Inhibitors of tau protein

Abstract

The present invention relates to a compound for preventing or treating tauopathy and a pharmaceutical composition thereof.

Description

Technical Field

[0001] The present invention relates to a compound for preventing or treating tau protein disease and a pharmaceutical composition thereof. Background Art

[0002] Tau is a microtubule-associated protein found primarily in the axons and dendrites of neurons, where it plays a role in microtubule organization and regulation. 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 granulomatosis (AGD), and frontotemporal dementia with parkinsonism-17 (FTDP-17). 1 The neuropathological hallmarks of AD are extracellular amyloid plaques composed of amyloid-β (Aβ), and intracellular inclusions of aggregated tau protein known as neurofibrillary tangles (NFTs). 2

[0003] The spread of aggregated tau pathology correlates with the degree of clinical dementia in AD. 3,4 Tau pathology is thought to spread throughout the brain in a typical manner: aggregates of tau first form in the nucleus coelicolor and spread through the entorhinal cortex and neocortex, which are adjacent to the hippocampus. 5 Therefore, potential therapeutic strategies for AD include targeting tau aggregation and spread. In addition, tau aggregation can be observed at least 20 years before the onset of clinical symptoms of AD, suggesting that inhibiting this process may be a preventive measure against AD. 6,7

[0004] According to the terminology proposed by the Common Alzheimer's Disease and Related Dementias Research Ontology (CADRO), pharmaceutical agents developed to target amyloid plaques or tau aggregates in AD are classified as "disease-modifying therapies (DMTs)" for AD. 8 Among DMTs in the AD drug development pipeline over the past 5 years, Aβ-directed therapies have accounted for the majority, while tau-based therapies have lagged behind. 9 Notable examples of Aβ-targeted DMTs are Aducanumab and Lecanemab, which were approved by the U.S. Federal Drug Administration (FDA) in 2021 and 2023, respectively. However, some AD patients treated with these two antibodies experienced brain hemorrhage, suggesting a potentially low risk-benefit ratio. Many clinical trials of other Aβ-directed drugs have failed, primarily due to lack of efficacy. 6In the case of tau-based DMTs, progress has been made in identifying small molecules that disrupt tau aggregation. However, to date, no tau-based DMTs have received FDA approval. Therefore, there is a need for drugs that can prevent or treat tauopathies. Summary of the invention

[0005] The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for preventing or treating tauopathy.

[0006]

[0007] in

[0008] G is Best

[0009] X is N or CH, preferably N;

[0010] Y is N or CH, preferably N;

[0011] L is Best

[0012] A is N or CR; preferably CR, more preferably CH;

[0013] E is N or CR 1 ; CR is preferred 1 , more preferably CH;

[0014] R is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano or -O(C1-C6)alkyl, preferably hydrogen;

[0015] R 1 is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano or -O(C1-C6)alkyl, preferably hydrogen;

[0016] R 2 is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano, -O(C1-C6)alkyl or difluoromethoxy, preferably -O(C1-C6)alkyl;

[0017] R 3 for or -NR 8 R 9 , preferably

[0018] R 4 is hydrogen, -(C1-C6)alkyl or -(C3-C6)cycloalkyl;

[0019] R 5 is hydrogen, fluorine or chlorine;

[0020] R 6 is hydrogen, fluorine or chlorine;

[0021] R 7 is hydrogen or -(C1-C6)alkyl;

[0022] R 8 is hydrogen or -(C1-C6)alkyl;

[0023] R 9 is hydrogen or -(C1-C6)alkyl;

[0024] R 10 is hydrogen or -(C1-C6)alkyl;

[0025] R 11 is hydrogen, -(C1-C6)alkyl or -CH2CON((C1-C6)alkyl)2;

[0026] R 12 is hydrogen or -(C1-C6)alkyl;

[0027] wherein optionally,

[0028] One or more hydrogen atoms may be replaced by deuterium atoms;

[0029] One or more carbon atoms may be 11 C isotope substitution;

[0030] One or more nitrogen atoms may be replaced by the corresponding 13 N-isotope substitution;

[0031] One or more fluorine atoms may be replaced by the corresponding 18 F-isotope substitution.

[0032] In another embodiment, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof

[0033]

[0034] in

[0035] G is Best

[0036] X is N or CH, preferably N;

[0037] Y is N or CH, preferably N;

[0038] L is Best

[0039] A is N or CR; preferably CR, more preferably CH

[0040] E is N or CR 1 ; CR is preferred 1 , more preferably CH

[0041] R is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano or -O(C1-C6)alkyl, preferably hydrogen;

[0042] R 1 is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano or -O(C1-C6)alkyl, preferably hydrogen;

[0043] R 2 is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano, -O(C1-C6)alkyl or difluoromethoxy, preferably -O(C1-C6)alkyl;

[0044] R 3 for or -NR 8 R 9 , preferably

[0045] R 4 is hydrogen, -(C1-C6)alkyl or -(C3-C6)cycloalkyl;

[0046] R 5 is hydrogen, fluorine or chlorine;

[0047] R 6 is hydrogen, fluorine or chlorine;

[0048] R 7 is hydrogen or -(C1-C6)alkyl;

[0049] R 8 is hydrogen or -(C1-C6)alkyl, preferably -(C1-C6)alkyl;

[0050] R 9 is hydrogen or -(C1-C6)alkyl, preferably -(C1-C6)alkyl;

[0051] R 10 is hydrogen or -(C1-C6)alkyl;

[0052] R 11 is hydrogen, -(C1-C6)alkyl or -CH2CON((C1-C6)alkyl)2;

[0053] R12 is hydrogen or -(C1-C6)alkyl;

[0054] Provided that the compound is not selected from

[0055]

[0056]

[0057]

[0058]

[0059] wherein optionally,

[0060] One or more hydrogen atoms may be replaced by deuterium atoms;

[0061] One or more carbon atoms may be 11 C isotope substitution;

[0062] One or more nitrogen atoms may be replaced by the corresponding 13 N isotope substitution;

[0063] One or more fluorine atoms may be replaced by the corresponding 18 F isotope substitution.

[0064] The compounds of the present invention have been found to covalently bind to Cys291 and Cys322 of tau in vitro and inhibit tau aggregation. It has been further demonstrated that the compounds disrupt in situ tau aggregation in cells in a Drosophila model of tau overexpression, as well as tau-induced pathology in vivo (see Fig.12 ).

[0065] In another embodiment, the present invention relates to a pharmaceutical composition for preventing or treating tauopathy, comprising a compound according to formula (I), (Ia) and / or (Ib) and at least one pharmaceutically acceptable carrier.

[0066] The compounds or pharmaceutical compositions of the present invention are also used to prevent or treat tauopathies 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), globular glial tauopathy (GGT), tauopathy with hippocampal 4-repeat tau immunoreactive spherical inclusions, limbic dominant neuronal inclusions 4R Tauopathies (LNT), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), argyrophilic grain disease (AGD), Huntington's disease, glial globular tauopathy, senile neuro-astrocytic tauopathy, familial behavioral variant frontotemporal dementia associated with astrocyte-dominant tauopathy, amyotrophic lateral sclerosis (ALS), spinocerebellar ataxia type 11, spinal muscular atrophy (SMA), tauopathies associated with progressive ataxia and palatal tremor, cerebral amyloid angiopathy (CAA), IgLON5 antibody-associated tauopathy, trisomy 21-associated Alzheimer's disease (Down syndrome), vascular dementia, cerebral amyloid Vascular disease, Gerstmann-Straussler-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, Parkinson's disease-Guam dementia, Guadeloupe Parkinson's disease, Kosaka-Shibayama disease, postencephalitic parkinsonism, SYNJ1 (PARK20) early-onset recessive parkinsonism with nigral tauopathy associated with seizures and dystonia, multiple system atrophy, tauopathies associated with familial parkinsonism and progressive respiratory failure, TARDBP gene mutations (I383;P112H), neuronal-glial tauopathy in limbic-dominant neuron-glial tauopathy, neuronal 4R tauopathy in fatal familial insomnia (PRNP D178N mutation), tauopathy associated with ADCY5 dyskinesia, tauopathy in chronic temporal lobe epilepsy, neurodegenerative disease with brain iron accumulation (NBIA), tauopathy in NBIA PANK2 and WDR45 gene mutations, NBIA Tauopathies in PLA2G6 mutations, tauopathies in NBIA associated with autosomal dominant mitochondrial membrane protein-associated neurodegeneration (MPAN), neurofibrillary pretangles and neurofibrillary tangles in human immunodeficiency virus (HIV)-negative opioid abusers, tauopathies associated with acquired immune deficiency syndrome (AIDS), diffuse neurofibrillary tangles with calcification, progressive ataxia and palatal tremor, Parkinson's disease associated with SLC9A6, tauopathies associated with mutations in the SPG7 gene, striatal 4R associated with X-linked parkinsonism spasticity (ATP6AP2) Tauopathies, tauopathies associated with hereditary spastic paraplegia associated with the SPAST gene, autism, autism spectrum disorder, retinal tauopathy, West Nile encephalomyelitis, TTBK2 gene-associated spinocerebellar ataxia 11, herpes simplex encephalitis, tangled dementia (TOD), tau astrogliopathy associated with aging (ARTAG), hippocampal tauopathy, tauopathy associated with subacute sclerosing panencephalitis (SSPE), FTLD-C9ORF72, Christianson syndrome, vacuolar tauopathy, lytico-bodig disease, ganglioglioma and gangliocytoma, meningioangiomatosis, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, neuronal ceroid lipofuscinosis, myotonic dystrophy , Fukuyama congenital muscular dystrophy, hemimerical encephaly, focal cortical dysplasia, Wolcott-Rallison syndrome, primary lateral sclerosis, progressive freezing of gait, PSP with Parkinson's disease, Richardson syndrome, non-fluent / grammatical variant of primary progressive aphasia, semantic variant of primary progressive aphasia, language reduction variant of primary progressive aphasia, primary progressive apraxia of speech, amnestic Alzheimer's disease, preferably Alzheimer's disease (AD), Pick's disease (PiD), progressive supranuclear palsy (PSP), cortical basal 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). ; BRIEF DESCRIPTION OF THE DRAWINGS

[0067] Figure 1: Osimertinib, compound 2 (AZ7550) and compound 3 (AZ5104) inhibited in vitro seed aggregation / fibrosis of full-length tau in a dose-dependent manner. (A) Effects of osimertinib, compound 2 and 3 on ThT fluorescence curves of 2N4R tau in different formulations. Within 3 days, tau aggregation reached an equilibrium state, corresponding to maximum ThT fluorescence. At this point, the aggregation mixture containing osimertinib was precipitated 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) was approximately 80-90% lower than the control without fibrosis (lane 2). (C) At a molar ratio of 1:10 (tau: osimertinib), osimertinib significantly increased the amount of soluble tau. The error bars correspond to the standard deviation of the band intensities from two replicates. Statistically significant differences were determined by one-way ANOVA analysis and Dunnett's multiple comparison test, p<0.033 (*).

[0068] Figure 2 : In vitro seeding fibrillization assay of 2N4R tau in the presence of osimertinib and 2.5% (v / v) dimethyl sulfoxide (DMSO). Osimertinib has low solubility in water without cosolvents or counterions, which interferes with fluorescence-based aggregation assays. To avoid this problem, aggregation assays and other experiments can be performed in the presence of the cosolvent DMSO. At a concentration of 2.5% (v / v) DMSO, there is strong inhibitory activity of osimertinib on tau aggregation.

[0069] Figure 3 :With cysteine-free tau mutant (tau C291S，C322S ) aggregation, osimertinib mesylate more effectively inhibited wild-type tau (tau wt ) and 3R tau (tau 3R ) aggregation. wt , tau 3R or 2N4Rtau C291S，C322S Aggregation / fibrillation of in vitro seeding was performed by incubation with osimertinib mesylate (tau:compound molar ratio of 1:3 and 1:10) and monitoring the increase in ThT fluorescence over time (A). The normalized Tm values ​​at 1:3 and 1:10 molar ratios did not show a clear dose dependence (B). However, at 1:3 and 1:10 molar ratios of tau wt and tau 3R Statistically lower normalized Span values ​​were observed under tau C291S，C322S0.002 (**), p≤0.0002 (***), and p≤0.0001 (****).

[0070] Figure 4 : Osimertinib, compound 2 (AZ7550 hydrochloride) and compound 3 (AZ5104) partially dissolve 2N4Rtau aggregates in vitro. (A) ThT fluorescence curves monitoring tau aggregation. Tau fibrillization reaches a saturation point in about 40 hours, and then compounds or buffer (reference) are added to tau fibrils (tau monomer: compound molar ratio 1:3 and 1:10). The experiment was performed in duplicate. (B) The addition of compounds resulted in a decrease in ThT fluorescence intensity, indicating that tau aggregates were partially dissolved. Statistically significant differences were determined by one-way ANOVA and Dunnett's multiple comparison test. Significant differences corresponding to p≤0.0002 (***) and p≤0.0001 (****) are shown. (C) After incubation with the compounds for 1 day, transmission electron micrographs of tau-compound mixtures show the presence of fibrils, indicating that the compounds did not completely dissolve the fibrils. The scale bar is 500nm. (D) SDS-PAGE analysis of precipitated tau-compound mixtures shows an increase in the amount of soluble tau after incubation of tau fibrils with compound 3 (AZ5104) and compound 2 (AZ7550 hydrochloride (1:10 tau:compound)). The dashed lines correspond to one standard deviation above the reference value.

[0071] Figure 5 : NMR spectra showing the binding of osimertinib mesylate, compound 2 (AZ7550 hydrochloride) and compound 3 (AZ5104) to tau. 15 N-labeled 2N4R tau (18 μM tau, 50 mM sodium phosphate, pH 6.8) was incubated at 37°C for 16 h in the presence or absence of compound, and SOFAST-heteronuclear multiple quantum coherence (HMQC) spectra were recorded at 5°C. (A) Overlapping spectra of 2N4R tau with (black) and without (grey) a 100-fold molar excess of osimertinib mesylate. (B) A magnified version of the boxed area from (A) shows peaks from residues belonging to the N-terminal region of tau with significant broadening. Chemical shift perturbations (CSPs) above ˜0.002 ppm and signal broadening induced by compound addition (I / I0<1) indicate that the compound binds to the monomeric form of tau. As seen in the case of compound 3 (AZ5104), the I / I0 values ​​are greater than 1, indicating that the compound solubilizes tau oligomers formed during the 16-hour incubation, thus resulting in an increase in the NMR signal intensity.

[0072] Figure 6 : Electrospray ionization mass spectrometry (ESI-MS) confirmed the covalent modification of cysteine ​​(244-372, 13818 Da) in tau by osimertinib (500 Da), compound 2 (AZ7550) without hydrochloride (486 Da) and compound 3 (AZ5104) (486 Da).

[0073] Figure 7 : MS / MS analysis of osimertinib adducts in tau. Full-length 2N4R tau was incubated with osimertinib mesylate (tau:compound molar ratio of 1:5) at 37°C overnight, trypsin digested, and the trypsin digest analyzed by LC-MS / MS. Peptide mass spectra corresponding to osimertinib-modified C291 (A) and C322 (B) confirm covalent modification of cysteine ​​in tau. (C) In total, approximately 350 tau peptide spectrum matches showed osimertinib-modified cysteine ​​sites for osimertinib-treated tau samples. (D) Negative control corresponding to compound-free tau incubated at 37°C shows a small number of false positive peptide spectrum matches.

[0074] Figure 8 : The binding epitope of osimertinib to monomeric tau was determined and compared with the epitopes of compound 2 (AZ7550 hydrochloride) and compound 3 (AZ5104). Monomeric 2N4R tau (10 μM and 20 μM) was added to the compound (250 μM in 50 mM sodium phosphate buffer, containing 2.5% v / v DMSO, pH 6.8, 37°C), inducing chemical shift perturbations (CSPs) in the compound ID NMR spectrum, which allowed identification of aliphatic and aromatic protons (AB) involved in binding to monomeric tau. The CSPs were graded and localized to the chemical structure of osimertinib (C). Large CSPs represent the most likely binding epitopes for osimertinib.

[0075] Fig. 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 with 100 mM NaCl and 2.5% (v / v) DMSO at pH 6.8, 37°C. 1D was obtained in an interleaved manner. 1 HNMR STD spectra, alternating between off-resonance illumination at 60 ppm and resonant illumination at -2 ppm. The control 1D spectrum is shown in grey, the difference spectrum in black. (DF) Measurement of STD effect (I STD) and normalized to the maximum STD signal (indolemethyl protons set to 100%). (GI) Protons surrounding the chemical structures of osimertinib, compound 2 (AZ7550 hydrochloride), and compound 3 (AZ5104) showing the STD effect. These protons represent the tau fibrillization binding epitope of each compound.

[0076] Fig.10 Osimertinib inhibits tau seeding aggregation in HEK biosensor cells. P301L Lysates of HEK cells seeded with fibril-seeded Tau aggregates induced aggregate formation in the cells. Cells were incubated with drugs (0.25 μM or 7.5 μM) for 22 hours in the presence of lipofectamine. Cells incubated with higher concentrations of osimertinib showed reduced aggregate formation.

[0077] Fig.11 : Comparison of the inhibitory activity of osimertinib, compound 2 (AZ7550) hydrochloride and compound 3 (AZ5104) on tau aggregation in HEK biosensor cells. As described above (see Fig.10 ), cell assays were performed using different batches of HEK cells (each dose response curve corresponds to one batch of cells), and the number of cells with aggregates (grey) was quantified relative to the negative control represented by cells treated with vehicle (DMSO). The total amount of cells (black) remained close to 100% until the compound concentration was about 5.0 μM, indicating low toxicity in the concentration range of 0-5.0 μM. The half-maximal inhibitory concentration (IC) was determined by curve fitting 50 ). Control compounds such as rociletinib and EGCG showed much lower inhibitory activity.

[0078] Fig.12 : Osimertinib mesylate and compound 2 (AZ7550) mesylate alleviate tau-induced degeneration in Drosophila eyes. (AC) Drosophila with (GMR-Gal4 / WT tau) and without (GMR-Gal4 / +) tau expression were treated with osimertinib mesylate or compound 2 (AZ7550) mesylate from larval to adult stages. The percentage of degeneration was assessed from microscopic images of Drosophila eyes. Five independent parallel experiments were performed on different flies on different days for osimertinib mesylate and compound 2 (AZ7550) mesylate. Statistically different groups were determined by unpaired t-test, p≤0.05 (*) p≤0.01 (**), p≤0.001 (***), p≤0.0001 (****). (D) Representative SEM micrographs of flies treated with osimertinib showing significantly reduced signs of degeneration compared to control GMR-Gal4 / WT tau.

[0079] Fig.13 : The 2N4R tau aggregation inhibitory activity of exemplary compounds was compared 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) and repeated at least 3 times. For each condition, the midpoint T of the aggregation curve was quantified and normalized. m , and it is a measure of the degree of aggregation delay. Among the compounds tested, only compound 31 with an -OCH2- linking group (L) showed significant aggregation delay and was significantly more effective than compound 1 and all other compounds with L=-NH-. This demonstrates the advantage of modifying the linker structure to increase anti-aggregation activity. Null hypothesis testing was performed by one-way ANOVA with Dunnett's multiple comparisons. Significant differences corresponding to p≤0.0001 (****) are shown.

[0080] Fig.14 : Comparison between the inhibitory activity of osimertinib derivatives designated as compound 13 and compound 9 on tau aggregation in HEK biosensor cells. The number of cells with aggregates (black circles) was quantified relative to the negative control represented by cells treated with vehicle (DMSO). The total amount of cells (grey triangles) remained close to 100% until the compound concentration was about 4.0 μM, indicating low toxicity in the concentration range of 0-4.0 μM. The half maximal inhibitory concentration (IC) was determined by curve fitting. 50 ). DETAILED DESCRIPTION

[0081] definition

[0082] The term "alkyl" refers to a monovalent group of a saturated straight 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, isopentyl, n-hexyl, isohexyl, sec-hexyl, n-heptyl, isoheptyl, n-octyl, 2-ethyl-hexyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, etc.

[0083] "Pharmaceutically acceptable salts" include salts formed with pharmaceutically acceptable acids or bases. Pharmaceutically acceptable acids include inorganic acids and organic 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 alkali metal (e.g., sodium or potassium) and alkaline earth metal (e.g., calcium or magnesium) hydroxides and organic bases, such as alkylamines, arylalkylamines and heterocyclic amines.

[0084] ****

[0085] Unless otherwise indicated, the term "at least" before a series of elements is understood to refer to each element in the series. Those skilled in the art will recognize, or be able to determine, many equivalents of the specific embodiments of the invention described herein using no more than routine experimentation. These equivalents are also included in the present invention.

[0086] The term "and / or" as used herein includes the meanings of "and", "or" and "all or any other combinations of the elements connected by the term".

[0087] Throughout the specification and in the claims that follow, unless the context requires otherwise, the word "comprise" and variations such as "include" and "comprising" should be understood to imply the inclusion of stated integers or steps or groups of integers or steps, but not the exclusion of any other integers or steps or groups of integers or steps. When used herein, the term "comprising" may be replaced with the terms "containing" or "including", or sometimes when used herein, with the term "having". When used herein, "consisting of excludes any elements, steps or ingredients not specified.

[0088] The term "including" means "including but not limited to." "Including" and "including but not limited to" are used interchangeably.

[0089] ****

[0090] Compound

[0091] The compounds of the present invention are defined by the following formula (I):

[0092]

[0093] in

[0094] G is Best More preferred

[0095] X is N or CH, preferably N;

[0096] Y is N or CH, preferably N;

[0097] L is Best

[0098] A is N or CR; preferably CR, more preferably CH;

[0099] E is N or CR 1 ; CR is preferred 1 , more preferably CH;

[0100] Preferably, only one or two of A, E, X and Y are N; A and E are not N at the same time, and A and Y are not N at the same time;

[0101] R is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano or -O(C1-C6)alkyl, preferably hydrogen;

[0102] R 1 is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano or -O(C1-C6)alkyl, preferably hydrogen;

[0103] R 2 is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano, -O(C1-C6)alkyl or difluoromethoxy, preferably -O(C1-C6)alkyl;

[0104] R 3 for or -NR 8 R 9 , preferably

[0105] R 4 is hydrogen, -(C1-C6)alkyl or -(C3-C6)cycloalkyl;

[0106] R 5 is hydrogen, fluorine or chlorine;

[0107] R 6 is hydrogen, fluorine or chlorine;

[0108] R 7 is hydrogen or -(C1-C6)alkyl;

[0109] R 8 is hydrogen or -(C1-C6)alkyl;

[0110] R 9 is hydrogen or -(C1-C6)alkyl;

[0111] R 10 is hydrogen or -(C1-C6)alkyl;

[0112] R 11 is hydrogen, -(C1-C6)alkyl or -CH2CON((C1-C6)alkyl)2;

[0113] R 12 is hydrogen or -(C1-C6)alkyl;

[0114] wherein optionally,

[0115] One or more hydrogen atoms are replaced by deuterium atoms;

[0116] One or more carbon atoms are 11 C isotope substitution;

[0117] One or more nitrogen atoms are replaced by the corresponding 13 N isotope substitution;

[0118] One or more fluorine atoms are replaced by the corresponding 18 F isotope substitution;

[0119] or a pharmaceutically acceptable salt thereof.

[0120] In another embodiment, at least one of the following conditions applies:

[0121] i) R is fluorine, chlorine or cyano;

[0122] ii) R 1 is fluorine, methoxy;

[0123] iii) R 2 is hydrogen, fluorine, chlorine, cyano or methyl;

[0124] iv) R 3 is pyrrolidinyl, piperidinyl, 4-methylpiperidinyl or 2-methylpyrrolidinyl;

[0125] v) L is -OCH2-

[0126] In another embodiment, the compound is not selected from

[0127]

[0128]

[0129]

[0130]

[0131]

[0132] In a further embodiment

[0133] G is

[0134] X is selected from N;

[0135] Y is selected from N;

[0136] L is

[0137] A is selected from N or CR;

[0138] E is selected from N or CR 1 ;

[0139] R is hydrogen;

[0140] R 1 is hydrogen;

[0141] R 2 is -O(C1-C6)alkyl;

[0142] R 3 for

[0143] R 4 is hydrogen or -(C1-C6)alkyl;

[0144] R 5 is hydrogen;

[0145] R 6 is hydrogen;

[0146] R 8 is hydrogen or -(C1-C6)alkyl, preferably -(C1-C6)alkyl;

[0147] R 9 is hydrogen or -(C1-C6)alkyl, preferably -(C1-C6)alkyl;

[0148] R 10 is hydrogen or -(C1-C6)alkyl;

[0149] In a further embodiment,

[0150] G is

[0151] X is selected from N;

[0152] Y is selected from N;

[0153] L is

[0154] A is selected from N or CR;

[0155] E is selected from N or CR 1 ;

[0156] R is hydrogen;

[0157] R 1 is hydrogen;

[0158] R 2 is -OMe;

[0159] R 3 for

[0160] R 4 is methyl;

[0161] R 5 is hydrogen;

[0162] R 6 is hydrogen;

[0163] R 8 is hydrogen or methyl;

[0164] R 9 is methyl;

[0165] R 10 It is methyl.

[0166] In a further embodiment, the compound is according to formula (Ia or Ib)

[0167]

[0168] in

[0169] A is N or CR; preferably CR, more preferably CH;

[0170] E is N or CR; preferably CR, more preferably CH;

[0171] X is N, CH, preferably CH;

[0172] R is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano, -O(C1-C6)alkyl, preferably hydrogen;

[0173] L is Best

[0174] R 1 Selected from alkylamines, non-aromatic heterocycles, partially or fully deuterated alkylamines, and partially or fully deuterated non-aromatic heterocycles, including Best

[0175] R 2is hydrogen, fluorine, -(C1-C6)alkyl, preferably hydrogen;

[0176] R 3 is hydrogen, -(C1-C6)alkyl, -(C3-C6)cycloalkyl, preferably methyl;

[0177] R 4 is hydrogen, -(C1-C6)alkyl, preferably hydrogen;

[0178] R 5 is hydrogen, -(C1-C6)alkyl, preferably hydrogen;

[0179] R 6 is hydrogen, -(C1-C6)alkyl, preferably hydrogen;

[0180] R 7 is hydrogen, fluorine, -O(C1-C6)alkyl, -(C1-C6)alkyl, preferably hydrogen;

[0181] R 8 is hydrogen, -(C1-C6)alkyl, preferably hydrogen;

[0182] R 9 is hydrogen, -(C1-C6)alkyl, preferably -(C1-C6)alkyl;

[0183] R 10 is hydrogen, -(C1-C6)alkyl, preferably -(C1-C6)alkyl;

[0184] R 11 is hydrogen, -(C1-C6)alkyl, preferably -(C1-C6)alkyl;

[0185] R 12 is -(C1-C6)alkyl, -(C3-C6)cycloalkyl, preferably -(C1-C6)alkyl:

[0186] The condition is A = N and Can not exist in the same compound at the same time.

[0187] wherein optionally,

[0188] One or more hydrogen atoms are replaced by deuterium atoms;

[0189] One or more carbon atoms are 11 C isotope substitution;

[0190] One or more nitrogen atoms are replaced by the corresponding 13 N isotope substitution;

[0191] One or more fluorine atoms are replaced by the corresponding 18 F isotope substitution.

[0192] Specific exemplary compounds are listed in Table 1.

[0193] Table 1: List of compounds

[0194]

[0195]

[0196]

[0197]

[0198]

[0199]

[0200]

[0201]

[0202]

[0203]

[0204]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210] Medical Applications

[0211] The compounds according to formula (I), (Ia) and (Ib) as described above or the pharmaceutical compositions of the present invention are used for preventing or treating tauopathy.

[0212] In general, tauopathies belong to a class of diseases associated with pathological aggregation of tau protein in neurofibrillary cords, anterior horns, neurofibrils or neurofibrillary tangles in the nervous system. Tau protein undergoes several post-translational modifications, including phosphorylation, acetylation, ubiquitination and myristoylation, among which phosphorylation is the major modification of tau and its cellular functions. In tauopathies, tau aggregates are found to be extensively phosphorylated at several serine, threonine or tyrosine residues, and acetylated and ubiquitinated at several lysine residues 11, 12, 13, 14. Hyperphosphorylation and acetylation of tau reduce its ability to bind microtubules, thereby interfering with cargo transport and inducing pathogenic accumulation of tau in cells.

[0213] Thus, the term tauopathy includes both loss-of-function effects on microtubules and gain-of-function effects of toxic tau species. In detail, the consequences of tau hyperphosphorylation and acetylation, and potentially other post-translational modifications, include not only its loss of function in binding and stabilizing microtubules, but also the acquisition of neurotoxic properties through tau aggregation into neuropil threads, anterior horns, neurofibrils, or neurofibrillary tangles.

[0214] Preferably, the prevention or treatment of tauopathy 15, 16 with the compounds of formula (I) according to the invention includes the etiological treatment of tauopathy.

[0215] Preferably, the prevention or treatment of tauopathy comprises inhibiting the cellular activity of tau by binding a compound of formula (I), (Ia) or (Ib) to tau. Binding to tau may comprise covalent binding of a compound of formula (I) to tau, preferably, the binding comprises a thiol-Michael addition of a -SH group of tau to an acrylamide group in the compound of formula (I).

[0216] Preferably, the tauopathy is a dementia-associated tauopathy.

[0217] Preferably, the tauopathy is selected from 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, tauopathies with hippocampal 4-repeat tau immunoreactive spherical inclusions, limbic predominant neuronal inclusion 4R tauopathy (LNT), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) 17 , Argyrophilic Grain Disease (AGD) 17 Huntington's disease 17 , glial globular tauopathy, senile neuro-astrocytic tauopathy, familial behavioral variant frontotemporal dementia associated with astrocyte-dominant tauopathy, amyotrophic lateral sclerosis (ALS) 18 , spinocerebellar ataxia type 11, spinal muscular atrophy (SMA), tauopathy associated with progressive ataxia and palatal tremor, cerebral amyloid angiopathy (CAA), IgLON5 antibody-associated tauopathy, trisomy 21-associated Alzheimer's disease (Down syndrome), vascular dementia, cerebral amyloid angiopathy, Gerstmann-Straussler-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 , Parkinson's disease-Guam dementia 17 , Guadeloupe Parkinson's disease 17 ,Kosaka-Shibayama disease 17 , postencephalitis parkinsonism 17 , SYNJ1 (PARK20) early-onset recessive parkinsonism with nigral tauopathy associated with seizures and dystonia, multiple system atrophy, tauopathy associated with familial Parkinson's disease and progressive respiratory failure, limbic-dominant neuronal-glial tauopathy in TARDBP gene mutations (I383; P112H), neuronal 4R tauopathy (PRNP D178N mutation) in fatal familial insomnia, tauopathy associated with ADCY5 movement disorders, tauopathy in chronic temporal lobe epilepsy, neurodegenerative disorder with brain iron accumulation (NBIA), tauopathy in NBIA PANK2 and WDR45 gene mutations, tauopathy in NBIA PLA2G6 mutations, tauopathy in NBIA associated with autosomal dominant mitochondrial membrane protein-associated neurodegeneration (MPAN), neurofibrillary pretangles and neurofibrillary tangles in human immunodeficiency virus (HIV)-negative opioid abusers 20 , tauopathy associated with acquired immunodeficiency syndrome (AIDS) 20, diffuse neurofibrillary tangles with calcifications, progressive ataxia and palatal tremor, Parkinson's disease associated with SLC9A6, tauopathy associated with mutations in the SPG7 gene, striatal 4R tauopathy associated with X-linked parkinsonism spasticity (ATP6AP2), tauopathy associated with hereditary spastic paraplegia associated with the SPAST gene, autism, autism spectrum disorder, retinal tauopathy 21 , West Nile encephalomyelitis, TTBK2 gene-related spinocerebellar ataxia 11, herpes simplex encephalitis, tau astrogliosis (TOD), aging-related tau astrogliosis (ARTAG) 17 , hippocampal tauopathy, tauopathy associated with subacute sclerosing panencephalitis (SSPE), FTLD-C9ORF72, Christianson syndrome, vacuolar tauopathy, lytico-bodig disease, gangliogliomas and gangliocytomas, meningioangiomatosis, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, neuronal ceroid lipofuscinosis, myotonic dystrophy, Fukuyama congenital muscular dystrophy, hemimegalencephaly, focal cortical dysplasia, Wolcott-Rallison syndrome, primary lateral sclerosis, progressive freezing of gait, PSP with Parkinson's disease, Richardson syndrome, nonfluent / grammatical variant of primary progressive aphasia, semantic variant of primary progressive aphasia, reduced speech variant of primary progressive aphasia, primary progressive apraxia of speech, amnestic Alzheimer's disease.

[0218] More preferred are Alzheimer's disease (AD), Pick's disease (PiD), progressive supranuclear palsy (PSP), cortical basal degeneration (CBD), chronic traumatic encephalopathy (CTE), argyrophilic tauopathies (AGD), frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD) and primary age-related tauopathy (PART).

[0219] Drug composition and dosage

[0220] The pharmaceutical composition of the present invention comprises a compound for preventing or treating tau disease and at least one pharmaceutically acceptable carrier.

[0221] "Carrier" refers to a diluent, adjuvant, excipient or medium used together with the therapeutic agent. Such a pharmaceutical carrier can be a sterile liquid, such as water and oil, including liquids of petroleum, animal, plant or synthetic origin, including but not limited to peanut oil, soybean oil, mineral oil, sesame oil, etc. When the pharmaceutical composition is administered orally, water is a preferred carrier. When the pharmaceutical composition is administered intravenously, saline and glucose aqueous solution are preferred carriers. Saline solution and glucose aqueous solution and glycerol solution are preferably used as liquid carriers for injection 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, glyceryl monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene glycol, water, ethanol, etc. If necessary, the composition can also contain a small amount of wetting agent or emulsifier, or pH buffer. These compositions can take the form of solution, suspension, emulsion, tablet, pill, capsule, powder, sustained release formulations, etc. The composition can be made into suppositories together with traditional binders and carriers such as triglycerides. Oral formulations can include standard carriers, such as pharmaceutical grade aqueous hydroxypropyl methylcellulose, low-substituted hydroxypropyl cellulose, mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. 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 a purified form, and an appropriate amount of carrier to provide a form for appropriate administration to the patient. The preparation should be suitable for the mode of administration.

[0222] In one embodiment, the pharmaceutical composition is a tablet comprising mannitol, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, sodium stearyl fumarate. Preferably, the coating of the tablet comprises polyvinyl alcohol, titanium dioxide, polyethylene glycol 3350, talc, yellow iron oxide (E 172), red iron oxide (E172), black iron oxide.

[0223] Preferably, the pharmaceutical composition is administered parenterally or orally, preferably orally.

[0224] Typically, out of 100% (for a pharmaceutical preparation / composition), the amount of active ingredient (in particular, the amount of the compound of the present invention, optionally together with other therapeutically active agents, if present in a pharmaceutical preparation / composition) will be in the range of about 0.01% to about 99%, preferably about 0.1% to about 70%, most preferably about 1% to about 30%, with the remainder preferably consisting of one or more pharmaceutically acceptable excipients.

[0225] In unit dosage form and / or when administered alone or for treatment, the amount of active ingredient, e.g., a compound of the invention, can be from about 0.1 mg to about 400 mg (e.g., from about 1 mg to about 400 mg, such as from about 10 mg to about 90 mg or from about 40 mg to 80 mg or from about 80 mg to 400 mg) per unit, administration, or treatment. In certain embodiments, a suitable amount of such active ingredient can be calculated using the mass or body surface area of ​​an individual, including amounts of about 1 mg / kg to 10 mg / kg (e.g., from about 2 mg / kg to 5 mg / kg), or about 1 mg / m 2 About 400 mg / m 2 (e.g. about 3 mg / m 2 About 350 mg / m 2 or about 10 mg / m 2 About 200 mg / m 2 ).

[0226] In one embodiment, the pharmaceutical composition does comprise further therapeutically active ingredients, for example in a combination therapy.In one embodiment, the pharmaceutical composition does not comprise an allosteric EGFR (epidermal growth factor receptor) inhibitor.

[0227] Embodiments of the present invention

[0228] Chemical synthesis, in vitro assays, and bioassay methods

[0229] The following abbreviations may be used:

[0230] DMSO-Dimethyl sulfoxide

[0231] NMR - Nuclear Magnetic Resonance

[0232] HMQC-Heteronuclear Multiple Quantum Coherence

[0233] TOCSY - Total Correlation Spectroscopy

[0234] NOESY - Nuclear Ostwald Effect Spectroscopy

[0235] STD - Saturation Transfer Difference

[0236] ThT-Thioflavin T

[0237] TCEP-(Tris(2-carboxyethyl)phosphine)

[0238] HEK - Human Embryonic Kidney

[0239] GFP - Green Fluorescent Protein

[0240] SDS-PAGE - sodium dodecyl sulfate polyacrylamide gel electrophoresis

[0241] TFA-Trifluoroacetic acid

[0242] DIPEA-N,N-Diisopropylethylamine

[0243] FCC-Fast Column Chromatography

[0244] THF-Tetrahydrofuran

[0245] DMF-N,N-dimethylformamide

[0246] Synthesis / Preparation of Compounds

[0247] N-[2-(2-dimethylaminoethylmethylamino)-4-methoxy-5-[[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino]phenyl]prop-2-enamide (1) is referred to as "osimertinib", and N-[2-(2-dimethylaminoethylmethylamino)-4-methoxy-5-[[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino]phenyl]prop-2-enamide methanesulfonate is referred to as "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 (Massachusetts, USA, catalog number O-7200), and osimertinib mesylate was purchased from MedChem Express (catalog number HY-15772A).

[0248] 4-Methoxy-5-[[4-(1-methyl-1H-indol-3-yl)-2-pyrimidinyl]amino]-2-[methyl[2-(methylamino)ethyl]amino]phenyl]-2-acrylamide (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-acrylamide 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-acrylamide (2) methanesulfonate. Compound 2 is formed as a metabolite of osimertinib in vivo (reference: J. Med. Chem. 2014, 57, 8249-8267). Compound 2 hydrochloride (catalog number HY-B0794A) and compound 2 methanesulfonate (catalog number HY-B0794B) were purchased from MedChemExpress.

[0249] N-[2-[[2-(Dimethylamino)ethyl]methylamino]-5-[[4-(1H-indol-3-yl)-2-pyrimidinyl]amino]-4-methoxyphenyl]-2-acrylamide (3). Compound 3 was purchased from MedChemExpress (Catalog No. HY-B0793).

[0250] N-[5-[[4-(1-cyclopropyl-1H-indol-3-yl)-2-pyrimidinyl]amino]-2-[[2-(dimethylamino)ethyl]methylamino]-4-methoxyphenyl]-2-acrylamide (13) was purchased from Hoelzel Biotech (Germany, catalog number HS-10296, CAS number 1899921-05-1).

[0251] N-[2-[[2-(Dimethyloxyamino)ethyl]methylamino]-4-methoxy-5-[[4-(1-methyl-1H-indol-3-yl)-2-pyrimidinyl]amino]phenyl-2-acrylamide (14) was purchased from Biozol Diagnostika (Germany, catalog number 0702275, CAS number 1975982-94-5).

[0252] N-[2-[[2-(Dimethylamino)ethyl]methylamino]-5-[[4-(7-fluoro-1-methyl-1H-indol-3-yl)-2-pyrimidinyl]amino]-4-methoxyphenyl]-2-acrylamide (4). CAS No. 1883592-81-1.

[0253] N-[2-[[2-(Dimethylamino)ethyl]methylamino]-5-[[4-(5-fluoro-1-methyl-1H-indol-3-yl)-2-pyrimidinyl]amino]-4-methoxyphenyl]-2-acrylamide (5). CAS No. 1903753-67-2.

[0254] The syntheses of compounds 4 and 5 are described in Bioinorganic & Medicinal Chemistry (2017), 25, 1, 4553-4559.

[0255] 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-acrylamide (6) CAS No. 2050521-74-7.

[0256] The synthesis of compound 6 is described in Bioinorganic & Medicinal Chemistry (2018), 26, 23-24, 6135-6145.

[0257] N-[2-[[2-(Dimethylamino)ethyl]methylamino]-4-methoxy-5-[[4-(3-methyl-1H-indol-1-yl)-2-pyrimidinyl]amino]phenyl]-2-acrylamide (21) CAS No. 1835666-87-9. The synthesis of compound 21 is described in European Journal of Medicinal Chemistry (2017), 135, 12-23.

[0258] Synthesis of compounds 7, 8, 27 and 28

[0259]

[0260] Scheme 1: The general scheme for the synthesis of compounds 7, 8, 27 and 28 is shown above.

[0261]

[0262] In 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 was added to an aqueous mixture of tripotassium phosphate and the catalyst tetrakis(triphenylphosphine)palladium and incubated overnight. The crude product was evaporated to dryness, bound to silica powder, and then purified by flash silica chromatography with a solvent gradient of 0-20% methanol / dichloromethane. The pure fractions containing 3-(2-chloropyridin-4-yl)-1-methyl-1H-indole were evaporated to dryness.

[0263]

[0264] Subsequently, 3-(2-chloropyridin-4-yl)-1-methyl-1H-indole is mixed with an equimolar amount of 4-fluoro-3-nitroaniline dissolved in 2-pentanol. 4-Methylbenzenesulfonic acid is added to the mixture, which is then heated to 105° C. for several hours. The mixture is cooled to room temperature, the crude product is washed with 2-pentanol, and subjected to flash silica gel chromatography or recrystallization to obtain pure fractions of N-(4-fluoro-3-nitrophenyl)-4-(1-methyl-1H-indol-3-yl)pyridin-2-amine.

[0265]

[0266] Any 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. "Base" corresponds to a pyrrolidinyl, piperidinyl, 4-methylpiperidinyl or 2-methylpyrrolidinyl substituent. The mixture is heated at 140°C for 1 hour in a microwave, after which the crude product is cooled to room temperature and subjected to ion exchange chromatography. Flash silica gel chromatography is then performed using a 0-4% 7M ammonia / methanol in dichloromethane gradient. The pure fractions containing the desired base-substituted nitrobenzyl ring are evaporated to dryness.

[0267]

[0268] The base-substituted nitrobenzene intermediate was dissolved in ethanol / water and refluxed for 2 hours in the presence of ammonium chloride. The crude product was subjected to ion exchange chromatography followed by flash silica gel chromatography using eluting solvent (7M ammonia / methanol). The pure fractions containing the desired base-substituted aniline product were evaporated to dryness.

[0269]

[0270] In the final reaction step, a solution of acryloyl chloride in dichloromethane was added to a stirred solution of the base-substituted aniline compound dissolved in dichloromethane containing DIPEA and cooled to 4°C. The reaction was allowed to proceed for 1.5 hours and quenched by diluting 5-fold with dichloromethane and then washing with saturated aqueous sodium bicarbonate. The resulting hydrophobic (organic) layer was separated from the aqueous layer and subjected to flash chromatography on silica with a solvent gradient of 0-4% 7M ammonia / methanol in dichloromethane. The pure fractions were evaporated to dryness. The compound had the formula:

[0271]

[0272] 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.

[0273] Synthesis of compounds 17, 18, 29, and 30

[0274]

[0275] Scheme 2: General scheme for the synthesis of compounds 17, 18, 29, 30

[0276]

[0277] As described in Bioconjugate Chemistry 2020, 31 (2), 224-228, 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. Alternatively, the compound can 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, and the desired product is then purified by flash silica gel chromatography.

[0278]

[0279] The base substituted nitrophenylmethanol intermediate is prepared by reacting any amine base precursor ("H-base") selected from pyrrolidine, 2-methylpyrrolidine, piperidine with 1-bromo-4-[[[(1,1-dimethylethyl)dimethylsilyl]oxy]methyl]-2-nitrobenzene and DIPEA in 2,2,2-trifluoroethanol solvent at 140° C. for one hour. The crude product is subjected to flash silica gel chromatography and the pure fractions are evaporated to dryness.

[0280]

[0281] In the presence of cesium carbonate, the base-substituted nitrophenylmethanol intermediate is reacted with commercially available 3-(2-chloropyrimidin-4-yl)-1-methyl-1H-indole (CAS No. 1032452-86-0). The reaction is carried out in dimethylformamide solvent at 100° C. for 5 hours, and the crude product is cooled to room temperature and subjected to flash silica gel chromatography to obtain the desired fraction having an -OCH2- bond between the pyrimidine ring and the nitrobenzyl ring.

[0282]

[0283] The -OCH2-connected base-substituted nitrobenzene intermediate was dissolved in ethanol / water and refluxed for 2 hours in the presence of ammonium chloride. The crude product was subjected to ion exchange chromatography and then flash silica gel chromatography using an eluting solvent (7M ammonia / methanol). The pure fractions containing the desired product (-OCH2-connected base-substituted aniline compound) were evaporated to dryness.

[0284]

[0285] In the final reaction step, a solution of acryloyl chloride in dichloromethane was added to a stirred solution of the -OCH2- linked, base-substituted aniline intermediate dissolved in dichloromethane containing DIPEA and cooled in an ice / water bath. The reaction was allowed to proceed for 1.5 hours, diluted 5-fold with dichloromethane, and then quenched by washing with saturated aqueous sodium bicarbonate. The resulting hydrophobic (organic) layer was separated from the aqueous layer and subjected to silica flash chromatography using a 0-4% 7M ammonia / methanol in dichloromethane gradient. The pure fractions were evaporated to dryness. The compound has the formula:

[0286]

[0287] 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.

[0288] Synthesis of compounds 31, 32, 19, and 20

[0289]

[0290] Scheme 3: General scheme for the synthesis of compounds 31, 32, 19, and 20.

[0291]

[0292] 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 was added to an aqueous mixture of tripotassium phosphate and the catalyst tetrakis(triphenylphosphine)palladium and incubated overnight. The crude product was evaporated to dryness, bound to silica powder, and then purified by flash silica chromatography with a solvent gradient of 0-20% methanol / dichloromethane. The pure fractions containing 3-(2-chloropyridin-4-yl)-1-methyl-1H-indole were evaporated to dryness.

[0293]

[0294] The basic substituted nitrophenylmethanol intermediate was prepared using the first two reactions described in Scheme 2, and the basic substituted nitrophenylmethanol intermediate was reacted with 3-(2-chloropyridin-4-yl)-1-methyl-1H-indole in the presence of cesium carbonate. The reaction was carried out in dimethylformamide solvent at 100°C for 5 hours. The crude product was cooled to room temperature and subjected to flash silica chromatography to obtain the desired fraction having an -OCH2- bond between the pyridine ring and the nitrobenzene ring.

[0295]

[0296] The -OCH2-linked base-substituted nitrobenzene intermediate was dissolved in ethanol / water and refluxed for 2 hours in the presence of ammonium chloride. The crude product was subjected to ion exchange chromatography followed by flash silica chromatography using eluting solvent (7M ammonia / methanol). The pure fractions containing the desired product (-OCH2-linked base-substituted aniline compound) were evaporated to dryness.

[0297]

[0298] In the final reaction step, a solution of acryloyl chloride in dichloromethane was added to a stirred solution of -OCH2-linked, base-substituted aniline intermediate dissolved in dichloromethane containing DIPEA and cooled to 4°C. The reaction was allowed to proceed for 1.5 hours and quenched by diluting 5-fold with dichloromethane followed by washing with saturated aqueous sodium bicarbonate. The resulting hydrophobic (organic) layer was separated from the aqueous layer and subjected to flash chromatography on silica with a solvent gradient of 0-4% 7M ammonia / methanol in dichloromethane. The pure fractions were evaporated to dryness. The compound had the formula:

[0299]

[0300] 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.

[0301] NMR spectroscopy

[0302] Compound stocks for NMR experiments were prepared by dissolving the powder directly in NMR buffer (50 mM sodium phosphate buffer, pH 6.8, 0.01% NaN3) or as 10 mM compounds in deuterated DMSO.

[0303] To obtain residue-specific information about the interaction between Tau and the compound, the 2N4R Tau was monitored during the titration with increasing amounts of compound. 1 H- 15 N related spectrum. [U- 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) D2O, and compound concentrations corresponded to a molar ratio of Tau: compound of 1:10 and 1:100. All NMR samples were incubated overnight (~16 hours) at 37°C before spectral acquisition. The assignment of tau amides was taken 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 assignment was obtained using a tau sample prepared in the presence of a reducing agent, some amide resonances belonging to the tau repeat region (near C291 and C322) were not clearly transferred to the SOFAST-HMQC spectrum obtained under oxidative conditions. Therefore, in order to obtain almost complete backbone assignments in the repeat region, [U- 13 C, 15 3D HNCA, 3D DHNCOCA and 3D CBCACONH spectra of the N-4R Tau (K18 construct) sample. The weighted normalized chemical shift perturbation (CSP) was calculated as CSP = √(0.5[(ΔδH) 2 +(ΔδN) 2 For each titration point, the NMR signal intensity ratio I / I0 (where I is the peak intensity of the Tau amide backbone resonance in the presence of the compound and I0 is the peak intensity in the absence of the compound) was calculated.

[0304] To obtain the assignments of non-labile 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 for each compound: 1D 1 H-NMR, 2D with 80 ms mixing time 1H- 1 H TOCSY and 2D with 200 ms mixing time 1 H- 1 H NOESY. Experiments were performed at 25 °C and 37 °C using a Bruker 800 MHz or Bruker 600 MHz spectrometer equipped with a triple resonance cryoprobe.

[0305] To determine the binding epitope of the compounds on monomeric 2N4R Tau, 1D 1 H-NMR titrations, where spectra of osimertinib, AZ7550 hydrochloride and AZ5104 (250 μM compound) were recorded with unlabeled 2N4R Tau (0, 10 and 20 μM). NMR titration samples were prepared in 50 mM sodium phosphate at pH 6.8 with 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.

[0306] To determine the binding epitope of the compounds for fibrillar Tau, 1D saturation transfer difference (STD) experiments were performed with osimertinib, AZ7550 hydrochloride, and AZ5104 in the presence of sonicated 2N4R Tau fibrils (~100 nm fibril length). 2N4R Tau fibrils were prepared using the aggregation method described in the in vitro assay section 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 at pH 6.8 with 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 spectra were acquired in an interleaved manner. 1 H-NMR STD spectrum, alternating between off-resonance radiation at 60 ppm and on-resonance radiation at -2 ppm.

[0307] NMR spectra were processed with 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).

[0308] In vitro assays

[0309] In vitro assay 1: naïve and seeded Tau aggregation assays

[0310] Aggregation of 2N4R Tau was performed using a previously published cofactor-free aggregation protocol with some modifications (reference: Chakraborty, P. et al. Co-factor-free aggregation of tau into seeding-competent RNA-sequestering amyloid fibrils. Nat. Commun. 12, 4231 (2021)). Aggregation experiments used Tau protein stock prepared in 25mM HEPES pH 7.4, 1mM TCEP. Stock solutions of compounds were prepared in aggregation buffer (25mM HEPES, 10mM KCl, 5mM MgCl2, pH 7.2) or DMSO (10mM compound).

[0311] To prepare Tau fibril seeds by initial aggregation, 2N4R Tau with a monomer concentration of 25 μM was diluted in aggregation buffer. After dilution of the mother solution, the working concentration of TCEP in the aggregation mixture was <25 μM. Thioflavin T (ThT) was added to the mixture to a final concentration of 50 μM. In the wells of a 96-well microplate (Greiner Bio-One), a total volume of 100 μL of the Tau-ThT mixture was incubated in the presence of two polytetrafluoroethylene beads. The microplate was subjected to an implemented procedure in a Tecan Spark plate reader, which included incubation at 37°C, interval double-orbital oscillation, and ThT fluorescence emission measurement. The excitation wavelength of ThT was set to 430 nm and the emission wavelength was set to 485 nm. The details of the plate reader program have been previously disclosed (reference: Chakraborty, P. et al. Co-factor-free aggregation of tau into seeding-competent RNA-sequestering amyloid fibrils. Nat. Commun. 12, 4231 (2021)). After about 3-6 days of incubation, ThT fluorescence emission reaches a maximum value, indicating that Tau fibrillization has reached a saturation point. The mixture containing fibrils collected from the microplate is used as a seeding mixture for subsequent aggregation assays.

[0312] Seeded aggregation assays were performed as described above, except that 1.0% of the well volume was occupied by the seeding mixture which was sonicated for 1 minute. The assays were performed in the presence and absence of compound and the working concentration range was 0-250 μM compound.

[0313] Analysis of fluorescence curves was performed using Graphpad PRISM version 8. Fluorescence data were fitted with a sigmoid function. For each sigmoid curve, the value corresponding to half-maximal ThT fluorescence was determined. TM Statistically significant differences were determined by one-way ANOVA and Dunnett's multiple comparison test or unpaired t-test.

[0314] The seeding aggregation protocol was also applicable to the C291S / C322S mutants of 3R Tau and 2N4R Tau.

[0315] In vitro test 2: Tau pelleting test

[0316] Aggregation assays were performed as described in the in vitro assay section 1. Aggregation mixtures were collected and ultracentrifuged (55000 rpm, using a JLA 100.3 rotor, Optima MAX-XP) for 30 minutes to separate soluble (supernatant) and insoluble (precipitate) fractions. Soluble fractions were analyzed by SDS-PAGE using 12% or 15% acrylamide separating gels and 5% acrylamide stacking gels. The band intensity of soluble Tau was quantified by ImageLab (Bio-Rad Laboratories).

[0317] Negative stain transmission electron microscopy

[0318] The fibril samples were negatively stained using glow-discharged 400 mesh carbon-coated copper grids. Images were acquired on a Talos L120C transmission electron microscope (Thermo Fisher, Eindhoven, The Netherlands) after staining with 1% uranyl acetate solution.

[0319] Mass spectrometry analysis method

[0320] Mass spectrometry analysis 1: ESI-MS

[0321] The analysis was performed using a 4R Tau construct (K18 construct, 13818 Da) containing residues 244-372 of 2N4R Tau. The compound (330 μM) was incubated with 4R Tau (65 μM) at 37°C in 10 mM ammonium bicarbonate, pH 7.0 for 16 hours. The compound / 4R Tau mixture was injected into an LC system (ACQUITY) coupled to a single quadrupole mass detection system (SQDetector 2, Waters) equipped with a diode array detector (DAD). The DAD spectrum covers 210-400 nm wavelengths. 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 in water, buffer B is 0.1% TFA in acetonitrile). Covalent modification of 4R Tau is equivalent to mass addition of 500 Da (osimertinib) and 486 Da (AZ7550 or AZ5104).

[0322] Mass spectrometry 2: MS / MS

[0323] Compounds (330 μM) were incubated with 2N4R Tau (65 μM) in buffer (25 mM HEPES, 10 mM KCl, 5 mM MgCl2, pH 7.2) at 37°C for 16 h and then stored at 4°C until in-gel digestion.

[0324] As previously described, trypsin (Sigma Aldrich) was used for in-gel digestion and peptides were extracted for mass spectrometry analysis (reference: 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). The extracted peptides were resuspended in 2% acetonitrile and 0.05% TFA and analyzed with an Orbitrap Exploris 480 mass spectrometer (Thermo Fisher). The mass spectrometer used an emission voltage of 2100 kV and an ion transfer tube temperature of 300°C. A full scan was recorded in the range of 350 to 1600 m / z with a mass resolution of 15000. The peptides were fragmented in a collision cell set at an energy of 28%. Tandem mass spectra were acquired using the scan range mode "Define First Mass" and an isolation window of 1.6 m / z with a mass resolution of 15,000.

[0325] Biological test methods

[0326] Biological Test 1: HEK Biosensor Cell Tau Seeding Assay

[0327] Compound stocks were prepared as 10 mM solutions of compound in DMSO.

[0328] The human tau repeat domain (RD) carrying the P301L / V337M mutation and fused to a C-terminal GFP tag (named TauRD LM HEK293T biosensor cells expressing P301L human full-length Tau and C-terminally tagged with GFP were engineered to detect Tau seeding in cells (reference: Liu, S., Hossinger, A., Heumüller, 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 C-terminally tagged with GFP were incubated with pre-sonicated hTau 40 P301L The cells were harvested and incubated with HEK TauRD LM In further experiments with -GFP cells, the resulting cell lysates were used to seed Tau aggregates.

[0329] In a typical seeding test, HEK cells were incubated with cell lysate, compound (0-40 μM) and lipofectamine for 22-48 hours. Negative control experiments without compound (vehicle DMSO only) and with cell lysate were performed in parallel.

[0330] Dose response curves were constructed by quantifying the number of cells aggregated at each compound concentration. The curves were fitted with a sigmoid function using Graphpad PRISM version 8. The inhibitory concentration (IC) of each compound at 50% activity was determined. 50 ).

[0331] Biological test 2: Drosophila neurodegeneration model

[0332] Drug treatment of fruit flies:

[0333] Stock solutions of each compound were prepared by dissolving 10 mg of compound in 100 μl of 100% DMSO. Osimertinib, osimertinib mesylate and AZ 7550 mesylate were orally administered to flies by mixing them with food. The final concentration of each compound used to treat fruit flies was based on the recommended concentration for human use (reference concentration) based on body weight. We used three concentrations for each compound. One was equal to the reference concentration and two were higher than the reference concentration. The final concentrations of the different compounds mixed with fly food were: osimertinib mesylate, 5 μM, 10 μM and 15 μM; osimertinib, 10 μM; AZ7550 mesylate, 10 μM and 20 μM. After hatching, first instar larvae were transferred to food containing one of the compounds. The food mixed with the desired concentration of drug was replaced with fresh food every 48 hours.

[0334] Optical Microscope:

[0335] After mounting the fly heads on a glass slide using double-sided tape, optical microscopy was performed. Whole-eye images were captured with the help of an Infinity Analyze camera connected to an Olympus SZX7 microscope with a halogen light source. The total eye surface area and the degraded eye surface area were marked and calculated using Infinity Analyze software (Lumenera). The percentage of degraded area was calculated and plotted using GraphPad Prism8.

[0336] Scanning Electron Microscope:

[0337] After eclosion, treated and control flies (2-3 days old) were fixed in 1% formaldehyde for two hours, followed by a series of dehydration steps in 25%, 50%, 75%, and 100% ethanol for 12 hours each. Flies were stored in 100% ethanol and dried with CPD before mounting on the spines for imaging. Eye images were captured at 150X and analyzed using Infinity Analyze software.

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Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof for preventing or treating tauopathy, in G is Best X is N or CH, preferably N; Y is N or CH, preferably N; L is Best A is N or CR; preferably CR, more preferably CH; E is N or CR 1 ; CR is preferred 1 , more preferably CH; R is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano or -O(C1-C6)alkyl, preferably hydrogen; R 1 is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano or -O(C1-C6)alkyl, preferably hydrogen; R 2 is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano, -O(C1-C6)alkyl or difluoromethoxy, preferably -O(C1-C6)alkyl; R 3 for or -NR 8 R 9 , preferably R 4 is hydrogen, -(C1-C6)alkyl or -(C3-C6)cycloalkyl; R 5 is hydrogen, fluorine or chlorine; R 6 is hydrogen, fluorine or chlorine; 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)2; R 12 is hydrogen or -(C1-C6)alkyl; wherein optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms are 11 C isotope substitution; One or more nitrogen atoms are replaced by the corresponding 13 N isotope substitution; One or more fluorine atoms are replaced by the corresponding 18 F isotope substitution.

2. A compound of formula (I) or a pharmaceutically acceptable salt thereof in G is Best X is selected from N or CH; Y is selected from N or CH; L is Best A is selected from N or CR; E is selected from N or CR 1 ; R is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano or -O(C1-C6)alkyl, preferably hydrogen; R 1 is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano or -O(C1-C6)alkyl, preferably hydrogen; R 2 is hydrogen, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano, -O(C1-C6)alkyl or difluoromethoxy, preferably -O(C1-C6)alkyl; R 3 for or -NR 8 R 9 , preferably R 4 is hydrogen, -(C1-C6)alkyl or -(C3-C6)cycloalkyl; R 5 is hydrogen, fluorine or chlorine; R 6 is hydrogen, fluorine or chlorine; 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)2; R 12 is hydrogen or -(C1-C6)alkyl; Provided that the compound of formula (I) is not selected from wherein optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms are 11 C isotope substitution; one or more nitrogen atoms are replaced by the corresponding 13 N isotope substitution; one or more fluorine atoms are replaced by the corresponding 18 F isotope substitution.

3. Compounds of formula (Ia or Ib) in 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, fluorine, chlorine, bromine, -(C1-C6)alkyl, cyano, -O(C1-C6)alkyl, preferably hydrogen; L is Best R 1 Selected from alkylamines, non-aromatic heterocycles, partially or fully deuterated alkylamines, and partially or fully deuterated non-aromatic heterocycles, including Best R 2 is hydrogen, fluorine, chlorine, bromine, cyano, -(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, fluorine, -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: The condition is A = N and Can not exist in the same compound at the same time. wherein optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms are 11 C isotope substitution; One or more nitrogen atoms are replaced by the corresponding 13 N isotope substitution; One or more fluorine atoms are replaced by the corresponding 18 F isotope substitution.

4. The compound for use according to claim 1 or the compound according to claim 2, wherein in formula (I) G is X is selected from N; Y is selected from N; A is selected from N or CR; E is selected from N or CR 1 ; R is hydrogen; R 1 is hydrogen; R 2 is methoxy; R 3 for 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 It is hydrogen or -(C1-C6)alkyl.

5. The compound for use according to claim 1, or the compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein in formula (I), G is X is selected from N; Y is selected from N; A is selected from N or CR; E is selected from N or CR 1 ; R is hydrogen; R 1 is hydrogen; R 2 is methoxy; R 3 for 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; wherein optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms are 11 C isotope substitution; One or more nitrogen atoms are replaced by the corresponding 13 N isotope substitution; One or more fluorine atoms are replaced by the corresponding 18 F isotope substitution.

6. The compound for use according to claims 1, 4, 5 and the compound according to claims 2, 4 and 5, wherein only one or two of A, E, X and Y are N; only one of A and E is N, and only one of A and Y is N.

7. The compound for use according to claims 1 and 3 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from wherein optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms are 11 C isotope substitution; One or more nitrogen atoms are replaced by the corresponding 13 N isotope substitution; One or more fluorine atoms are replaced by the corresponding 18 F isotope substitution.

8. The compound for use according to claim 7, wherein the compound is selected from wherein optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms are 11 C isotope substitution; One or more nitrogen atoms are replaced by the corresponding 13 N isotope substitution.

9. A compound selected from the following or a pharmaceutically acceptable salt thereof, wherein optionally, One or more hydrogen atoms are replaced by deuterium atoms; One or more carbon atoms are 11 C isotope substitution; one or more nitrogen atoms are replaced by the corresponding 13 N isotope substitution; one or more fluorine atoms are replaced by the corresponding 18 F isotope substitution.

10. A pharmaceutical composition for preventing or treating tauopathy, comprising a compound of formula (I), formula (Ia) or formula (Ib) according to claims 1 to 9 and at least one pharmaceutically acceptable carrier.

11. A compound for use according to claims 1 and 4 to 8, or a pharmaceutical composition for use according to claim 10, wherein the prevention or treatment of tau disease comprises the etiological treatment of the tau disease, or a compound according to claims 2, 3, 4 to 6 and 9, for the prevention or treatment of tau disease comprising the etiological treatment of tau disease.

12. A compound for use according to any one of claims 1, 4 to 8, 10, or a pharmaceutical composition for use according to claim 10 or 11, or the compound, wherein the prevention or treatment of tauopathy comprises inhibiting the cellular activity of tau by binding the compound according to formula (I) to tau, or a compound according to claims 2, 3, 4 to 6 and 9, which comprises inhibiting the cellular activity of tau by binding a compound according to formula (I), formula (Ia) or formula (Ib) for the prevention or treatment of tauopathy.

13. A compound for use according to claims 1, 4 to 8 and 10 to 12, or a pharmaceutical composition for use according to claims 10 to 12, wherein the binding to tau comprises covalent binding of a compound of formula (I) to tau, preferably the binding comprises thiol-Michael addition of a -SH group of tau to an acrylamide group in a compound of formula (I), or a compound according to claims 2, 3, 4 to 6 and 9, comprising use for preventing or treating tauopathy by inhibiting the cellular activity of tau by binding to a compound of formula (I), (Ia) or (Ib), wherein the binding to tau comprises covalent binding of a compound of formula (I) to tau, preferably the binding comprises thiol-Michael addition of a -SH group of tau to an acrylamide group in a compound of formula (I), (Ia) or (Ib).

14. A compound for use according to claims 1, 4 to 8 and 10 to 13, or a pharmaceutical composition for use according to claims 10 to 13, wherein the tauopathy is a dementia-related tauopathy, or a compound according to claims 2, 3, 4 to 6 and 9 for use in preventing or treating a dementia-related tauopathy.

15. A compound for use according to claims 1, 4 to 8 and 10 to 14, or a pharmaceutical composition for use according to claims 10 to 14, or a compound for the prevention or treatment of tauopathy according to claims 2, 3, 4 to 6 and 9, wherein 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), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), Pick's disease (PiD), dementia with Lewy bodies (DLB), progressive supranuclear palsy (PSP), globular glial tauopathy (GGT), tauopathy with hippocampal 4-repeat tau immunoreactive spherical inclusions, limbic predominant neuronal inclusions 4R Tauopathies (LNT), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), argyrophilic grain disease (AGD), Huntington's disease, glial globular tauopathy, senile neuro-astrocytic tauopathy, familial behavioral variant frontotemporal dementia associated with astrocyte-dominant tauopathy, amyotrophic lateral sclerosis (ALS), spinocerebellar ataxia type 11, spinal muscular atrophy (SMA), tauopathies associated with progressive ataxia and palatal tremor, cerebral amyloid angiopathy (CAA), IgLON5 antibody-associated tauopathy, trisomy 21-associated Alzheimer's disease (Down syndrome), vascular dementia, cerebral amyloid Vascular disease, Gerstmann-Straussler-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, Parkinson's disease-Guam dementia, Guadeloupe Parkinson's disease, Kosaka-Shibayama disease, postencephalitic parkinsonism, SYNJ1 (PARK20) early-onset recessive parkinsonism with nigral tauopathy associated with seizures and dystonia, multiple system atrophy, tauopathies associated with familial parkinsonism and progressive respiratory failure, TARDBP gene mutations (I383;P112H), neuronal-glial tauopathy in limbic-dominant neuron-glial tauopathy, neuronal 4R tauopathy in fatal familial insomnia (PRNP D178N mutation), tauopathy associated with ADCY5 dyskinesia, tauopathy in chronic temporal lobe epilepsy, neurodegenerative disease with brain iron accumulation (NBIA), tauopathy in NBIAPANK2 and WDR45 gene mutations, NBIA Tauopathies in PLA2G6 mutations, tauopathies in NBIA associated with autosomal dominant mitochondrial membrane protein-associated neurodegeneration (MPAN), neurofibrillary pretangles and neurofibrillary tangles in human immunodeficiency virus (HIV)-negative opioid abusers, tauopathies associated with acquired immune deficiency syndrome (AIDS), diffuse neurofibrillary tangles with calcification, progressive ataxia and palatal tremor, Parkinson's disease associated with SLC9A6, tauopathies associated with mutations in the SPG7 gene, striatal 4R associated with X-linked parkinsonism spasticity (ATP6AP2) Tauopathies, tauopathies associated with hereditary spastic paraplegia associated with the SPAST gene, autism, autism spectrum disorder, retinal tauopathy, West Nile encephalomyelitis, TTBK2 gene-associated spinocerebellar ataxia 11, herpes simplex encephalitis, tangled dementia (TOD), tau astrogliopathy associated with aging (ARTAG), hippocampal tauopathy, tauopathy associated with subacute sclerosing panencephalitis (SSPE), FTLD-C9ORF72, Christianson syndrome, vacuolar tauopathy, osteolytic encephalopathy (lytico-bodig disease), gangliogliomas and gangliocytomas, meningioangiomatosis, lead encephalopathy, tuberous sclerosis, pantothenate kinase-associated neurodegeneration, neuronal ceroid lipofuscinosis, myotonic dystrophy, Fukuyama congenital muscular dystrophy, hemimerical encephaly, focal cortical dysplasia, Wolcott-Rallison syndrome, primary lateral sclerosis, progressive freezing of gait, PSP with Parkinson's disease, Richardson syndrome, nonfluent / grammatical variant of primary progressive aphasia, semantic variant of primary progressive aphasia, language reduction variant of primary progressive aphasia, primary progressive apraxia of speech, amnestic Alzheimer's disease,; Preferred are Alzheimer's disease (AD), Pick's disease (PiD), progressive supranuclear palsy (PSP), cortical basal degeneration (CBD), chronic traumatic encephalopathy (CTE), argyrophilic tauopathies (AGD), frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), Parkinson's disease (PD) and primary age-related tauopathy (PART).

16. The pharmaceutical composition for use according to claims 10 to 15, wherein the pharmaceutical composition is administered parenterally or orally, preferably orally.

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