6)alkyl; R 3 teeth TIFF2025515141000006.tif132170, preferably TIFF2025515141000007.tif23128; R 4 is hydrogen, -(C1-C6)alkyl, or -(C3-C6)cycloalkyl; R 5 is hydrogen, fluoro, or chloro; R 6 is hydrogen, fluoro, or chloro; R 7 is hydrogen or -(C1-C6)alkyl; R 8 is hydrogen or -(C1-C6)alkyl; R 9 is hydrogen or -(C1-C6)alkyl; R 10 is hydrogen or -(C1-C6)alkyl; R 11is hydrogen, -(C1-C6)alkyl, or -CH2CON((C1-C6)alkyl); R 12 is hydrogen or -(C1-C6)alkyl; where, optionally, One or more hydrogen atoms may be replaced with deuterium atoms; One or more carbon atoms may be 11 may be replaced by C isotopes; The nitrogen atom or atoms may be selected from the group consisting of the corresponding 13 may be substituted with N isotopes; The one or more fluoro atoms may be 18 It may be replaced by F isotopes.

[0007] In a further aspect, the present invention relates to a compound of formula (I) The present invention relates to a compound of formula (I) or a pharma- ceutically acceptable salt thereof, During the ceremony, G is TIFF2025515141000009.tif25128, preferably TIFF2025515141000010.tif24128; X is N or CH, preferably N; Y is N or CH, preferably N; L is TIFF2025515141000011.tif12128, preferably TIFF2025515141000012.tif12128; A is N or CR; preferably CR, more preferably CH; E is N or CR 1 ;Preferably CR 1 , more preferably CH; R is hydrogen, fluoro, chloro, bromo, -(C1-C6)alkyl, cyano, or -O(C1-C6)alkyl, preferably hydrogen; R 1is hydrogen, fluoro, chloro, bromo, -(C1-C6)alkyl, cyano, or -O(C1-C6)alkyl, preferably hydrogen; R 2 is hydrogen, fluoro, chloro, bromo, -(C1-C6)alkyl, cyano, -O(C1-C6)alkyl, or difluoromethoxy, preferably -O(C1-C6)alkyl; R 3 teeth TIFF2025515141000013.tif125164, preferably TIFF2025515141000014.tif23128; R 4 is hydrogen, -(C1-C6)alkyl, or -(C3-C6)cycloalkyl; R 5 is hydrogen, fluoro, or chloro; R 6 is hydrogen, fluoro, or chloro; R 7 is hydrogen or -(C1-C6)alkyl; R 8 is hydrogen or -(C1-C6)alkyl, preferably -(C1-C6)alkyl; R 9 is hydrogen or -(C1-C6)alkyl, preferably -(C1-C6)alkyl; R 10 is hydrogen or -(C1-C6)alkyl; R 11 is hydrogen, -(C1-C6)alkyl, or -CH2CON((C1-C6)alkyl); R 12 is hydrogen or -(C1-C6)alkyl; However, the compound is as follows: not selected from the group consisting of TIFF2025515141000015.tif193166TIFF2025515141000016.tif188166TIFF2025515141000017.tif192166TIFF2025515141000018.tif64166; where, optionally, One or more hydrogen atoms may be replaced with deuterium atoms; One or more carbon atoms may be 11 may be replaced by C isotopes; The nitrogen atom or atoms may be selected from the group consisting of the corresponding 13 may be substituted with N isotopes; The one or more fluoro atoms may be 18 It may be replaced by F isotopes.

[0008] Compounds of the invention have been shown to covalently bind to Cys291 and Cys322 of tau and inhibit tau aggregation in vitro. Additionally, the compounds have been shown to disrupt tau aggregation in situ in cells and disrupt tau-induced pathology in vivo in a tau-overexpressing Drosophila model (see FIG. 12).

[0009] In a further aspect, the present invention is directed to a pharmaceutical composition comprising a compound of Formula (I), (Ia), and / or (Ib) and at least one pharma- ceutically acceptable carrier for use in the prevention or treatment of a tauopathy.

[0010] The compounds or pharmaceutical compositions of the present invention are further for use in the prevention or treatment of a tauopathy selected from the group consisting of Alzheimer's disease, frontotemporal dementia, primary age-related tauopathy (PART), familial British dementia (FBD), familial Danish dementia (FDD), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), Pick's disease (PiD), dementia with Lewy bodies (DLB), progressive supranuclear palsy (PSP), glioglobular tauopathy (GGT), tauopathy with hippocampal 4-repeat tau immunoreactive spherical inclusions, limbic-predominant neuronal inclusion body 4R tauopathy. tauopathy (LNT), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), argyrophilic grain disease (AGD), Huntington's disease, glioglobular tauopathy, neuroastroglial tauopathy variant in elderly, familial behavioural variant frontotemporal dementia associated with astrocyte-predominant tauopathytauopathy), amyotrophic lateral sclerosis (ALS), spinocerebellar ataxia type 11, spinal muscular atrophy (SMA), progressive ataxia and palatal tremor-associated tauopathy, cerebral amyloid angiopathy (CAA), IgLON5 antibody-associated tauopathy, trisomy 21-related Alzheimer's disease (Down's syndrome), vascular dementia, cerebral amyloid angiopathy, Gerstmann-Sträussler-Scheinker disease (GSS), Creutzfeldt-Jakob disease, Fatal familial insomnia, kuru, Niemann-Pick disease type C related tauopathy, nodding syndrome, non-Guam motor neuron disease with neurofibrillary tangles, Parkinson's disease (PD), Parkinson's disease with dementia, Parkinsonism-dementia in Guam, Guadeloupian parkinsonism, Kosaka-Shibayama disease, postencephalitic parkinsonism, SYNJ1 (PARK20) early-onset recessive parkinsonism with seizures and dystonia-associated nigral tau pathology (SYNJ1 (PARK20) early-onset recessive form of parkinsonism with seizures and dystonia associated nigral tau pathology, tau pathology associated with multiple system atrophy, familial parkinsonism and progressive respiratory failure, limbic-predominant neuro-glial tau pathology in TARDBP mutation (I383;P112H), neuronal 4R tau pathology in fatal familial insomnia (PRNP D178N mutation), tau pathology associated with ADCY5 dyskinesia, tau pathology in chronic temporal lobe epilepsy, neurodegeneration with intracranial iron deposition (NBIA), tau pathology in NBIA PANK2 and WDR45 mutations, tau pathology in NBIA PLA2G6 mutations, tau pathology in NBIA associated with autosomal dominant mitochondrial membrane protein-associated neurodegeneration (MPAN), pretangles and neurofibrillary tangles in HIV-negative opiate abusers pretangles and neurofibrillary tangles in human immunodeficiency virus (HIV)-negative opiatetau pathology associated with chronic myelopathy (CMM), diffuse neurofibrillary tangles with calcifications, progressive ataxia and palatal tremor, SLC9A6-associated parkinsonism, tau pathology associated with SPG7 gene mutations, striatal 4R tau pathology associated to X-linked parkinsonism with spasticity ATP6AP2), tau pathology associated with SPAST-related hereditary spastic paraplegia, autism, autism spectrum disorder, retinal tauopathy, West Nile encephalomyelitis, TTBK2-related spinocerebellar degeneration11, herpes simplex encephalitis, neurofibrillary tangle dementia (TOD), age-related tau astrogliopathy (ARTAG), hippocampal tauopathy, subacute sclerosing panencephalitis (SSPE)-associated tauopathy, FTLD-C9ORF72, Christianson syndrome, vacuolar tauopathy tauopathy), Ritiko-Bodig disease, ganglioglioma and gangliocytoma, meningioangiomatosis, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, neuronal ceroid lipofuscinosis, myotonic dystrophy, Fukuyama-type congenital muscular dystrophy, unilateral megalencephaly, focal cortical dysmorphism, Walcott-Rallison syndrome, primary lateral sclerosis, progressive freezing of gait, PSP with parkinsonism, Richardson syndrome, non-fluent / agrammatic primary progressive aphasia aphasia, semantic primary progressive aphasia, logopenic primary progressive aphasia, primary progressive apraxia of speech, amnesic Alzheimer's disease, preferably Alzheimer's disease (AD), Pick's disease (PiD), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), chronic traumatic encephalopathy (CTE), argyrophilic grain disease (AGD), frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD), as well as primary age-related tauopathy (PART). [Brief description of the drawings]

[0011] [Figure 1]In vitro seeding aggregation / fibrillization of full-length tau is dose-dependently inhibited by osimertinib, compound 2 (AZ7550) and compound 3 (AZ5104). (A) Effect of various formulations of osimertinib, compound 2 and compound 3 on the ThT fluorescence curves of 2N4R tau. Within 3 days, tau aggregation reached an equilibrium state corresponding to maximum ThT fluorescence. At this point, the aggregation mixture containing osimertinib was pelleted and the amount of soluble tau in the mixture was quantified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). A representative SDS-PAGE gel is shown in B. The amount of soluble tau in the absence of osimertinib (lane 3) is approximately 80-90% lower than the no-fibrillization control (lane 2). (C) At a molar ratio of 1:10 (tau:osimertinib), osimertinib significantly increases the amount of soluble tau. Error bars correspond to the standard deviation of band intensities from two replicates. Statistical significance was determined by one-way ANOVA with Dunnett's multiple comparison test, p<0.033(*). [Diagram 2] In vitro seeding fibrillation assay of 2N4R tau in the presence of osimertinib and 2.5% (v / v) dimethyl sulfoxide (DMSO). Osimertinib without cosolvent or counterion is poorly soluble in water, which interferes with the fluorescence-based aggregation assay. To circumvent this issue, aggregation assays and other experiments may be performed in the presence of the cosolvent DMSO. At a DMSO concentration of 2.5% (v / v), there is a strong inhibitory activity of osimertinib against tau aggregation. [Diagram 3]Osimertinib mesylate inhibits the aggregation of wild-type tau (tauwt) and 3R tau (tau3R) more effectively than that of cysteine-free tau mutants (tauC291S, C322S). In vitro seeded aggregation / fibrillization was performed by incubating monomeric 2N4R tauwt, tau3R or 2N4R tauC291S, C322S with osimertinib mesylate (tau:compound molar ratios of 1:3 and 1:10) and the increase in ThT fluorescence was monitored over time (A). Normalized Tm values ​​for the 1:3 and 1:10 molar ratios show no clear dose-dependence (B). However, statistically lower normalized Span values ​​were observed at 1:3 and 1:10 molar ratios for tau wt and tau3R, but not for tau C291S and C322S, indicating the involvement of cysteine ​​residues in the binding to the compounds (C). Statistical significance was determined by one-way unpaired t-test. Significant differences corresponding to p ≤ 0.002 (**), p ≤ 0.0002 (***), and p ≤ 0.0001 (****) are indicated. [Figure 4] Osimertinib, compound 2 (AZ7550 hydrochloride), and compound 3 (AZ5104) partially dissolve 2N4R tau aggregates in vitro. (A) ThT fluorescence curves monitoring tau aggregation. Tau fibrillization reached a saturation point within approximately 40 h, and then compounds or buffer (reference) were added to tau fibrils (tau monomer:compound molar ratios of 1:3 and 1:10). Experiments were performed in duplicate. (B) Addition of compounds reduces ThT fluorescence intensity, indicating partial dissolution of tau aggregates. Statistical significance was determined by one-way ANOVA with Dunnett's multiple comparison test. Significant differences correspond to p ≤ 0.0002 (***) and p ≤ 0.0001 (****). (C) After 1 day of incubation with compounds, transmission electron micrographs of tau-compound mixtures show the presence of fibrils, indicating that the compounds do not completely dissolve the fibrils. Scale bar is 500 nm. (D) SDS-PAGE analysis of pelleted tau-compound mixtures shows increased amounts of soluble tau after incubation of tau fibrils with compound 3 (AZ5104) and compound 2 (AZ7550 hydrochloride (1:10 tau:compound)). The dotted line corresponds to one standard deviation above the reference value. [Figure 5A] NMR spectroscopy reveals the binding of osimertinib mesylate, compound 2 (AZ7550 hydrochloride), and compound 3 (AZ5104) to tau. 15N-labeled 2N4R tau (18 μM tau in 50 mM sodium phosphate pH 6.8) was incubated for 16 h at 37 °C in the presence or absence of compounds, and then SOFAST-heteronuclear multiple quantum coherence (HMQC) spectra were recorded at 5 °C. (A) Overlay spectra of 2N4R tau with (black) and without (gray) a 100-fold molar excess of osimertinib mesylate. (B) Enlargement of the boxed region in (A) shows the significantly broadened peaks of residues belonging to the N-terminal region of tau. Chemical shift perturbations (CSPs) above about 0.002 ppm as well as signal broadening (I / I0<1) induced by the addition of the compounds suggest binding of the compounds to the monomeric form of tau. I / I0 values ​​above 1 suggest that the compounds dissolve tau oligomers formed during the 16-h incubation period, as seen in the case of compound 3 (AZ5104), leading to an increase in NMR signal intensity. [Figure 5B] See legend to Figure 5A. [Figure 5C] See legend to Figure 5A. [Figure 6] Electrospray ionization mass spectrometry (ESI-MS) confirms covalent modification of cysteines of tau (244-372, 13818 Da) by osimertinib (500 Da), compound 2 (AZ7550) without the hydrochloride salt (486 Da), and compound 3 (AZ5104) (486 Da). [Figure 7A] MS / MS analysis of osimertinib adducts of tau. Full-length 2N4R tau was incubated with osimertinib mesylate (1:5 molar ratio of tau:compound) at 37 °C overnight and subjected to tryptic digestion, and the tryptic digest was analyzed by LC-MS / MS. Peptide mass spectra corresponding to osimertinib-modified C291 (A) and C322 (B) confirm the covalent modification of cysteines in tau. [Figure 7B] See legend to Figure 7A. [Figure 7C] MS / MS analysis of osimertinib adducts in tau. Full-length 2N4R tau was incubated with osimertinib mesylate (1:5 molar ratio of tau:compound) at 37 °C overnight and subjected to tryptic digestion, and the tryptic digest was analyzed by LC-MS / MS. (C) Overall, approximately 350 tau peptide spectral matches indicate osimertinib-modified cysteine ​​sites in osimertinib-treated tau samples. [Figure 7D] MS / MS analysis of osimertinib adducts of tau. Full-length 2N4R tau was incubated with osimertinib mesylate (1:5 molar ratio of tau:compound) at 37 °C overnight, subjected to tryptic digestion, and the tryptic digest was analyzed by LC-MS / MS. (D) The negative control corresponding to tau incubated at 37 °C without compound shows a small number of false positive peptide spectral matches. [Figure 8A] Determination of the binding epitope of osimertinib on monomeric tau compared to those of compound 2 (AZ7550 hydrochloride) and compound 3 (AZ5104). Addition of monomeric 2N4R tau (10 μM and 20 μM) to the compounds (250 μM / 50 mM sodium phosphate buffer containing 2.5% (v / v) DMSO, pH 6.8, 37 °C) induced chemical shift perturbations (CSPs) in the 1D NMR spectra of the compounds, allowing the identification of aliphatic and aromatic protons involved in binding with monomeric tau (A-B). The CSPs were localized and mapped to the chemical structure of osimertinib (C). The larger CSPs are likely to be the binding epitopes of osimertinib. [Figure 8B] See legend to Figure 8A. [Figure 8C] See legend to Figure 8A. [Figure 9]Determination of the tau fibril binding epitope of osimertinib by saturation transfer difference (STD) NMR spectroscopy. Tau fibrils were incubated with a 48-fold excess of osimertinib (A), compound 2 (AZ7550 hydrochloride) (B), or compound 3 (AZ5104) (C) in 50 mM sodium phosphate containing 100 mM NaCl and 2.5% (v / v) DMSO, pH 6.8, at 37 °C. 1D 1H NMR STD spectra were acquired in an interleaved fashion alternating between off-resonance irradiation at 60 ppm and on-resonance irradiation at -2 ppm. Control 1D spectra are shown in grey and difference spectra in black. (D-F) The strength of the STD effect (ISTD) was measured and normalized with respect to the maximum STD signal (indole methyl protons set as 100%). (G-I) Chemical structures of osimertinib, compound 2 (AZ7550 hydrochloride), and compound 3 (AZ5104) enclose protons that exhibit STD effects. These protons represent the tau fibril-binding epitopes of each compound. [Figure 10] Osimertinib inhibits tau seeding aggregation in HEK biosensor cells. Intracellular aggregate formation was induced by lysates of HEK cells containing tau aggregates seeded by tau P301L fibrils. Cells were incubated with the drug (0.25 μM or 7.5 μM) for 22 h in the presence of lipofectamine. Cells incubated with higher concentrations of osimertinib showed reduced aggregate formation. [Figure 11] Comparison of tau aggregation inhibitory activity in HEK biosensor cells of osimertinib, compound 2 (AZ7550) hydrochloride, and compound 3 (AZ5104). Cell assays were performed as previously described (see Figure 10) with different batches of HEK cells (each dose-response curve corresponds to one batch of cells), and the number of cells with aggregates (gray) was quantified relative to the negative control, which was cells treated with vehicle (DMSO). The total amount of cells (black) remained close to 100% until the compound concentration was approximately 5.0 μM, indicating low toxicity within the concentration range of 0-5.0 μM. The half-maximal inhibitory concentration (IC50) was determined by curve fitting. Control compounds such as rociletinib and EGCG showed much lower inhibitory activity. [Figure 12A] Osimertinib mesylate and Compound 2 (AZ7550) mesylate rescue tau-induced degeneration in Drosophila eyes. (A-C) Tau-expressing (GMR-Gal4 / WT tau) and non-expressing (GMR-Gal4 / +) flies were treated with osimertinib mesylate or Compound 2 (AZ7550) mesylate from larval to adult stages. Percent degeneration was assessed from microscopic images of Drosophila eyes. Five independent replicates were performed with different flies on different days for both osimertinib mesylate and Compound 2 (AZ7550) mesylate. Statistically different groups were determined by independent t-test, p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***), p ≤ 0.0001 (****). [Figure 12B] See legend to Figure 12A. [Figure 12C] See legend to Figure 12A. [Figure 12D] Osimertinib mesylate and compound 2 (AZ7550) mesylate rescue tau-induced degeneration in the Drosophila eye. (D) Representative SEM micrographs of Drosophila treated with osimertinib show visually reduced signs of degeneration compared to controls with GMR-Gal4 / WT tau. [Figure 13] Comparison of 2N4R tau aggregation inhibitory activity of exemplary compounds to compound 1 (osimertinib). In vitro seeding fibrillation assays of 2N4R tau were performed in the absence (reference) or presence of exemplary compounds, using a 1:3 tau:compound molar ratio and 2.5% (v / v) dimethyl sulfoxide (DMSO), with at least three replicates. The midpoint of the aggregation curve, Tm, was quantified and normalized for each condition, which is a measure of the degree of aggregation retardation. Of the compounds tested, only compound 31 with -OCH2- linker group (L) showed significant aggregation retardation, which is significantly more effective than compound 1 and all other compounds with L = -NH-. This indicates the advantage of modifying the linker structure to enhance anti-aggregation activity. Rejection tests were performed by one-way ANOVA with Dunnett's multiple comparisons. Significant differences of p ≤ 0.0001 (****) are indicated. [Figure 14] Comparison of tau aggregation inhibitory activity in HEK biosensor cells of osimertinib derivatives designated compound 13 and compound 9. The number of cells with aggregates (black circles) was quantified relative to the negative control, cells treated with vehicle (DMSO). The total amount of cells (grey triangles) remained close to 100% until compound concentrations reached approximately 4.0 μM, indicating low toxicity within the concentration range of 0-4.0 μM. The half-maximal inhibitory concentration (IC50) was determined by curve fitting. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Detailed Description of the Invention definition The term "alkyl" refers to a monoradical of a saturated straight chain or branched hydrocarbon. Preferably, the alkyl group contains 1 to 6 carbon atoms, i.e., 1, 2, 3, 4, 5, 6 carbon atoms, more preferably 1 to 4 carbon atoms, and most preferably 1 carbon atom. Exemplary alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, 1,2-dimethyl-propyl, isoamyl, n-hexyl, isohexyl, sec-hexyl, n-heptyl, isoheptyl, n-octyl, 2-ethyl-hexyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, and the like.

[0013] "Pharmaceutically acceptable salts" include salts with pharmaceutically acceptable acids or bases. Pharmaceutically acceptable acids include both inorganic acids, such as hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid, hydroiodic acid and nitric acid, and organic acids, such as citric acid, fumaric acid, maleic acid, malic acid, mandelic acid, ascorbic acid, oxalic acid, succinic acid, tartaric acid, benzoic acid, acetic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid or p-toluenesulfonic acid. Pharmaceutically acceptable bases include hydroxides of alkali metals (e.g., sodium or potassium) and alkaline earth metals (e.g., calcium or magnesium) and organic bases, such as alkylamines, arylalkylamines and heterocyclic amines.

[0014] Unless otherwise specified, the term "at least" preceding a series of elements is understood to refer to every element in the series. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the present invention.

[0015] Whenever used in this specification, the term "and / or" includes the meaning of "and", "or" and "all or any other combination of the elements connected by the term".

[0016] Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", are understood to mean the inclusion of a recited integer or step or group of integers or steps, but not the exclusion of any other integer or step or group of integers or steps. As used herein, the term "comprising" can be replaced with the terms "containing" or "including", or, as used herein, with the term "having". As used herein, "consisting of" excludes any element, step, or ingredient not specified.

[0017] The term "including" means "including but not limited to." "Including" and "including but not limited to" are used interchangeably.

[0018] compound The compound of the present invention is represented by the following formula (I): TIFF2025515141000019.tif41128 or a pharma- ceutically acceptable salt thereof: During the ceremony, G is TIFF2025515141000020.tif25128, preferably TIFF2025515141000021.tif25128, more preferably TIFF2025515141000022.tif24128; X is N or CH, preferably N; Y is N or CH, preferably N; L is TIFF2025515141000023.tif15128, preferably TIFF2025515141000024.tif14128; A is N or CR; preferably CR, more preferably CH; E is N or CR 1 ;Preferably CR 1 , more preferably CH; Preferably, only one or two of A, E, X, and Y are N; A and E are not both N, and A and Y are not both N; R is hydrogen, fluoro, chloro, bromo, -(C1-C6)alkyl, cyano, or -O(C1-C6)alkyl, preferably hydrogen; R 1 is hydrogen, fluoro, chloro, bromo, -(C1-C6)alkyl, cyano, or -O(C1-C6)alkyl, preferably hydrogen; R 2 is hydrogen, fluoro, chloro, bromo, -(C1-C6)alkyl, cyano, -O(C1-C6)alkyl, or difluoromethoxy, preferably -O(C1-C6)alkyl; R 3 teeth TIFF2025515141000025.tif132170, preferably TIFF2025515141000026.tif23128; R 4 is hydrogen, -(C1-C6)alkyl, or -(C3-C6)cycloalkyl; R 5 is hydrogen, fluoro, or chloro; R 6 is hydrogen, fluoro, or chloro; R 7 is hydrogen or -(C1-C6)alkyl; R 8 is hydrogen or -(C1-C6)alkyl; R 9 is hydrogen or -(C1-C6)alkyl; R 10 is hydrogen or -(C1-C6)alkyl; R 11is hydrogen, -(C1-C6)alkyl, or -CH2CON((C1-C6)alkyl); R 12 is hydrogen or -(C1-C6)alkyl; where, optionally, one or more hydrogen atoms are replaced with deuterium atoms; One or more carbon atoms may be 11 It is replaced by C isotopes; The nitrogen atom or atoms may be selected from the group consisting of the corresponding 13 Replaced with N isotopes; The one or more fluoro atoms may be 18 It has been replaced by the F isotope.

[0019] In further embodiments, at least one of the following conditions applies: (i) R is fluoro, chloro, or cyano; (ii)R 1 is fluoro, methoxy; (iii)R 2 is hydrogen, fluoro, chloro, cyano, or methyl; (iv) R 3 is pyrrolidinyl, piperidinyl, 4-methylpiperidinyl, or 2-methylpyrrolidinyl; (v) L is -OCH2-.

[0020] In a further embodiment, the compound is TIFF2025515141000027.tif193168TIFF2025515141000028.tif189168TIFF2025515141000029.tif192168TIFF2025515141000030.tif178168.

[0021] In further aspects G is TIFF2025515141000031.tif24128; X is selected from N; Y is selected from N; L is TIFF2025515141000032.tif8128; A is selected from N or CR; E is N or CR 1 Selected from; R is hydrogen; R 1 is hydrogen; R 2 is -O(C1-C6)alkyl; R 3 teeth TIFF2025515141000033.tif23128; R 4 is hydrogen or -(C1-C6)alkyl; R 5 is hydrogen; R 6 is hydrogen; R 8 is hydrogen or -(C1-C6)alkyl, preferably -(C1-C6)alkyl; R 9 is hydrogen or -(C1-C6)alkyl, preferably -(C1-C6)alkyl; R 10 is hydrogen or -(C1-C6)alkyl.

[0022] In a further embodiment, G is TIFF2025515141000034.tif24128; X is selected from N; Y is selected from N; L is TIFF2025515141000035.tif8128; A is selected from N or CR; E is N or CR 1 Selected from; R is hydrogen; R 1 is hydrogen; R 2 is -OMe; R 3 teeth TIFF2025515141000036.tif23128; R 4 is methyl; R 5 is hydrogen; R 6 is hydrogen; R 8 is hydrogen or methyl; R 9 is methyl; R 10 is methyl.

[0023] In a further embodiment, the compound has formula (Ia or Ib): TIFF2025515141000037.tif73128TIFF2025515141000038.tif75128 is a compound of: During the ceremony, A is N or CR; preferably CR, more preferably CH; E is N or CR; preferably CR, more preferably CH; X is N, CH, preferably CH; R is hydrogen, fluoro, chloro, bromo, -(C1-C6)alkyl, cyano, -O(C1-C6)alkyl, preferably hydrogen; L is TIFF2025515141000039.tif12128, preferably TIFF2025515141000040.tif10128; R 1 teeth, TIFF2025515141000041.tif104163, preferably TIFF2025515141000042.tif29128, selected from the group of alkylamines, non-aromatic heterocycles, partially or fully deuterated alkylamines, and partially or fully deuterated non-aromatic heterocycles; R 2is hydrogen, fluoro, -(C1-C6)alkyl, preferably hydrogen; R 3 is hydrogen, -(C1-C6)alkyl, -(C3-C6)cycloalkyl, preferably methyl; R 4 is hydrogen, -(C1-C6)alkyl, preferably hydrogen; R 5 is hydrogen, -(C1-C6)alkyl, preferably hydrogen; R 6 is hydrogen, -(C1-C6)alkyl, preferably hydrogen; R 7 is hydrogen, fluoro, -O(C1-C6)alkyl, -(C1-C6)alkyl, preferably hydrogen; R 8 is hydrogen, -(C1-C6)alkyl, preferably hydrogen; R 9 is hydrogen, -(C1-C6)alkyl, preferably -(C1-C6)alkyl; R 10 is hydrogen, -(C1-C6)alkyl, preferably -(C1-C6)alkyl; R 11 is hydrogen, -(C1-C6)alkyl, preferably -(C1-C6)alkyl; R 12 is -(C1-C6)alkyl, -(C3-C6)cycloalkyl, preferably -(C1-C6)alkyl; where A=N and TIFF2025515141000043.tif12128 never exist simultaneously in the same compound. where, optionally, one or more hydrogen atoms are replaced with deuterium atoms; One or more carbon atoms may be 11 It is replaced by C isotopes; The nitrogen atom or atoms may be selected from the group consisting of the corresponding 13 Replaced with N isotopes; The one or more fluoro atoms may be 18It has been replaced by the F isotope.

[0024] Certain exemplary compounds are listed in Table 1.

[0025] (Table 1) List of compounds TIFF2025515141000044.tif195169TIFF2025515141000045.tif246169TIFF2025515141000046.tif199170TIFF20255151410 00047.tif197169TIFF2025515141000048.tif205170TIFF2025515141000049.tif243170TIFF2025515141000050.tif243170 TIFF2025515141000051.tif189170TIFF2025515141000052.tif202170TIFF2025515141000053.tif231170TIFF20255151410 00054.tif208170TIFF2025515141000055.tif216170TIFF2025515141000056.tif202170TIFF2025515141000057.tif203170

[0026] Medical Applications The compounds of formulae (I), (Ia) and (Ib) above or the pharmaceutical compositions of the present invention are for use in the prevention or treatment of tauopathies.

[0027] In general, tauopathies belong to a class of diseases associated with pathological aggregation of tau protein in thread-like structures, pretangles, neurofibrillaries or glial fibrillary tangles in the nervous system. Tau protein undergoes several post-translational modifications including phosphorylation, acetylation, ubiquitination and myristoylation, with phosphorylation being the major modification of tau and its cellular functions. In tauopathies, tau aggregates are found to be extensively phosphorylated on several serine, threonine or tyrosine residues and acetylated and ubiquitinated on several lysine residues. 11、12、13、14Hyperphosphorylation and acetylation of tau reduces its ability to bind to microtubules, thereby impeding cargo transport and inducing the pathogenic accumulation of tau within cells.

[0028] Thus, the term tauopathy encompasses both loss-of-function effects on microtubules and gain-of-function effects of toxic tau species. In particular, the consequences of tau hyperphosphorylation and acetylation, and potentially other post-translational modifications, include not only the loss of its function to bind and stabilize microtubules, but also the gain of neurotoxicity due to tau aggregation into thread-like structures, pretangles, neurofibrillary or glial fibrillary tangles.

[0029] Preferably, the prevention or treatment of tauopathies with the compounds of formula (I) of the present invention. 15、16 This includes treating the cause of tauopathy.

[0030] Preferably, the prevention or treatment of tauopathy comprises inhibiting the cellular activity of tau by binding of a compound of formula (I), (Ia) or (Ib) to tau. Binding to tau may comprise covalent binding of a compound of general formula (I) to tau, preferably the binding comprises a thiol-Michael addition of an -SH group of tau to an acrylamide group in a compound of formula (I).

[0031] Preferably, the tauopathy is a dementia-associated tauopathy.

[0032] Preferably, the tauopathy is selected from the group consisting of: Alzheimer's disease, frontotemporal dementia, primary age-related tauopathy (PART), familial British dementia (FBD), familial Danish dementia (FDD), chronic traumatic encephalopathy (CTE). 15 , progressive supranuclear palsy (PSP) 15 , Corticobasal degeneration (CBD) 15 , Pick's disease (PiD) 15 , Dementia with Lewy Bodies (DLB) 15 , progressive supranuclear palsy (PSP) 15 , Globular glial tauopathy (GGT) 15, tauopathy with hippocampal 4-repeat tau-immunoreactive globular inclusions, limbic-predominant neuronal inclusion 4R tauopathy (LNT), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) 17 , silvery granulosis (AGD) 17 , Huntington's disease 17 , glioglobular tauopathy, neuroastroglial tauopathy variants in elderly people, familial behavioral disorder-associated frontotemporal dementia with astrocyte-predominant tauopathy, amyotrophic lateral sclerosis (ALS) 18 , spinocerebellar ataxia type 11, spinal muscular atrophy (SMA), progressive ataxia and palatal tremor-associated tauopathy, cerebral amyloid angiopathy (CAA), IgLON5 antibody-associated tauopathy, trisomy 21-associated Alzheimer's disease (Down's syndrome), vascular dementia, cerebral amyloid angiopathy, Gerstmann-Sträussler-Scheinker disease (GSS), Creutzfeldt-Jakob disease, fatal familial insomnia, kuru, Niemann-Pick disease type C-associated tauopathy, nodding syndrome, non-Guam motor neuron disease with neurofibrillary tangles, Parkinson's disease (PD) 19 , Parkinson's disease with dementia 17 , Parkinsonism-Dementia in Guam 17 , Guadeloupe-type parkinsonism 17 , Kosaka-Shibayama disease 17 , postencephalitic parkinsonism 17, SYNJ1 (PARK20) early-onset recessive parkinsonism with seizures and dystonia-associated nigral tau pathology, tau pathology associated with multiple system atrophy, familial parkinsonism and progressive respiratory failure, limbic-predominant neuro-glial tau pathology in TARDBP mutations (I383;P112H), neuronal 4R tau pathology in fatal familial insomnia (PRNP D178N mutation), tau pathology associated with ADCY5 dyskinesia, tau pathology in chronic temporal lobe epilepsy, neurodegeneration with intracranial iron deposition (NBIA), tau pathology in NBIA PANK2 and WDR45 mutations, tau pathology in NBIA PLA2G6 mutations, tau pathology in NBIA associated with autosomal dominant mitochondrial membrane protein-associated neurodegeneration (MPAN), pretangles and neurofibrillary tangles in human immunodeficiency virus (HIV)-negative opiate abusers 20 , Tau pathology associated with acquired immune deficiency syndrome (AIDS) 20 , diffuse neurofibrillary tangles with calcification, progressive ataxia and palatal tremor, SLC9A6-related parkinsonism, tau pathology associated with SPG7 gene mutations, striatal 4R tau pathology associated with X-linked parkinsonism with spasticity (ATP6AP2), tau pathology associated with SPAST gene-related hereditary spastic paraplegia, autism, autism spectrum disorder, retinal tauopathy 21 , West Nile encephalomyelitis, TTBK2 gene-related spinocerebellar degeneration 11, herpes simplex encephalitis, neurofibrillary tangle dementia (TOD), age-related tau astrogliopathy (ARTAG) 17, hippocampal tauopathy, subacute sclerosing panencephalitis (SSPE)-associated tauopathy, FTLD-C9ORF72, Christianson syndrome, vacuolar tauopathy, Ritiko-Bodig disease, ganglioglioma and gangliocytoma, meningioangiomatosis, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, neuronal ceroid lipofuscinosis, myotonic dystrophy, Fukuyama-type congenital muscular dystrophy, unilateral megalencephaly, focal cortical dysmorphism, Walcott-Rallison syndrome, primary lateral sclerosis, progressive freezing of gait, PSP with parkinsonism, Richardson syndrome, non-fluent / agrammatic primary progressive aphasia, semantic primary progressive aphasia, logopenic primary progressive aphasia, primary progressive apraxia of speech, amnesic Alzheimer's disease.

[0033] More preferably, Alzheimer's disease (AD), Pick's disease (PiD), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), chronic traumatic encephalopathy (CTE), argyrophilic grain disease (AGD), frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD) and primary age-related tauopathy (PART).

[0034] Pharmaceutical Compositions and Dosages The pharmaceutical compositions of the present invention comprise a compound for use in the prevention or treatment of a tauopathy and at least one pharma- ceutically acceptable carrier.

[0035] "Carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered. Such pharmaceutical carriers can be sterile liquids, such as, but not limited to, water and oils, such as those of petroleum, animal, vegetable, or synthetic origin, such as, but not limited to, peanut oil, soybean oil, mineral oil, sesame oil, and the like. When the pharmaceutical composition is administered orally, water is the preferred carrier. When the pharmaceutical composition is administered intravenously, saline and aqueous dextrose solutions are the preferred carriers. Saline solutions and aqueous dextrose and glycerol solutions are preferably used as liquid carriers for injectable solutions. Suitable pharmaceutical excipients include aqueous hydroxypropyl methylcellulose, low-substituted hydroxypropyl cellulose, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like. The compositions may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents, if desired. These compositions may take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained release formulations, and the like. The compositions may be formulated as suppositories, with traditional binders and carriers, such as triglycerides. Oral formulations may contain standard carriers, such as pharmaceutical grade aqueous hydroxypropylmethylcellulose, low-substituted hydroxypropylcellulose, mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, and the like. Examples of suitable pharmaceutical carriers are described in ''Remington's Pharmaceutical Sciences'' by EW Martin. Such compositions will contain a therapeutically effective amount of the therapeutic agent, preferably in purified form, together with an appropriate amount of carrier, so as to provide the form for proper administration to the patient. The formulation should suit the mode of administration.

[0036] In one embodiment, the pharmaceutical composition is a tablet comprising mannitol, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, sodium stearyl fumarate.Preferably, the tablet coating comprises polyvinyl alcohol, titanium dioxide, macrogol 3350, talc, iron oxide yellow (E 172), iron oxide red (E 172), iron oxide black.

[0037] Preferably, the pharmaceutical composition is applied parenterally or orally, preferably orally.

[0038] Generally, out of 100% (for a pharmaceutical formulation / composition), the amount of active ingredient (particularly the amount of a compound of the present invention, optionally together with other therapeutically active agents if present in the pharmaceutical formulation / composition) will range from about 0.01% to about 99%, preferably from about 0.1% to about 70%, most preferably from about 1% to about 30%, with the remainder preferably being made up of one or more pharma- ceutically acceptable excipients.

[0039] The amount of active ingredient, e.g., a compound of the invention, in a unit dosage form and / or when administered to an individual or used in a therapy may range from about 0.1 mg to about 400 mg (e.g., about 1 mg to about 400 mg, e.g., about 10 mg to about 90 mg or about 40 mg to 80 mg or about 80 mg to 400 mg) per unit, administration or therapy. In certain embodiments, suitable amounts of such active ingredients may be calculated using the mass or body surface area of ​​the individual and may range from about 1 mg / kg to 10 mg / kg (e.g., about 2 mg / kg to 5 mg / kg), or about 1 mg / m 2 ~about 400mg / m 2 Between (approx. 3 mg / m 2 ~about 350mg / m 2 Between or about 10 mg / m 2 ~about 200mg / m 2 This includes the amount of

[0040] In one embodiment, the pharmaceutical composition comprises an additional therapeutically active ingredient, for example in a combination therapy, hi one embodiment, the pharmaceutical composition does not comprise an allosteric EGFR (epidermal growth factor receptor) inhibitor. EXAMPLES

[0041] Example of the invention Chemical synthesis, in vitro assay, and biological assay procedures The following abbreviations may be used: DMSO - Dimethyl sulfoxide NMR-Nuclear Magnetic Resonance HMQC - Heteronuclear Multiple Quantum Coherence TOCSY-Total Correlation Spectroscopy NOESY - Nuclear Overhauser Effect Spectroscopy STD-Saturation Transfer Difference ThT-Thioflavin T TCEP-(Tris(2-carboxyethyl)phosphine) HEK - Human Embryonic Kidney GFP - Green Fluorescent Protein SDS-PAGE - Sodium dodecyl sulfate polyacrylamide gel electrophoresis TFA - Trifluoroacetic acid DIPEA-N,N-Diisopropylethylamine FCC-Flash Column Chromatography THF - Tetrahydrofuran DMF-N,N-Dimethylformamide

[0042] Compound synthesis / preparation N-[2-(2-dimethylaminoethylmethylamino)-4-methoxy-5-[[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino]phenyl]prop-2-enamide (1) is called "osimertinib" and N-[2-(2-dimethylaminoethylmethylamino)-4-methoxy-5-[[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino]phenyl]prop-2-enamide mesylate is called "osimertinib mesylate". The synthesis of osimertinib and osimertinib mesylate is reported in J. Med. Chem. (2014), 57, 8249-8267. Osimertinib was purchased from LC Laboratories (Massachussetts, USA, catalog number O-7200), and osimertinib mesylate was purchased from MedChemExpress (catalog number HY-15772A).

[0043] 4-Methoxy-5-[[4-(1-methyl-1H-indol-3-yl)-2-pyrimidinyl]amino]-2-[methyl[2-(methylamino)ethyl]amino]phenyl]-2-propenamide (2) is referred to as "AZ7550", N-[4-methoxy-5-[[4-(1-methyl-1H-indol-3-yl)-2-pyrimidinyl]amino]-2-[methyl[2-(methylamino)ethyl]amino]phenyl]-2-propenamide hydrochloride (2) and N-[4-methoxy-5-[[4-(1-methyl-1H-indol-3-yl)-2-pyrimidinyl]amino]-2-[methyl[2-(methylamino)ethyl]amino]phenyl]-2-propenamide (2) mesylate. Compound 2 is produced in vivo as a metabolite of osimertinib (Reference: J. Med. Chem. 2014, 57, 8249-8267). Compound 2 hydrochloride (catalog number HY-B0794A) and compound 2 mesylate (catalog number HY-B0794B) were purchased from MedChemExpress.

[0044] N-[2-[[2-(dimethylamino)ethyl]methylamino]-5-[[4-(1H-indol-3-yl)-2-pyrimidinyl]amino]-4-methoxyphenyl]-2-propenamide (3). Compound 3 was purchased from MedChemExpress (catalog number HY-B0793).

[0045] N-[5-[[4-(1-cyclopropyl-1H-indol-3-yl)-2-pyrimidinyl]amino]-2-[[2-(dimethylamino)ethyl]methylamino]-4-methoxyphenyl]-2-propenamide (13) was purchased from Hoelzel Biotech (Germany, catalog number HS-10296, CAS number 1899921-05-1).

[0046] N-[2-[[2-(dimethyloxidoamino)ethyl]methylamino]-4-methoxy-5-[[4-(1-methyl-1H-indol-3-yl)-2-pyrimidinyl]amino]phenyl-2-propenamide (14) was purchased from Biozol Diagnostika (Germany, catalog number 0702275, CAS number 1975982-94-5).

[0047] N-[2-[[2-(dimethylamino)ethyl]methylamino]-5-[[4-(7-fluoro-1-methyl-1H-indol-3-yl)-2-pyrimidinyl]amino]-4-methoxyphenyl]-2-propenamide (4). CAS number 1883592-81-1.

[0048] N-[2-[[2-(dimethylamino)ethyl]methylamino]-5-[[4-(5-fluoro-1-methyl-1H-indol-3-yl)-2-pyrimidinyl]amino]-4-methoxyphenyl]-2-propenamide (5). CAS number 1903753-67-2.

[0049] The synthesis of compounds 4 and 5 is described in Bioinorganic & Medicinal Chemistry (2017), 25, 1, 4553-4559.

[0050] N-[2-[[2-(dimethylamino)ethyl]methylamino]-4-methoxy-5-[[4-(1-methyl-1H-pyrrolo[3,2-b]pyridin-3-yl)-2-pyrimidinyl]amino]phenyl]-2-propenamide (6) CAS number 2050521-74-7.

[0051] The synthesis of compound 6 is described in Bioorganic & Medicinal Chemistry (2018), 26, 23-24, 6135-6145.

[0052] N-[2-[[2-(dimethylamino)ethyl]methylamino]-4-methoxy-5-[[4-(3-methyl-1H-indol-1-yl)-2-pyrimidinyl]amino]phenyl]-2-propenamide (21) CAS number 1835666-87-9. The synthesis of compound 21 is described in European Journal of Medicinal Chemistry (2017), 135, 12-23.

[0053] Synthesis of compounds 7, 8, 27, and 28 TIFF2025515141000058.tif85157 Scheme 1: A general scheme for the synthesis of compounds 7, 8, 27, and 28 is shown above. TIFF2025515141000059.tif28128In the first step, a mixture of 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole and 4-bromo-2-chloropyridine in toluene is added to an aqueous mixture of tripotassium phosphate and catalytic tetrakis(triphenylphosphine)palladium and incubated overnight. The crude product is evaporated to dryness, bound to silica powder, and then purified by flash silica chromatography using a solvent gradient of 0-20% methanol in dichloromethane. The pure fractions containing 3-(2-chloropyridin-4-yl)-1-methyl-1H-indole are evaporated to dryness.

[0054] TIFF2025515141000060.tif36128The 3-(2-chloropyridin-4-yl)-1-methyl-1H-indole is then combined with an equimolar amount of 4-fluoro-3-nitroaniline dissolved in 2-pentanol. 4-Methylbenzenesulfonic acid is added to the mixture, followed by heating at 105°C for several hours. The mixture is cooled to room temperature and the crude product is washed with 2-pentanol and subjected to flash silica chromatography or recrystallization to obtain a pure fraction of N-(4-fluoro-3-nitrophenyl)-4-(1-methyl-1H-indol-3-yl)pyridin-2-amine.

[0055] TIFF2025515141000061.tif39136 Either an amine base precursor ("H-base") selected from pyrrolidine, 2-methylpyrrolidine, piperidine, or 4-methylpiperidine is added to a suspension of N-(4-fluoro-3-nitrophenyl)-4-(1-methyl-1H-indol-3-yl)pyridin-2-amine and N,N-diisopropylethylamine (DIPEA) in 2,2,2-trifluoroethanol. The "base" corresponds to a pyrrolidinyl, piperidinyl, 4-methylpiperidinyl, or 2-methylpyrrolidinyl substituent. The mixture is heated in a microwave at 140 °C for 1 h, after which the crude product is cooled to room temperature and subjected to ion exchange chromatography. Flash silica chromatography is then performed using a solvent gradient of 0-4% 7M ammonia / methanol in dichloromethane. Pure fractions containing the desired base-substituted nitrobenzene ring are evaporated to dryness.

[0056] TIFF2025515141000062.tif27128 The base-substituted nitrobenzene intermediate is dissolved in ethanol / water and refluxed in the presence of ammonium chloride for 2 hours. The crude product is subjected to ion exchange chromatography followed by flash silica chromatography using elution solvent (7M ammonia / methanol). Pure fractions containing the desired base-substituted aniline product are evaporated to dryness.

[0057] TIFF2025515141000063.tif25128In the final reaction step, acryloyl chloride in dichloromethane is added to a stirred solution of the base-substituted aniline compound dissolved in dichloromethane containing DIPEA and cooled to 4°C. The reaction is carried out for 1.5 hours and quenched by dilution 5-fold with dichloromethane and then washed with saturated aqueous sodium bicarbonate. The resulting hydrophobic (organic) layer is separated from the aqueous layer and subjected to silica flash chromatography using a solvent gradient of 0-4% 7M ammonia / methanol in dichloromethane. Pure fractions are evaporated to dryness. The compound has the formula: TIFF2025515141000064.tif24128

[0058] For compound 27, the "base" is pyrrolidinyl. For compound 7, the "base" is piperidinyl. For compound 28, the "base" is 4-methylpiperidinyl. For compound 8, the "base" is 2-methylpyrrolidinyl.

[0059] Synthesis of compounds 17, 18, 29, and 30 TIFF2025515141000065.tif95168 Scheme 2: General scheme for the synthesis of compounds 17, 18, 29, and 30 The intermediate 1-bromo-4-[[[(1,1-dimethylethyl)dimethylsilyl]oxy]methyl]-2-nitrobenzene is prepared by reacting 4-bromobenzoic acid with (1,1-dimethylethyl)dimethylsilyl chloride as described in TIFF2025515141000066.tif52128Bioconjugate Chemistry 2020, 31(2), 224-228. Alternatively, the compound may be prepared by adding (1,1-dimethylethyl)dimethylsilyl chloride to a mixture of (4-bromo-3-nitrophenyl)methanol and imidazole in dichloromethane. The reaction is carried out at room temperature overnight, after which the desired product is purified by flash silica chromatography.

[0060] TIFF2025515141000067.tif41128 Prepare base-substituted nitrophenylmethanol intermediates by reacting any of the amine base precursors ("H-base") selected from pyrrolidine, 2-methylpyrrolidine, piperidine with 1-bromo-4-[[[(1,1-dimethylethyl)dimethylsilyl]oxy]methyl]-2-nitrobenzene and DIPEA in 2,2-trifluoroethanol solvent at 140 °C for 1 h. Subject the crude product to flash silica chromatography and evaporate pure fractions to dryness.

[0061] TIFF2025515141000068.tif35128 The base-substituted nitrophenylmethanol intermediate is reacted with commercially available 3-(2-chloropyrimidin-4-yl)-1-methyl-1H-indole (CAS number 1032452-86-0) in the presence of cesium carbonate. The reaction is carried out in dimethylformamide solvent at 100°C for 5 hours. The crude product is cooled to room temperature and subjected to flash silica chromatography to obtain the desired fraction with -OCH2- linkage between the pyrimidine ring and the nitrobenzene ring.

[0062] TIFF2025515141000069.tif36128-OCH2-linked base-substituted nitrobenzene intermediate is dissolved in ethanol / water and refluxed in the presence of ammonium chloride for 2 hours. The crude product is subjected to ion exchange chromatography followed by flash silica chromatography using elution solvent (7M ammonium / methanol). Pure fractions containing the desired product (-OCH2-linked base-substituted aniline compound) are evaporated to dryness.

[0063] TIFF2025515141000070.tif37128In the final reaction step, acryloyl chloride in dichloromethane is added to a stirred solution of -OCH2-linked base-substituted aniline intermediate dissolved in dichloromethane containing DIPEA and cooled in an ice-water bath. The reaction is carried out for 1.5 h and quenched by dilution 5-fold with dichloromethane followed by washing with saturated aqueous sodium bicarbonate. The resulting hydrophobic (organic) layer is separated from the aqueous layer and subjected to silica flash chromatography using a solvent gradient of 0-4% 7M ammonia / methanol in dichloromethane. Pure fractions are evaporated to dryness. The compound has the formula: TIFF2025515141000071.tif39128

[0064] For compound 17, the "base" is pyrrolidinyl. For compound 18, the "base" is piperidinyl. For compound 29, the "base" is 4-methylpiperidinyl. For compound 30, the "base" is 2-methylpyrrolidinyl.

[0065] Synthesis of compounds 31, 32, 19, and 20 TIFF2025515141000072.tif96163 Scheme 3: General scheme for the synthesis of compounds 31, 32, 19, and 20 TIFF2025515141000073.tif29128 A mixture of 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole and 4-bromo-2-chloropyridine in toluene is added to an aqueous mixture of tripotassium phosphate and catalytic tetrakis(triphenylphosphine)palladium and incubated overnight. The crude product is evaporated to dryness, bound to silica powder, and then purified by flash silica chromatography using a solvent gradient of 0-20% methanol in dichloromethane. The pure fractions containing 3-(2-chloropyridin-4-yl)-1-methyl-1H-indole are evaporated to dryness.

[0066] TIFF2025515141000074.tif34128 The base-substituted nitrophenylmethanol intermediate is prepared using the first two reactions described in Scheme 2. The base-substituted nitrophenylmethanol intermediate is reacted with 3-(2-chloropyridin-4-yl)-1-methyl-1H-indole in the presence of cesium carbonate. The reaction is carried out in dimethylformamide solvent at 100° C. for 5 hours. The crude product is cooled to room temperature and subjected to flash silica chromatography to obtain the desired fraction having an -OCH2- linkage between the pyridine ring and the nitrobenzene ring.

[0067] TIFF2025515141000075.tif37128-OCH2-linked base-substituted nitrobenzene intermediate is dissolved in ethanol / water and refluxed for 2 hours in the presence of ammonium chloride. The crude product is subjected to ion exchange chromatography followed by flash silica chromatography using elution solvent (7M ammonium / methanol). Pure fractions containing the desired product (-OCH2-linked base-substituted aniline compound) are evaporated to dryness.

[0068] TIFF2025515141000076.tif37128In the final reaction step, acryloyl chloride in dichloromethane is added to a stirred solution of -OCH2-linked base-substituted aniline intermediate dissolved in dichloromethane containing DIPEA and cooled to 4°C. The reaction is carried out for 1.5 hours and quenched by dilution 5-fold with dichloromethane followed by washing with saturated aqueous sodium bicarbonate. The resulting hydrophobic (organic) layer is separated from the aqueous layer and subjected to silica flash chromatography using a solvent gradient of 0-4% 7M ammonia / methanol in dichloromethane. Pure fractions are evaporated to dryness. The compound has the formula: TIFF2025515141000077.tif36128

[0069] For compound 31, the "base" is pyrrolidinyl. For compound 32, the "base" is piperidinyl. For compound 19, the "base" is 4-methylpiperidinyl. For compound 20, the "base" is 2-methylpyrrolidinyl.

[0070] NMR spectroscopy Stocks of compounds for NMR experiments were prepared either by dissolving the powder directly in NMR buffer (50 mM sodium phosphate buffer pH 6.8, 0.01% NaN3) or as 10 mM compound in deuterated DMSO.

[0071] To obtain residue-specific information on the interactions between tau and compounds, 1 H- 15 The N correlation spectrum was monitored during titration with increasing amounts of compound. 15 N]-tau 1 H- 15N SOFAST-HMQC (reference: Schanda, P., Kupce, E. & Brutscher, B. SOFAST-HMQC experiments for recording two-dimensional heteronuclear correlation spectra of proteins within a few seconds. J. Biomol. NMR 33, 199-211 (2005)) spectra were acquired at 5 °C on a Bruker 800 MHz spectrometer equipped with a triple-resonance cryoprobe. Tau samples (18–25 μM) were prepared in NMR buffer containing 10% (v / v) DO at compound concentrations corresponding to tau:compound molar ratios of 1:10 and 1:100. All NMR samples were incubated overnight (approximately 16 h) at 37 °C before spectral acquisition. The amide assignments of tau were obtained from a previous study (Reference: Mukrasch, M. et al. Structural polymorphism of 441-residue tau at single residue resolution. PLoS Biol. 7, e34 (2009)). However, because the previous assignments were obtained using tau samples prepared in the presence of reducing agents, some amide resonances belonging to the repeat regions of tau (near C291 and C322) were not clearly transferred to the SOFAST-HMQC spectrum obtained under oxidizing conditions. Therefore, to obtain a nearly complete assignment of the backbone of the repeat regions, we used [U- 13 C, 15 3D HNCA, 3D HNCOCA, and 3D CBCACONH spectra were recorded for the [N]-4R tau (K18 construct) sample. The weighted, normalized chemical shift perturbation (CSP) was calculated as CSP=√[0.5 [(Δδ H) 2 +(Δδ N) 2 The NMR signal intensity ratio, I / I0 (I is the peak intensity of the tauamide backbone resonance in the presence of compound and I0 is the peak intensity in the absence of compound) was calculated for each titration point.

[0072] To obtain assignments of stable protons in osimertinib, AZ7550 hydrochloride, and AZ5104 (250 μM compound in 50 mM sodium phosphate, pH 6.8, containing 2.5% (v / v) deuterated DMSO), the following NMR experiments were performed on each compound: 1D 1 H-NMR, 2D 1 H- 1 H TOCSY, mixing time 80 ms, and 2D 1 H- 1 H NOESY, mixing time 200 ms. Experiments were performed at 25 °C and 37 °C on a Bruker 800 MHz or Bruker 600 MHz spectrometer equipped with a triple resonance cryoprobe.

[0073] To determine the binding epitopes of the compounds on monomeric 2N4R tau, spectra of osimertinib, AZ7550 hydrochloride, and AZ5104 (250 μM compound) are recorded along with unlabeled 2N4R tau (0, 10, and 20 μM), 1D 1 H-NMR titrations were performed. NMR titration samples were prepared in 50 mM sodium phosphate, pH 6.8, containing 2.5% (v / v) deuterated DMSO. NMR spectra were recorded at 37 °C using a Bruker 800 MHz spectrometer equipped with a triple resonance cryoprobe.

[0074] To determine the binding epitopes of the compounds on fibrillar tau, 1D saturation transfer difference (STD) experiments were performed with osimertinib, AZ7550 hydrochloride, and AZ5104 in the presence of sonicated 2N4R tau fibrils (fibril length approx. 100 nm). 2N4R tau fibrils were prepared using the aggregation procedure described in In Vitro Assay 1 without Thioflavin T. The molar ratio of "free" compound to fibrils was 48:1 (corresponding to a total compound concentration of 250 μM and a total fibril concentration of 5 μM tau). NMR samples were prepared in 50 mM sodium phosphate, pH 6.8, containing 100 mM NaCl and 2.5% (v / v) DMSO. Spectra were recorded at 37 °C using a Bruker 700 MHz spectrometer equipped with a triple resonance cryoprobe. 1D 1H-NMR STD spectra were acquired in a mode alternating between off-resonance irradiation at 60 ppm and on-resonance irradiation at −2 ppm.

[0075] NMR spectra were processed using Topspin 3.6.2 (Bruker) and analyzed using NMRFAM-Sparky (Reference: Bioinformatics. 2015 Apr 15; 31(8):1325-7. Epub 2014 Dec 12 NMRFAM-SPARKY: enhanced software for biomolecular NMR spectroscopy).

[0076] In vitro assay procedures In vitro assay 1: de novo and seeded tau aggregation assays Aggregation of 2N4R tau was performed using a previously published cofactor-free aggregation protocol with minor modifications (Reference: Chakraborty, P. et al. Co-factor-free aggregation of tau into seeding-competent RNA-sequestering amyloid fibrils. Nat. Commun. 12, 4231 (2021)). For aggregation experiments, tau protein stocks prepared in 25 mM HEPES pH 7.4, 1 mM TCEP were used. Compound stocks were prepared in aggregation buffer (25 mM HEPES, 10 mM KCl, 5 mM MgCl2, pH 7.2) or DMSO (10 mM compound).

[0077] To prepare tau fibril species by de novo aggregation, 2N4R tau at a monomer concentration of 25 μM was diluted in aggregation buffer. The working concentration of TCEP in the aggregation mixture was <25 μM after dilution of the stock. To this mixture, thioflavin T (ThT) was added to a final concentration of 50 μM. A total of 100 μL of the tau-ThT mixture was incubated in the wells of a 96-well microplate (Greiner Bio-one) in the presence of two polytetrafluoroethylene beads. The microplate was incubated at 37 °C, shaken intermittently in a dual orbital motion, and subjected to a program implemented in a Tecan Spark plate reader that performed ThT fluorescence emission measurements. The excitation wavelength of ThT was set at 430 nm, and the emission wavelength was 485 nm. Details of this plate reader program have been published previously (Reference: Chakraborty, P. et al. Co-factor-free aggregation of tau into seeding-competent RNA-sequestering amyloid fibrils. Nat. Commun. 12, 4231 (2021)). After approximately 3–6 days of incubation, ThT fluorescence emission reached a maximum value, suggesting that tau fibrillization had reached a saturation point. The fibril-containing mixture recovered from the microplate was used as the seeding mixture for the subsequent aggregation assay.

[0078] Seeded aggregation assays were performed as described above, except that 1.0% of the well volume was the seeding mixture and sonicated for 1 min. Assays were performed in the presence and absence of compound, with working concentrations ranging from 0 to 250 μM compound.

[0079] Analysis of the fluorescence curves was performed using Graphpad PRISM version 8. Fluorescence data were fitted to a sigmoidal function. For each sigmoidal curve, the time corresponding to 50% of the maximum ThT fluorescence was calculated. TM and the span of fluorescence intensity was determined. Statistical significance was determined by one-way analysis of variance with Dunnett's multiple comparison test or unpaired t-test.

[0080] The seeding aggregation protocol described above was similarly applied to the C291S / C322S mutants of 3R tau and 2N4R tau.

[0081] In vitro assay 2: Tau pelleting assay Aggregation assays were performed as described in In Vitro Assay 1. The aggregation mixture was collected and ultracentrifuged (55000 rpm, JLA 100.3 rotor, Optima MAX-XP) for 30 min to separate the soluble (supernatant) and insoluble (pellet) fractions. The soluble fraction was analyzed by SDS-PAGE using 12% or 15% acrylamide separating gels and 5% acrylamide stacking gels. The band intensity of soluble tau was quantified with ImageLab (Bio-Rad Laboratories).

[0082] Negative staining transmission electron microscopy Fibril samples were negatively stained using glow-discharged 400-mesh carbon-coated copper grids, and images were acquired with a Talos L120C transmission electron microscope (Thermo Fisher, Eindhoven, The Netherlands) after staining with 1% uranyl acetate solution.

[0083] Mass spectrometry procedures Mass spectrometry 1:ESI-MS The analysis was performed with a 4R tau construct (K18 construct, 13818 Da) containing residues 244-372 of 2N4R tau. Compound (330 μM) was incubated with 4R tau (65 μM) for 16 h at 37 °C in 10 mM ammonium bicarbonate, pH 7.0. The compound / 4R tau mixture was injected into an LC system (ACQUITY) coupled to a single quadrupole mass detection system (SQ Detector 2, Waters) equipped with a diode array detector (DAD). The DAD spectrum covers wavelengths from 210 to 400 nm. The components of the compound-4R tau mixture were separated using a BioResolve RP mAb column (Waters) with a gradient of 5-95% buffer B (buffer A is 0.1% TFA / water and buffer B is 0.1% TFA / acetonitrile). Covalent modification of 4R tau corresponded to mass additions of 500 Da (osimertinib) and 486 Da (AZ7550 or AZ5104).

[0084] Mass spectrometry 2:MS / MS Compounds (330 μM) were incubated with 2N4R tau (65 μM) in buffer (25 mM HEPES, 10 mM KCl, 5 mM MgCl 2 , pH 7.2) at 37° C. for 16 h and then stored at 4° C. until in-gel digestion.

[0085] In-gel digestion with trypsin (Sigma Aldrich) and subsequent extraction of peptides for mass spectrometry analysis were performed as previously described (see Shevchenko, A., Tomas, H., Havlis, J., Olsen, JV, and Mann, M. (2006). In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nature protocols 1, 2856-2860). Extracted peptides were resuspended in 2% acetonitrile and 0.05% TFA and analyzed using an Orbitrap Exploris 480 Mass Spectrometer (Thermo Fisher). The mass spectrometer emitter voltage was 2100 kV and the ion transfer tube temperature was 300 °C. Full scans were recorded in the range 350–1600 m / z at a mass resolution of 15000. Peptides were fragmented in the collision cell at an energy setting of 28%. Tandem mass spectra were acquired in Scan Range Mode "Define First Mass" and an isolation window of 1.6 m / z with a mass resolution of 15,000.

[0086] Biological Assay Procedures Biological assay 1: HEK biosensor cell tau seeding assay Compound stock solutions were prepared as 10 mM compound in DMSO.

[0087] Human tau repeat domain (RD) carrying the P301L / V337M mutation and fused to a C-terminal GFP tag (TauRD LMHEK293T biosensor cells expressing human tau (termed hTau-GFP) were engineered to study tau seeding in cells (reference: Liu, S. Hossinger, A., Heumueller, SE. et al. Highly efficient intercellular spreading of protein misfolding mediated by viral ligand-receptor interactions. Nat Commun 12, 5739 (2021). https: / / doi.org / 10.1038 / s41467-021-25855-2). HEK cells expressing P301L human full-length tau and tagged with C-terminus GFP were incubated with pre-sonicated hTau 40 P301L The HEK TauRD was incubated with tau fibrils to induce tau aggregate formation. The cells were harvested and the resulting cell lysate was used to synthesize HEK TauRD. LM In further experiments tau aggregates were seeded with -GFP cells.

[0088] In a typical seeding assay, HEK cells were incubated with cell lysate, compound (0–40 µM), and lipofectamine for 22–48 h. Negative control experiments in the absence of compound (vehicle DMSO only) and in the presence of cell lysate were performed in parallel.

[0089] Dose-response curves were generated by quantifying the number of cells with aggregates for each compound concentration. The curves were fitted to a sigmoidal function using Graphpad PRISM version 8. The inhibitory concentration for 50% activity (IC 50 ) was determined for each compound.

[0090] Biological assay 2: Drosophila neurodegeneration model Drug treatment of Drosophila: Stock solutions of the individual compounds were prepared by dissolving 10 mg of compound in 100 μl of 100% DMSO. Osimertinib, osimertinib mesylate and AZ7550 mesylate were orally administered to the flies in their food. The final concentration of each compound used to treat the flies was derived based on the recommended concentration for humans (reference concentration) based on body weight. Three concentrations were used for each compound; one equal to the reference concentration and two higher than the reference concentration. The final concentrations of the different compounds mixed into the fly food were: osimertinib mesylate, 5 μM, 10 μM and 15 μM; osimertinib, 10 μM; AZ7550 mesylate, 10 μM and 20 μM. After hatching, the first instar larvae were transferred onto food containing one of the compounds. Food mixed with the required concentration of drug was replaced with fresh food every 48 h.

[0091] Optical microscopy: After the fly heads were fixed to a glass slide with double-sided sticky tape, light microscopy was performed. Whole-eye images were taken using an Infinity Analyze camera attached to an Olympus SZX7 microscope equipped with a halogen light source. The total and degenerated eye surface areas were marked and calculated using Infinity Analyze software (Lumenera corporation). The percentage of degenerated area was calculated and plotted using GraphPad Prism 8.

[0092] Scanning Electron Microscopy: After eclosion, treated and control flies (2-3 days old) were fixed in 1% formaldehyde for 2 hours, followed by serial dehydration in 25%, 50%, 75% and 100% ethanol for 12 hours each. Flies were stored in 100% ethanol and dried in CPD before being punctured and fixed for imaging. Eye images were taken at 150x and analyzed using Infinity Analyze software.

[0093] References TIFF2025515141000078.tif218166TIFF2025515141000079.tif247166TIFF2025515141000080.tif131165

Claims

1. Formula (I) for use in the prevention or treatment of tauopathy A pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof: During the ceremony, G is And; X is either N or CH; Y is either N or CH; L is And; A is N, CR, or CH; E is N, CR 1 , or CH; R is hydrogen, fluoro, chloro, bromo, -(C 1 -C 6 )alkyl, cyano, or -O(C 1 -C 6 ) is alkyl; R 1 is hydrogen, fluoro, chloro, bromo, -(C 1 -C 6 )alkyl, cyano, or -O(C 1 -C 6 )alkyl; R 2 is hydrogen, fluoro, chloro, bromo, -(C 1 -C 6 )alkyl, cyano, -O(C 1 -C 6 ) alkyl or difluoromethoxy; R 3 teeth And; R 4 is hydrogen, -(C 1 -C 6 )alkyl, or -(C 3 -C 6 )It is a cycloalkyl; R 5 is hydrogen, fluoro, or chloro; R 6 is hydrogen, fluoro, or chloro; R 7 is hydrogen or -(C 1 -C 6 ) is alkyl; R 8 is hydrogen or -(C 1 -C 6 ) is alkyl; R 9 is hydrogen or -(C 1 -C 6 ) is alkyl; R 10 is hydrogen or -(C 1 -C 6 ) is alkyl; R 11 is hydrogen, -(C 1 -C 6 ) alkyl, or -CH 2 CON((C 1 -C 6 )alkyl) 2 And; R 12 is hydrogen or -(C 1 -C 6 ) is alkyl; Here, optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms correspond 11 It has been replaced with a C isotope; One or more nitrogen atoms correspond 13 It is replaced with the N isotope; One or more fluoro atoms correspond 18 It is replaced with the fluorine isotope.

2. Equation (I) Compounds of or pharmaceutically acceptable salts thereof: During the ceremony, G is And; X is selected from N or CH; Y is selected from N or CH; L is And; A is selected from N or CR; E is N or CR 1 Selected from; R is hydrogen, fluoro, chloro, bromo, -(C 1 -C 6 )alkyl, cyano, or -O(C 1 -C 6 ) is alkyl; R 1 is hydrogen, fluoro, chloro, bromo, -(C 1 -C 6 )alkyl, cyano, or -O(C 1 -C 6 ) is alkyl; R 2 is hydrogen, fluoro, chloro, bromo, -(C 1 -C 6 )alkyl, cyano, -O(C 1 -C 6 ) alkyl or difluoromethoxy; R 3 teeth And; R 4 is hydrogen, -(C 1 -C 6 )alkyl, or -(C 3 -C 6 )It is a cycloalkyl; R 5 is hydrogen, fluoro, or chloro; R 6 is hydrogen, fluoro, or chloro; R 7 is hydrogen or -(C 1 -C 6 ) is alkyl; R 8 is hydrogen or -(C 1 -C 6 ) is alkyl; R 9 is hydrogen or -(C 1 -C 6 ) is alkyl; R 10 is hydrogen or -(C 1 -C 6 ) is alkyl; R 11 is hydrogen, -(C 1 -C 6 ) alkyl, or -CH 2 CON((C 1 -C 6 ) alkyl) 2 and; R 12 is hydrogen or -(C 1 -C 6 ) is alkyl; However, the compounds of formula (I) are as follows: Not selected from the group consisting of; Here, optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms correspond 11 It has been replaced with a C isotope; One or more nitrogen atoms correspond 13 It is replaced with the N isotope; One or more fluoro atoms correspond 18 It is replaced with the fluorine isotope.

3. Formula (Ia) or (Ib) Compounds of: During the ceremony, A is N, CR, or CH; E is N, CR, or CH; X is N or CH; R is hydrogen, fluoro, chloro, bromo, -(C 1 -C 6 )alkyl, cyano, -O(C 1 -C 6 )alkyl; L is And; R 1 teeth, Selected from the group consisting of alkylamines, non-aromatic heterocycles, partially or completely deuterated alkylamines, and partially or completely deuterated non-aromatic heterocycles; R 2 is hydrogen, fluoro, chloro, bromo, cyano, or -(C 1 -C 6 ) is alkyl; R 3 is hydrogen, -(C 1 -C 6 )alkyl, -(C 3 -C 6 ) Cycloalkyl or methyl; R 4 is hydrogen, or -(C 1 -C 6 ) is alkyl; R 5 is hydrogen, or -(C 1 -C 6 ) is alkyl; R 6 is hydrogen, or -(C 1 -C 6 ) is alkyl; R 7 is hydrogen, fluoro, -O(C 1 -C 6 )alkyl, or -(C 1 -C 6 ) is alkyl; R 8 is hydrogen, or -(C 1 -C 6 ) is an alkyl hydrogen; R 9 is hydrogen, or -(C 1 -C 6 )alkyl-(C 1 -C 6 ) is alkyl; R 10 is hydrogen, or -(C 1 -C 6 )alkyl-(C 1 -C 6 ) is alkyl; R 11 is hydrogen, or -(C 1 -C 6 )alkyl-(C 1 -C 6 ) is alkyl; R 12 ha-(C 1 -C 6 )alkyl, or -(C 3 -C 6 )It is a cycloalkyl; However, A = N and They do not exist simultaneously in the same compound; Here, optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms correspond 11 It has been replaced with a C isotope; One or more nitrogen atoms correspond 13 It is replaced with the N isotope; One or more fluoro atoms correspond 18 It is replaced with the fluorine isotope.

4. In formula (I), G is And; X is N; Y is N; A is N or; E is N or; R is hydrogen; R 1 is hydrogen; R 2 is a cytokine; R 3 teeth And; R 4 is hydrogen, or -(C 1 -C 6 ) is alkyl; R 5 is hydrogen; R 6 is hydrogen; R 8 is hydrogen, or -(C 1 -C 6 ) is alkyl; R 9 is hydrogen, or -(C 1 -C 6 ) is alkyl; R 10 is hydrogen, or -(C 1 -C 6 ) is alkyl, The pharmaceutical composition according to claim 1.

5. In formula (I), G is And; X is N; Y is N; A is N or CR; E is N, or CR 1 And; R is hydrogen; R 1 is hydrogen; R 2 is a cytokine; R 3 teeth And; R 4 is methyl; R 5 is hydrogen; R 6 is hydrogen; R 8 is hydrogen or methyl; R 9 is methyl; R 10 is methyl; Here, optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms correspond 11 It has been replaced with a C isotope; One or more nitrogen atoms correspond 13 It is replaced with the N isotope; One or more fluoro atoms correspond 18 It is replaced with the F isotope. The pharmaceutical composition according to claim 1.

6. The pharmaceutical composition according to claim 1, wherein only one or two of A, E, X, and Y is N; and only one of A and E is N, and only one of A and Y is N.

7. The pharmaceutical composition according to claim 1, wherein the compound is selected from the group consisting of the following: In the formula, optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms correspond 11 It has been replaced with a C isotope; One or more nitrogen atoms correspond 13 It is replaced with the N isotope; One or more fluoro atoms correspond 18 It is replaced with the fluorine isotope.

8. The pharmaceutical composition according to claim 7, wherein the compound is selected from the group consisting of the following: In the formula, optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms correspond 11 It has been replaced with a C isotope; One or more nitrogen atoms correspond 13 It has been replaced with the N isotope.

9. Compounds selected from the following group or pharmaceutically acceptable salts thereof: In the formula, optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms correspond 11 It has been replaced with a C isotope; One or more nitrogen atoms correspond 13 It is replaced with the N isotope; One or more fluoro atoms correspond 18 It is replaced with the fluorine isotope.

10. The pharmaceutical composition according to claim 1, comprising at least one pharmaceutically acceptable carrier.

11. The pharmaceutical composition according to claim 1, wherein the prevention or treatment of tauopathy includes treatment of the cause of said tauopathy.

12. The pharmaceutical composition according to claim 1, wherein the prevention or treatment of tauopathy comprises inhibiting the cellular activity of tau by binding a compound of formula (I) to tau.

13. The pharmaceutical composition according to claim 12, wherein the bond to tau comprises a covalent bond to tau of the compound of formula (I), or the bond to tau comprises the addition of a thiol-Michael group of the -SH group of tau to an acrylamide group in the compound of formula (I).

14. The pharmaceutical composition according to claim 1, wherein the tauopathy is dementia-related tauopathy.

15. The pharmaceutical composition according to claim 1, wherein the tauopathy is selected from the group consisting of the following: Alzheimer's disease, frontotemporal dementia, primary age-related tauopathy (PART), familial British dementia (FBD), familial Danish dementia (FDD), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), Pick's disease (PiD), Lewy body dementia (DLB), progressive supranuclear palsy (PSP), glial tauopathy (GGT), tauopathy with hippocampal 4-repeat tau immunoreactive spherical inclusions, limbic-predominant neuronal inclusion body 4R tauopathy. Tauopathy (LNT), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), argyrophilic granulopathy (AGD), Huntington's disease, glial globular tauopathy, neuronal astrogenic tauopathy variant in the elderly, familial behavioral variant frontotemporal dementia associated with astrocyte-predominant tauopathy Tauopathy, amyotrophic lateral sclerosis (ALS), spinocerebellar ataxia type 11, spinal muscular atrophy (SMA), progressive ataxia and palatal tremor-related tauopathy, cerebral amyloid angiopathy (CAA), IgLON5 antibody-related tauopathy, Alzheimer's disease associated with trisomy 21 (Down syndrome), vascular dementia, cerebral amyloid angiopathy, Gerstmann-Streussler-Scheinker disease (GSS), Creutzfeldt-Jakob disease, Fatal familial insomnia, Kuru, Niemann-Pick disease type C associated tauopathy, nodding syndrome, non-Guam type motor neuron disease with neurofibrillary tangles, Parkinson's disease (PD), Parkinson's disease with dementia, Guam parkinsonism-dementia, Guadeloupe type parkinsonism, Kosaka-Shibayama disease, post-encephalitis parkinsonism, SYNJ1 (PARK20) early-onset recessive parkinsonism with seizure and dystonia-associated substantia nigra tau pathology (SYNJ1 (PARK20)Early-onset recessive form of parkinsonism with seizures and dystonia associated nigral tau pathology), multiple system atrophy, tau pathology associated with familial parkinsonism and progressive respiratory failure, limbic-dominant neuro-glial tau pathology in TARDBP gene mutation (I383; P112H), neuronal 4R tau pathology in fatal familial insomnia (PRNP D178N mutation), tau pathology associated with ADCY5 dyskinesia, tau pathology in chronic temporal lobe epilepsy, neurodegenerative diseases with intracranial iron deposition (NBIA), tau pathology in NBIA PANK2 and WDR45 gene mutations, tau pathology in NBIA PLA2G6 mutation, tau pathology in NBIA associated with autosomal dominant mitochondrial membrane protein-associated neurodegeneration (MPAN), neuropil threads pretangles and neurofibrillary tangles in human immunodeficiency virus (HIV)-negative opioid abusers. Neurofibrillary tangles in human immunodeficiency virus (HIV)-negative opiate abusers, tau pathology associated with acquired immunodeficiency syndrome (AIDS), diffuse neurofibrillary tangles with calcification, progressive ataxia and palatal tremor, SLC9A6-associated parkinsonism, tau pathology associated with SPG7 gene mutations, striatal 4R tau pathology associated with X-linked parkinsonism with spasticity Spasticity: ATP6AP2), tau pathology associated with SPAST gene-associated hereditary spastic paraplegia, autism, autism spectrum disorder, retinal tauopathy, West Nile encephalomyelitis, TTBK2 gene-associated spinocerebellar degeneration11, herpes simplex encephalitis, neurofibrillary tangle-type senile dementia (TOD), age-related tauastropathy (ARTAG), hippocampal tauopathy, subacute sclerosing panencephalitis (SSPE)-associated tauopathy, FTLD-C9ORF72, Christianson syndrome, vacuolar tauopathyTauopathy), Ritico-Bodig disease, ganglioglioma and gangliocytoma, meningeal hemangioma, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, neuronal ceroid lipofuscinosis, myotonic dystrophy, Fukuyama-type congenital muscular dystrophy, unilateral megacephaly, focal cortical dysplasia, Walcott-Larrisson syndrome, primary lateral sclerosis, progressive freezing of gait, PSP with parkinsonism, Richardson syndrome, non-fluent / agrammatic primary progressive aphasia, semantic primary progressive aphasia, logopenic primary progressive aphasia, primary progressive apraxia of speech, and amnesic Alzheimer's disease.

16. The pharmaceutical composition according to claim 10, which is applied parenterally or orally.

17. In formula (I), G is And; X is N; Y is N; A is N or CR; E is N, or CR 1 And; R is hydrogen; R 1 is hydrogen; R 2 is a cytokine; R 3 teeth And; R 4 is hydrogen, or -(C 1 -C 6 ) is alkyl; R 5 is hydrogen; R 6 is hydrogen; R 8 is hydrogen, or -(C 1 -C 6 ) is alkyl; R 9 is hydrogen, or -(C 1 -C 6 ) is alkyl; R 10 is hydrogen, or -(C 1 -C 6 ) is alkyl, The compound according to claim 2 or a pharmaceutically acceptable salt thereof.

18. In formula (I), G is And; X is N; Y is N; A is N or CR; E is N, or CR 1 And; R is hydrogen; R 1 is hydrogen; R 2 is a cytokine; R 3 teeth And; R 4 is methyl; R 5 is hydrogen; R 6 is hydrogen; R 8 is hydrogen or methyl; R 9 is methyl; R 10 It is methyl, Here, optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms correspond 11 It has been replaced with a C isotope; One or more nitrogen atoms correspond 13 It is replaced with the N isotope; One or more fluoro atoms correspond 18 It is replaced with the F isotope. The compound according to claim 2 or a pharmaceutically acceptable salt thereof.

19. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein only one or two of A, E, X, and Y are N; or only one of A and E is N, and only one of A and Y is N.

20. A pharmaceutical composition comprising a compound of formula (I) according to claim 2 or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of tauopathy.

21. A pharmaceutical composition comprising a compound of formula (Ia) or (Ib) according to claim 3 or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of tauopathy.

22. The pharmaceutical composition according to claim 20, wherein the prevention or treatment of tauopathy includes treatment of the cause of said tauopathy.

23. The pharmaceutical composition according to claim 21, wherein the prevention or treatment of tauopathy includes treatment of the cause of said tauopathy.

24. The pharmaceutical composition according to claim 20, wherein the prevention or treatment of tauopathy comprises inhibiting the cellular activity of tau by binding a compound of formula (I) to tau.

25. The pharmaceutically active composition according to claim 21, wherein the prevention or treatment of tauopathy comprises inhibiting the cellular activity of tau by binding a compound of formula (Ia) or (Ib) to tau.

26. The pharmaceutically active composition according to claim 20, wherein the prevention or treatment of tauopathy comprises inhibiting the cellular activity of tau by binding a compound of formula (I) to tau, or comprising covalent bonding of a compound of formula (I) to tau, or comprising thiol-Michael addition of the -SH group of tau to the acrylamide group in the compound of formula (I).

27. The pharmaceutically active composition according to claim 21, wherein the prevention or treatment of tauopathy comprises inhibiting the cellular activity of tau by binding a compound of formula (I) to tau, or comprising covalent bonding of a compound of formula (I) to tau, or comprising thiol-Michael addition of the -SH group of tau to the acrylamide group in the compound of formula (I).

28. The pharmaceutical composition according to claim 20, wherein the tauopathy is dementia-related tauopathy.

29. The pharmaceutical composition according to claim 21, wherein the tauopathy is dementia-related tauopathy.

30. The pharmaceutical composition according to claim 20, wherein the tauopathy is selected from the group consisting of the following: Alzheimer's disease, frontotemporal dementia, primary age-related tauopathy (PART), familial British dementia (FBD), familial Danish dementia (FDD), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), Pick's disease (PiD), Lewy body dementia (DLB), progressive supranuclear palsy (PSP), glial tauopathy (GGT), tauopathy with hippocampal 4-repeat tau immunoreactive spherical inclusions, limbic-predominant neuronal inclusion body 4R tauopathy. Tauopathy (LNT), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), argyrophilic granulopathy (AGD), Huntington's disease, glial globular tauopathy, neuronal astrogenic tauopathy variant in the elderly, familial behavioral variant frontotemporal dementia associated with astrocyte-predominant tauopathy Tauopathy, amyotrophic lateral sclerosis (ALS), spinocerebellar ataxia type 11, spinal muscular atrophy (SMA), progressive ataxia and palatal tremor-related tauopathy, cerebral amyloid angiopathy (CAA), IgLON5 antibody-related tauopathy, Alzheimer's disease associated with trisomy 21 (Down syndrome), vascular dementia, cerebral amyloid angiopathy, Gerstmann-Streussler-Scheinker disease (GSS), Creutzfeldt-Jakob disease, Fatal familial insomnia, Kuru, Niemann-Pick disease type C associated tauopathy, nodding syndrome, non-Guam type motor neuron disease with neurofibrillary tangles, Parkinson's disease (PD), Parkinson's disease with dementia, Guam parkinsonism-dementia, Guadeloupe type parkinsonism, Kosaka-Shibayama disease, post-encephalitis parkinsonism, SYNJ1 (PARK20) early-onset recessive parkinsonism with seizure and dystonia-associated substantia nigra tau pathology (SYNJ1 (PARK20)Early-onset recessive form of parkinsonism with seizures and dystonia associated nigral tau pathology), multiple system atrophy, tau pathology associated with familial parkinsonism and progressive respiratory failure, limbic-dominant neuro-glial tau pathology in TARDBP gene mutation (I383; P112H), neuronal 4R tau pathology in fatal familial insomnia (PRNP D178N mutation), tau pathology associated with ADCY5 dyskinesia, tau pathology in chronic temporal lobe epilepsy, neurodegenerative diseases with intracranial iron deposition (NBIA), tau pathology in NBIA PANK2 and WDR45 gene mutations, tau pathology in NBIA PLA2G6 mutation, tau pathology in NBIA associated with autosomal dominant mitochondrial membrane protein-associated neurodegeneration (MPAN), neuropil threads pretangles and neurofibrillary tangles in human immunodeficiency virus (HIV)-negative opioid abusers. Neurofibrillary tangles in human immunodeficiency virus (HIV)-negative opiate abusers, tau pathology associated with acquired immunodeficiency syndrome (AIDS), diffuse neurofibrillary tangles with calcification, progressive ataxia and palatal tremor, SLC9A6-associated parkinsonism, tau pathology associated with SPG7 gene mutations, striatal 4R tau pathology associated with X-linked parkinsonism with spasticity Spasticity: ATP6AP2), tau pathology associated with SPAST gene-associated hereditary spastic paraplegia, autism, autism spectrum disorder, retinal tauopathy, West Nile encephalomyelitis, TTBK2 gene-associated spinocerebellar degeneration11, herpes simplex encephalitis, neurofibrillary tangle-type senile dementia (TOD), age-related tauastropathy (ARTAG), hippocampal tauopathy, subacute sclerosing panencephalitis (SSPE)-associated tauopathy, FTLD-C9ORF72, Christianson syndrome, vacuolar tauopathyTauopathy), Ritico-Bodig disease, ganglioglioma and gangliocytoma, meningeal hemangioma, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, neuronal ceroid lipofuscinosis, myotonic dystrophy, Fukuyama-type congenital muscular dystrophy, unilateral megacephaly, focal cortical dysplasia, Walcott-Larrisson syndrome, primary lateral sclerosis, progressive freezing of gait, PSP with parkinsonism, Richardson syndrome, non-fluent / agrammatic primary progressive aphasia, semantic primary progressive aphasia, logopenic primary progressive aphasia, primary progressive apraxia of speech, and amnesic Alzheimer's disease.

31. The pharmaceutical composition according to claim 21, wherein the tauopathy is selected from the group consisting of the following: Alzheimer's disease, frontotemporal dementia, primary age-related tauopathy (PART), familial British dementia (FBD), familial Danish dementia (FDD), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), Pick's disease (PiD), Lewy body dementia (DLB), progressive supranuclear palsy (PSP), glial tauopathy (GGT), tauopathy with hippocampal 4-repeat tau immunoreactive spherical inclusions, limbic-predominant neuronal inclusion body 4R tauopathy. Tauopathy (LNT), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), argyrophilic granulopathy (AGD), Huntington's disease, glial globular tauopathy, neuronal astrogenic tauopathy variant in the elderly, familial behavioral variant frontotemporal dementia associated with astrocyte-predominant tauopathy Tauopathy, amyotrophic lateral sclerosis (ALS), spinocerebellar ataxia type 11, spinal muscular atrophy (SMA), progressive ataxia and palatal tremor-related tauopathy, cerebral amyloid angiopathy (CAA), IgLON5 antibody-related tauopathy, Alzheimer's disease associated with trisomy 21 (Down syndrome), vascular dementia, cerebral amyloid angiopathy, Gerstmann-Streussler-Scheinker disease (GSS), Creutzfeldt-Jakob disease, Fatal familial insomnia, Kuru, Niemann-Pick disease type C associated tauopathy, nodding syndrome, non-Guam type motor neuron disease with neurofibrillary tangles, Parkinson's disease (PD), Parkinson's disease with dementia, Guam parkinsonism-dementia, Guadeloupe type parkinsonism, Kosaka-Shibayama disease, post-encephalitis parkinsonism, SYNJ1 (PARK20) early-onset recessive parkinsonism with seizure and dystonia-associated substantia nigra tau pathology (SYNJ1 (PARK20)Early-onset recessive form of parkinsonism with seizures and dystonia associated nigral tau pathology), multiple system atrophy, tau pathology associated with familial parkinsonism and progressive respiratory failure, limbic-dominant neuro-glial tau pathology in TARDBP gene mutation (I383; P112H), neuronal 4R tau pathology in fatal familial insomnia (PRNP D178N mutation), tau pathology associated with ADCY5 dyskinesia, tau pathology in chronic temporal lobe epilepsy, neurodegenerative diseases with intracranial iron deposition (NBIA), tau pathology in NBIA PANK2 and WDR45 gene mutations, tau pathology in NBIA PLA2G6 mutation, tau pathology in NBIA associated with autosomal dominant mitochondrial membrane protein-associated neurodegeneration (MPAN), neuropil threads pretangles and neurofibrillary tangles in human immunodeficiency virus (HIV)-negative opioid abusers. Neurofibrillary tangles in human immunodeficiency virus (HIV)-negative opiate abusers, tau pathology associated with acquired immunodeficiency syndrome (AIDS), diffuse neurofibrillary tangles with calcification, progressive ataxia and palatal tremor, SLC9A6-associated parkinsonism, tau pathology associated with SPG7 gene mutations, striatal 4R tau pathology associated with X-linked parkinsonism with spasticity Spasticity: ATP6AP2), tau pathology associated with SPAST gene-associated hereditary spastic paraplegia, autism, autism spectrum disorder, retinal tauopathy, West Nile encephalomyelitis, TTBK2 gene-associated spinocerebellar degeneration11, herpes simplex encephalitis, neurofibrillary tangle-type senile dementia (TOD), age-related tauastropathy (ARTAG), hippocampal tauopathy, subacute sclerosing panencephalitis (SSPE)-associated tauopathy, FTLD-C9ORF72, Christianson syndrome, vacuolar tauopathyTauopathy), Ritico-Bodig disease, ganglioglioma and gangliocytoma, meningeal hemangioma, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, neuronal ceroid lipofuscinosis, myotonic dystrophy, Fukuyama-type congenital muscular dystrophy, unilateral megacephaly, focal cortical dysplasia, Walcott-Larrisson syndrome, primary lateral sclerosis, progressive freezing of gait, PSP with parkinsonism, Richardson syndrome, non-fluent / agrammatic primary progressive aphasia, semantic primary progressive aphasia, logopenic primary progressive aphasia, primary progressive apraxia of speech, and amnesic Alzheimer's disease.

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