Compounds, compositions, and method of use to inhibit TAU protein and alpha-synuclein aggregation

Abstract

Compounds comprising a triazole scaffold, compositions comprising same, and the use of such compounds and compositions to inhibit tubulin-associated unit (tau) protein and alpha-synuclein (α-syn) protein aggregation, including neurofibrillary tangles (NFTs) and Lewy bodies.

Description

70948-02COMPOUNDS, COMPOSITIONS, AND METHODOF USE TO INHIBIT TAU PROTEIN AND ALPHA-SYNUCLEIN AGGREGATIONCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority from U. S. Appl. No. 63 / 723,835, filed November 22, 2024, which is incorporated by reference as if fully set forth herein.STATEMENT OF GOVERNMENT SUPPORT

[0002] This invention was made with government support under AG071985 awarded by the National Institutes of Health, The government has certain rights in the invention.TECHNICAL FIELD

[0003] This disclosure relates to compounds comprising a triazole scaffold, compositions comprising same, and the use of such compounds and compositions to inhibit tubulin-associated unit (tau) protein and alpha-synuclein (a-syn) protein aggregation, including, but not limited to, neurofibrillary tangles (NFTs), associated with tauopathies (e.g., Alzheimer’s disease, Downs syndrome, progressive supranuclear palsy, and traumatic brain injury) and Lewy bodies, associated with synucleinopathies (e.g., Parkinson’s disease, dementia with Lewy bodies (DLB), and multiple system atrophy (MSA)).BACKGROUND

[0004] Neurodegenerative diseases like Alzheimer's disease (AD) and Parkinson's disease (PD) are characterized by the abnormal accumulation of misfolded proteins.’ These proteins, which are normally harmless, undergo structural changes that cause them to aggregate into compact insoluble fibrils.2As these fibrils build up in the brain, they disrupt normal cellular processes, leading to progressive neuronal damage and dysfunction.2'3The resulting impairment in brain activity manifests in the cognitive and motor symptoms associated with these devastating chronic diseases.4AD is characterized by distinct pathological features that disrupt normal brain function. Among these, amyloid plaques and neurofibrillary tangles (NFTs) are particularly significant.5Amyloid plaques consist of clusters of amyloid-beta proteins (predominantly fragments Api-40 and Api-42) that accumulate outside neurons, while NFTs are formed from70948-02tau proteins that have undergone abnormal modifications.6, 7These NFTs develop inside neurons and disrupt their structural integrity. However, NFTs have also been detected in the synaptic cleft.8Together, these plaques and NFTs interfere with the brain's ability to transmit signals effectively, leading to the gradual decline in cognitive abilities associated with AD. Although amyloid-beta plaques develop early in the disease, research shows that cognitive decline is more strongly correlated with the spread of NFTs throughout the brain.9, 10NFTs arise from the accumulation of tau proteins, which normally play a critical role in stabilizing microtubules within neurons. However, when tau proteins become hyperphosphorylated, they lose their ability to bind to microtubules and get detached, leading to the formation of NFTs and contributing to neuronal dysfunction and cell death.10, 11When tau proteins detach from microtubules, they become prone to interacting with other tau molecules.12These interactions lead to the formation of small oligomers, which gradually assemble into larger structures with a P-sheet configuration. These aggregates grow in size and complexity, eventually forming tau fibrils that develop into NFTs.13,14NFTs first appear in the transentorhinal cortex, a region in the temporal lobe, before spreading to other critical areas of the brain such as the hippocampus and, eventually, the neocortex.15This spread of NFTs is a key driver of the cognitive decline seen in AD, as they progressively damage the brain's neuronal networks.16

[0005] PD prominently features the accumulation of abnormal alpha-synuclein (a-syn) aggregates known as Lewy bodies (LBs) within neuronal cells.17Central to this pathologic effect is a-syn, which normally exists as a soluble, largely monomeric entity in the brain. Under certain conditions, a-syn undergoes various post-translational modifications, such as phosphorylation, acetylation, glycation, and ubiquitination, which are, at least in part, disease-associated. Moreover, the transition from an unstructured form to one that is more folded and richer in alphahelices has been proposed to be an early event in disease-relevant a-syn dyshomeostasis.18This and other conformational changes are thought to facilitate the protein's aggregation into dimers and oligomers. Over time, such oligomers combine into larger, insoluble aggregates that constitute LBs.19The accumulation of these structures likely disrupts normal neuronal function by impairing neurotransmitter signaling and triggering pathways that contribute to neuronal cell death.20

[0006] Current therapeutic approaches for AD and PD primarily focus on managing symptoms rather than targeting the underlying pathophysiological mechanisms. This770948-02symptomatic treatment often provides only short-term benefits and does not significantly enhance long-term health outcomes. In view of the foregoing, it is an object of the present disclosure to provide multitarget agents, with the aim of inhibiting the aggregation processes of a-syn and tau proteins. This and other objects and advantages, as well as inventive features, will be apparent from the description provided herein.SUMMARY

[0007] Provided is a compound of formula I:Rwherein R is 1-3 substituents, which are independently selected from -O-C1-C6alkyl, N(C1-C3alkyl)2, halo, -NO2, and -OH, or a pharmaceutically acceptable salt or hydrate thereof. In embodiments, -O-C1-C6alkyl is -O-methyl. In embodiments, C1-C3alkyl is methyl. In embodiments, halo is Cl, F, or Br. In embodiments, the compound is a compound of formula II:or a pharmaceutically acceptable salt or hydrate thereof;wherein:3a R1= H, R2= OCH3, R3= H;3b R1= OCH3, R2= OCH3, R3= H;3c R1- OCII3, R2H. R3- OCH3;3d R1= OCH3, R2= OCH3, R3= OCH3;3e R1= H, R2= N(CH3)2, R3= H;31‘R1=== H, R Cl, R3- II;3g R1= Cl, R2= H, R3= Cl;3h R’ - H, R2- NO2, R3- II;4a R1= OH, R2= OH, R3= H;4b R’ = OH, R2= H, R3= OH;70948-024c R1= OH, R2= OH, R3= OH.

[0008] In an embodiment, the compound is compound 3e (5-(l -(4-Dimethylaminophenyl)- l / 7-l,2,3-triazol-4-yl)-l / 7-indole). In an embodiment, the compound is compound 4b (4-[4-(l / f- indol-5-yl)-l / f-l,2,3-triazol-l-yl]benzene-l,2-diol). In an embodiment, the compound is compound 4d (5-[4-(1H-indol-5-yl)-1H-1,2,3-triazol-1-yl]benzene-1,2,3-triol).

[0009] In view of the above, also provided is a pharmaceutical composition comprising an above-described compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier.

[0010] Also in view of the above, provided is a method of inhibiting alpha-synuclein (a- syn) protein fibril formation in a subject having, or at risk for, a-syn protein fibril formation. The method comprises administering to the subject an effective amount of (i) an above-described compound, or a pharmaceutically acceptable salt or hydrate thereof, or (ii) a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier. In embodiments, the subject has, or is at risk for, Parkinson’s disease or dementia with Lewy bodies (DLB).. In embodiments, the compound is compound 4d (5-[4-(lH-indol-5-yl)-lH-l,2,3-tnazol-l-yl]benzene-l,2,3-triol), or a pharmaceutically acceptable salt or hydrate thereof, or a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier, and the compound also inhibits a-syn oligomers.

[0011] Further provided is a method of inhibiting tubulin-associated unit (tau) protein (isoforms 0N4R, 1N4R, 2N4R, 0N3R, 1N3R, 2N3R) oligomer formation in a subject having, or at risk for, tau (isoforms 0N4R, 1N4R, 2N4R, 0N3R, 1N3R, 2N3R) oligomer formation.. The method comprises administering to the subject an effective amount of (i) an above-described compound, or a pharmaceutically acceptable salt or hydrate thereof, or (ii) a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier. In embodiments, the compound is compound 4d (5-[4-(1H-indol-5-yl)-1H-1,2,3-triazol-1-yl]benzene-1,2,3-triol), or a pharmaceutically acceptable salt or hydrate thereof, or a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier.70948-02FIGURES

[0012] Fig. 1. Design of the synthesized compounds as dual anti-fibrillar agents. Fig.1A. Anti-fibrillar agents I24and II22containing indole scaffold previously published from Dr. Fortin’s laboratory. Fig. IB. Polyphenol catechin (-)-epi-gallocatechin-3-gallate (EGCG). Fig.1C. The newly designed compounds.

[0013] Fig. 2. Synthetic procedure for the target compounds 3a-h, and 4b-d (Scheme 1). Reagents and conditions: (a) NaNCh, NaNs, HCl:H₂O (1:1), 0-5°C, 3 h, 85-100%; HC≡Si(CH3)3; (b) (i), PdCl2(PPh3)2, Cui, TEA, DMF, r.t., 5-8 h; (n) aq. NaOH, MeOH, r.t., 1 h, 72% (over the two steps); (c) Na ascorbate, CuSO₄·5H₂O, THF: H2O (1:1), r.t., 71-91%; (d)BBr3, DCM, 0°C, overnight, 73%.

[0014] Fig. 3. Kinetics of a-syn formation with several triazole based-compound and dose-response of the lead, compound 4d. Fig.3A. Thioflavin T (ThT) kinetic curves illustrating the fibrillation process of a-syn (2 pM) with the triazole compounds (100 pM). The control condition included only the vehicle (0.25% DMSO). For each experimental condition, triplicate measurements were taken at ten consecutive time points during the plateau phase of the fibrillation process. Fig. 3B. Monomeric a-syn (6 μM) was subjected to variable concentrations of compound 4d to achieve the dose-response curve. The Log EC50 indicated was obtained from the Log(agonist) versus normalized response correlation as calculated with Prism GraphPad. The dose-response curve exhibited a clear, dose-dependent linear relationship, indicating the compound’s potency in modulating the biological response across the tested concentration range of 6.25, 12.5, 25, 50, and 100 pM at 37 hours of incubation.

[0015] Fig. 4. Lead 4d, a polyphenol triazole-based compound, reduced the aggregation of tau non-phosphorylated and phosphorylated 4R isoform by about 50%. The ThS fluorescence curves of 4d illustrate the aggregation kinetics of (Fig. 4A) tau 0N4R (6 μM), (Fig.4B) tau 2N4R (12 pM), and (Fig.4C) p-tau 1N4R (6 μM) in PBS-treated with chelex beads. The solution was supplemented with 150 μM heparin, 5 mM dithiothreitol (DTT), 40 pM ThS, and 10 mg / ml arachidonic acid in order to induce the aggregation. The control condition contained 0.25% DMSO, while compound 4d was tested at a concentration of 100 pM. The data represented by each curve are an average of three independent replicates.

[0016] Fig. 5. Lead compound 4d exhibited a concentration-dependent inhibitory effect on a-syn oligomerization. Fig. 5A. Compound 4d effectively prevented the formation of70948-02a-syn oligomers induced by Tris(bipyridyl)ruthenium(II) chloride (Ru(bpy)3) and ammonium persulfate under brief light exposure (1 s) in the PICUP cross-linking assay. For this assay, a-syn (30 pM) was cross-linked in the presence of 100 pM of the lead compound. In the control sample, which contained 0.25% DMSO without the test compound, higher molecular weight a-syn oligomers were clearly observed on Coomassie blue-stained polyacrylamide gels. In contrast, additional control samples that were either not exposed to light or lacked the crosslinking agent (Ru(bpy)3) did not show any cross-linked products. Fig. 5B. Monomeric a-syn (30 pM) was subjected to varying concentrations of the lead compound followed by PICUP crosslinking assay. Samples were loaded on a 16% polyacrylamide SDS-Page gel electrophoresis and stained with Coomassie blue to reveal the cross-linking products. The concentration-dependent a-syn anti- oligomer effect of lead compound 4d was confirmed. As the concentration of compound 4d decreased, there was a corresponding increase in the intensity of the high molecular weight bands (corresponding to oligomer). DMSO (0.25%) was used as a control, showing no significant impact on oligomer formation. The percentage of oligomer reduction is calculated by dividing the oligomer pixel density of the lane with compound treatment by the oligomer pixel density of the control lane (without the compound) and then multiplied by 100. The resulting value is then subtracted from 100 to determine the reduction.

[0017] Fig. 6. Lead compound 4d exhibited a tau 0N4R anti-oligomer effect at high micromolar concentration. The tau 0N4R high molecular weight band corresponding to oligomer was observed above 180 kDa after loading samples subjected to the PICUP cross¬ linking assay. Tau (6 μM) in the presence of Tris(bipyridyl)ruthenium(II)chloride (Ru(bpy)3) and ammonium persulfate was cross-linked under light exposure at a duration of 15 seconds. The percentage of oligomer reduction is calculated as indicated in the Fig. 5 legend.

[0018] Fig.7. Transmission electron microscopy (TEM) evaluation demonstrated that compound 4d significantly inhibited a-syn fibril formation. a-Syn (6 μM) was incubated with 0.25% DMSO as the control (CTRL) and treated with compound 4d at concentrations of 12.5 and 100 pM. After an incubation period of approximately 37 hours, the samples were visualized using TEM. Scale bars in the images represent 200 nm.

[0019] Fig. 8. Lead compound 4d reduced the formation of tau 2N4R fibril formation as validated by transmission electron microscopy (TEM). TEM was employed to directly assess the effect of compound 4d on the monomeric tau isoform 2N4R (tested at 12 pM). Samples70948-02were treated with either DMSO (0.25%) as a control or 100 pM of compound 4d and incubated at 37°C for 5 days. Scale bars correspond to 200 nm, with images captured at 40k magnification.

[0020] Fig. 9. Compound 4d and EGCG disaggregate and prevent the formation of amyloid beta 1-40 (Aβ1-40) fibrils. Fig. 9A. The Thioflavin T fibril formation kinetics of 21 pM Api-40 was assessed in the presence of compound 4d and EGCG at 25, 50, and 100 pM concentrations. ThT was used at a final concentration of 40 pM. Fig. 9B. The maximum fluorescence intensity in percentage was obtained at the end of the kinetics (approximately at 86 h) to evaluate the effects of compound 4d on Api-40 fibril formation in comparison with the control (0.25% DMSO). Fig. 9C. Similarly the maximum fluorescence intensity in percentage were plotted for the EGCG treatment and the control (0,25% DMSO), For those two histograms, compound 4d and EGCG treatment were statistically significantly different compared to the control (0,25% DMSO) at p < 0,001 using the one-way ANOVA, Dunnet’s post-hoc test.

[0021] Fig. 10. Compound 4d exhibited a moderate disaggregation effect in comparison to EGCG, as confirmed by transmission electron microscopy (TEM). The disaggregation effect of compound 4d was compared to EGCG. EGCG resulted non fibrillar structures. The non-treated control presented dense plaques comprised of fibrils. Amyloid beta (0.593 ± 0.095 mg / mL) obtained from the brain of an Alzheimer's disease patient were incubated with DMSO (1.5%; referred to as " CTRL"); compound 4d (at 50 pM); or EGCG (at 50 pM) for 5 days at 37 °C before preparation of formvar / carbon supported copper grids and direct visualization by TEM. Pictures were acquired at 40K. The scale bars correspond to 200 nm.

[0022] Fig. 11. Formation of a-Syn inclusion was inhibited by 4d. M17D cells expressing the inclusion-prone aS-3K:: YFP fusion protein (dox-inducible) were treated with 0.1% DMSO (vehicle; “0 pM”) as well as 1.25, 2.5, 5, 10, 20 and 40 pM of compounds 4b and 4d at t = 24 h after plating. aS-3K:: YFP expression was induced with doxycycline at t = 48 h. Fig. 11A. Incucyte- based analysis of punctate YFP signals relative to 0.1% DMSO was done at t = 96 h (N = 3 independent experiments, n = 6-18 individual wells total (0 pM, n = 18; 40, 20 and 10 pM, n = 6; 5, 2.5 and 1.25 pM n = 12). Fig. 11B. Same as panel A, but confluence fold changes relative to DMSO vehicle (0 pM) were plotted. Fig. 11C. Representative IncuCyte images of reporter cells treated with vehicle vs 40, 20, 10 and 5 pM compound 4b and 4d (t = 96 h), green channel. Arrows indicate aS-rich TTP-positive inclusions. Scale bar, 50 pm. All data are presented as fold-70948-02changes relative to DMSO control + / - standard deviation. One-way AN OV A, Dunnett’s post- hoc test; *, p < 0.05; ****, p < 0.0001; ns, non-significant.DESCRIPTION

[0023] A novel group of polyphenolic triazole-linked indole compounds was designed and synthesized to curtail the aggregation of a-synuclein (a-syn) and tubulin-associated unit (tau) proteins, which are closely linked to the physiopathology of PD and AD, respectively. A Thioflavin T (ThT) fluorescence assay was used to measure fibril formation and assess the antiaggregation effects of various compounds. To further validate these findings, transmission electron microscopy (TEM) was employed as a direct method to visualize the impact of these compounds on fibril morphology. Inhibition of oligomer formation was evaluated using photo¬ induced cross-linking of unmodified proteins (PICUP), enabling the detection of early protein aggregation events. During fibril formation assays, three compounds (3e, 4b, 4d) demonstrated superior inhibitory activity as assessed by ThT fluorescence and TEM imaging. Among them, the trihydroxy derivative, compound 4d, showed a highly significant anti-fibrillar activity as depicted by decreasing the thioflavin- T (ThT) fluorescence to lower than 5% against a-syn and to 50% towards 2N4R tau. Transmission electron microscope (TEM) analyses of 4d revealed powerful inhibitory activities against dense fibrils of both proteins which were consistent with their ThT assay results. In addition, 4d was able to inhibit the early-stage oligomer formation of a-syn protein in a dose-dependent manner. 4d reduced tau 0N4R oligomers at high micromolar concentration. Finally, 4d obstructed the a-syn inclusion formation in M17D neuroblastoma reporter cells. Thus, compound 4d was identified as the most promising candidate overall,

[0024] In view of the foregoing, provided is a compound of formula I:R htyNA-%, A Jwherein R is 1-3 substituents, which are independently selected from -O-C1-C6alkyl, N(C1-C6alkyl)2, halo, -NO2, and -OH, or a pharmaceutically acceptable salt or hydrate thereof. In embodiments, -O-C1-C6alkyl is -O-methyl. In embodiments, C1-C6alkyl is methyl. In embodiments, halo is Cl, F, or Br. In embodiments, the compound is a compound of formula II:70948-02or a pharmaceutically acceptable salt or hydrate thereof;wherein:3a R1= H, R2= OCH3, R3= H;3b R1= OCH3, R2= OCH3, R3= H;3c R1= OCH3, R2= H, R3= OCH3;3d R1= OCH3, R2= OCH3, R3= OCH3;3e R1= H, R2= N(CH3)2, R3= H;3f R1= H, R2= Cl, R3= H;3g R1= Cl, R2= H, R3= Cl;3h R1= H, R2= NO?., R3= H;4a R1= OH, R2= OH, R3= H;4b R1= OH, R2= H, R3= OH;4c R1= OH, R2= OH, R3= OH.

[0025] In an embodiment, the compound is compound 3e (5-( 1 -(4-Dimethylaminophenyl)- lH-l,2,3-triazol-4-yl)-lH-indole). In an embodiment, the compound is compound 4b (4-[4-(lH- indol-5-yl)-1H-1,2,3-triazol-1-yl]benzene-1,2-diol). In an embodiment, the compound is compound 4d (5-[4-(lH-indol-5-yl)-lH-l,2,3-triazol-l-yl]benzene-l,2,3-triol).

[0026] Provided are also compounds of formula (I):bk-Nwherein:each R4is independently alkyl (e.g., Ci-Ce alkyl, such as C1-C3 alkyl and CII3), optionally substituted aryl or optionally substituted heteroaryl;each R5is independently H, halo, alkyl, alkoxy, aryl, aryloxy, nitro, cyano, carboxy or acyl; X1is CR5or N;Z is NR6, O or S, wherein R6is H, alkyl or alkenyl; and70948-02n is 1, 2 or 3;or a pharmaceutically acceptable salt or hydrate thereof. In embodiments, the compound of formula (I) is a compound of formula (la):N^Nwherein:each R7is independently H, hydroxy, halo, alkyl, alkoxy, aryl, aryloxy, nitro, cyano, carboxy or acyl;each R5is independently H, alkyl, alkoxy, halo, aryl, aryloxy, amino, nitro or cyano;Z is NR6, O or S, wherein R6is H, alkyl or alkenyl;X1is CR5or N;n is 1, 2 or 3; andm is 1, 2, or 3;or a pharmaceutically acceptable salt or hydrate thereof. In other embodiments, the compound of formula (I) is a compound of formula (lb):wherein:each R7is independently H, hydroxy, halo, alkyl, alkoxy, aryl, aryloxy, nitro, cyano, carboxy or acyl;each R5is independently H, alkyl, alkoxy, halo, aryl, aryloxy, amino, nitro or cyano;X1is CR5or N;Z is NR6, O or S, wherein R6is H, alkyl or alkenyl;n is 1, 2 or 3; andm is 1, 2, or 3;or a pharmaceutically acceptable salt or hydrate thereof. In still other embodiments, the compound of formula (I) is a compound of formula (lb):70948-02wherein:each R7is independently H, hydroxy, halo, alkyl, alkoxy, aryl, aryloxy, nitro, cyano, carboxy or acyl;R' is H, alkyl, alkoxy, halo, aryl, aryloxy, amino, nitro or cyano;X1is CR5or N;Z is NR6, O or S, wherein R6is H, alkyl or alkenyl;n is 1, 2 or 3; andm is 1, 2, or 3;or a pharmaceutically acceptable salt or hydrate thereof.

[0027] In any of the compounds of formulae (I) and (la)-(Ic), X1is CR5. In addition, or alternatively, Z can be NR6, wherein R6can be H. Alternatively, in any of the compounds of formulae (I) and (la)-(Ic), Z can be S. Alternatively, in any of the compounds of formulae (I) and (la)-(Ic), Z can be 0. In sum, in any of the compounds of formulae (I) and (la)-(Ic), the group:

[0028] Alternatively, in any of the compounds of formulae (I) and (la)-(Ic), X1can be N. In addition, Z can be NR6, wherein R6can be H. Alternatively, in any of the compounds of formulae (I) and (la)-(Ic), Z can be S, Alternatively, in any of the compounds of formulae (I) and (la)-(Ic), Z can be O. In sum, in any of the compounds of formulae (I) and (la)-(Ic), the group:70948-02

[0029] In any of the compounds of formulae (I) and (la)-(Ic), the group:N=: NH f 1Z,R4can be:R4R4N=N ^-NH R4-M’ I Hl4 I hl II z z v? or. This applies equally to instances where R4is a group of the formula:m

[0030] In any of the compounds of formulae (I) and (la)-(Ic) n can be I. Alternatively, n can be 2.Alternatively, n can be 3. In addition, or alternatively, m, where applicable, can be 1. / Xlternatively, m can be 2. Alternatively, m can be 3. Any combination of n and m is contemplated herein, such as instances where n is 1 and m is 1; n is 1 and m is 2; n is 1 and m is 3; n is 2 and m is 1; n is 2 and m is 2; n is 2 and m is 3; n is 3 and m is 1; n is 3 and m is 2; and n is 3 and m is 3.

[0031] In any of the compounds of formulae (I) and (la)-(lc), each R3can independently be I I. Alternatively, each R5can independently be alkyl, such as Ci-Cg alkyl, such as Ci-Cb alkyl, C1-C3 alkyl, C1-C2. alkyl, and CH?,.

[0032] In any of the compounds of formulae (I) and (la)-(Ic), one or more R7can independently be -O-Ci-Ce alkyl, amino (e.g., alkylamino, such as N(CI-C3 alkyl)?.), halo, -NO?., and -OH (hydroxy). Thus, for example, one or more R7can independently be -O-C1-C4 alkyl, -O-C1-C3 alkyl, -O-C1-C2 alkyl or -OCH3. In another example, one or more R7is amino, such as alkylamino wherein each alkyl is C1-C3 alkyl, such as methyl. In still another example, one or more R7is halo, such as Cl, F, or Br. In yet other examples, one or more R7is -OH. In examples, m is 1 and R7is - O-C1-C4 alkyl, -O-C1-C3 alkyl, -O-C1-C2 alkyl or -OCH3. In other examples, m is 2 and each R7is independently -O-C1-C4 alkyl, -O-C1-C3 alkyl, -O-C1-C2 alkyl or -OCH3 In still other examples, m is 3 and each R7is independently -O-C1-C4 alkyl, -O-C1-C3 alkyl, -O-C1-C2 alkyl or -OCH3. In examples, m is 1 and R7is -OH. In other examples, m is 2 and each R7is -OH. In still other70948-02examples, m is 3 and each R7is -OH. In examples, m is 1 and R7is halo, such as Cl or F. In one example, m is 1 and R7is Cl. In another example, m is 1 and R7is F. In other examples, m is 2 and each R7is independently halo, such as Cl or F. In examples m is 2 and each R7is Cl. In examples m is 2 and each R7is F. In still other examples, m is 1 and R7is amino, such as dialkylamino like -N(CH3)2. In still other examples, m is 1 and R7is NO2. In yet other examples, m is 1 and R7is CN.

[0033] Compounds of the formulae (I) and (la)-(Ic) include compounds of formula:N-Nor a pharmaceutically acceptable salt or hydrate thereof, wherein n and R5are defined herein. In embodiments, n is i and R5is H.

[0034] Compounds of the formulae (I) and (la)-(Ic) include compounds of formula:70948-02or a pharmaceutically acceptable salt or hydrate thereof, wherein n and R5are defined herein. In embodiments, n is 1 and R5is H

[0035] Compounds of the formulae (I) and (la)-(Ic) include compounds of formula:N-N70948-02or a pharmaceutically acceptable salt or hydrate thereof, wherein n and R5are defined herein. In embodiments, n is 1 and R5is H.

[0036] Compounds of the formulae (I) and (la)-(Ic) include compounds of formula:70948-02or a pharmaceutically acceptable salt or hydrate thereof, wherein n and R5are defined herein. In embodiments, n is 1 and R5is H.70948-02or a pharmaceutically acceptable salt or hydrate thereof, wherein n and R' are defined herein. In embodiments, n is 1 and R5is H.

[0038] Compounds of the formulae (I) and (la)-(Ic) include compounds of formula:or a pharmaceutically acceptable salt or hydrate thereof, wherein n and R5are defined herein. In embodiments, n is 1 and R5is H.

[0039] The above compounds include isotopic variants and compounds in which one or more hydrogen atoms have been substituted with deuterium. The compounds may contain one or more chiral centers or may otherwise be capable of existing as multiple stereoisomers. In one embodiment, the compounds are not limited to any particular stereochemical requirement, and that the compounds, and compositions, methods, uses, and medicaments that include them may be optically pure, or may be any of a variety of stereoisomeric mixtures, including racemic and other mixtures of enantiomers, other mixtures of diastereomers, and the like. Such mixtures of70948-02stereoisomers may include a single stereochemical configuration at one or more chiral centers, while including mixtures of stereochemical configuration at one or more other chiral centers.

[0040] Similarly, the compounds may include geometric centers, such as cis, trans isomers, diastereomers, enantiomers, and E and Z double bonds. In another embodiment, the compounds are not limited to any particular geometric isomer requirement, and that the compounds, and compositions, methods, uses, and medicaments that include them may be pure, or may be any of a variety of geometric isomer mixtures. Such mixtures of geometric isomers may include a single configuration at one or more double bonds and chiral carbons, while including mixtures of geometry at one or more other double bonds and chiral carbons.

[0041] The term “salts” and “pharmaceutically acceptable salts” refer to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic groups such as amines; and alkali or organic salts of acidic groups such as carboxylic acids. Pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, and nitric; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, and isethionic, and the like.

[0042] Pharmaceutically acceptable salts can be synthesized from the parent compound, which contains a basic or acidic moiety, by conventional chemical methods. In some instances, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts are found in Remington’s Pharmaceutical Sciences, 18th ed., Mack Publishing Company, Easton, Pa., 1990, the disclosure of which is hereby incorporated by reference.70948-02

[0043] The term “solvate” means a compound, or a salt thereof, that further includes a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. Where the solvent is water, the solvate is a hydrate.

[0044] The terms “substituted,” “substituent,” and “functional group” refer to a group that can be or is substituted onto a molecule or onto another group (e.g., on an aryl or an alkyl group). Examples of substituents include, but are not limited to, a halogen (e.g., F, Cl, Br, and I), OR, OC(O)N(R)2, CN, NO, NO2, ONO2, azido, CFa, OCF3, R, O (oxo), S (thiono), C(O), S(O), methylenedioxy, ethylenedioxy, N(R)2, SR, SOR, SO2R, SO2N(R)2, SO3R, -(CH2)o-2P(0)(OR)2, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R, C(O)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2)O-2N(R)C(0)R, (CH2)O-2N(R C(0)OR, (CH2)O-2N(R)N(R)2, N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R)2, N(R)SO2R, N(R)SO2N(R)2, N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R)2, N(R)C(S)N(R)2, N(COR)COR, N(OR)R, C(=NH)N(R)2, C(O)N(OR)R, or C(=NOR)R wherein each R can be, independently, hydrogen, alkyl, acyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroarylalkyl, wherein any alkyl, acyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroaryl alkyl or two R groups bonded to a nitrogen atom or to adjacent nitrogen atoms can together with the nitrogen atom or atoms form a heterocyclyl, which can be mono- or independently multi-substituted.

[0045] The term “alkyl” as used herein refers to substituted or unsubstituted straight chain and branched mono- or divalent alkyl groups and cycloalkyl groups having from 1 to 40 carbon atoms (C1-C40), 1 to about 20 carbon atoms (C1-C20), 1 to 12 carbons (C1-C12), 1 to 8 carbon atoms (Ci-Cs), or, in some embodiments, from 1 to 6 carbon atoms (Ci-Ce). Examples of straight chain alkyl groups include those with from 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and 2,2- dimethylpropyl groups. As used herein, the term “alkyl” encompasses n-alkyl, isoalkyl, and ante- isoalkyl groups as well as other branched chain forms of alkyl. Representative substituted alkyl groups can be substituted one or more times with any of the groups listed herein, for example, ammo, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0046] The term “alkenyl” as used herein refers to substituted or unsubstituted straight chain and branched mono- or divalent alkenyl groups and cycloalkenyl groups having at least one double bond and having from 1 to 40 carbon atoms (C1-C40), 1 to about 20 carbon atoms (C1-C20),70948-021 to 12 carbons (C1-C12), 1 to 8 carbon atoms (Ci-Cs), or, in some embodiments, from 1 to 6 carbon atoms (Ci-Ce). Examples of straight chain alkenyl groups include those with from 1 to 8 carbon atoms such as -CH=CH-, -CH=CHCH3, and -CH2CH=CHCH2- groups, wherein the double bonds can have an E- or Z-configuration. And when there are multiple bonds, each double bond can, independently, have an E- or a Z-configuration. Examples of branched alkenyl groups include, but are not limited to, -CH=C(CH3)- and CH2C=CH(CH3) groups. Representative substituted alkenyl groups can be substituted one or more times with any of the groups listed herein, for example, ammo, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0047] The term “cycloalkyl” as used herein refers to substituted or unsubstituted cyclic alkyl groups such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, the cycloalkyl group can have 3 to about 8-12 ring members, whereas in other embodiments the number of ring carbon atoms range from 3 to 4, 5, 6, or 7. Cycloalkyl groups can have any number of carbon atoms, e.g, 3 to 8 carbon atoms (Cs-Cs), 3 to 6 carbon atoms (C3-C0), and 4 to 8 carbon atoms (Cr-Cs). Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like.

[0048] The term “cycloalkylalkyl” as used herein refers to substituted or unsubstituted alkyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group as defined herein is replaced with a bond to a cycloalkyl group as defined herein. Representative cycloalkylalkyl groups include, but are not limited to, cyclopentylalkyl.

[0049] The term “alkylcycloalkyl” as used herein refers to substituted or unsubstituted cycloalkyl groups as defined herein in which a hydrogen of a cycloalkyl group as defined herein is replaced with a bond to an alkyl group as defined herein. Representative alkylcycloalkyl groups include, but are not limited to, alkylcyclopropyl.

[0050] The term “acyl” as used herein refers to a group containing a carbonyl moiety wherein the group is bonded via the carbonyl carbon atom. The carbonyl carbon atom is also bonded to another carbon atom, which can be part of a substituted or unsubstituted alkyl, aryl, aralkyl cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl group or the like. In the special case wherein the carbonyl carbon atom is bonded to a hydrogen, the group is a “formyl” group, an acyl group as the term is defined herein. An acyl group can70948-02include 0 to about 12-40, 6-10, 1-5 or 2-5 additional carbon atoms bonded to the carbonyl group. An acryloyl group is an example of an acyl group. An acyl group can also include heteroatoms within the meaning here. A nicotinoyl group (pyridyl-3-carbonyl) is an example of an acyl group within the meaning herein. Other examples include acetyl, benzoyl, phenylacetyl, pyridylacetyl, cinnamoyl, and acryloyl groups and the like. When the group containing the carbon atom that is bonded to the carbonyl carbon atom contains a halogen, the group is termed a “haloacyl” group. An example is a trifluoroacetyl group.

[0051] The term “aryl” as used herein refers to substituted or unsubstituted cyclic aromatic hydrocarbons that do not contain heteroatoms m the ring. Thus, aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments, aryl groups contain about 6 to about 14 carbons (C6-C14) or from 6 to 10 carbon atoms (C6-C10) in the ring portions of the groups. Aryl groups can be unsubstituted or substituted, as defined herein. “Aryl” and the phrase “aryl group” includes fused ring species including those that include fused aromatic and non-aromatic groups. Accordingly, “aryl” and the phrase “aryl group” include groups of the formula:Osubstituted or unsubstituted, such as hydroxy substituted.

[0052] Representative substituted aryl groups can be mono-substituted or substituted more than once, such as, but not limited to, 2-, 3-, 4-, 5-, or 6-substituted phenyl or 2-8 substituted naphthyl groups, which can be substituted with carbon or non-carbon groups such as those listed herein.

[0053] The terms “aralkyl” and “arylalkyl” refer to alkyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to an aryl group as defined herein. Representative aralkyl groups include benzyl and phenylethyl groups and fused (cycloalkylaryl)alkyl groups such as 4-ethyl-indanyl. Aralkenyl groups are alkenyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to an aryl group as defined herein.70948-02

[0054] “Heterocycloalkyl” is a saturated or partially unsaturated non-aromatic monocyclic, bicyclic, tricyclic or polycyclic ring system that has from 3 to 14 members, such as 6 to 10 and 3 to 6 members, in which 1 to 3 carbon atoms in the ring are replaced by heteroatoms of O, S or N. The ring heteroatoms can also include oxidized S or N, such as sulfinyl, sulfonyl, and N-oxides of a tertiary ring nitrogen. A heterocycloalkyl can be fused to another ring system, such as with an aryl or heteroaryl of 5-6 ring members. The point of attachment of the heterocycloalkyl ring is at a carbon or heteroatom such that a stable ring is retained. Examples of heterocycloalkyl groups include without limitation morpholino, tetrahydrofuranyl, dihydropyridinyl, piperidinyl, pyrrolidinyl, piperazinyl, dihydrobenzofuryl, and dihydroindolyl. A heterocycloalkyl group can be unsubstituted or optionally substituted with one or more substituents as described herein.

[0055] “Heteroaryl,” alone or in combination with any other moiety described herein, is a monocyclic aromatic ring structure containing 6 to 10, such as 5 or 6 ring atoms, or a bicyclic aromatic group having 8 to 10 atoms, containing one or more, such as 1-4, 1-3, or 1-2, heteroatoms independently selected from the group consisting of O, S, and N. Heteroaryl is also intended to include oxidized S or N, such as sulfinyl, sulfonyl and N-oxide of a tertiary ring nitrogen. N-methyl, N-aryl, N-substituted to include A carbon or heteroatom is the point of attachment of the heteroaryl ring structure such that a stable compound is produced. Examples of heteroaryl groups include, but are not limited to, pyridinyl, pyridazinyl, pyrazinyl, quinoxalyl, indolizinyl, benzo[b]thienyl, quinazolinyl, purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, pyrazolyl, oxazolyl, thiazolyl, thienyl, isoxazolyl, oxathiadiazolyl, isothiazolyl, tetrazolyl, imidazolyl, triazolyl, furanyl, benzofuryl, and indolyl. A heteroaryl group can be unsubstituted or optionally substituted with one or more substituents as described herein.

[0056] The term “alkoxy” refers to an oxygen atom connected to an alkyl group, including a cycloalkyl group, as are defined herein. Examples of linear alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like. Examples of branched alkoxy include, but are not limited to, isopropoxy, sec- butoxy, tert- butoxy, isopentyloxy, isohexyloxy, and the like. Examples of cyclic alkoxy include, but are not limited to, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. An alkoxy group can include one to about 12-20 or about 12-40 carbon atoms bonded to the oxygen atom, can further include double or triple bonds, and can also include heteroatoms. For example, an70948-02allyloxy group is an alkoxy group within the meaning herein. A methoxyethoxy group is also an alkoxy group within the meaning herein, as is a methylenedioxy group in a context where two adjacent atoms of a structure are substituted therewith.

[0057] The terms “amine,” “amine group,” “amino,” and “amino group” refer to a substituent of the form -NH2, -NHR, -NR2, or -NRA, wherein each R is defined herein, and protonated forms of each, except for -NRA, which cannot be protonated. Accordingly, any compound substituted with an amino group can be viewed as an amine. An “amino group” within the meaning herein can be a primary, secondary, tertiary, or quaternary amino group.

[0058] An “alkylamino” group includes a monoalkylamino, dialkylamino, and trialkylamino group. An example of a “alkylamino” is -NH-alkyl and -N(alkyl)2.

[0059] An example of a “cycloalkylamino” group is -NH-cycloalkyl and -N(cycloalkyl)2.

[0060] An example of a “cycloalkyl heterocycloammo” group is -NH-(heterocyclo cycloalkyl), wherein the heterocyclo group is attached to the nitrogen and the cycloalkyl group is attached to the heterocyclo group,

[0061] / Xn example of a “heterocyclo cycloamino” group is -NH-(cycloalkyl heterocycle), wherein the cycloalkyl group is attached to the nitrogen and the heterocyclo group is attached to the cycloalkyl group.

[0062] The term “amido” refers to a group of the formula -C(O)NR, wherein R is defined herein.

[0063] The terms “halo,” “halogen,” and “halide” group, by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

[0064] The term “haloalkyl” group includes mono-halo alkyl groups, poly-halo alkyl groups, wherein all halo atoms can be the same or different, and per-halo alkyl groups, wherein all hydrogen atoms are replaced by halogen atoms, such as fluoro. Examples of haloalkyl include trifluoromethyl, 1, 1 -dichloroethyl, 1,2-dichloroethyl, l,3-dibromo-3,3-difluoropropyl, perfluorobutyl, -CF(CH3)2 and the like.

[0065] Further, the formulae include and represent not only all pharmaceutically acceptable salts of the compounds, but also include any and all hydrates and / or solvates of the compound formulae or salts thereof. It is to be appreciated that certain functional groups, such as the hydroxy, amino, and like groups form complexes and / or coordination compounds with water70948-02and / or various solvents, in the various physical forms of the compounds. Accordingly, the above formulae are to be understood to include and represent those various hydrates and / or solvates. In each of the foregoing and following embodiments, it is also to be understood that the formulae include and represent each possible isomer, such as stereoisomers and geometric isomers, both individually and in any and all possible mixtures. In each of the foregoing and following embodiments, it is also to be understood that the formulae include and represent any and all crystalline forms, partially crystalline forms, and non-crystalline and / or amorphous forms of the compounds.

[0066] The above compounds, and pharmaceutically acceptable salts and solvates, such as hydrates, thereof, can be synthesized in accordance with methods known in the art and exemplified herein. See, e.g., Experimental Section.

[0067] The compounds can be used to inhibit the aggregation of proteins prone to aggregate in a state of disease. The ability of such compounds to inhibit aggregation can include, but is not limited to, inhibition of oligomer formation, inhibition of fibril formation, and / or inhibition of the formation of a-syn inclusions, “Aggregation” and its derivatives are used herein to refer to all such inhibitory actions for ease of reference. Proteins prone to aggregate include, but are not limited to, islet amyloid polypeptide, amyloid-P, a-synuclein (a-syn), tubulin associated unit (tau), and transthyretin. The tau can be tau isoform 0N4R, 1N4R, 2N4R, 0N3R, 1N3R, and / or 2N3R with and without post-translational changes (such as phosphorylation). The compounds described herein can be used to inhibit the aggregation of a-syn. Diseases involving protein aggregation include, but are not limited to, AA amyloidosis, Alzheimer's disease, monoclonal immunoglobulin light-chain amyloidosis, Huntington's disease, Parkinson's disease, Creutzfeldt-Jacob disease, prion disorders, amyotrophic lateral sclerosis, type 2 diabetes, or transthyretin amyloidosis. The compounds described herein can be used to inhibit the aggregation of a-syn in a subject having, or at risk for, Alzheimer’s disease, dementia with Lewy bodies (DLB), or multiple system atrophy (MSA). The compounds described herein also can be used to inhibit the formation of a-syn inclusions in a subject with a neuroblastoma. The compounds also can be used to inhibit islet amyloid polypeptide (IAPP) fibril formation.

[0068] The compounds can be used to inhibit tau protein aggregation in tauopathies. Tauopathies are a group of disorders that result from abnormal tau phosphorylation, abnormal levels of tau, abnormal tau splicing, and mutations in the tau gene, for example.70948-02N eurodeg enerative diseases have been classified based on this protein accumulation. Tauopathies encompass more than 20 clinicopathological conditions, including Alzheimer’s disease (AD), which is the most common tauopathy. Other tauopathies include, but are not limited to, familial AD, primary age-related tauopathy (PART), Creutzfeldt-Jacob disease, dementia pugilistica, Gerstmann-Straussler-Scheinker disease (GSS), inclusion-body myositis, cortico-basal degeneration (CBD), Picks disease (PiD), progressive supranuclear palsy (also known as Steele, Richardson, and Olszewski disorder), Down syndrome, Parkinsonism with dementia, myotonic dystrophy, prion protein cerebral amyloid angiopathy, traumatic brain injury (TBI), amyotrophic lateral sclerosis (ALS), Parkinsonism-dementia complex of Guam, non-Guamanian motor neuron disease with neurofibrillary tangles, argyrophilic grain disease, diffuse neurofibrillary tangles with calcification, frontotemporal dementia with Parkinsonism linked to chromosome 17 (FTDP- 17), Hal ler-vorden- Spatz disease, multiple system atrophy (MS A), Niemann-Pick disease type C, pallido-ponto-nigral degeneration, progressive subcortical gliosis, progressive supranuclear palsy (PSP), subacute sclerosing panencephalitis, tangle predominant dementia, postencephalitic Parkinsonism, myotonic dystrophy, subacute sclerosis panencephalopathy, mutations in L. RRK2, chronic traumatic encephalopathy (CTE), familial British dementia, familial Danish dementia, other frontotemporal lobar degenerations, Guadeloupean Parkinsonism, neurodegeneration with brain iron accumulation, SLC9A6-related mental retardation, white matter tauopathy with globular glial inclusions, epilepsy, Lewy body dementia (LBD), mild cognitive impairment (MCI), multiple sclerosis, Parkinson’s disease, HIV-related dementia, adult onset diabetes, senile cardiac amyloidosis, glaucoma, ischemic stroke, psychosis m AD, Huntington’s disease, and prion diseases with tangles. The majority of neurodegenerative diseases are characterized by the deposition of insoluble protein in cells of the neuromuscular system.

[0069] The compounds can be formulated as pharmaceutical compositions comprising a pharmaceutically acceptable carrier using methods well-known in the art. “Carrier” is used generically herein to refer to pharmaceutically acceptable carriers, diluents, adjuvants, and excipients. See, e.g., Remington: The Science and Practice of Pharmacy, 23rd edition, October 30, 2020, Adeboye Adejare, ed. The term "pharmaceutically acceptable carrier" is art-recognized and refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting any subject composition or component thereof. Each carrier must be "acceptable" in70948-02the sense of being compatible with the subject composition and its components and not injurious to the patient. Some examples of materials, which may serve as pharmaceutically acceptable carriers, include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) algmic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

[0070] In view of the foregoing, also provided is a pharmaceutical composition. The composition comprises an above-described compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier.

[0071] Also in view of the above, provided is a method of inhibiting alpha-synuclein (a- syn) protein fibril formation in a subject having, or at risk for, a-syn protein fibril formation. The method comprises administering to the subject an effective amount of (i) an above-described compound, or a pharmaceutically acceptable salt or hydrate thereof, or (ii) a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier. In embodiments, the subject has, or is at risk for, Parkinson’s disease or dementia with Lewy bodies (DLB). In embodiments, the compound is compound 4d (5-[4-(l / / -indol-5-yl)-l / / -l,2,3-triazol-l-yl]benzene-l,2,3-triol), or a pharmaceutically acceptable salt or hydrate thereof, or a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier, and the compound also inhibits a-syn oligomers.

[0072] Further provided is a method of inhibiting tubulin-associated unit (tau) protein (isoforms 0N4R, 1N4R, 2N4R, 0N3R, 1N3R, 2N3R) oligomer formation in a subject having, or at risk for, tau (isoforms 0N4R, 1N4R, 2N4R, 0N3R, 1N3R, 2N3R) oligomer formation. The method comprises administering to the subject an effective amount of (i) an above-described compound, or a pharmaceutically acceptable salt or hydrate thereof, or (ii) a pharmaceutical70948-02composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier. In embodiments, the compound is compound 4d (5- [4- (117-indol-5-yl)-177-l,2,3-triazol-l-yl]benzene-l,2,3-triol), or a pharmaceutically acceptable salt or hydrate thereof, or a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier.

[0073] The terms “inhibit,” “inhibiting,” “inhibited,” or “inhibition” (with respect to a disease or condition) is an approach for obtaining beneficial or desired results including and preferably clinical results and includes, but is not limited to, one or more of the following: improving a condition associated with a disease, curing a disease, lessening severity of a disease, delaying progression of a disease, alleviating one or more symptoms associated with a disease, increasing the quality of life of one suffering from a disease, prolonging survival and / or prophylactic or preventative treatment.

[0074] The term “administering” includes all means of introducing the compounds and compositions described herein to the patient, including, but are not limited to, oral (po), intravenous (iv), intramuscular (im), subcutaneous (sc), transdermal, inhalation, buccal, ocular, sublingual, vaginal, rectal, and the like. The compounds and compositions may be administered m unit dosage forms and / or formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles.

[0075] An “effective amount” refers to any amount that is sufficient to achieve a desired biological effect. Combined with the teachings provided herein, by choosing among the various active conjugates or compounds and weighing factors such as potency, relative bioavailability, patient body weight, severity of adverse side-effects and mode of administration, an effective prophylactic or therapeutic treatment regimen can be planned which does not cause substantial unwanted toxicity and yet is effective to treat the particular subject. The effective amount for any particular application can vaiy depending on such factors as the disease or condition being treated, the particular compound being administered, the size of the subject, or the severity of the disease or condition. One of ordinary skill in the art can empirically determine the effective amount of a particular compound and / or other therapeutic agent without necessitating undue experimentation. A maximum dose can be used, that is, the highest safe dose according to some medical judgment. Multiple doses per day can be used to achieve appropriate systemic levels of compounds.70948-02Appropriate systemic levels can be determined by, for example, measurement of the patient’s peak or sustained plasma level of the drug. “Dose” and “dosage” are used interchangeably herein.

[0076] Generally, daily oral doses of a compound are, for human subjects, from about 0.01 milligrams / kg per day to 1,000 milligrams / kg per day. Oral doses in the range of 0.5 to 50 milligrams / kg, in one or more administrations per day, can yield therapeutic results. Dosage can be adjusted appropriately to achieve desired drug levels, local or systemic, depending upon the mode of administration. For example, intravenous administration can vary from one order to several orders of magnitude lower dose per day. If the response in a subject is insufficient at such doses, even higher doses (or effective higher doses by a different, more localized delivery route) can be employed to the extent that patient tolerance permits. Multiple doses per day are contemplated to achieve appropriate systemic levels of the compound.

[0077] Any suitable route of administration can be used in the above methods. Examples include, but are not limited to, oral, parenteral, intravenous, intracranial, intracerebroventricular, and intracerebral. An effective amount can be determined by one of ordinary skill in the art using dosage range determining methods known in the art. Typically, a physician (or veterinarian for non-human subjects) will determine the actual dosage, which will be most suitable for an individual subject. The specific dose level and frequency of dosage for an individual may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, gender, diet, mode and time of administration, rate of excretion, other administered drugs, and the severity of the particular condition. The compound / compositions described herein can be administered with other biologically active compounds as appropriate.

[0078] The disclosure also relates to the following enumerated Embodiments, which are listed in no particular order of importance:

[0079] Embodiment 1. A compound of formula (I):Hhf JRtc(R5(i)wherein:70948-02each R4is independently alkyl, aryl or heteroaryl (e.g, optionally substituted aryl and optionally substituted heteroaryl);each R5is independently H, halo, alkyl, alkoxy, aryl, aryloxy, nitro, cyano, carboxy or acyl; X1is CR5or N;Z is NR6, O or S, wherein R6is H, alkyl or alkenyl; andn is 1, 2 or 3;or a pharmaceutically acceptable salt or hydrate thereof.

[0080] Embodiment 2. The compound of Embodiment 1, wherein the compound of formula (I) is a compound of formula (la):N=Nwherein:each R7is independently H, hydroxy, halo, alkyl, alkoxy, aryl, aryloxy, nitro, cyano, carboxy or acyl;each R5is independently H, alkyl, alkoxy, halo, aryl, aryloxy, amino, nitro or cyano;Z is NR6, O or S, wherein R6is H, alkyl or alkenyl;X1is CR5or N;n is 1, 2 or 3; andm is 1, 2, or 3;or a pharmaceutically acceptable salt or hydrate thereof.

[0081] Embodiment 3. The compound of Embodiment 1, wherein the compound of formula (I) is a compound of formula (lb):wherein:each R7is independently H, hydroxy, halo, alkyl, alkoxy, aryl, aryloxy, nitro, cyano, carboxy or acyl;70948-02each R5is independently H, alkyl, alkoxy, halo, aryl, aryloxy, amino, nitro or cyano;X1is CR5or N;Z is NR6, O or S, wherein R6is H, alkyl or alkenyl;n is 1, 2 or 3; andm is 1, 2, or 3;or a pharmaceutically acceptable salt or hydrate thereof.

[0082] Embodiment 4. The compound of Embodiment 1, wherein the compound of formula (I) is a compound of formula (lb):wherein:each R7is independently H, hydroxy, halo, alkyl, alkoxy, aryl, aryloxy, nitro, cyano, carboxy or acyl;each R5is independently H, alkyl, alkoxy, halo, aryl, aryloxy, amino, nitro or cyano;X1is CR5or N;Z is NR6, O or S, wherein R6is H, alkyl or alkenyl;n is 1, 2 or 3; andm is 1, 2, or 3;or a pharmaceutically acceptable salt or hydrate thereof.

[0083] Embodiment 5. The compound of any of Embodiments 1-4, wherein X1is CR5.

[0084] Embodiment 6. The compound of any of Embodiments 1-5, wherein Z is NR6.

[0085] Embodiment 7. The compound of Embodiment 6, wherein R6is H.

[0086] Embodiment 8. The compound of any of Embodiments 1-5, wherein Z is S.

[0087] Embodiment 9. The compound of any of Embodiments 1-5, wherein Z is O.

[0088] Embodiment 10. The compound of any of Embodiments 1-4, wherein X1is N.

[0089] Embodiment 11. The compound of Embodiment 10, wherein Z is NR6.

[0090] Embodiment 12. The compound of Embodiment 11, wherein R6is H.

[0091] Embodiment 13. The compound of Embodiment 10, wherein Z is S.

[0092] Embodiment 14. The compound of Embodiment 10, wherein Z is O.70948-02

[0093] Embodiment 15. The compound of Embodiment 5, wherein each R7is independently alkyl.

[0094] Embodiment 16. The compound of Embodiment 15, wherein each R7is independently - O-C₁-C₆ alkyl, N(C₁-C₃ alkyl)₂, halo, -NO₂, and -OH, or a pharmaceutically acceptable salt or hydrate thereof.

[0095] Embodiment 17. The compound of Embodiment 16, wherein -O-C₁-C₆ alkyl is -O-methyl.

[0096] Embodiment 18. The compound of Embodiment 16, wherein C1-C3 alkyl is methyl.

[0097] Embodiment 19, The compound of Embodiment 16, wherein halo is Cl, F, or Br,

[0098] Embodiment 20. The compound of any one of Embodiments 1-19, wherein the compound is a compound of formula:N=N N=NN-N70948-02or a pharmaceutically acceptable salt or hydrate thereof, wherein n and R5are defined in Embodiment 1.

[0100] Embodiment 21. The compound of any one of Embodiments 1-20, wherein the compound is a compound of formula:or a pharmaceutically acceptable salt or hydrate thereof.70948-02

[0101] Embodiment 22. The compound of Embodiment 20 or 21, wherein the compound is (5- (l-(4-Dimethylaminophenyl)-l -l,2,3-triazol-4-yl)-l -indole) or a pharmaceutically acceptable salt or hydrate thereof and has the formula:N— Nor a pharmaceutically acceptable salt or hydrate thereof.

[0102] Embodiment 23. The compound of Embodiment 20 or 21, wherein the compound is (4- [4-(l / f-mdol-5-yl)-lJ7-I,2,3-triazol-l-yl]benzene-l,2-diol) or a pharmaceutically acceptable salt or hydrate thereof and has the formula:N=Nor a pharmaceutically acceptable salt or hydrate thereof.

[0103] Embodiment 24. The compound of Embodiment 20 or 21, wherein the compound is (5- [4-(l / f-indol-5-yl)-lK-l,2,3-triazol-l-yl]benzene-l,2,3-triol) or a pharmaceutically acceptable salt or hydrate thereof and has the formula:N-NHO ~ hi 1jy -x oj nOHor a pharmaceutically acceptable salt or hydrate thereof.

[0104] Embodiment 25. A pharmaceutical composition comprising a compound of any one of Embodiments 1-24, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier.

[0105] Embodiment 26. A method of inhibiting alpha-synuclein (a-syn) protein fibril formation in a subject having, or at risk for, a-syn protein fibril formation, which method comprises administering to the subject an effective amount of (i) a compound of any one of Embodiments 1-24, or a pharmaceutically acceptable salt or hydrate thereof, or (ii) a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier.70948-02

[0106] Embodiment 27. The method of Embodiment 26, wherein the subject has, or is at risk for, Parkinson’s disease or dementia with Lewy bodies (DLB).

[0107] Embodiment 28. The method of Embodiment 26 or 27, wherein the compound is the compound of Embodiment 24, or a pharmaceutically acceptable salt or hydrate thereof, or a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier, and the compound also inhibits a-syn oligomers.

[0108] Embodiment 29. A method of inhibiting tubulin-associated unit (tau) protein (isoforms 0N4R, 1N4R, 2N4R, 0N3R, 1N3R, 2N3R) oligomer formation in a subject having, or at risk for, tau (isoforms 0N4R, 1N4R, 2N4R, 0N3R, 1N3R, 2N3R) oligomer formation, which method comprises administering to the subject an effective amount of (i) a compound of any one of Embodiments 1-24, or a pharmaceutically acceptable salt or hydrate thereof, or (ii) a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier.

[0109] Embodiment 30. The method of Embodiment 29, wherein the compound is the compound of Embodiment 24, or a pharmaceutically acceptable salt or hydrate thereof, or a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier.EXAMPLESDesign of the prepared compounds

[0110] The indole scaffold has been identified and elaborated in several significantly active anti-fibrillar compounds against different prone-to-aggregate proteins (compounds 1 and II, Fig.1A).21’24Thanks to their strong hydrogen- bonding capabilities, electron-donating characteristics, and stability, the triazoles can interact with multiple biological targets in neurodegeneration,23’26such as inhibiting the formation of amyloid plaques and preventing neurotoxicity.2' Therefore, triazoles are believed to be promising candidates for preventing or reducing the formation of toxic protein aggregates associated with neurodegenerative diseases. The polyphenolic green tea catechin, (-)-epi-gallocatechin-3-gallate (EGCG), revealed powerful anti-fibrillar and anti- oligomeric properties towards a-syn and tau deposits (Fig. IB).28-22Thus, the prepared compounds were designed to have the indole moiety connected to polyphenols using the triazole70948-02linker (Fig. 1C). In addition, the importance of the polyphenolic groups was evaluated by exchanging them with electron-rich or electron-deficient groups, such as methoxy and dimethylamino groups, or nitro and chloro substituents, respectively.Chemistry

[0111] The synthesis of the designed compounds is outlined in Scheme 1 (Fig.2). Briefly, different azides (la-h) were simply prepared by diazotization of the corresponding anilines with sodium nitrite and hydrochloric acid followed by reaction with sodium azide. The commercially available 5-iodo-l / f-indole reacted with ethynyltrimethyl silane in the presence of a palladium catalyst and base. The crude product was then subjected to hydrolysis with sodium hydroxide to get the 5-ethynyl derivative, 2, in a good yield. The targeted compounds 3a-h were successfully achieved by Hiiisgen Cu(I)-mediated [3 + 2] click cycloaddition reaction between the substituted aromatic azides (la-h) and the terminal alkyne (2). Finally, demethylation of the di- or trimethoxy groups was performed by reacting 3b-d with boron tribromide yielding the corresponding di- or tri-phenols, 4b-d, respectively.General Characterizations

[0112] / All reagents and solvents were commercially available (Sigma Aldrich, St. Louis, MO; Thermo Scientific (formerly Alfa Aesar), Waltham, MA; Matrix Scientific, Columbia, SC; Ambeed, Arlington Hts, IL) and were used without further purification. Thin-layer chromatography (TLC) was used to monitor the reaction progress. Organic solutions were dried over anhydrous magnesium sulfate. The solvents were evaporated on a Buchi rotavapor R-100 equipped with a Buchi V-100 vacuum controller. The nuclear magnetic resonance (NMR) spectra were recorded on a Bruker spectrometer at a frequency of 500 MHz for ’H and at 126 MHz for13C. Proton chemical shifts are reported m parts per million (ppm) with the solvent reference relative to tetramethyl silane (TMS) employed as the internal standard (CDC13, 87.26; DMSO-d6 8 2.54). The multiplicities of NMR signals are designated as s (singlet), d (doublet), dd (double doublet), t (triplet), q (quartet), m (multiplet, for unresolved lines). High-resolution mass spectrometry (HRMS) of the compounds was carried out on Advion Mass Spectrometer (Advion Expression CMS) at the Analytical Mass Spectrometry Facility within the Purdue Institute for Drug Discovery. Uncorrected melting points (m.p.) were scored using an electrothermal apparatus (Barnstead International, Dubuque, Iowa, USA).General procedure for the synthesis of substituted azido benzene (la-d)70948-02

[0113] The appropriate aniline derivative (1.0 eq.) was dissolved in a 10 mL mixture of cone. HC1 and water (1:1) at 0-5°C. Sodium nitrite (1.5 eq.) was gradually added over 5 minutes and the reaction mixture was stirred for 30 minutes. Sodium azide (2 eq.) was then added carefully, and the reaction mixture continuously stirred for 2-3 hours. After completion, the mixture was extracted with dichloromethane (3 × 15 mL), filtered over anhydrous magnesium sulfate, and concentrated in vacuo to give the desired compounds. Some azides were needed for further purification using silica gel column chromatography (hexane / ethyl acetate 10:1 v / v) yielding the pure compounds in excellent yields (85-100%).

[0114] l-Azido-4-methoxybenzene (la). 87%, white solid; 1H NMR (500 MHz, CDC13) 86.96 (d,.7 = 9.0 Hz, 2H), 6.89 (d,.7= 9.0 Hz, 2H), 3.79 (s, 3H).13C NMR (126 MHz, CDCh) 8 157.0, 132.3, 120.0, 115.1, 55.5.

[0115] l-Azido-3,4-dimethoxybenzene (lb). 92%, white solid; 'H NMR (500 MHz, CDCh) 86.60 (d, 7 = 8.6 Hz, 1H), 6.35 (dd, J= 8.6, 2.6 Hz, 1H), 6.29 (d,.7 = 2.7 Hz, 1H), 3.63 (d,.7=2.7Hz, 6H).13C NMR (126 MHz, CDCh) 8149.9, 146.4, 132.5, 111.9, 110.2, 103.0, 55.8, 55.6.

[0116] l-Azido-3,5-dimethoxybenzene (1c). 96%, white solid;NMR (500 MHz, CDCh) 86.24 (t, J --- 2.2 Hz, 1H), 6.17 (d, J --- 2.3 Hz, 2H), 3.76 (s, 6H).13C NMR (126 MHz, CDC13) 8 161.7, 141.9, 97.5, 97.2, 55.3.

[0117] l-Azido-3,4,5-trimethoxybenzene (Id). 100%, white solid; 1H NMR (500 MHz, CDC13) 86.04 (s, 2H), 3.65 (s, 6H), 3.62 (s, 3H).13C NMR (126 MHz, CDCh) 8 153.9, 135.4, 96.2, 60.6, 55.8.

[0118] l-Azido-4-diniethylamino benzene (le). 97%, white solid; 'H NMR (500 MHz, CDCh) 87.16 - 6.88 (m, 2H), 6.79 (s, 2H), 2.95 (s, 6H).

[0119] l-Azido-4-chIorobenzene (If). 97%, pale grey solid;]H NMR (500 MHz, CDCh) 87.31 (d, J= 8.8 Hz, 2H), 6.96 (d, J= 8.8 Hz, 2H).!3C NMR (126 MHz, CDCh) 8 138.7, 130.2, 129.9, 120.3.

[0120] l-Azido-3,5-dichlorobenzene (lg).85%, white solid;]H NMR (500 MHz, CDCh?) 8 7.10 (t, J= 1.8 Hz, 1H), 6.89 (d, J= 1.9 Hz, 2H).!3C NMR (126 MHz, CDCh) 8 142.4, 136.0, 125.1, 117.7.70948-02

[0121] l-Azido-4-nitrobenzene (Ih). 88%, white solid;NMR (500 MHz, DMSO) 8 8.19 (d, J = 9.1 Hz, 2H), 7.28(d, 9.1 Hz, 2H).13C NMR (126 MHz, DMSO) 8 147.1, 144.5, 125.9, 120.5, 40.1, 39.9, 39.8.Synthesis 5-Ethynyl-LH- indole (2).

[0122] 5-Iodo-lH-indole (1 eq.), bis(triphenylphosphine) palladium(II) dichloride (0.05 eq.), cuprous iodide (0.05 eq.), and triethyl amine (2 eq) were added to DMF (2 mL) at room temperature under a N2 atmosphere. Ethynyltrimethylsilane was slowly added, and the reaction was stirred for 5-8 h at room temperature. The reaction mixture was filtered through celite pad and the organic layer was concentrated in vacuo. The residue was then treated with 0.2 M NaOH (2 ml.) in methanol (5 mL) at room temperature for another 1 h. The reaction mixture was poured into water and extracted with ethyl acetate (3 x 20 mL), filtered over anhydrous magnesium sulfate, and concentrated. The desired compound was further purified by column chromatography using hexane and ethyl acetate volume ratio of 5: 1 (v / v).

[0123] 5-EthynyI-177-indoIe (2). 72%, white solid; ’H NMR (500 MHz, CDCh) 88.15 (s, IH), 7.89 (dd,.7= 1.6, 0.8 Hz, IH), 7.37 (d,.7= 1.6 Hz, IH), 7.34 - 7.27 (m, IH), 7.21 - 7.16 (m, IH), 6.59-6.53 (m, IH), 3.08 (s, IH).13C NMR(126MHz, CDCh) 8135.7, 127.7, 126.0, 125.4, 125.4, 113.1, 111.3, 102.9, 85.5, 74.9.General procedure for the synthesis of 5-(l-(substituted phenyI)-17Z-l,2,3-triazol-4-yi)-lJI-indoie (3a-3h) and 4-(benzo[ / >jthiophen-5-yI)-l-(3,4,5-triniethoxyphenyl)-l / / -l,2 3-triazole by click chemistry (6)

[0124] The azide derivative (1 eq.) and the acetylene derivative (e.g. 2, 1.2 eq.) were dissolved in a mixture of THF and water (1:1) at room temperature. Then, sodium ascorbate (0.1 eq. ) and Copper sulfate (0.05 eq.) were added, and the reaction was stirred and followed by TLC. The reaction mixture was then partitioned between ethyl acetate and water. The organic layers were collected, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure, The desired triazole was afforded by column chromatography using hexane / ethyl acetate at excellent yields.

[0125] 5-(l-(4-MethoxyphenyI)-177-l,2,3-triazoI-4-yl)-LH-indoIe (3a). 84% white solid; ’H NMR (500 MHz, CDCh) 88.29 (s, IH), 8.19 (dd, J= 1.6, 0.9 Hz, IH), 8.10 (s, IH), 7.77 (dd, J= 8.5, 1.6 Hz, IH), 7.74 - 7.68 (m, 2H), 7.52 - 7.45 (m, IH), 7.26 (d, J= 1.5 Hz, IH), 7.08 -70948-027.02 (m, 2H), 6.65 - 6.59 (m, 1H), 3.88 (s, 3H).l3C NMR (126 MHz, CDCh) 5 159.7, 149.6, 135.9, 130.8, 128.3, 125.0, 122.4, 122.2, 120.5, 118.3, 117.1, 114.8, 111.5, 103.1, 55.7.

[0126] 5-(l-(3,4-DimethoxyphenyI)-l / T-l,2,3-triazoI-4-yl)-lH-indole (3b). 83% white solid; NMR (500 MHz, CDCh) 58.40 (s, 1H), 8.22 -8.16 (m, 1H), 8.11 (s, 1H), 7.76 (dd, J = 8.4, 1.7 Hz, 1H), 7.47 (d, J= 8.4 Hz, 1H), 7.43 (d, J= 2.5 Hz, 1H), 7.26 - 7.21 (m, 2H), 6.97 (d, J= 8.6 Hz, 1H), 6.65 - 6.58 (m, 1H), 3.98 (s, 3H), 3.95 (s, 3H).13C NMR (126 MHz, CDCh) 5 149.8, 149.7, 149.3, 136.0, 130.9, 128.3, 125.2, 122.2, 120.4, 118.2, 117.3, 112.4, 111.6, 111.2, 105.0, 102.9, 56.2, 56.2.

[0127] 5-(l-(3,5-DimethoxyphenyI)-17 / -l,2,3-triazoI-4-yl)-lH-indole (3c). 91% white solid;!H NMR (500 MHz, CDCh) 8 8.35 (s, 1H), 8.21 - 8.16 (m, 1H), 8.14 (s, 1H), 7.76 (dd, J = 8.4, 1.7 Hz, 1H), 7.47 (d, J= 8.4 Hz, 1H), 7.26 - 7.24 (m, 1H), 6.98 (d, J= 2.2 Hz, 2H), 6.64 - 6.59 (m, 1H), 6.52 (t, J =2.2 Hz, 1H), 3.88 (s, 6H).13C NMR (126 MHz, CDCh) 8 161.5, 149.7, 138.8, 136.0, 128.3, 125.1, 122.1, 120.5, 118.3, 117.0, 111.6, 103.1, 100.4, 98.9, 55.8.

[0128] 5-(1-(3,4,5-TrimethoxyphenylI)-1H-1,2,3-triazol-4-yl)-1H-indole (3d). 88% white solid;jH NMR (500 MHz, CDCh) 88.42 (s, 1H), 8.19 (s, 1H), 8.13 (s, 1H), 7.76 (dd, J = 8.5, 1.7 Hz, 1H), 7.47 (d, J === 8.4 Hz, 1 II), 7.26 - 7.24 (m, 1 H), 7.02 (s, 2H), 6.65 - 6.57 (m, 1H), 3.95 (s, 6H), 3.91 (s, 3H).13C NMR (126 MHz, CDCh) 8153.9, 149.7, 138.2, 136.0, 133.2, 128.3, 125.1, 122.1, 120.4, 118.3, 117.2, 111.6, 103.1, 98.5, 61.1, 56.5.

[0129] 5-(l-(4-DimethylaminophenyI)-l / / -l,2,3-triazol-4-yI)-17 / -indoIe (3e). 79% white solid;1H NMR (500 MHz, CDCh) 88.25 (s, 1H), 8.19 (s, 1H), 8.08 (s, 1H), 7.77 (dd, J = 8.4, 1.6 Hz, 1H), 7.67 (d, J = 8.6 Hz, 2H), 7.48 (d, J = 8.5 Hz, 1H), 7.25 (s, 1H), 6.96 (s, 2H), 6.65 - 6.53 (m, 1H), 3.06 (s, 6H).]3C NMR (126 MHz, CDCh) 8 144.4, 135.9, 131.2, 126.8, 126.4, 125.8, 125.0, 121.9, 120.6, 118.3, 117.0, 111.5, 103.2, 29.7.

[0130] 5-(l-(4-ChlorophenyI)-lfir-l,2,3-ti'iazol-4-yl)-lHr-iiidole (3f). 71% pale grey solid, m.p.: 261.6 - 262.4°C.; ’H NMR (500 MHz, DMSO) 811.22 (s, 1H), 9.21 (s, 1H), 8.13 (d, J= 1.7 Hz, 1H), 8.04 - 7.97 (m, 2H), 7.73 - 7.65 (m, 3H), 7.52 - 7.46 (m, 1H), 7.39 (t, J = 2.7 Hz, 1H), 6.55 - 6.45 (m, 1H).!3C NMR (126 MHz, DMSO) 8 149.6, 136.4, 136.1, 133.2, 130.4, 128.4, 126.8, 121.9, 121.6, 119.6, 118.8, 117.6, 112.4, 102.0.

[0131] 5-(l-(3,5-DichIorophenyI)-lH-l,23Hriazol-4-yI)-lH-indoIe (3g). 71% white solid; ’H NMR (500 MHz, CDCh) 88.32 (s, 1H), 8.22 - 8.15 (m, 1H), 8.14 (s, 1H), 7.80 - 7.67 (m, 2H), 7.58 (d, J = 1.9 Hz, 1H), 7.52 - 7.39 (m, 2H), 6.74 (d, J = 1.8 Hz, 1H), 6.66 - 6.57 (m,70948-02IH).i3C NMR(126 MHz, CDCh) 5150.3, 138.5, 136.3, 136.0, 129.2, 128.4, 125.2, 123.8, 121.5, 120.4, 118.7, 118.5, 116.4, 111.6, 111.1, 103.2.

[0132] 5-(1-(4-NitrophenylI)-1H-1,2,3-triazol-4-yl)-1H-indole (3h). 75% white solid; 'H NMR (500 MHz, CDCh) 5 8.29 (s, IH), 8.26 - 8.22 (m, 1H), 8.10 - 8.05 (m, 1H), 7.88 (d, J = 1.7 Hz, IH), 7.63 - 7.57 (m, 2H), 7.43 - 7.33 (m, 2H), 6.97 (dd, J= 8.4, 1.8 Hz, 1H), 6.68 - 6.58 (m, IH), 6.58 - 6.48 (m, IH).i3C NMR (126 MHz, CDCh) 8 135.8, 134.0, 126.4, 125.9, 125.3, 125.2, 124.8, 122.5, 121.7, 113.4, 112.0, 111.2, 103.3, 103.1.General procedure for the demethylation of (poly)methoxy derivatives (4b-d)

[0133] The methoxy substrate (1.00 equiv.) was dissolved in DCM (5.0 mL) at room temperature and cooled to 0°C on an ice bath. Boron tribromide (5.00 equiv. for each methoxy functionality) was added dropwise at 0°C over 15 min. Then, the reaction mixture was stirred at 0°C and allowed to warm to room temperature gradually overnight. Upon completion, the reaction was quenched with saturated NH4CI and extracted with ethyl acetate. The organic layer was dried with anhydrous magnesium sulfate and concentrated under a vacuum. The crude mixture was purified by column chromatography (DCM: methanol; 9:1 v / v).

[0134] 4-[4-(lZ / -indoI-5-yl)-l / -l,2,3-triazol-l-yljbenzene-l,2-dioI (4b). ^I NMR (500 MHz, Acetone) 8 10.34 (s, IH), 8.69 (s, IH), 8.54 (s, IH), 8.42 (s, IH), 8.19 (d, J - 0.9 Hz, IH), 7.76 (dd, J = 8.4, 1.6 Hz, IH), 7.50 (d, J === 8.4 Hz, IH), 7.46 (d, J = 2.6 Hz, HI), 7.39 - 7.34 (m, IH), 7.26 (dd, J - 8.5, 2.6 Hz, IH), 7.01 (d, J = 8.5 Hz, HI), 6.53 (ddd, J = 3.0, 1.9, 0.9 Hz, IH).13C NMR (126 MHz, Acetone) 8 149.2, 145.9, 145.5, 136.3, 130.4, 128.5, 125.6, 122.4, 119.6, 117.4, 117.4, 115.6, 111.7, 111.6, 108.1, 101.9.

[0135] 5-[4-(1H-indol-5-yl)-1H-1,2,3-triazol-1-yl]benzene-1,3-diol (4c). ’H NMR (500 MHz, MeOD) 88.63 (s, IH), 8.09 (d, J = 0.9 Hz, IH), 7.65 (dd, J = 8.4, 1.7 Hz, IH), 7.52 - 7.41 (m, IH), 7.27 (d, J = 3.2 Hz, IH), 6.83 (dd, J = 4.5, 2.1 Hz, 2H), 6.51 (dd, J = 3.1, 0.9 Hz, IH), 6.38 (t, J = 2.1 Hz, IH).]3C NMR (126 MHz, MeOD) 8159.5, 149.7, 138.5, 136.5, 128.4, 125.3, 120.9, 119.2, 117.7, 117.5, 111.3, 102.4, 101.5, 98.7.

[0136] 5-[4-(lH-indoi-5-yi)-lH-l,2,3-triazol-l-yI]benzeiie-l,2 -trioi (4d). IH NMR (500 MHz, DMSO) 8 11.17 (s, IH), 9.43 (s, 2H), 8.90 (s, IH), 8.58 (s, IH), 8.10 (s, IH), 7.67 (dd, J = 8.4, 1.7 Hz, IH), 7.45 (d, J = 8.4 Hz, IH), 7.36 (d, J = 3.2 Hz, IH), 6.84 (s, 2H), 6.47 (d, J = 3.1 Hz, IH).!3C NMR (126 MHz, DMSO) 8 149.0, 147.1, 136.3, 133.9, 128.9, 128.4, 126.6, 122.0, 119.6, 118.5, 117.5, 112.2, 101.9, 100.0.70948-02Biophysical EvaluationThioflavin T or Thioflavin S (ThT, or ThS) Fluorescence Assays

[0137] The effects of the synthesized compounds on a-syn fibril formation were tested using a Thioflavin- T (ThT) fluorescence assay. The experiment was set up in a 96-well microplate. Compounds were added to each well to reach a final concentration of 100 pM, along with ThT, which was also at 100 pM. a-Syn (obtained from rPeptide) was then added to achieve a final concentration of 2 or 6 μM, using a stock solution of 277 pM dissolved in 20 mM Tris- HC1 (pH 7.4). The assay buffer included 10 mMPBS (pH 7.4), 300 mMNaCl, 0.5 mM SDS, and a 3 mm borosilicate bead in each well. The plate was loaded into a Synergy HT multi-mode microplate reader (BioTek, Winooski, VT) and maintained at 37°C. ThT fluorescence was monitored by measuring excitation at 440 nm and emission at 485 nm. Data were collected every 20 minutes, with the plate being shaken for 10 seconds before each reading. The fibril formation process was monitored for > 37 hours. Samples were tested in triplicate.

[0138] For the Thioflavin S (ThS) assay, a black 384-well flat-bottom microplate (Brand, ref 784076, medium binding) was employed. PBS (pH 7.4) from Gibco (catalog number 10010- 023), treated with chelex beads (Biosciences, BTNM-0024), was added to each well to reach a final volume of 10 pL. The 2N4R tau peptide (obtained from rPeptide), 0N4R (recombinant protein produced in the laboratory), or p-tau (recombinant protein produced in the laboratory) was then introduced to achieve a final concentration of 12 pM, followed by the addition of DTT to 5 mM. The test compound was added to each well at a final concentration of 100 pM, and heparin was included at 150 pM. Arachidonic acid was then added to the mixture to reach a final concentration of 0.092 pg / mL (A0781, Fischer TCI). Thioflavin S (ThS) was incorporated at a final concentration of 40 pM to monitor fibril formation kinetics. Fluorescence intensity was recorded every 5 minutes, with a 30-second agitation step preceding each measurement,

[0139] To assess the ability of compounds to inhibit a-syn fibrillation, fibril formation was monitored using Thioflavin T (ThT) fluorescence intensity over time. ThT is known to bind to p- sheet structures present in amyloid-like fibrils, which restricts the rotation of the molecule and increases its fluorescence signal.33In our study, we incubated each compound (100 pM) with a- syn monomers (6 μM) for > 27 h. The results were reported as the average of the maximum fluorescence intensity at the plateau phase where the fibrils are mature, and the disaggregation and elongation are in equilibrium. The percentages are compared to evaluate the compound70948-02effects on fibril formation. Throughout the incubation period, fluorescence was measured every 20 min to monitor changes in intensity. The introduction of compounds Sa, 3b, 3c, 3d, 3g, and 3h led to an increase in fluorescence. Conversely, compounds 3e, 4b, and 4d reduced fluorescence intensity by > 80% compared to the control, where monomeric a-syn was incubated with DMSO at 0.25%. This suggests that these specific compounds have the potential to inhibit a-syn fibril formation. The maximum ThT fluorescence intensities in percentage resulting from a-syn incubated with compounds 3e, 4b, and 4d were 11.8 ± 3.7%, 14.3 ± 6.3%, and 3.9 ± 1.7%, respectively (Table 1). Interestingly, the triazoles with electron-rich substituent such as polyphenol or phenyldimethylamino groups result in drastic reduction of ThT fluorescence. One triazole with polyphenol compound, 4c, andone with the nitro (electron-deficient group) substituted aromatic, 3f, resulted in a moderate reduction of ThT fluorescence intensity of about 25 to 48%, respectively (Table 1).CMtr:Table 1. The anti-fibrillary activity of the newly synthesized triazoles (100 μM) on a-syn fibril formation (2 μM). The average of three triplicate results was taken from the maximum thioflavin T (ThT) fluorescence intensity at the plateau phase of the kinetics of aggregation. The average of the maximum of fluorescence is expressed as a percentage with the standard error of the mean (SEM). Control consisted of 0.25% DMSO. ThT results over 100% refer to the incapacity at reducing a-syn fibril formation.a-syn a-syn Compound CompoundRi R? Rs ThT Rs R? Rs ThT ID ID% %3a H OCH₃ H 3g Cl H Cl >100>100OCH3OCH3H >100 3h H NO2H >100 3b70948-0214.3 ± 3c OCH3H OCH3>100 4b OH OH H6.377.1 ± 3d OCH3OCH3OCH3>100 4c OH H OH2.211.8 ±3e H N(CH3)2H 4d OH OH OH 3.9 ± 1.73.754.0 ±3f H Cl H1.5

[0140] The kinetics of fibril formation of the best triazole compounds are presented in Fig.3A. Compounds 3a, 3c-d, and 3g-h were excluded from the kinetic analysis curves because their ThT fluorescence values significantly exceeded those of the control. Compounds 3a, 3b, 3c, 3d, 3g, and 3h, resulted in ThT data that were over 100% (Table 1). These compounds were not able to reduce the fibril formation. They most likely have increased the formation of fibrils. Following the initial ThT assay, a dose-response curve was generated for compound 4d (Fig. 3B). To validate the inhibition of a-syn aggregation, the ThT assay was repeated with two concentrations of protein (2 and 6 μM). Compound 4d was tested at incremental increase of concentrations. For the a-syn tested at 6 μM, the analysis using Prism software revealed a Log(agonist) versus normalized response correlation with a variable slope, yielding a LogEC₅₀ value of 14.55 ± 1.97 (Fig. 3B) At a lower concentration of a-syn (2 pM), compound 4d LogEC₅₀ value corresponded to 6.22 ± 1.23. The result shows the varying a-syn anti-aggregation effect of compound 4d in the dose-response profile.

[0141] Additionally, 4d was tested at 100 pM resulting in an approximate molar ratio protein: compound of 1: 8 (12 pM tau 2N4R) and 1: 16 (6 μM for tau 0N4R and p-tau 1 N4R). The anti-aggregation effect of the best lead (i.e. 4d) on the 4R tau isoforms (0N4R, 2N4R) and the phosphorylated tau isoform 1N4R was assessed using thioflavin S (ThS) (Fig. 4). The kinetics of fibril formation of the non-phosphorylated and phosphorylated tau isoform 4R demonstrated a reduction of approximately 50% in fibril formation, indicating the potential efficacy of compound 4d in inhibiting the aggregation of tau proteins.Photo-Induced Cross-linking of Unmodified Protein (PICUP) assay70948-02

[0142] Monomeric a-syn (obtained from rPeptide) and tau 0N4R (produced in the laboratory) were diluted to 30 pM and 6 μM in PBS (pH 7.4), respectively. To initiate the crosslinking reaction, 2 μL of a 300 μM tris(bipyridine)ruthenium(II) complex solution and 2 μL of 6 mM ammonium persulfate were added to each tube. The PCR tubes were placed inside a dark enclosure. Control samples, which did not contain the Ru(bpy) complex, consisted of 0.25% DMSO. Light exposure was applied using a 53 W (120 V) incandescent bulb: a-syn was exposed for 1 s, and tau 0N4R was exposed for 15 s. After light exposure, 15% 2-mercaptoethanol was added to stop the free radicals. The samples were heated at 95°C for 10 min on a heating block. The samples were then deposited into a 16% SDS-PAGE gel and, after electrophoresis, stained with Coomassie Blue. The final gels were scanned using the G: BOX F3 imaging system (Syngene), When necessary, high molecular weight band pixels were measured using image- J.

[0143] The formation of oligomers from amyloid-like fibril forming proteins is a crucial event in the development of protein-misfolding diseases.34These oligomers are generated at the early-stage of the kinetics of aggregation. They form pore-like structures capable to disturb the integrity of lipid membranes of organelles and cells. The stress response generated by these oligomers may trigger apoptosis. These intermediate species participate to the generation of further aggregated protein. Therefore, these oligomers are known for their cytotoxic properties and play a crucial role in the spread of further misfolded protein and the progression of the disease.34, 35In our earlier research, the Photo-induced Cross-linking of Unmodified Proteins (PICUP) assay was used to generate stable a-syn and tau oligomers / '1’23This technique provided a means to test the effectiveness of various compounds in inhibiting or mitigating the formation of these pathological oligomers. Within the experimental condition used herein, the PICUP assays allow for the formation of oligomer at 35-40 kDa for syn and >180 kDa for tau 0N4R. The proteins are treated with different compound and subjected readily (without any incubation time) to the cross-linking reagents and incandescent light exposure for the generation of the oligomer formation.

[0144] The impact of compounds on the oligomerization of a-syn and tau 0N4R isoform was explored. Initially, the three best triazole-based compounds were evaluated at inhibiting a- syn fibril formation (i.e., 3e, 4b, and 4d) at a concentration of 100 μM against 30 μM of a-syn to identify their potential anti-oligomerization effects (Fig. 5A). Among the tested compounds, compound 4d demonstrated the highest efficacy in preventing a-syn oligomerization while 4b70948-02and 3e had little effect as indicated by the percentage of reduction of oligomer formation. The most effective anti-oligomer compound was further analyzed to determine its dose-response relationship with regard to inhibiting a-syn oligomer formation. Compound 4d was tested at variable concentrations ranging from ~3 to 100 μM. Compound 4d at 100 μM reduced 99 to 92% and at 50 μM reduced 52% of a-syn oligomer located between 35-45 kDa. The dose- response studies of the lead compound indicated concentration-dependent inhibitory effects on a-syn oligomerization as shown in Fig. 5B.

[0145] Prior experience suggests that the reduction of tau aggregation, particularly the oligomer formation, is more challenging in comparison with the reduction of a-syn aggregation. Taking this difficulty into consideration, the tau anti-oligomer effect was assessed using the isoform 0N4R (at 6 μM) and a higher concentration of the lead compound 4d (Fig. 6). The high molecular weight bands resulting from the cross-linking reaction were located above the 180 kDa benchmark. Pixel density measurements of each band were executed. A reduction of 39%, 56%, and 71% of the tau 0N4R oligomer formation was obtained with 100, 200, and 300 μM of compound 4d, respectively, in comparison with the condition without compound. The monomeric band was more abundant with the treated tau 0N4R with 100, 200, and 300 μM of compound 4d Transmission Electron Microscope (TEM) analysis

[0146] TEM analysis was carried out according to established protocols.21'24, 39Tau isoform 2N4R (12 pM, purchased from rPeptide), along with the compounds of interest and control with 0.25% DMSO, was incubated in a reaction mixture containing 10 mM PBS (pH 7.4), 150 μM heparin, 5 mM dithiothreitol (DTT), and 10 mg / mL arachidonic acid for 5 days at 37°C. For a-syn, the samples used in the ThT fibril formation assay were also analysed by TEM. Concerning the disaggregation assays, a procedure previously published was followed using amyloid-p plaques isolated from AD brain and treated with different compounds at a concentration of 50 μM for 5 days at 37°C prior to TEM analysis.22Protein determination was performed with the Lowry method using a bovine serum albumin (BSA) standard curve. For all different experiments, a small volume of sample (10 pL aliquot) was deposited on a 400-mesh Formvar-carbon-coated copper grid (Electron Microscopy Sciences, Hatfield, PA) and incubated for 1 min before being washed with distilled water. The grid was then air-dried, followed by a 1- minute staining with 1% uranyl acetate. Grids were dried with filter paper prior to visualization with the JEOL 1400 Flash electron microscope (Japan) operating at 100 kV.70948-02

[0147] To explore the influence of compound 4d on the ultrastructural morphology of a- syn and tau 2N4R fibrils, TEM analyses were conducted. Fig. 7 presents TEM images of a-syn fibrils collected after the ThT assay from a 96-well plate. The images depict fibrils formed under untreated conditions and those exposed to compound 4d at 12.5 and 100 μM. The untreated control, which included 0.25% DMSO, displayed numerous fibrils, and the same was observed with the compound treatment at 12.5 μM. In contrast, the sample treated with 100 pM compound 4d showed a significant decrease in fibril density, corroborating the inhibitory effect observed in the ThT assay. In the TEM images of tau 2N4R (Fig. 8), compound 4d demonstrated the ability to prevent the formation of fibrils. Tau 2N4R was co-incubated with either compound 4d (100 μM), EGCG (100 μM) or DMSO (0.25%; 'CTRL') for five days prior to imaging. In this experiment, EGCG was used as a positive control due to its proven ability to inhibit the aggregation of misfolded proteins, such as tau fibrils.29'31The known response of EGCG provides a reference for comparison, allowing us to assess the degree of inhibition of tau 2N4R aggregation under treatment with the best compound. The sample treated with compound 4d showed a significant decrease in fibril formation, highlighting the compound’s enhanced inhibitory effect compared to the control condition (0.25% DMSO).Disaggregation assay using amyloid-P plaques extracted from AD brains

[0148] The conceptualization of the triazole-based compounds in this study was designed using the indole moiety (as utilized in our prior work)21'24and polyphenol moiety of EGCG. EGCG is known to disaggregate pre-formed aS and amyloid-beta (Aβ) fibrils.8Next, we investigated the disaggregation effect of compound 4d because EGCG demonstrated disaggregation effects on several amyloid-like fibrils. The anti-aggregation and disaggregation effects of compound 4d was explored first with the recombinant amyloid beta 1-40 (Api-40). Api-40 was pre-treated only one hour with 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP) and exhibited some aggregates at the beginning of the kinetics (Fig. 9A). Aβ1-40 was used at 21 μM and treated with ranging concentration of each compound. The curves obtained from EGCG and 4d demonstrated the disaggregation and anti-aggregation effect on the amyloid beta fragment, resulting in low percentage of ThT (Fig. 9A). The maximum fluorescence intensity in percentage were obtained from the last ten data at the end of the kinetics for each compound (Figs. 9B-9C). Compound 4d and EGCG reduced the percentage of ThT fluorescence significantly.70948-02

[0149] From prior experience, general inhibitors of aggregation often exhibit disaggregation activity on mature fibrils. We aimed to confirm the disaggregation effect of compound 4d. The recombinant amyloid-like fibrils are slightly different than the amyloid-like fibrils isolated from disease patients. For this reason, the disaggregation effect of lead compound 4d was assessed using Ap plaques extracted from AD patients (ex vivo). Protein concentration was measured with the Lowry method, and samples corresponded to 0.593 ± 0.095 mg / mL. The AP plaque-rich solution was incubated with 50 pM of compound 4d or EGCG. The non-treated control showed fibril-dense rich plaques (Fig. 10). In comparison to the control (0.25% DMSO), the materials resulting from compound 4d were less defined and fibril-sparse. EGCG treatment resulted in less defined extravesicular-like materials supporting the reversal of fibrils into presumably nontoxic species,a-Synuclein Inclusion-Forming Neuroblastoma Cell Experiment

[0150] As in prior studies, doxycycline-inducible M17D-TR / aS-3K:: YFP neuroblastoma cells were utilized to assess the anti-inclusion effect of compounds.21"24, 37, 39For the assay, cells were initially seeded at 30,000 cells per well in 96-well plates. After a 24 h incubation period, compounds were added, and the aS-3K:: YFP transgene expression was induced 24 h later by adding doxycycline to the media at a final concentration of 1 μg / mL. Continuous imaging of the cells was performed using the Incucyte Zoom 2000 system (Essen Biosciences), capturing both green fluorescence and bright field images. The assessment of inclusion formation and cell growth was conducted 48 hours post-induction (96 hours after initial plating). The Incucyte analysis settings for “Inclusions” were adjusted with a Fixed Threshold at 50 GCU, Edge Split with Sensitivity set to 100, and Cleanup parameters including Hole Fill set at 10 μm2and Adjust Size at 0 pixels. The filtering criteria included a maximum Area of 50 μm2, a minimum Mean Intensity of 60, and a minimum Integrated Intensity of 2000. Cell confluence was measured using the 'Cells' processing settings with a Segmentation Adjustment of 0.7, Cleanup parameters set to 0, and Filters applying a minimum Area of 345 μm2

[0151] To assess how different compounds influence a-syn inclusion formation, ‘3KY19’ M17D neuroblastoma cells were used.37These cells are engineered to express the aS3K:: YFP fusion protein when subjected to doxycycline (dox) treatment. The setup was designed to mimic the pathology of familial Parkinson's disease, particularly focusing on the E46K mutation. To intensify the disease model, a triple mutation (E35K + E46K + E61K), known as aS3K, was70948-02incorporated that readily forms distinct round inclusions in the cytoplasm of cultured cells. This model serves as an effective tool for studying disease mechanisms and evaluating the antiinclusion effect of various compounds.21’243839The cells were treated with different concentrations of compounds 4b and 4d, starting 24 h after they were plated. 4b was selected because of its lack of anti-oligomer effect. Compound 4b had a slightly weaker anti-fibrillar effect in comparison with compound 4d. Inclusion formation was initiated 48 h after plating via the addition of doxycycline to initiate the expression of the fusion protein. Inclusion formation was quantified at 96 h to determine the dose-dependent effects of compounds 4b and 4d. Compound 4d markedly decreased a-syn inclusion formation at concentrations of 2.5, 5, 10, 20, and 40 pM, with the most notable reduction observed at 40 pM (Figs. 11A and 11C). Conversely, compound 4b did not reduce a-syn inclusion levels. The cell confluence was unaffected during the treatment period (Fig. 11B).

[0152] In summary, phenolic indoles connected to a triazole linker were designed and synthesized to prevent the fibrils of a-syn and tau aggregates. Using a ThT assay, the triphenolic derivative 4d induced a significant loss in the fluorescence intensity of about 95% with a-syn aggregates and approximately 50% with tau (0N4R, 2N4R) and p-tau (1N4R). Moreover, 4d exhibited an a-syn and tau (0N4R) anti-oligomer activity at high micromolar concentration employing the PICUP assay. The TEM analyses validated the ability of 4d to reduce the fibril length and density of both a-syn and 2N4R. In addition, the cell-permeable compound 4d robustly prevented the formation of a-syn inclusion in a cell-based assay. Compound 4d demonstrated the ability to disaggregate A fibrils. Further structural modification is ongoing to target early-stage oligomers exclusively.References

[0153] (1) Irvine, G. B.; El-Agnaf, O. M.; Shankar, G. M.; Walsh, D. M. Protein aggregation in the brain: the molecular basis for Alzheimer's and Parkinson’s diseases. Mol Med 2008, 14 (7-8), 451-464. DOI: 10.2119 / 2007-00100. Irvine from NLM Medline

[0154] (2) Peng, C.; Trojanowski, J. Q.; Lee, V. M. Protein transmission in neurodegenerative disease. Nat Rev Neurol 2020, 16 (4), 199-212. DOI: 10.1038 / s41582-020- 0333-7 from NLM Medline.

[0155] (3) Sweeney, P.; Park, H.; Baumann, M.; Dunlop, J.; Frydman, J.; Kopito, R.; McCampbell, A.; Leblanc, G; Venkateswaran, A.; Nurmi, A.; et al. Protein misfolding in70948-02neurodegenerative diseases: implications and strategies. Transl Neurodegener 2017, 6, 6. DOI: 10.1186 / s40035-017-0077-5 from NLM PubMed-not-MEDLINE.

[0156] (4) Waddington-Cruz, M.; Ackermann, E. J.; Polydefkis, M.; Heitner, S. B.; Dyck, P. J.; Barroso, F. A.; Wang, A. K.; Berk, J. L.; Dyck, P. J. B.; Monia, B. P.; et al. Hereditary transthyretin amyloidosis: baseline characteristics of patients in the NEURO-TTR trial. Amyloid 2018, 25 (3), 180-188. DOI: 10.1080 / 13506129.2018.1503593 From NLM Medline.

[0157] (5) Price, J. L.; Morris, J. C Tangles and plaques in nondemented aging and "preclinical" Alzheimer's disease. Ann Neurol 1999, 45 (3), 358-368. DOI: 10.1002 / 1531- 8249(199903)45:3<358::aid-anal2>3.0.co;2-x from NLM Medline.

[0158] (6) Rajmohan, R.; Reddy, P. H. Amyloid-Beta and Phosphorylated Tau Accumulations Cause Abnormalities at Synapses of Alzheimer's disease Neurons. J Alzheimers Dis 2017, 57 (4), 975-999. DOI: 10.3233 / JAD-160612 from NLM Medline.

[0159] (7) Gulisano, W.; Maugeri, D.; Baltrons, M. A.; Fa, M.; Amato, A.; Palmeri, A.; D'Adamio, L.; Grassi, C.; Devanand, D. P.; Honig, L. S.; et al. Role of Amyloid-beta and Tau Proteins in Alzheimer's Disease: Confuting the Amyloid Cascade. J Alzheimers Dis 2018, 64 (si), S611-S631. DOI: 10.3233 / JAD-179935 from NLM Medline.

[0160] (8) Buxbaum, J. N.; Dispenzieri, A.; Eisenberg, D. S.; Fandrich, M.; Merlin, G.; Saraiva, M. J. M.; Sekijima, Y.; Westermark, P. Amyloid nomenclature 2022: update, novel proteins, and recommendations by the International Society of Amyloidosis (ISA) Nomenclature Committee. Amyloid 2022, 29 (4), 213-219. DOI: 10.1080 / 13506129.2022.2147636 from NLM Medline.

[0161] (9) Metaxas, A.; Kempf, S. J. Neurofibrillary tangles in Alzheimer's disease: elucidation of the molecular mechanism by immunohistochemistry and tau protein phosphoproteomics. Neural Regen Res 2016, 11 (10), 1579-1581. DOI: 10.4103 / 1673-5374.193234 from NLM PubMed-not-MEDLINE.

[0162] (10) Braak, H.; Alafuzoff, I.; Arzberger, T.; Kretzschmar, H.; Del Tredici, K. Staging of Alzheimer disease-associated neurofibrillary pathology using paraffin sections and immunocytochemistry. Acta Neuropathol 2006, 112 (4), 389-404. DOI: 10.1007 / s00401-006- 0127-z from NLM Medline.70948-02

[0163] (11) Rawat, P.; Sehar, U.; Bisht, J.; Selman, A.; Culberson, J.; Reddy, P. H.Phosphorylated Tau in Alzheimer's Disease and Other Tauopathies. Int J Mol Sci 2022, 23 (21). DOI: 10.3390 / ijms232112841 from NLM Medline.

[0164] (12) Mietelska-Porowska, A.; Wasik, U.; Goras, M.; Filipek, A.; Niewiadomska, G. Tau protein modifications and interactions: their role in function and dysfunction. Int J Mol Sci 2014, 15 (3), 4671-4713. DOI: 10.3390 / ijms15034671 from NLM Medline.

[0165] (13) Diociaiuti, M.; Bonanni, R.; Cariati, I.; Frank, C.; D'Arcangelo, G. Amyloid Prefibrillar Oligomers: The Surprising Commonalities in Their Structure and Activity. Int J Mol Sci 2021, 22 (12). DOI: 10.3390 / ijms22126435 from NLM Medline.

[0166] (14) Mandelkow, E. M.; Mandelkow, E. Biochemistry and cell biology of tau protein in neurofibrillary degeneration. Cold Spring Harb Perspect Med 2012, 2 (7), a006247. DOI: 10.1101 / cshperspect.a006247 from NLM Medline.

[0167] (15) Serrano-Pozo, A.; Frosch, M. P.; Masliah, E.; Hyman, B. T. Neuropathological alterations in Alzheimer disease. Cold Spring Harb Perspect Med 2011, 1 (1), a006189. DOI:10.1101 / cshperspect.a006189 from NLM Medline.

[0168] (16) Ohm, D. T.; Fought, A. J.; Martersteck, A.; Coventry, C.; Sridhar, J.; Gefen, T.; Weintraub, S.; Bigio, E.; Mesulam, M. M.; Rogalski, E.; et al. Accumulation of neurofibrillary tangles and activated microglia is associated with lower neuron densities in the aphasic variant of Alzheimer's disease. Brain Pathol 2021, 31 (1), 189-204. DOI: 10.1111 / bpa.12902 from NLM Medline.

[0169] (17) Wakabayashi, K.; Tanji, K.; Mori, F.; Takahashi, H. The Lewy body in Parkinson's disease: molecules implicated in the formation and degradation of alpha-synuclein aggregates. Neuropathology’ 2007, 27 (5), 494-506. DOI: 10.1111 / j.1440-1789.2007.00803.x from NLM Medline.

[0170] (18) Manzanza, N. O.; Sedlackova, L.; Kalaria, R. N. Alpha-Synuclein Post- translational Modifications: Implications for Pathogenesis of Lewy Body Disorders. Front Aging Neurosci 2021, 13, 690293. DOI: 10.3389 / fnagi.2021.690293 from NLM PubMed-not- MEDLINE.

[0171] (19) Zamel, J.; Chen, J.; Zaer, S.; Harris, P. D.; Drori, P.; Lebendiker, M.; Kalisman, N.: Dokholyan, N. V.; Lerner, E. Structural and dynamic insights into alpha-synuclein70948-02dimer conformations. Structure 2023, 31 (4), 411-423 e416. DOI: 10.1016 / j.str.2023.01.011 from NLM Medline.

[0172] (20) Saramowicz, K.; Siwecka, N.; Galita, G.; Kucharska-Lusina, A.; Rozpedek- Kaminska, W.; Majsterek, I. Alpha-Synuclein Contribution to Neuronal and Glial Damage in Parkinson's Disease. Int J Mol Sci 2023, 25 (1). DOI: 10.3390 / ijms25010360 from NLM Medline.

[0173] (21) Ramirez, E.; Ganegamage, S. K.; Elbatrawy, A. A.; Alnakhala, H.; Shimanaka, K.; Tripathi, A.; Min, S.; Rochet, J. C.; Dettmer, U.; Fortin, J. S. 5-Nitro-l,2-benzothiazol-3- amine and N-Ethyl-1-[(ethylcarbamoyl)(5-nitro-1,2-benzothiazol-3-yl)amino]formamide Modulate alpha-Synuclein and Tau Aggregation. ACS Omega 2023, 8 (22), 20102-20115. DOI: 10.1021 / acsomega.3c02668 from NLM PubMed-not-MEDLINE.

[0174] (22) Ramirez, E.; Ganegamage, S. K.; Min, S.; Patel, H.; Ogunware, A.; Plascencia-Villa, G.; Alnakhala, H.; Shimanaka, K.; Tripathi, A.; Wang, K. W.; et al. Evaluation of N- and O-Linked Indole Triazines for a Dual Effect on alpha-Synuclein and Tau Aggregation. ACS Chem Neurosci 2023, 14 (21), 3913-3927. DOI: 10.1021 / acschemneuro.3c00464 from NLM Medline.

[0175] (23) Ramirez, E.; Min, S.; Ganegamage, S. K.; Shimanaka, K.; Sosa, M. G.; Dettmer, U.; Rochet, J. C; Fortin, J. S. Discovery of 4-aminoindole carboxamide derivatives to curtail alpha-synuclein and tau isoform 2N4R oligomer formation. Results Chem 2023, 5. DOI: 10.1016 / j.rechem.2023.100938 from NLM PubMed-not-MEDLINE.

[0176] (24) Elbatrawy, A. A.; Ademoye, T. A.; Alnakhala, H.; Tripathi, A.; Zami, A.; Ostafe, R.; Dettmer, U.; Fortin, J. S. Discovery of small molecule benzothiazole and indole derivatives tackling tau 2N4R and alpha-synuclein fibrils. Bioorg Med Chem 2024, 100, 117613. DOI: 10.1016 / j.bmc.2024.117613 from NLM Medline.

[0177] (25) Dheer, D.; Singh, V.; Shankar, R. Medicinal attributes of 1,2,3-triazoles: Current developments. Bioorg Chem 2017, 71, 30-54. DOI: 10.1016 / j.bioorg.2017.01.010 from NLM Medline.

[0178] (26) Qu, W.; Kung, M. P.; Hou, C.; Oya, S.; Kung, H. F. Quick assembly of 1,4- diphenyltriazoles as probes targeting beta-amyloid aggregates in Alzheimer's disease. J Med Chem 2007, 50 (14), 3380-3387. DOI: 10.1021 / jm0704671 from NLM Medline.

[0179] (27) Das, S.; Smid, S. D. Identification of dibenzyl imidazolidine and triazole acetamide derivatives through virtual screening targeting amyloid beta aggregation and70948-02neurotoxicity in PCT2 cells. Eur J Med Chem 2017, 130, 354-364. DOI: 10.1016 / j.ejmech.2017.02.057 from NLM Medline.

[0180] (28) Bieschke, J.; Russ, J.; Friedrich, R P.; Ehrnhoefer, D. E.; Wobst, EL; Neugebauer, K; Wanker, E. E. EGCG remodels mature alpha-synuclein and amyloid-beta fibrils and reduces cellular toxicity. Proc Natl Acad Sci U S A 2010, 107 (17), 7710-7715. DOI: 10.1073 / pnas.0910723107 From NLM Medline.(29) Sonawane, S. K.; Chidambaram, H.; Boral, D; Gorantla, N. V; Balmik, A. A.; Dangi, A.; Ramasamy, S.; Marelli, U. K.; Chinnathambi, S. EGCG impedes human Tau aggregation and interacts with Tau. Sci Rep 2020, 10 (1), 12579. DOI: 10.1038 / s41598-020-69429-6 from NLM Medline.

[0181] (30) Yang, Z.; Yao, Y.; Zhou, Y.; Li, X.; Tang, Y.; Wei, G. EGCG attenuates alpha- synuclein protofibril-membrane interactions and disrupts the protofibril. Int J Biol Macromol 2023, 230, 123194. DOI: 10.1016 / j. ijbiomac.2023.123194 from NLM Medline.

[0182] (31) Goncalves, P, B.; Sodero, A. C. R.; Cordeiro, Y. Green Tea EpigaHocatechin- 3-gallate (EGCG) Targeting Protein Misfolding in Drug Discovery for Neurodegenerative Diseases. Biomolecules 2021, 11 (5). DOI: 10,3390 / bioml 1050767 from NLM Medline.

[0183] (32) Martins, G. F.; Nascimento, C.; Galamba, N, Mechanistic Insights into Polyphenols' Aggregation Inhibition of alpha-Synuclein and Related Peptides. ACS Chem Neurosci 2023, 74 (10), 1905-1920. DOI: 10.1021 / acschemneuro.3c00162 from NLM Medline.

[0184] (33) Biancalana, M.; Makabe, K.; Koide, A.; Koide, S. Molecular mechanism of thioflavin- T binding to the surface of beta-rich peptide self-assemblies. J Mol Biol 2009, 385 (4), 1052-1063. DOI: 10.1016 / j.jmb.2008.11.006 from NLM Medline.

[0185] (34) Rinauro, D. J.; Chiti, F.; Vendruscolo, M.; Limbocker, R. Misfolded protein oligomers: mechanisms of formation, cytotoxic effects, and pharmacological approaches against protein misfolding diseases. Mol Neurodegener 2024, 19 (1), 20. DOI: 10.1186 / sl3024-023- 00651-2 from NLM Medline.

[0186] (35) Wells, C.; Brennan, S.; Keon, M.; Ooi, L. The role of amyloid oligomers m neurodegenerative pathologies. Int J Biol Macromol 2021, 181, 582-604. DOI: 10.1016 / j. ijbiomac.2021.03.113 from NLM Medline.

[0187] (36) Yao, Y.; Tang, Y.; Wei, G. Epigallocatechin Gallate Destabilizes alpha- Synuclein Fibril by Disrupting the E46-K80 Salt-Bridge and Inter-protofibril Interface. ACS70948-02Chem Neurosci 2020, 11 (24), 4351-4361. DOI: 10.1021 / acschemneuro.0c00598 from NLM Medline.

[0188] (37) Terry-Kantor, E.; Tripathi, A.; Imberdis, T.; LaVoie, Z. M.; Ho, G. P. H.; Selkoe, D.; Fanning, S.; Ramalingam, N.; Dettmer, U. Rapid Alpha-Synuclein Toxicity in a Neural Cell Model and Its Rescue by a Stearoyl-CoA Desaturase Inhibitor. Int J Mol Sci 2020, 21 (15). DOI: 10.3390 / ijms21155193 from NLM Medline.

[0189] (38) Imberdis, T.; Negri, J.; Ramalingam, N.; Terry-Kantor, E.; Ho, G. P. H.; Fanning, S.; Stirtz, G; Kim, T. E.; Lexy, O A.; Young-Pearse, T. L.; et al. Cell models of lipid- rich alpha-synuclein aggregation validate known modifiers of alpha-synuclein biology and identify stearoyl-CoA desaturase. Proc Natl Acad Sci USA 2019, 116 (41), 20760-20769. DOI: 10.1073 / pnas.1903216116 from NLM Medline.

[0190] (39) Fortin, J, S.; Shimanaka, K.; Saraswati, A, P.; Liu, M.; Wang, K. W.; Hagar, H. T.; Maity, S.; Ganegamage, S. K; Ellsworth, E.; Counts, S. E.; et al, Anti-fibrillization effects of sulfonamide derivatives on alpha-synuclein and hyperphosphorylated tau isoform 1N4R. J Mol Struci 2022, 1267. DOI: 10.1016 / j.molstruc.2022.133574 from NLM PubMed-not-MEDLINE.

[0191] (40) Sui, D.; Xu, X.; Ye, X.; Liu, M.; Mianecki, Mi; Rattanasinchai, C.; Buehl, C. Deng, X.; Kuo, M H. Protein interaction module- assisted function X (PIMAX) approach to producing challenging proteins including hyperphosphorylated tau and active CDK5 / p25 kinase complex. Mol Cell Proteomics 2015, 14 (1), 251-262. DOI: 10.1074 / mcp. Ol 14.044412 from NLM Medline.

[0192] (41) Liu, M.; Dexheimer, T.; Sui, D.; Hovde, S.; Deng, X.; Kwok, R.; Bochar, D. A.; Kuo, M. H. Hyperphosphorylated tau aggregation and cytotoxicity modulators screen identified prescription drugs linked to Alzheimer's disease and cognitive functions. Sci Rep 2020, 10 (1), 16551. DOI: 10.1038 / s41598-020-73680-2 from NLM Medline.

[0193] All patents, patent application publications, journal articles, textbooks, and other publications mentioned in the specification are indicative of the level of skill of those in the art to which the disclosure pertains. All such publications are incorporated herein by reference to the same extent as if each individual publication were specifically and individually indicated to be incorporated by reference.

[0194] The invention illustratively described herein may be suitably practiced in the absence of any element(s) or limitation(s), which is / are not specifically disclosed herein. Thus,70948-02for example, each instance herein of any of the terms "comprising," "consisting essentially of," and "consisting of may be replaced with either of the other two terms. Likewise, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, references to "the method" includes one or more methods and / or steps of the type, which are described herein and / or which will become apparent to those ordinarily skilled in the art upon reading the disclosure.

[0195] The term “about,” when referring to a number or a numerical value or range (including, for example, whole numbers, fractions, and percentages), means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error) and thus the numerical value or range can vary between 1% and 15% of the stated number or numerical range (e.g., + / - 5 % to 15% of the recited value) provided that one of ordinary skill in the art would consider equivalent to the recited value (e.g., having the same function or result). The term “substantially” can allow for a degree of variability in a value or range, for example, within 90%, within 95%, or within 99% of a stated value or of a stated limit of a range,

[0196] The terms and expressions, which have been employed, are used as terms of description and not of limitation. In this regard, where certain terms are defined under " Definitions" and are otherwise defined, described, or discussed elsewhere in the " Detailed Description," all such definitions, descriptions, and discussions are intended to be attributed to such terms. There also is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof. Furthermore, while subheadings, e.g., " Definitions," are used in the " Detailed Description," such use is solely for ease of reference and is not intended to limit any disclosure made in one section to that section only; rather, any disclosure made under one subheading is intended to constitute a disclosure under each and every other subheading.

[0197] It is recognized that various modifications are possible within the scope of the claimed invention. Thus, although the present invention has been specifically disclosed in the context of preferred embodiments and optional features, those skilled in the art may resort to modifications and variations of the concepts disclosed herein. Such modifications and variations are considered within the scope of the invention as claimed herein.

Claims

70948-02WHAT IS CLAIMED IS:

1. A compound of formula (I):wherein:each R4is independently alkyl, optionally substituted aryl or optionally substituted heteroaryl; each R5is independently H, halo, alkyl, alkoxy, aryl, aryloxy, nitro, cyano, carboxy or acyl; X1is CR5or N;Z is NR6, O or S, wherein R6is H, alkyl or alkenyl; andn is 1, 2 or 3;or a pharmaceutically acceptable salt or hydrate thereof.

2. The compound of claim 1, wherein the compound of formula (1) is a compound of formula (la):wherein:each R7is independently H, hydroxy, halo, alkyl, alkoxy, aryl, aryloxy, nitro, cyano, carboxy or acyl;each R5is independently H, alkyl, alkoxy, halo, aryl, aryloxy, amino, nitro or cyano;Z is NR6, O or S, wherein R6is H, alkyl or alkenyl;X1is CR5or N;n is 1, 2 or 3; andm is 1, 2, or 3;or a pharmaceutically acceptable salt or hydrate thereof.70948-023. The compound of claim 1, wherein the compound of formula (I) is a compound of formula (lb):wherein:each R7is independently H, hydroxy, halo, alkyl, alkoxy, aryl, aryloxy, nitro, cyano, carboxy or acyl;each R5is independently H, alkyl, alkoxy, halo, aryl, aryloxy, amino, nitro or cyano;X1is CR5or N;Z is NR6, 0 or S, wherein R6is H, alkyl or alkenyl;n is 1, 2 or 3; andm is 1, 2, or 3;or a pharmaceutically acceptable salt or hydrate thereof.

4. The compound of claim 1, wherein the compound of formula (1) is a compound of formula (lb):wherein:each R7is independently H, hydroxy, halo, alkyl, alkoxy, aryl, aryloxy, nitro, cyano, carboxy or acyl;each R5is independently H, alkyl, alkoxy, halo, aryl, aryloxy, amino, nitro or cyano;X1is CR5or N;Z is NR6, O or S, wherein R6is H, alkyl or alkenyl;n is 1, 2 or 3; andm is 1, 2, or 3;or a pharmaceutically acceptable salt or hydrate thereof.70948-025. The compound of any of claims 1-4, wherein X1is CR5.

6. The compound of claim 5, wherein Z is NR6.

7. The compound of claim 6, wherein R6is H8. The compound of any of claims 1-4 or claim 5, wherein Z is S.

9. The compound of any of claims 1 -4 or claim 5, wherein Z is O,10. The compound of any of claims 1 -4, wherein X1is N.

11. The compound of claim 10, wherein Z is NR6.

12. The compound of claim 11, wherein R6is H13. The compound of claim 10, wherein Z is S.

14. The compound of claim 10, wherein Z is 0.

15. The compound of claim 5, wherein each R7is independently alkyl.

16. The compound of claim 13, wherein each R7is independently -O-C1-C6alkyl, N(C1-C3alkyl)2, halo, -NO2, and -OH, or a pharmaceutically acceptable salt or hydrate thereof.

17. The compound of claim 16, wherein -O-Ci-C’6 alkyl is -O-methyl.

18. The compound of claim 16, wherein C1-C3 alkyl is methyl.

19. The compound of claim 16, wherein halo is Cl, F, or Br.70948-02or a pharmaceutically acceptable salt or hydrate thereof, wherein n and R4are defined in claim 1.

21. The compound of claim 1, wherein the compound is a compound of formula:70948-02or a pharmaceutically acceptable salt or hydrate thereof.

22. The compound of claim 20 or 21, wherein the compound is (5-(l-(4-Dimethylaminophenyl)- l / f-l,2,3-triazol-4-yl)-l / f-indole) or a pharmaceutically acceptable salt or hydrate thereof and has the formula:N-Nor a pharmaceutically acceptable salt or hydrate thereof.

23. The compound of claim 20 or 21, wherein the compound is (4-[4-(l / / -indol-5-yl)-lH-l,2,3- triazol-l-yl]benzene-l,2-diol) or a pharmaceutically acceptable salt or hydrate thereof and has the formula:or a pharmaceutically acceptable salt or hydrate thereof.70948-0224. The compound of claim 20 or 21, wherein the compound is (5-[4-(l / / -indol-5-yl)-l / / -l,2,3-triazol-l-yl]benzene-l,2,3-tnol) or a pharmaceutically acceptable salt or hydrate thereof and has the formula:N-NOHor a pharmaceutically acceptable salt or hydrate thereof.

25. A pharmaceutical composition comprising a compound of any of claims 1-4, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier.

26. A method of inhibiting aipha-synuclein (a-syn) protein fibril formation in a subject having, or at risk for, a-syn protein fibril formation, which method comprises administering to the subject an effective amount of (i) a compound of any of claims 1-4 or 24, or a pharmaceutically acceptable salt or hydrate thereof, or (ii) a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable earner.

27. The method of claim 26, wherein the subject has, or is at risk for, Parkinson’s disease or dementia with Lewy bodies (DLB).

28. The method of claim 26, wherein the compound is the compound of claim 24, or a pharmaceutically acceptable salt or hydrate thereof, or a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier, and the compound also inhibits a-syn oligomers.

29. A method of inhibiting tubulin-associated unit (tau) protein (isoforms 0N4R, 1N4R, 2N4R, 0N3R, 1N3R, 2N3R) oligomer formation in a subject having, or at risk for, tau (isoforms 0N4R, 1N4R, 2N4R, 0N3R, 1N3R, 2N3R) oligomer formation, which method comprises administering to the subject an effective amount of (i) a compound of any one of claims 1-4 or 24, or a pharmaceutically acceptable salt or hydrate thereof, or (ii ) a pharmaceutical composition70948-02comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier.

30. The method of claim 29, wherein the compound is the compound of claim 24, or a pharmaceutically acceptable salt or hydrate thereof, or a pharmaceutical composition comprising the compound, or a pharmaceutically acceptable salt or hydrate thereof, and a pharmaceutically acceptable carrier.

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