Benzothiazole and indole derivative compounds, compositions, and uses thereof for dual inhibition of TAU 2n4r

Abstract

Compounds of formula (I), pharmaceutical compositions of formula (I), and methods of use of compounds of formula (I) in the inhibition of α-syn protein aggregation, tau protein aggregation, oligomer formation, and / or fibril formation in a subject having, or at risk for, said aggregation / formation.

Description

[0001] BENZOTHIAZOLE AND INDOLE DERIVATIVE COMPOUNDS, COMPOSITIONS,

[0002] AND USES THEREOF FOR DUAL INHIBITION OF TAU 2N4R

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS

[0004] This application claims the benefit of U.S. Provisional Application No. 63 / 626,045, filed January 28, 2024, which is incorporated by reference as if fully set forth herein.

[0005] STATEMENT OF GOVERNMENT SUPPORT

[0006] This invention was made with government support under contracts AG070447 and AG071985 awarded by the National Institutes of Health. The government has certain rights in the invention.

[0007] TECHNICAL FIELD

[0008] The present disclosure relates to benzothioazole and indole derivative compounds having structural features characteristic of N744 and Anlel38b compounds, compositions comprising same, and methods of use in the dual inhibition of tau 2N4R and a-synuclein fibril aggregation. N744 is a cyanine dye also known as 3,3‘-bis(P-hydroxyethyl)-9-ethyl-5,5‘- dimethoxythiacarbocyanine iodide and having the structure:

[0009] Anlel38b is also known as 3-(l,3-benzodioxol-5-yl)-5-(3-bromophenyl)-lH-pyrazole and

[0010] Emrusolmin and has the structure:

[0011] BACKGROUND

[0012] The most prevalent neurodegenerative disorders impacting millions of patients and their caregivers annually are Alzheimer’s disease (AD) and Parkinson’s disease (PD). Classic AD symptoms include dementia, memory decline or loss, coherent speech difficulty, and general cognitive impairment. While in PD, the symptoms are mainly motorial, including bradykinesia, tremor, and postural instability; however, nonmotor signs, such as depression and sleep disturbance, have also been experienced in some patients. The amyloid cascade hypothesis suggested that progressive deposition of amyloid- / ? (A / ?) protein is the pivotal hallmark in AD, and all the attendant pathological events, such as neurofibrillary tangles (NFTs), formation of hyperphosphorylated tau protein, neuritic and glial cytopathology, and eventually dementia, directly result from this deposition. However, recent findings emphasized that microtubule-associated protein tau (MAPT), hyperphosphorylated tau, and NFTs are more strongly linked with the degree of cognitive decline than with A / ? aggregates. Moreover, compounds targeting aggregates of Aβ showed no meaningful results in clinical trials. In addition to AD, tau is significantly featured in frontotemporal dementia with Parkinsonism (FTDP-17) as well as in Pick’s disease, Down’s syndrome, and progressive supranuclear palsy (PSP), collectively termed tauopathies. Accordingly, tau has gained a lot of attention as a therapeutic target for AD and other tauopathies in the past few decades. Tau is present in six isoforms in the human brain and are categorized by the absence or the presence of a 29-amino- acid, N-terminal domain (ON, IN, and 2N) and the number of microtubule-binding C-terminal repeats (3R and 4R Importantly, the 4R isoforms have faster aggregation rates and consequently a higher propensity to produce neurodegeneration than the 3R variants. The benzothiazole cyanine dye N744 was identified as a strong tau aggregation inhibitor. N744 was found not only to inhibit the full-length (2N4R) tau aggregates (IC5o = 300 nM) but also to induce remarkably the disaggregation of preformed filaments.

[0013] In contrast, aberrant accumulation of intracellular proteinaceous aggregates of a- synuclein (a-syn) is the main histopathological characteristic of PD and dementia with Lewy bodies (DLB). a-Syn is a presynaptic, acidic protein that plays potential roles in the management of synaptic vesicle release and neuronal survival. Under pathological conditions, misfolding of a-syn forms Lewy bodies (LBs) in the neurons. Braak and other colleagues have identified the correlation between LBs and disease progression. Recently, the diphenyl pyrazole Anlel38b has been synthesized as a disease-modifying therapy for PD by inhibiting the formation of a-syn pathological aggregates (77% inhibition) in vitro as well as in in vivo settings. Fluorescence measurements of Anlel38b showed that significant fluorescence change occurred upon incubation with a-syn fibrils but not with monomers.

[0014] As a corollary, there is a noticeable interplay between tau and LBs of a-syn as both aggregates coprecipitate in some brains of AD, PD, and DLB patients. Moreover, tau and a- syn synergistically facilitate the formation of each other’s aggregates in mouse models. Accordingly, there is an urgent need to design and prepare small molecule agents targeting both protein aggregates to manage the progression of neurodegeneration. In view of the foregoing, it is an object of the present disclosure to provide such small molecule agents. This and other objects and advantages, as well as inventive features, will be apparent from the detailed description provided herein.

[0015] SUMMARY

[0016] Provided is a compound of formula I: wherein Ar is 4-amino-indole, 5-amino-indole, 6-amino-indole, or 7-amino-indole, Rlais hydrogen, C1-C6 alkyl (e.g., methyl), hydroxyl, C1-C6 alkoxy (e.g., methoxy), di-hydroxy, di- C1-C6 alkyl-hydroxy (e.g., di-methoxy), tri-hydroxy, tri- C1-C6 alkyl-hydroxy (e.g., trimethoxy), a halo group, or a nitro group, R3is hydrogen, hydroxyl, C1-C6 alkoxy (e.g., methoxy), tri-hydroxy, or tri- C1-C6 alkyl-hydroxy (e.g., tri-methoxy), a halo group, or a nitro group, and R2ais hydrogen, hydroxyl, C1-C6 alkoxy (e.g., methoxy), di-hydroxy, di- C1-C6 alkyl-hydroxy (e.g., di-methoxy), tri-hydroxy, tri- C1-C6 alkyl-hydroxy (e.g., tri-methoxy), a halo group, or a nitro group, or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester). Ar can be optionally substituted with a C1-C6 alkyl or a halo group. The halo group can be F, Cl, Br, or I. In some embodiments, the halo group can be I. In some embodiments, Ar is 4-amino-indole, Rlais hydrogen and R3ais a nitro group. In other embodiments, Ar is 6-amino-indole, Rlais hydrogen, and R3ais a nitro group. Such a compound can inhibit oligomer formation, fibril formation, or both.

[0017] Also provided is a pharmaceutical composition comprising at least one compound of the disclosure or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester) and a pharmaceutically acceptable carrier.

[0018] In some embodiments, the pharmaceutical composition comprises the compound in which Ar is 4-amino-indole, Rlais hydrogen, and R3ais a nitro group. In other embodiments, the pharmaceutical composition comprises the compound in which Ar is 6-amino-indole, Rlais hydrogen, and R3ais a nitro group.

[0019] Also provided is a compound of formula II: wherein RAis 4-cyano or 4-nitro, R4ais C1-C6 alkyl-hydroxy (e.g., methyoxy), hydroxyl, or C(O)O-alkyl, and R5ais hydrogen, nitro, amino, C1-C6 alkyl amino, or C1-C6 di-alkyl amino, or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester). In some embodiments, RAis 4-cyano, R4ais methoxy, and R5ais hydrogen. In some embodiments, RAis 4-nitro, R4ais methoxy, and R5ais hydrogen. Such compounds have anti-fibrillar activity.

[0020] In view of the above, also provided is a method of inhibiting a-synuclein (a-syn) protein aggregation, tau isoforms 0N4R, 1N4R, 2N4R, 0N3R, 1N3R, and / or 2N3R with and without post-translational changes (such as phosphorylation) protein aggregation, or both in a subject having, or at risk for, said protein aggregation. The method comprises administering to the subject a pharmaceutical composition, e.g., comprising a compound of Formula I, in an amount effective to inhibit a-syn protein aggregation, tau isoform 2N4R protein aggregation, or both, whereupon said protein aggregation is inhibited in the subject having, or at risk for, said protein aggregation. The subject can have, or be at risk for, Alzheimer’s disease. The subject can have, or be at risk for, Parkinson’s disease or dementia with Lewy bodies (DLB). The subject can have neuroblastoma, in which case the method inhibits the formation of a-syn inclusions, such as when a pharmaceutical composition comprising a compound in which Ar is 6-amino-indole, Rlais hydrogen, and R2ais a nitro group.

[0021] Also in view of the above, provided is a method of inhibiting fibril formation in a subject having, or at risk for, fibril formation. The method comprises administering to the subject an above-described pharmaceutical composition (comprising a compound of Formula II) in an amount effective to inhibit fibril formation, whereupon fibril formation is inhibited in the subject having, or at risk for, said fibril formation.

[0022] FIGURES

[0023] Fig. 1 Design of methoxy benzothiazole and indole derivatives as dual 2N4R tau and a-syn aggregation inhibitors. Fig. 2 Synthesis of the amide or the sulfonamide counterparts of N744 and Anlel38b using the corresponding benzoyl chloride or benzene sulfonyl chloride, respectively, in the presence of anhydrous potassium carbonate and pyridine. Reagents and conditions: (a) DCM, r.t., 3-12 h, 74-93%; (b) anhydrous K2CO3, pyridine, r.t., 8-10 h, 61-90%.

[0024] Fig- 3 Further modification of the 2-iodo and the 4-nitro sulfonamides and their urea analogs by replacing the 4-MBT with 4-, 5-, or 6-amino indole (Al) using the same reaction conditions. Reagents and conditions: (a) DCM, r.t., 3-12 h, 79-100%; (b) anhydrous K2CO3, pyridine, r.t., 3-12 h, 73-86%.

[0025] Fig- 4 Thioflavin-T (ThT) kinetic curves of compounds 46, 47, and 48 (100 pM) when tested with: (A) a-syn (2 pM); and (B) dose-dependent inhibition of varying concentrations (3.125, 6.25, 12.5, 25, 50, and 100 pM) of compound 46 on a-syn (2 pM) fibril formation using ThT fluorescence assay. For each concentration triplicate data were gathered at the plateau phase from five consecutive time points. The error bars represent the individual standard error of mean (SEM) for each condition.

[0026] Fig. 5 ThT kinetic curves of compounds 46, 47, 48, and EGCG (100 pM) when tested with: A. u-syn (2 pM) in 10 mM PBS buffer (pH 7.4), supplemented with 0.5 mM SDS and 300 mM NaCl with 20 pM of ThT (average of triplicate using 96 well plate format); B. tau 2N4R (6 pM) with 1.5 pM heparin, 5 mM DTT, and 40 pM ThS in 10 mM PBS (average of duplicate using 384 well plate format). Compounds reduced both u-syn and tau 2N4R fibril formation at a similar degree than EGCG.

[0027] Fig- 6 Inhibition of the a-syn oligomer formation induced by the photo-induced cross-linking of unmodified protein (PICUP) assay. In the PICUP experiment, a-syn (60 pM) was crosslinked with varying concentrations of compounds 46, 47, and 48 (50, 100, and 200 pM). The optical density of the high molecular-weight bands represents a-syn oligomer, while the lower bands represent the protein monomers. In particular, compound 46 remarkably inhibited the development of a-syn oligomer-corresponding high molecular- weight bands between 35 and 45 kDa. The control consisted of no light exposure and no Tris(2,2'-bipyridyl)ruthenium(II) chloride (Ru(bpy)3, a cross-linking agent). Fig. 7A Inhibition of oligomer formation induced by the PICUP assay on the tau isoform 2N4R. The protein (10 pM) was exposed to different concentrations of compounds 46, 47, and 48 (50 pM, 100 pM, and 200 pM). None of the compounds demonstrated the ability to suppress the formation of tau oligomers. The control consisted of no light exposure and no Tris(2,2'- bipyridyl)ruthenium(II) chloride (Ru(bpy)3, a cross-linking agent).

[0028] Fig. 7B Inhibition of oligomer formation induced by the PICUP assay on the tau isoform 0N4R. The protein (10 pM) was exposed to different concentrations of compounds 46, 47, and 48 (50 pM, 100 pM, and 200 pM). None of the compounds demonstrated the ability to suppress the formation of tau oligomers. The control consisted of no light exposure and no Tris(2,2'- bipyridyl)ruthenium(II) chloride (Ru(bpy)3, a cross-linking agent).

[0029] Fig. 8 Transmission electron microscopy (TEM) analysis of compounds 46, 47, and 48 on the inhibition of mature fibrils of a-syn (2 pM, upper panel) and 2N4R tau (6 pM, lower panel). The unfolded protein was incubated with: (A)-(B) DMSO (0.25%); (C)-(D) compound 46 (100 pM); (E)-(F) compound 47 (100 pM); or (G)-(H) compound 48 (100 pM). Incubation time consisted of ~68 hours and 72 hours at 37° C for a-syn and 2N4R tau, respectively. Scale bars: 200 nm, magnification: 40K.

[0030] Figs. 9A-D M17D cells expressing the inclusion-prone a-Syn-3K::YFP fusion protein (dox- inducible) treated with 0.1% DMSO (vehicle; “0 pM”) as well as 1.25, 2.5, 5, 10, 20 and 40 pM of compounds 46 (Fig. 9A), 47 (Fig. 9B) and 48 (Fig. 9C) at t = 24 hours after plating. Cells were induced with doxycycline at t = 48 hours. Incucyte-based analysis of punctate YFP signals relative to 0.1% DMSO was done at t = 96 hours (N = 2 independent experiments, n = 6-12 individual wells total (0 pM, n = 12; 40, 20 and 10 pM, n = 6; all other concentrations, n = 12). Plot of confluence fold changes relative to DMSO vehicle (0 pM). (Fig. 9D) Representative IncuCyte images of reporter cells treated with vehicle vs 5 pM compound 46, 47, and 48 (t = 96 hours), green channel. Arrows indicate aS-rich YFP-positive inclusions. Scale bar, 50 pm. All data are presented as fold-changes relative to DMSO control + 1- standard deviation. One-way ANOVA, Dunnett’s post-hoc test; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001. Fig. 10 Inhibition of the a-syn oligomer formation induced by the PICUP assay. In the PICUP experiment, a-syn (60 pM) was cross-linked with 100 pM of compounds 32 - 35, 42, and 67. The optical density of the high molecular-weight bands represent a-syn oligomer while the lower bands represent the protein monomers. The control comprises no light and no Tris(2,2'- bipyridyl)ruthenium(II) chloride (Ru(bpy)3, a cross-linking agent).

[0031] Figs. 11A-E Compounds 33, 35, 42 and 67 prevent a-synuclein inclusion formation. 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 67 (Fig. HA), 35 (Fig. 11B), 42 (Fig. 11C) and 33 (Fig. 11D) at t = 24 h after plating. Cells were induced with doxycycline at t = 48 hours. Incucyte-based analysis of punctate YFP signals relative to 0.1% DMSO was done at t = 96 h (N = 2 independent experiments, n = 6-12 individual wells total (0 pM, n = 12; 40, 20 and 10 pM, n = 6; all other concentrations, n = 12). Plot of confluence fold changes relative to DMSO vehicle (0 pM). (Fig. HE) Representative IncuCyte images of reporter cells treated with vehicle vs 40 pM compound 67, 35, 42 and 33 (t = 96 h), green channel. Arrows indicate aS-rich YFP-positive inclusions. Scale bar, 50 pm. All data are presented as fold-changes relative to DMSO control + / - standard deviation. Oneway ANOVA, Dunnett’s post-hoc test; *, p < 0.05; **, p < 0.01; ***, p < 0.001.

[0032] DESCRIPTION

[0033] The present disclosure is based on the judicious design and synthesis of benzothiazole and indole derivatives of N744 and Anlel38b by simple single-step reactions to reduce the 2N4R tau and a-syn aggregates. In particular, the indole derivatives with urea linker, namely compounds 46 and 48, displayed the most promising results as a dual aggregation inhibitor for 2N4R tau and a-syn fibrils. Compounds 46 and 48 significantly decreased the Thioflavin-T (ThT) fluorescence to below 20% against both protein fibrils. In addition, compound 46 induced a noticeable inhibitory activity against a-syn oligomers, but not with the 2N4R tau oligomers. The use of transmission electron microscopy (TEM) revealed the powerful antiaggregation properties of compounds 46 and 48 on 2N4R tau and a-syn aggregates. In addition, compound 48 was highly effective in reducing a-syn inclusion using M17D cells.

[0034] In view of the above, provided is a compound of formula I: wherein Ar is 4-amino-indole, 5-amino-indole, 6-amino-indole, or 7-amino-indole, Rlais hydrogen, C1-C6 alkyl (e.g., methyl), hydroxyl, C1-C6 alkoxy (e.g., methoxy), di-hydroxy, di- C1-C6 alkyl-hydroxy (e.g., di-methoxy), tri-hydroxy, tri- C1-C6 alkyl-hydroxy (e.g., trimethoxy), a halo group, or a nitro group, R3ais hydrogen, hydroxyl, C1-C6 alkoxy (e.g., methoxy), tri-hydroxy, or tri- C1-C6 alkyl-hydroxy (e.g., tri-methoxy), a halo group, or a nitro group, and R2ais hydrogen, hydroxyl, C1-C6 alkoxy (e.g., methoxy), di-hydroxy, di- C1-C6 alkyl-hydroxy (e.g., di-methoxy), tri-hydroxy, tri- C1-C6 alkyl-hydroxy (e.g., tri-methoxy), a halo group, or a nitro group, or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester). Ar can be optionally substituted with a C1-C6 alkyl or a halo group. The halo group can be F, Cl, Br, or I. In some embodiments, the halo group can be I. In some embodiments, Ar is 4-amino-indole, Rlais hydrogen and R3ais a nitro group. In other embodiments, Ar is 6-amino-indole, Rlais hydrogen, and R3ais a nitro group. Such a compound can inhibit oligomer formation, fibril formation, or both.

[0035] Also, provided is a compound of formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester) wherein: the indole group is optionally substitued (e.g., amino substituted indole, such as NH2- substittuted indole); and

[0036] Rlb-R3bare each independently alkyl (e.g., methyl or CF3, such as mono-, di-, and tri- CF3), amino (e.g., methyl or dimethylamino), halo (e.g., fluoro or chloro, mono-, di-, and trifluoro and mono-, di-, and tri-chloro), alkoxy (e.g., methoxy, such as mono-, di-, and tri- methoxy) or OH (e.g., mono-, di-, and tri-OH); such as:

[0037] an mono- and di-alkylated versions of the amino

[0038] (NH2) group, such as demethylated versions of the amino group.

[0039] Also provided is a compound of formula II: wherein RAis 4-cyano or 4-nitro, R4ais C1-C6 alkyl-hydroxy (e.g., methyoxy), hydroxyl, or C(O)O-alkyl, and R5ais hydrogen, nitro, amino, C1-C6 alkyl amino, or C1-C6 di-alkyl amino, or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester). In some embodiments, RAis 4-cyano, R4ais methoxy, and R5ais hydrogen. In some embodiments, R is 4-nitro, R4ais methoxy, and R5ais hydrogen. Such compounds have anti-fibrillar activity.

[0040] Also provided are compounds of the formulae la, lb, and Ic: la lb Ic or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein Y1is: and Y2is wherein:

[0041] Z1is N or CH;

[0042] Z2is C, CH or N;

[0043] Z3is N or NRa;

[0044] Z1is O, S, or NRb; provided that Y1is not unsubstituted indolyl when Y2is: one of R'-R4is fluoro, chloro, NO2, C1-C2 alkyl or methoxy (OCH3);

[0045] Raand Rbare each independently H or optionally substituted C1-C20 alkyl; and

[0046] R'-R8and R1-R3are each independently -H, halogen, -OH, -NH2, -NO2, -CN, -CF3, -COOH,

[0047] -N3, -SO3H, -PO3H2, C1-C20 alkyl, C1-C20 hydroxyalkyl, C1-C20 alkylamino, C1-C20 dialkylamino, C1-20 alkoxy, C(0)0-( C1-C20) alkyl, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, or 3- to 15-membered heterocyclyl. In one example, each of R'-R8and R1-R3can each be independently substituted with one or more of halogen, -OH, C1-20 alkoxy, -NH2, -NO2, -CN, -CF3, -COOH, -N3, -SO3H, -PO3H2. In one example, Z1is N. Alternatively, or in addition, Z2is S. Alternatively, or in addition, Y1is: . Alternatively, or in addition, Y2is:

[0048] Compounds of the formula (la) include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0049] Z1is N or CH;

[0050] Z3is O, S, or NRa; provided that Z1is not CH when Z3is NRa;

[0051] Rais H or optionally substituted C1-20 alkyl;

[0052] R'-R3and R5-R8are each independently -H, halogen, -CN, -CF3, -NH2, -NO2, optionally substituted C1-20 alkyl or optionally substituted C1-20 alkoxy.

[0053] Compounds of the formula (la) also include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0054] Z1is N or CH;

[0055] Z3is O, S, or NRa; provided that Z1is not CH when Z3is NRa;

[0056] Rais H or optionally substituted alkyl;

[0057] Zris O, S, or NRb;

[0058] Raand Rbare each independently H or optionally substituted C1-20 alkyl; and R1-R3are each independently H, halogen, -OH, -NIL, -NO2, -CN, -COOH, -Ns,-SO3H, - PO3H2 C1-C20 alkyl, alkoxy, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, C3-C12 cycloalkenyl, or 3- to 15-membered heterocyclyl. In one example, each of R5-R8and R1-R3can each be independently substituted with one or more of halogen, -OH, C1-20 alkoxy, -NH2, -NO2, -CN, -CF3, -COOH, -N3, -SO3H, -PO3H2.

[0059] Compounds of the formula (lb) include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0060] Z1is N or CH;

[0061] Z3is O, S, or NRa;

[0062] Rais H or optionally substituted C1-20 alkyl; and

[0063] RkR8are each independently H, halogen, -CF3,-CN, -NO2, or alkoxy.

[0064] Compounds of the formula (lb) also include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0065] Z1is N or CH;

[0066] Z3is O, S, or NRa;

[0067] Rais H or optionally substituted alkyl;

[0068] Zris O, S, or NRb;

[0069] Raand Rbare each independently H or optionally substituted C1-20 alkyl; and

[0070] R1-R3are each independently H, halogen, -OH, -NH2, -NO2, -CF3, -CN, -COOH, -Ns, - SO3H, -PO3H2, C1-C20 alkyl, alkoxy, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, C3- C12 cycloalkenyl, or 3- to 15-membered heterocyclyl. In one example, each of R5-R8and R1- R3can each be independently substituted with one or more of halogen, -OH, C1-20 alkoxy, - NH2, -NO2, -CN, -CF3, -COOH, -N3, -SO3H, -PO3H2.

[0071] Compounds of the formula (lb) also include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0072] Y1is 4-amino-indole, 5-amino-indole, 6-amino-indole, or 7-amino-indole;

[0073] R1is -H, halogen, -OH, C1-C6 alkyl, C1-C6 alkyl-OH, C1-C6 alkoxy, or -NO2;

[0074] R2is -H, halogen, -OH, C1-C6 alkyl-OH, C1-C6 alkoxy, or -NO2; and

[0075] R3 is -H, halogen, -OH, C1-C6 alkyl-OH, C1-C6 alkoxy, or -NO2. In one example, Y1is optionally substituted with a C1-C6 alkyl or a halogen. In one example, the halogen can be I. In various other examples:

[0076] Y1is 4-amino-indole;

[0077] R1is -H, and

[0078] R3 is -NO2; or

[0079] Y1is 6-amino-indole,

[0080] R1is -H, and

[0081] R3 is -NO2.

[0082] Compounds of the formula (Ic) include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0083] Z1is N or CH

[0084] Z3is O, S, or NRa

[0085] Rais H or optionally substituted C1-20 alkyl; and

[0086] R'-R? are each independently H, halogen, -CF3, -CN, -NO2, or alkoxy.

[0087] Compounds of the formula (Ic)also include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0088] Z1is N or CH;

[0089] Z3is O, S, or NRa;

[0090] Z1is O, S, or NRb;

[0091] Raand Rbare each independently H or optionally substituted C1-20 alkyl; and

[0092] R1-R3is H, halogen, -OH, -NH2, -NO2, -CF3, -CN, -COOH, -N3, -SO3H, -PO3H2, C1-C20 alkyl, C1-C20 alkoxy, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, C3-C12 cycloalkenyl, or 3- to 15-membered heterocyclyl. In one example, each of R5-R8and R1-R3can each be independently substituted with one or more of halogen, -OH, C1-20 alkoxy, -NH2, -NO2, -CN, -CF3, -COOH, -N3, -SO3H, -PO3H2.

[0093] Compounds of the formula (Ic)also include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0094] R2and R3are each independently -H, -CN or -NO2;

[0095] R4is C1-C6 hydroxyalkyl, -OH, or C(0)0-(C1-C20) alkyl, and

[0096] R5-R8are each independently -H, -NO2, -NH2, C1-C6 alkylamino, or C1-C6 dialkylamino, Compounds of the formula (Ic)also include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein

[0097] R2and R3are each independently -H, -CN or -NO2;

[0098] R4is C1-C6 hydroxyalkyl, -OH, or C(0)0-(C1-C20) alkyl, and

[0099] R6-R8are each independently -H, -NO2, -NH2, C1-C6 alkylamino, or C1-C6 dialkylamino. In various examples:

[0100] R2or R3is -CN, R4is -OCH3, and

[0101] R5is -H; or

[0102] R2or R3is -NO2,

[0103] R4is -OCH3, and

[0104] R5is -H.

[0105] Compounds contemplated herein include:

[0106] or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester).

[0107] The disclosure also provides a method of inhibiting a-synuclein (a-syn) protein aggregation, tau isoform 2N4R protein aggregation, or both in a subject having, or at risk for, said protein aggregation, comprising administering to the subject at least one compound of the disclosure in an amount effective to inhibit a-syn protein aggregation, tau isoform 2N4R protein aggregation, or both, whereupon said protein aggregation is inhibited in the subject having, or at risk for, said protein aggregation. In one example, the at least one compound of the disclosure for use in the method of inhibiting a-synuclein (a-syn) protein aggregation, tau isoform 2N4R protein aggregation, or both in a subject having, or at risk for, said protein aggregation includes a (e.g., at least one) compound of the formulae la, lb, and / or Ic: la lb Ic or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein Y1is: and Y2is wherein:

[0108] Z1is N or CH;

[0109] Z2is C, CH or N;

[0110] Z3is N or NRa;

[0111] Z1is O, S, or NRb; provided that Y1is not unsubstituted indolyl when Y2is: one of R'-R4is fluoro, chloro, I-NO2, C1-C2 alkyl or methoxy (OCH3);

[0112] Raand Rbare each independently H or optionally substituted C1-C20 alkyl; and

[0113] RkR8and R1-R3are each independently -H, halogen, -OH, -NH2, -NO2, -CN, -CF3, -COOH,

[0114] -N3, -SO3H, -PO3H2, C1-C20 alkyl, C1-C20 hydroxyalkyl, C1-C20 alkylamino, C1-C20 dialkylamino, C1-20 alkoxy, C(0)0-(C1-C20) alkyl, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, or 3- to 15-membered heterocyclyl. In one example, each of R'-R8and R1-R3can each be independently substituted with one or more of halogen, -OH, C1-20 alkoxy, -NH2,

[0115] -NO2, -CN, -CF3, -COOH, -N3, -SO3H, -PO3H2. In one example,.

[0116] Compounds of the formula (la) include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0117] Z1is N or CH;

[0118] Z3is O, S, or NRa; provided that Z1is not CH when Z3is NRa;

[0119] Rais H or optionally substituted C1-20 alkyl;

[0120] R'-R3and R5-R8are each independently -H, halogen, -CN, -CF3, -NH2, -NO2, optionally substituted C1-20 alkyl or optionally substituted C1-20 alkoxy.

[0121] Compounds of the formula (la) also include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0122] Z1is N or CH;

[0123] Z3is O, S, or NRa; provided that Z1is not CH when Z3is NEC;

[0124] Rais H or optionally substituted alkyl;

[0125] Z1is O, S, or NRb;

[0126] Raand Rbare each independently H or optionally substituted C1-20 alkyl; and

[0127] R1-R3are each independently H, halogen, -OH, -NEL, -NO2, -CN, -COOH, -Ns,-SO3H, - PO3H2 C1-C20 alkyl, alkoxy, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, C3-C12 cycloalkenyl, or 3- to 15-membered heterocyclyl. En one example, each of R5-R8and R1-R3can each be independently substituted with one or more of halogen, -OH, C1-20 alkoxy, -NH2, -NO2, -CN, -CF3, -COOH, -N3, -SO3H, -PO3H2.

[0128] Compounds of the formula (Eb) include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0129] Z1is N or CH;

[0130] Z3is O, S, or NRa;

[0131] Rais H or optionally substituted C1-20 alkyl; and

[0132] RkR8are each independently H, halogen, -CF3,-CN, -NO2, or alkoxy.

[0133] Compounds of the formula (Eb) also include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0134] Z1is N or CH;

[0135] Z3is O, S, or NRa;

[0136] Rais H or optionally substituted alkyl;

[0137] Zris O, S, or NRb;

[0138] Raand Rbare each independently H or optionally substituted C1-20 alkyl; and

[0139] R1-R3are each independently H, halogen, -OH, -NEL, -NO2, -CF3, -CN, -COOH, -Ns, -

[0140] SO3H, -PO3H2, C1-C20 alkyl, alkoxy, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, C3- C12 cycloalkenyl, or 3- to 15-membered heterocyclyl. In one example, each of R5-R8and R1- R3can each be independently substituted with one or more of halogen, -OH, C1-20 alkoxy, - NH2, -NO2, -CN, -CF3, -COOH, -N3, -SO3H, -PO3H2.

[0141] Compounds of the formula (lb) also include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0142] Y1is 4-amino-indole, 5-amino-indole, 6-amino-indole, or 7-amino-indole;

[0143] Ri is -H, halogen, -OH, C1-C6 alkyl, C1-C6 alkyl-OH, C1-C6 alkoxy, or -NO2;

[0144] R2is -H, halogen, -OH, C1-C6 alkyl-OH, C1-C6 alkoxy, or -NO2; and

[0145] R3 is -H, halogen, -OH, C1-C6 alkyl-OH, C1-C6 alkoxy, or -NO2. In one example, Y1is optionally substituted with a C1-C6 alkyl or a halogen. In one example, the halogen can be I. In various other examples:

[0146] Y1is 4-amino-indole;

[0147] R1is -H, and

[0148] R3 is -NO2; or

[0149] Y1is 6-amino-indole,

[0150] R1is -H, and

[0151] R3 is -NO2.

[0152] Compounds of the formula (Ic) include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0153] Z1is N or CH

[0154] Z3is O, S, or NRa

[0155] Rais H or optionally substituted C1-20 alkyl; and

[0156] R'-R8are each independently H, halogen, -CF3, -CN, -NO2, or alkoxy.

[0157] Compounds of the formula (Ic)also include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0158] Z1is N or CH;

[0159] Z3is O, S, or NRa;

[0160] Z1is O, S, or NRb;

[0161] Raand Rbare each independently H or optionally substituted C1-20 alkyl; and

[0162] R1-R3is H, halogen, -OH, -NH2, -NO2, -CF3, -CN, -COOH, -N3, -SO3H, -PO3H2, C1-C20 alkyl, C1-C20 alkoxy, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, C3-C12 cycloalkenyl, or 3- to 15-membered heterocyclyl. In one example, each of R5-R8and R1-R3can each be independently substituted with one or more of halogen, -OH, C1-20 alkoxy, -NH2, -NO2, -CN, -CF3, -COOH, -N3, -SO3H, -PO3H2.

[0163] Compounds of the formula (Ic)also include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0164] R2and R3are each independently -H, -CN or -NO2;

[0165] R4is C1-C6 hydroxyalkyl, -OH, or C(0)0-(C1-C20) alkyl, and

[0166] R5-R8are each independently -H, -NO2, -NH2, C1-C6 alkylamino, or C1-C6 dialkylamino, Compounds of the formula (Ic)also include compounds of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein

[0167] R2and R3are each independently -H, -CN or -NO2; R4is C1-C6 hydroxyalkyl, -OH, or C(0)0-(C1-C20) alkyl, and

[0168] R6-R8are each independently -H, -NO2, -NH2, C1-C6 alkylamino, or C1-C6 dialkylamino. In various examples:

[0169] R2or R3is -CN,

[0170] R4is -OCH3, and

[0171] R5is -H; or

[0172] R2or R3is -NO2,

[0173] R4is -OCH3, and

[0174] R5is -H.

[0175] In one example, the at least one compound of the disclosure for use in the method of inhibiting a-synuclein (a-syn) protein aggregation, tau isoform 2N4R protein aggregation, or both in a subject having, or at risk for, said protein aggregation include compounds of the formulae:

[0176] or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester). 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 of stereoisomers 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.

[0177] 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.

[0178] The terms “salts” and “pharmaceutically acceptable salts” refer to derivatives of the 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.

[0179] 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, 17th ed., Mack Publishing Company, Easton, Pa., 1985, the disclosure of which is hereby incorporated by reference for its teachings regarding same.

[0180] The term “solvate” means a compound, or a salt or ester thereof (e.g., methyl or ethyl ester), 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.

[0181] The above compounds, and pharmaceutically acceptable salts and solvates thereof, can be synthesized in accordance with methods known in the art and exemplified herein. See, e.g., Example 2 and Figs. 1 and 2.

[0182] 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. The compounds described herein can be used to inhibit the aggregation of tau isoform 2N4R and 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.

[0183] 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, 23rdedition, October 30, 2020, Adeboye Adejare, ed. Accordingly, further provided is a pharmaceutical composition comprising a compound of formula I and a pharmaceutically acceptable carrier. Still further provided is a method of inhibiting a-synuclein (a-syn) protein aggregation, tau isoform 2N4R protein aggregation, or both in a subject having, or at risk for, said protein aggregation. The method comprises administering to the subject a pharmaceutical composition, e.g., comprising a compound of formula I, in an amount effective to inhibit a-syn protein aggregation, tau isoform 2N4R protein aggregation, or both, whereupon said protein aggregation is inhibited in the subject having, or at risk for, said protein aggregation. The subject can have, or be at risk for, Alzheimer’s disease. Over half of patients with Alzheimer’s disease not only have tau protein aggregation but also aggregated a-synuclein (a-syn)- containing Lewy bodies (LB). The subject can have, or be at risk for, Parkinson’s disease or dementia with Lewy bodies (DLB). The subject can have neuroblastoma, in which case the method inhibits the formation of a-syn inclusions, such as when a pharmaceutical composition comprising a compound in which Ar is 6-amino-indole, R1is hydrogen, and R2is a nitro group.

[0184] Even still further provided is a method of inhibiting fibril formation in a subject having, or at risk for, fibril formation. The method comprises administering to the subject an abovedescribed pharmaceutical composition (comprising a compound of Formula II) in an amount effective to inhibit fibril formation, whereupon fibril formation is inhibited in the subject having, or at risk for, said fibril formation.

[0185] 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.

[0186] The terms “substituted,” ’’substituent,” and “functional group” refer to the replacement of one or more hydrogen atoms on a molecule or group (e.g., on an aryl or an alkyl group) with another atom or group. Examples of substituents include, but are not limited to, a halogen (e.g., F, Cl, Br, and I), -OR, -OC(O)N(R)(R'), -CN, -NO, -NO2, -ONO2, -N3, -CF3, -OCF3, -R, =0 (oxo), =S (thiono), -C(O), -S(O), methylenedioxy, ethylenedi oxy, -N(R)(R'), -SR, -SOR, - SO2R, -SO2N(R)(R'), -SO3R, -(CH2)0-2P(O)(OR)(OR'), -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)(R'), -OC(O)N(R)(R'), - C(S)N(R)(R'), -(CH2)O-2N(R)C(0)R', -(CH2)O-2N(R)C(0)OR', -(CH2)O-2N(R)N(R')(R"), - N(R)N(R')C(O)R", -N(R)N(R')C(O)OR", -N(R)N(R')CON(R")(R"'), -N(R)SO2R', -

[0187] N(R)SO2N(R')(R"), -N(R)C(O)OR', -N(R)C(O)R', N(R)C(S)R', -N(R)C(O)N(R')(R"), - N(R)C(S)N(R')(R"), -N(COR)COR', -N(OR)R', -C(=NH)N(R)(R'), -C(O)N(OR)R', or - C(=NOR)R', wherein R, R', R", and R'" are each independently selected from hydrogen, alkyl, acyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroarylalkyl, and wherein R and R' when bonded to a nitrogen atom or to adjacent nitrogen atoms can together form a heterocyclyl, which can be mono- or multi-substituted.

[0188] The term “alkyl” as used herein refers to substituted or unsubstituted straight chain and branched mono- or divalent alkyl groups having from 1 to 40 carbon atoms (C1-C4o), 1 to about 20 carbon atoms (C1-C20), 1 to 12 carbons (C1-C12), 1 to 8 carbon atoms (Ci-Cs), and 1 to 6 carbon atoms ( C1-C6). The enumerated ranges also include all subranges, for example, Ci to C8 which includes C1-C2, C1-C3, C1-C4, C1-C5, C1-C6, C1-C7, C1-C8, C2-C3, C2-C4, C2-C5, C2- c6, C2-C7, C2-C8, C3-C4, C3-C5, C3-C6, C3-C7, C3-C8, c4-c5, C4-C6, C4-C7, C4-C8, C5-C6, C5- C7, C5-C8, C6-C7, C6-C8, and C7-C8. 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 substituents selected from amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0189] The term “alkenyl” as used herein refers to substituted or unsubstituted straight chain and branched mono- or divalent alkenyl groups having at least one double bond and having from 2 to 40 carbon atoms (C2-C4o), 2 to about 20 carbon atoms (C2-C20), 2 to 12 carbons (C2- C12), 2 to 8 carbon atoms (C2-C8), and 2 to 6 carbon atoms (C2-C6). The enumerated ranges also include all subranges, for example, C2 to C8 which includes C2-C3, C2-C4, C2-C5, C2-C6, C2-C7, C2-C8, C3-C4, C3-C5, C3-C6, C3-C7, C3-C8, c4-c5, C4-C6, C4-C7, C4-C8, C5-C6, C5-C7, C5-C8, C6-C7, C6-C8, and C7-C8. Examples of straight chain alkenyl groups include those with from 2 to 8 carbon atoms such as -CH=CH-, -CH=CHCH3, and -CEECE^CEICEE- 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(CHs)- and CH2C=CH(CHB) groups. Representative substituted alkenyl groups can be substituted one or more times with substituents selected from amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0190] The term "alkynyl" as used herein refers to substituted or unsubstituted straight chain and branched mono- or divalent alkynyl groups having at least one triple bond and having from 2 to 40 carbon atoms (C2-C4o), 2 to about 20 carbon atoms (C2-C20), 2 to 12 carbons (C2-C12), 2 to 8 carbon atoms (C2-C8), and 2 to 6 carbon atoms (C2-C6). The enumerated ranges also include all subranges, for example, C2 to Cs which includes C2-C3, C2-C4, C2-C5, C2-C6, C2-C7, C2-C8, C3-C4, C3-C5, C3-C6, C3-C7, C3-C8, c4-c5, C4-C6, C4-C7, C4-C8, C5-C6, C5-C7, C5-C8, C6-C7, Ce-Cs. Examples of straight chain alkynyl groups include those with from 2 to 8 carbon atoms such as -C=CH, -OCCH3, and -CH2OCCH2- groups. Examples of branched alkynyl groups include, but are not limited to, -C=C-CH-(CH3)2 and -CEhOC-CEE groups. Representative substituted alkynyl groups can be substituted one or more times with substituents selected from amino, cyano, carboxy, alkoxy, and halogen groups.

[0191] The term “alkoxy” as used herein refers to -O-alkyl groups wherein the alkyl portion is as defined above and includes groups having from 1 to 40 carbon atoms (Ci-C4o), 1 to about 20 carbon atoms (C1-C20), 1 to 12 carbons (C1-C12), 1 to 8 carbon atoms (Ci-Cs), and 1 to 6 carbon atoms ( C1-C6). The enumerated ranges also include all subranges, for example, Ci to C8which includes C1-C2, C1-C3, C1-C4, C1-C5, C1-C6, C1-C7, Ci-C8, C2-C3, C2-C4, C2-C5, C2- c6, C2-C7, C2-C8, C3-C4, C3-C5, C3-C6, C3-C7, C3-C8, c4-c5, C4-C6, C4-C7, C4-C8, C5-C6, C5- C7, C5-C8, C6-C7, C6-C8, and C7-C8. Examples include methoxy (-OCH3), ethoxy (-OCH2CH3), propoxy (-OCH2CH2CH3), and butoxy (-OCH2CH2CH2CH3) groups, as well as their branched isomers such as isopropoxy (-OCH(CH3)2 and tert-butoxy (O-C(CH3)3 groups. Representative substituted alkoxy groups can be substituted one or more times with substituents selected from amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0192] 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 (e.g. F, Cl, Br, or I).

[0193] The term “haloalkyl” as used herein refers to alkyl groups as defined above wherein one or more hydrogen atoms are replaced by halogen atoms (F, Cl, Br, or I). The haloalkyl groups can have from 1 to 40 carbon atoms (Ci-C4o), 1 to about 20 carbon atoms (C1-C20), 1 to 12 carbons (C1-C12), 1 to 8 carbon atoms (C1-C8), and 1 to 6 carbon atoms ( C1-C6). 1 to 40 carbon atoms (C1-C4o), 1 to about 20 carbon atoms (C1-C20), 1 to 12 carbons (C1-C12), 1 to 8 carbon atoms (Ci-Cs), and 1 to 6 carbon atoms ( C1-C6). The enumerated ranges also include all subranges, for example, Ci to Cs which includes C1-C2, C1-C3, C1-C4, C1-C5, C1-C6, C1-C7, Ci-C8, C2-C3, C2-C4, C2-C5, C2-C6, C2-C7, C2-C8, C3-C4, C3-C5, C3-C6, C3-C7, C3-C8, C4-C5, C4-C6, C4-C7, C4-C8, C5-C6, C5-C7, C5-C8, C6-C7, C6-C8, and C7-C8. Examples include fluoromethyl (CH2F), difluoromethyl (CHF2), trifluoromethyl (CF3), chloromethyl (-CH2CI), dichloromethyl (-CHCI2), trichloromethyl (-CCI3), 2-fluoroethyl (-CH2CH2F), 2,2- difluoroethyl (-CH2CF2H), 2,2,2-trifluoroethyl (-CH2CF3), 2-chloroethyl (-CH2CH2CI), 2,2- dichloroethyl (-CH2CCI2H), and 2,2,2-trichloroethyl (-CH2CCI3) As used herein, the term “haloalkyl” includes mono-halo alkyl groups and poly-halo alkyl groups, wherein all the halogen atoms can be the same or different, and per-halo alkyl groups, wherein all hydrogen atoms are replaced by halogen atoms, such as fluoro. Further examples of haloalkyls include 1,1 -di chloroethyl, 1,2-di chloroethyl, l,3-dibromo-3,3-difluoropropyl, perfluorobutyl, - CF(CH3)2and the like.

[0194] 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-C6), and 4 to 8 carbon atoms (C4-C8). The enumerated ranges also include all subranges, for example, C2 to Cs which includes C3-C4, C3- C5, C3-C6, C3-C7, C3-C8, C4-C5, C4-C6, C4-C7, C4-C8, C5-C6, C5-C7, C5-C8, C6-C7, C6-C8, and C7-C8. Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbomyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like.

[0195] The term “alkylcycloalkyl” 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. The alkylcycloalkyl groups can have from 4 to 40 total carbon atoms (C4-C4o), 4 to about 20 carbon atoms (C4-C2o), 4 to 10 carbons (C4-Cio), and 4 to 8 carbon atoms (C4-C8). The enumerated ranges also include all subranges, for example, C4to C10, which includes C4-C5, C4-Ce, C4-C?, C4-Cs, C5-C6, C5- C7, C5-C8, C6-C7, C6-C8, and C7-C8, C7-C9, C7-C10, C8-C9, Cs-Cio, and C9-C10. The alkyl portion can have from 1 to 6 carbon atoms ( C1-C6), and the cycloalkyl portion can have for example from 3 to 8 carbon atoms (C3-C8). Representative alkylcycloalkyl groups include, but are not limited to, methylcyclopropyl, methylcyclobutyl, methylcyclopentyl, methylcyclohexyl, ethylcyclopropyl, ethylcyclobutyl, ethylcyclopentyl, ethylcyclohexyl, and their higher homologs. Representative substituted alkylcycloalkyl groups can be substituted one or more times with substituents selected from amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0196] The term “cycloalkylalkyl” 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. The cycloalkylalkyl groups can have from 4 to 40 total carbon atoms (C4-C4o), 4 to about 20 carbon atoms (C4-C2o), 4 to 10 carbons (C4-Cio), and 4 to 8 carbon atoms (C4-C8). The enumerated ranges also include all subranges, for example, C4to Cio, which includes C4-C5, C4-C6, C4-C7, C4-C8, C5-C6, C5-C?, C5-C8, C6-C7, C6-C8, and C7-C8, C7-C9, C7-C10, C8-C9, C8-C10, and C9-C10. The alkyl portion can have for example from 1 to 6 carbon atoms ( C1-C6), and the cycloalkyl portion can have for example from 3 to 8 carbon atoms (C3-C8). Representative alkylcycloalkyl groups include, but are not limited to, methylcyclopropyl, methylcyclobutyl, methylcyclopentyl, methylcyclohexyl, ethylcyclopropyl, ethylcyclobutyl, ethylcyclopentyl, ethylcyclohexyl, and their higher homologs. Representative substituted alkylcycloalkyl groups can be substituted one or more times with substituents selected from amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0197] 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 can include 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, pyridyl acetyl, 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.

[0198] The term “heterocyclylcarbonyl” is an example of an acyl group that is bonded to a substituted or unsubstituted heterocyclyl group, as the term “heterocyclyl” is defined herein. An example of a heterocyclylcarbonyl group is a prolyl group, wherein the prolyl group can be a D- or an L-prolyl group.

[0199] The term “aryl” as used herein refers to substituted or unsubstituted cyclic aromatic hydrocarbons that do not contain heteroatoms in the ring. Thus, aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylyl, 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: , each of which can be substituted or unsubstituted, such as hydroxy substituted.

[0200] 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.

[0201] The terms “alkylaryl” 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.

[0202] The term "heteroaryl" as used herein refers to mono- and polycyclic aromatic groups containing from 5 to 15 ring atoms wherein at least one ring atom is a heteroatom selected from nitrogen (N), oxygen (O), and sulfur (S). Heteroaryl groups can be monocyclic (5- or 6- membered ring) or bicyclic (8-, 9-, or 10-membered ring). Examples of monocyclic heteroaryl groups include but are not limited to furanyl, thiophenyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, tetrazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, and triazinyl groups. Examples of bicyclic heteroaryl groups include but are not limited to benzofuranyl, benzothiophenyl, benzoxazolyl, benzothiazolyl, benzimidazolyl, indolyl, isoindolyl, indazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl, imidazo[l,2-a]pyridinyl, triazyolyl, tetrazolyl, benzoxazolinyl, thiazolyl, benzthiazolinyl, and benzimidazolinyl groups.

[0203] Examples of benzoxazolinyl groups include groups having the general formula:

[0204] Examples of benzthiazolinyl groups include groups having the general formula:

[0205] Examples of imidazo[l,2-a]pyridinyl groups include groups having the general formula:

[0206] Representative substituted heteroaryl groups can be optionally substituted one or more times with substituents selected from amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups. In some embodiments, the heteroaryl rings may be optionally substituted as defined above. In other embodiments, optional substitution includes N-H substitution of the heteroaryl ring. For example, the heteroaryl ring may be optionally substituted with alkyl, alkenyl, alkynyl, heterocyclyl, aryl, heteroaryl, or other substituents as described herein.

[0207] The term “heterocyclyl” or “heterocyclo” refers to substituted or unsubstituted aromatic and non-aromatic ring compounds containing 3 or more ring members, of which one or more (e.g., 1, 2 or 3) is a heteroatom such as, but not limited to, N, O, and S. Thus, a heterocyclyl can be a heterocycloalkyl or a heteroaryl or, if polycyclic, any combination thereof. In some embodiments, heterocyclyl groups include 3 to about 20 ring members, whereas other such groups have 3 to about 15 ring members. In some embodiments, heterocyclyl groups include heterocyclyl groups that include 3 to 8 carbon atoms (C3-C8), 3 to 6 carbon atoms (C3-C6), 3 to 5 carbon atoms (C3-C5) or 6 to 8 carbon atoms (Ce-Cs). A heterocyclyl group designated as a C2-heterocyclyl can be a 5 -membered ring with two carbon atoms and three heteroatoms, a 6- membered ring with two carbon atoms and four heteroatoms and so forth. Likewise, a C4- heterocyclyl can be a 5-membered ring with one heteroatom, a 6-membered ring with two heteroatoms, and so forth. The number of carbon atoms plus the number of heteroatoms equals the total number of ring atoms. Examples of heterocyclyl groups include, but are not limited to, azetidinyl, piperidynyl, piperazinyl, morpholinyl, pyrrolidinyl, or pyrrolidinone (e.g., the radical of pyrrolidin-2-one) groups.

[0208] A heterocyclyl ring can also include one or more double bonds, such as in the group

[0209] 3,6-dihydro-2H-pyran and 3,4-dihydro-2H-pyran, having the formula: , respectively, each of which may be substituted. Other representative heterocyclyl groups include, but are not limited to, tetrahydro-2H-thiopyran- 1,1 -di oxide, having the formula: , which may be substituted.

[0210] The term “heterocyclyl group” also includes fused ring species including those that include bicyclic groups. An example of a bicyclic heterocyclyl group is the 4a,5,6,7-tetrahydro- 4H-pyrrolo[l,2-d][l,3,4]oxadiazinyl group, having the general formula:

[0211] The term “heterocyclyl group” further includes fused ring species including fused aromatic and non-aromatic groups. Representative groups include, but are not limited to, tetrahydrobenzofuranyl, benzodi oxolyl, chromanyl (eg. a radical of 3,4-dihydro-2H- chromene), chromanonyl, indolinonyl, isoindolinonyl, and 4a,5,6,7-tetrahydro-4H- pyrrolof 1 ,2-d] [ 1 ,3 ,4]oxadiazinyl :

[0212] Examples of tetrahydrobenzofuranyl groups include groups having the general formula:

[0213] Examples of benzodi oxlyl groups include groups having the general formula:

[0214] Examples of chromanyl groups include groups having the general formula:

[0215] Examples of chromanonyl groups include groups having the general formula:

[0216] Examples of indolinonyl groups include groups having the general formula:

[0217] Examples of isoindolinonyl groups include groups having the general formula:

[0218] The aromatic and non-aromatic portion of the heterocyclyl group may both contain a heteroatom, for example, 3,4-dihydro-2H -1λ-l,8-naphthyridine, having the general formula:

[0219] The term “heterocyclylalkyl” refers to 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 heterocyclyl group as defined herein. Representative heterocyclylalkyl groups include, but are not limited to, furan-2-yl methyl, furan-3-yl methyl, pyridine-3-yl methyl, tetrahydrofuran-2- yl methyl, and indol-2-yl propyl.

[0220] The term “heterocyclylalkoxy” refers to 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 heterocyclyl group as defined herein and the alkyl group is attached to an oxygen. Representative heterocyclylalkoxy groups include, but are not limited to, -O- (CH2)qheterocyclyl, wherein q is an integer from 1 to 5. In some embodiments, heterocyclylalkoxy groups include -O-(CH2)qmorpholinyl such as -O-CH2CH2-morpholine.

[0221] The term “heteroarylalkyl” refers to alkyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to a heteroaryl group as defined herein.

[0222] 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, an allyloxy 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.

[0223] The terms “amine,” “amine group,” “amino,” and “amino group” refer to a substituent of the form -NH2, -NHR, -NR’R”, or -N+RR”R”’, wherein each R, R’, R” and R’” are defined herein, and protonated forms of each, except for -N+RR”R”’, 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.

[0224] An “alkylamino” group includes a monoalkylamino, dialkylamino, and trialkylamino group. An example of a “alkylamino” is -NH-alkyl and -N(alkyl)2.

[0225] An example of a “cycloalkylamino” group is -NH-cycloalkyl and -N(cycloalkyl)2.

[0226] An example of a “cycloalkyl heterocycloamino” group is -NH-(heterocyclo cycloalkyl), wherein the heterocyclo group is attached to the nitrogen and the cycloalkyl group is attached to the heterocyclo group.

[0227] An 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.

[0228] The term “amido” refers to a group of the formula -C(O)NR2, wherein R is defined herein.

[0229] The terms “treat,” “treating,” “treated,” or “treatment” (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. 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 vary 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. Appropriate 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.

[0230] 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.

[0231] A “therapeutically effective amount” (or “effective amount”) of a compound with respect to use in treatment, refers to an amount of the compound in a preparation which, when administered as part of a desired dosage regimen (to a mammal, such as a human) alleviates a symptom, ameliorates a condition, or slows the onset of disease conditions according to clinically acceptable standards for the disorder or condition to be treated or the cosmetic purpose, e.g., at a reasonable benefit / risk ratio applicable to any medical treatment.

[0232] For any compound a therapeutically effective amount can be initially determined from animal models. A therapeutically effective dose can also be determined from human data for compounds which have been tested in humans and for compounds which are known to exhibit similar pharmacological activities, such as other related active agents. Higher doses may be required for parenteral administration. The applied dose can be adjusted based on the relative bioavailability and potency of the administered compound. Adjusting the dose to achieve maximal efficacy based on the methods described above and other methods as are well-known in the art is well within the capabilities of the ordinarily skilled artisan.

[0233] The formulations can be administered in pharmaceutically acceptable solutions, which can routinely contain pharmaceutically acceptable concentrations of salt, buffering agents, preservatives, compatible carriers, adjuvants, and optionally other therapeutic ingredients. For use in therapy, an effective amount of the compound can be administered to a subject by any mode that delivers the compound to the desired surface. Administering a pharmaceutical composition can be accomplished by any means known to the skilled artisan. Routes of administration include, but are not limited to, intravenous, intramuscular, intraperitoneal, intravesical (urinary bladder), oral, subcutaneous, direct injection (for example, into a tumor or abscess), mucosal (e.g., topical to eye), inhalation, and topical.

[0234] For intravenous and other parenteral routes of administration, a compound can be formulated as a lyophilized preparation, as a lyophilized preparation of liposome-intercalated or -encapsulated active compound, as a lipid complex in aqueous suspension, or as a salt complex. Lyophilized formulations are generally reconstituted in suitable aqueous solution, e.g., in sterile water or saline, shortly prior to administration.

[0235] For oral administration, the compounds can be formulated readily by combining the active compound(s) with pharmaceutically acceptable carriers well-known in the art. Such carriers enable the compounds to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a subject to be treated. A pharmaceutical preparation for oral use can be obtained as a solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, and sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and / or polyvinyl pyrrolidone (PVP). If desired, disintegrating agents can be added, such as the cross-linked PVP, agar, or alginic acid or a salt thereof such as sodium alginate. Optionally the oral formulations can also be formulated in saline or buffers, e.g., EDTA for neutralizing internal acid conditions, or can be administered without any carriers.

[0236] Also contemplated are oral dosage forms of the compounds. The compounds can be chemically modified so that oral delivery of the derivative is efficacious. Generally, the chemical modification contemplated is the attachment of at least one moiety to the compound itself, where said moiety permits (a) inhibition of acid hydrolysis; and (b) uptake into the blood stream from the stomach or intestine. Also desired is the increase in overall stability of the compounds and increase in circulation time in the body. Examples of such moieties include polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, PVP and polyproline. Abuchowski and Davis, “Soluble Polymer-Enzyme Adducts,” In: Enzymes as Drugs, Hocenberg and Roberts, eds., Wiley- Interscience, New York, N.Y., pp. 367-383 (1981); Newmark et al., J Appl Biochem 4: 185-189 (1982). Other polymers that could be used are poly- 1,3 -di oxolane and poly-1, 3, 6-tioxocane. For pharmaceutical usage, as indicated above, polyethylene glycol moieties are suitable.

[0237] The location of release of a compound hereof can be the stomach, the small intestine (e.g., the duodenum, the jejunum, or the ileum), or the large intestine. One skilled in the art has available formulations, which will not dissolve in the stomach, yet will release the material in the duodenum or elsewhere in the intestine. The release can avoid the deleterious effects of the stomach environment, either by protection of the compound or by release of the compound beyond the stomach environment, such as in the intestine.

[0238] To ensure full gastric resistance a coating impermeable to at least pH 5.0 is essential. Examples of the more common inert ingredients that are used as enteric coatings are cellulose acetate trimellitate (CAT), hydroxypropylmethylcellulose phthalate (HPMCP), HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S, and shellac. These coatings can be used as mixed films.

[0239] A coating or mixture of coatings can also be used on tablets, which are not intended for protection against the stomach. This can include sugar coatings, or coatings which make the tablet easier to swallow. Capsules can consist of a hard shell (such as gelatin) for delivery of dry therapeutic (e.g., powder); for liquid forms, a soft gelatin shell can be used. The shell material of cachets could be thick starch or other edible paper. For pills, lozenges, molded tablets or tablet triturates, moist massing techniques can be used.

[0240] The compound can be included in the formulation as fine multi-particulates in the form of granules or pellets of particle size about 1 mm. The formulation of the material for capsule administration could also be as a powder, lightly compressed plugs or even as tablets. Therapeutic agent could be prepared by compression.

[0241] Colorants and flavoring agents may all be included. For example, the compound can be formulated (such as by liposome or microsphere encapsulation) and then further contained within an edible product, such as a refrigerated beverage containing colorants and flavoring agents.

[0242] One may dilute or increase the volume of the compound with an inert material. These diluents can include carbohydrates, especially mannitol, a-lactose, anhydrous lactose, cellulose, sucrose, modified dextrans and starch. Certain inorganic salts also can be used as fillers including calcium triphosphate, magnesium carbonate and sodium chloride. Some commercially available diluents are Fast-Flo, Emdex, STA-Rx 1500, Emcompress and Avicell.

[0243] Disintegrants can be included in the formulation of therapeutic agent into a solid dosage form. Materials used as disintegrates include, but are not limited to, starch, including the commercial disintegrant based on starch, Explotab. Sodium starch glycolate, Amberlite, sodium carboxymethylcellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acid carboxymethyl cellulose, natural sponge and bentonite may all be used. Another form of the disintegrant is the insoluble cationic exchange resin. Powdered gums can be used as disintegrants and as binders, and these can include powdered gums such as agar, Karaya or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants.

[0244] Binders can be used to hold the compound together to form a hard tablet and can include materials from natural products such as acacia, tragacanth, starch and gelatin. Others include methyl cellulose (MC), ethyl cellulose (EC) and carboxymethyl cellulose (CMC). PVP and hydroxypropylmethyl cellulose (HPMC) can both be used in alcoholic solutions to granulate therapeutic agent.

[0245] An anti -frictional agent can be included in the formulation of therapeutic to prevent sticking during the formulation process. Lubricants can be used as a layer between therapeutic agent and the die wall, and these can include, but are not limited to, stearic acid, including its magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils and waxes. Soluble lubricants can also be used, such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycol of various molecular weights, Carbowax 4000 and 6000.

[0246] Glidants, which can improve the flow properties of the drug during formulation and aid rearrangement during compression, can be added. The glidants can include starch, talc, pyrogenic silica and hydrated silicoaluminate.

[0247] To aid dissolution of therapeutic agent into the aqueous environment a surfactant can be added as a wetting agent. Surfactants can include anionic detergents, such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate. Cationic detergents, which can be used, include benzalkonium chloride and benzethonium chloride. Potential non-ionic detergents that can be included in the formulation as surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50 and 60, glycerol monostearate, polysorbate 40, 60, 65 and 80, sucrose fatty acid ester, methyl cellulose and carboxymethyl cellulose. These surfactants could be present in the formulation of the compound or derivative thereof either alone or as a mixture in different ratios.

[0248] Pharmaceutical preparations, which can be used orally, include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers can be added. Microspheres formulated for oral administration can also be used. Such microspheres have been well-defined in the art. All formulations for oral administration should be in dosages suitable for such administration.

[0249] For buccal administration, the compositions can take the form of tablets or lozenges formulated in conventional manner.

[0250] For topical administration, the compound can be formulated as solutions, gels, ointments, creams, suspensions, etc. as are well-known in the art. Systemic formulations include those designed for administration by injection, e.g., subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, as well as those designed for transdermal, transmucosal oral or pulmonary administration.

[0251] For administration by inhalation, compounds can be conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, tri chi orofluorom ethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges of e.g., gelatin for use in an inhaler or insufflator, can be formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0252] Also contemplated is pulmonary delivery of the compounds (or salts thereof). The compound is delivered to the lungs of a mammal while inhaling and traverses across the lung epithelial lining to the blood stream. Other reports of inhaled molecules include Adjei et al., Pharm Res 7:565-569 (1990); Adjei et al., Int J Pharmaceutics 63: 135-144 (1990) (leuprolide acetate); Braquet et al., J Cardiovasc Pharmacol 13(suppl. 5): 143-146 (1989) (endothelin-1); Hubbard et al., Annal Int Med 3:206-212 (1989) (al -antitrypsin); Smith et al., 1989, J Clin Invest 84: 1145-1146 (a- 1 -proteinase); Oswein et al., 1990, "Aerosolization of Proteins," Proceedings of Symposium on Respiratory Drug Delivery II, Keystone, Colorado, March, (recombinant hepatocyte growth hormone); Debs et al., 1988, J Immunol 140:3482-3488 (interferon-gamma and tumor necrosis factor alpha) and Platz et al., U.S. Pat. No. 5,284,656 (granulocyte colony stimulating factor; incorporated herein by reference). A method and composition for pulmonary delivery of drugs for systemic effect is described in U.S. Pat. No. 5,451,569 (specifically incorporated herein by reference for its disclosure regarding same), issued Sep. 19, 1995, to Wong et al.

[0253] Contemplated for use are a wide range of mechanical devices designed for pulmonary delivery of therapeutic products including, but not limited to, nebulizers, metered dose inhalers, and powder inhalers, all of which are familiar to those skilled in the art.

[0254] Nasal delivery of a pharmaceutical composition is also contemplated. Nasal delivery allows the passage of a pharmaceutical composition to the blood stream directly after administering therapeutic product to the nose, without the necessity for deposition of the product in the lung. Formulations for nasal delivery include those with dextran or cyclodextran.

[0255] The compounds, when it is desirable to deliver them systemically, can be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection can be presented in unit dosage form, e.g., in ampoules or in multidose containers, with an added preservative. The compositions can take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and / or dispersing agents.

[0256] Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active compounds can be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions can contain substances which increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension can also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.

[0257] Alternatively, the active compounds can be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

[0258] The compounds can also be formulated in rectal or vaginal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides. In addition to the formulations described above, a compound can also be formulated as a depot preparation. Such long-acting formulations can be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

[0259] The pharmaceutical compositions also can comprise suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.

[0260] Suitable liquid or solid pharmaceutical preparation forms are, for example, aqueous or saline solutions for inhalation, microencapsulated, encochleated, coated onto microscopic gold particles, contained in liposomes, nebulized, aerosolized, pelleted for implantation into the skin, or dried onto a sharp object to be scratched into the skin. The pharmaceutical compositions also include granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops or preparations with protracted release of active compounds, in whose preparation excipients and additives and / or auxiliaries such as disintegrants, binders, coating agents, swelling agents, lubricants, flavorings, sweeteners or solubilizers are customarily used as described above. The pharmaceutical compositions are suitable for use in a variety of drug delivery systems. For a brief review of methods for drug delivery, see Langer R, Science 249: 1527-1533 (1990).

[0261] The compound and optionally one or more other therapeutic agents can be administered per se (neat) or in the form of a pharmaceutically acceptable salt. When used in medicine the salts should be pharmaceutically acceptable, but non-pharmaceutically acceptable salts may conveniently be used to prepare pharmaceutically acceptable salts thereof. Such salts include, but are not limited to, those prepared from the following acids: hydrochloric, hydrobromic, sulphuric, nitric, phosphoric, maleic, acetic, salicylic, p-toluene sulphonic, tartaric, citric, methane sulphonic, formic, malonic, succinic, naphthalene-2-sulphonic, and benzene sulphonic. Also, such salts can be prepared as alkaline metal or alkaline earth salts, such as sodium, potassium or calcium salts of the carboxylic acid group.

[0262] Suitable buffering agents include acetic acid and a salt (1-2% w / v); citric acid and a salt (1-3% w / v); boric acid and a salt (0.5-2.5% w / v); and phosphoric acid and a salt (0.8-2% w / v). Suitable preservatives include benzalkonium chloride (0.003-0.03% w / v); chlorobutanol (0.3- 0.9% w / v); parabens (0.01-0.25% w / v); and thimerosal (0.004-0.02% w / v).

[0263] Pharmaceutical compositions contain an effective amount of a compound as described herein and optionally one or more other therapeutic agents included in a pharmaceutically acceptable carrier. The term “pharmaceutically acceptable carrier” means one or more compatible solid or liquid fillers, diluents or encapsulating substances, which are suitable for administration to a human or other vertebrate animal. The term “carrier” denotes an organic or inorganic ingredient, natural or synthetic, with which the active ingredient is combined to facilitate the application. The components of the pharmaceutical compositions also can be commingled with the compounds, and with each other, in a manner such that there is no interaction, which would substantially impair the desired pharmaceutical efficiency.

[0264] Therapeutic agent(s), including specifically, but not limited to, a compound, can be provided in particles. “Particles” means nanoparticles or microparticles (or in some instances larger particles) that can consist in whole or in part of the compound or the other therapeutic agent(s). The particles can contain therapeutic agent(s) in a core surrounded by a coating, including, but not limited to, an enteric coating. Therapeutic agent(s) also can be dispersed throughout the particles. Therapeutic agent(s) also can be adsorbed into the particles. The particles can be of any order release kinetics, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, and any combination thereof, etc. The particle can include, in addition to therapeutic agent(s), any of those materials routinely used in the art of pharmacy and medicine, including, but not limited to, erodible, nonerodible, biodegradable, or nonbiodegradable material or combinations thereof. The particles can be microcapsules which contain the compound in a solution or in a semi-solid state. The particles can be of virtually any shape.

[0265] Both non-biodegradable and biodegradable polymeric materials can be used in the manufacture of particles for delivering therapeutic agent(s). Such polymers can be natural or synthetic polymers. The polymer is selected based on the period of time over which release is desired. Bioadhesive polymers of particular interest include bioerodible hydrogels described in Sawhney et al., Macromolecules 26:5823-2787 (1993), the teachings of which are specifically incorporated by reference herein. These include polyhyaluronic acids, casein, gelatin, glutin, polyanhydrides, polyacrylic acid, alginate, chitosan, poly(methyl methacrylates), poly(ethyl methacrylates), poly(butylmethacrylate), poly(isobutyl methacrylate), poly(hexylmethacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly (isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate).

[0266] Therapeutic agent(s) can be contained in controlled-release systems. The term “controlled release” refers to any drug-containing formulation in which the manner and profile of drug release from the formulation are controlled. This refers to immediate as well as non- immediate release formulations, with non-immediate release formulations including, but not limited to, sustained release and delayed release formulations. The term “sustained release” (also referred to as “extended release”) refers to a drug formulation that provides for gradual release of a drug over an extended period of time, and that can result in substantially constant blood levels of a drug over an extended time period. The term “delayed release” refers to a drug formulation in which there is a time delay between administration of the formulation and the release of the drug therefrom. “Delayed release” may or may not involve gradual release of drug over an extended period of time, and thus may or may not be “sustained release.”

[0267] Use of a long-term, sustained-release implant can be particularly suitable for treatment of chronic conditions. “Long-term” release means that the implant is constructed and arranged to deliver therapeutic levels of the active ingredient for at least 7 days, and up to 30-60 days. Long-term sustained-release implants are well-known to those of ordinary skill in the art and include some of the release systems described above.

[0268] 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.

[0269] 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. The term “solvate” means a compound, or a salt or ester thereof (e.g., methyl or ethyl ester), 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.

[0270] The term “prodrug” means a derivative of a compound that can hydrolyze, oxidize, or otherwise react under biological conditions (in vitro or in vivo) to provide an active compound, particularly a compound of the invention. Examples of prodrugs include, but are not limited to, derivatives and metabolites of a compound of the invention that include biohydrolyzable moieties such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureides, and biohydrolyzable phosphate analogues. Specific prodrugs of compounds with carboxyl functional groups are the lower alkyl esters of the carboxylic acid. The carboxylate esters are conveniently formed by esterifying any of the carboxylic acid moieties present on the molecule. Prodrugs can typically be prepared using well-known methods, such as those described by Burger’s Medicinal Chemistry and Drug Discovery 6th ed. (Donald J. Abraham ed., 2001, Wiley) and Design and Application of Prodrugs (H. Bundgaard ed., 1985, Harwood Academic Publishers GmbH).

[0271] Further, in each of the foregoing and following embodiments, it is to be understood that 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 water and / 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.

[0272] 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" in the 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) alginic 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.

[0273] 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 in unit dosage forms and / or formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants, and vehicles.

[0274] Illustrative formats for oral administration include tablets, capsules, elixirs, syrups, and the like. Illustrative routes for parenteral administration include intravenous, intraarterial, intraperitoneal, epidural, intraurethral, intrasternal, intramuscular and subcutaneous, as well as any other art recognized route of parenteral administration.

[0275] Illustrative means of parenteral administration include needle (including microneedle) injectors, needle-free injectors and infusion techniques, as well as any other means of parenteral administration recognized in the art. Parenteral formulations are typically aqueous solutions, which may contain excipients such as salts, carbohydrates and buffering agents (preferably at a pH in the range from about 3 to about 9), but, for some applications, they may be more suitably formulated as a sterile non-aqueous solution or as a dried form to be used in conjunction with a suitable vehicle such as sterile, pyrogen-free water. The preparation of parenteral formulations under sterile conditions, for example, by lyophilization, may readily be accomplished using standard pharmaceutical techniques well-known to those skilled in the art. Parenteral administration of a compound is illustratively performed in the form of saline solutions or with the compound incorporated into liposomes. In cases where the compound, itself, is not sufficiently soluble to be dissolved, a solubilizer such as ethanol can be applied.

[0276] The dosage of each compound of the claimed combinations depends on several factors, including: the administration method, the condition to be treated, the severity of the condition, whether the condition is to be treated or prevented, and the age, weight, and health of the person to be treated. Additionally, pharmacogenomic (the effect of genotype on the pharmacokinetic, pharmacodynamic or efficacy profile of a therapeutic) information about a particular patient may affect the dosage regimen used.

[0277] In the methods the individual components of a co-administration, or combination, can be administered by any suitable means, contemporaneously, simultaneously, sequentially in either order, separately or in a single pharmaceutical formulation. Where the co-administered compounds or compositions are administered in separate dosage forms, the number of dosages administered per day for each compound may be the same or different. The compounds or compositions may be administered via the same or different routes of administration. The compounds or compositions may be administered according to simultaneous or alternating regimens, at the same or different times during the course of the therapy, concurrently in divided or single forms.

[0278] The term “therapeutically effective amount” refers to that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician, which includes alleviation of the symptoms of the disease or disorder being treated. In one aspect, the therapeutically effective amount is that which may treat or alleviate the disease or symptoms of the disease at a reasonable benefit / risk ratio applicable to any medical treatment. However, it is to be understood that the total daily usage of the compounds and compositions described herein may be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors, including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition employed; the age, body weight, general health, gender and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidentally with the specific compound employed; and like factors well-known to the researcher, veterinarian, medical doctor or other clinician of ordinary skill.

[0279] Depending upon the route of administration, a wide range of permissible dosages are contemplated, including doses falling in the range from about 1 pg / kg to about 1 g / kg. The dosages may be single or divided and may administered according to a wide variety of protocols, including q.d. (once a day), b.i.d. (twice a day), t.i.d. (three times a day), or even every other day, once a week, once a month, once a quarter, and the like. In each of these cases the described therapeutically effective amounts correspond to the instance of administration, or alternatively to the total daily, weekly, month, or quarterly dose, as determined by the dosing protocol.

[0280] An “effective amount” of any one or a mixture of the compounds can be determined by the attending diagnostician or physician by the use of known techniques and / or by observing results obtained under analogous circumstances. In determining the effective amount or dose, a number of factors are considered by the attending diagnostician or physician, including, but not limited to, the species of mammal, including human, its size, age, and general health, the specific disease or disorder involved, the degree of or involvement or the severity of the disease or disorder, the response of the individual patient, the particular compound administered, the mode of administration, the bioavailability characteristics of the preparation administered, the dose regimen selected, the use of concomitant medication, and other relevant circumstances.

[0281] The term “patient” includes human and non-human animals such as companion animals (dogs and cats and the like) and livestock animals. Livestock animals are animals raised for food production. The patient to be treated is preferably a mammal, in particular a human.

[0282] The term “Lewy body” refers to an abnormal intracellular inclusion primarily composed of aggregated, misfolded alpha-synuclein protein, often accompanied by ubiquitin and other cellular components. Lewy bodies are characteristic of certain neurodegenerative disorders, such as Parkinson’s disease and dementia with Lewy bodies, and are typically localized in brain regions associated with motor control, cognition, and other neurological functions.

[0283] The term “alpha-synuclein” (a-syn) refers to a small, soluble protein primarily expressed in the nervous system, where it is abundant in presynaptic terminals. Alpha- synuclein (a-syn) plays a role in synaptic vesicle trafficking and neurotransmitter release under normal physiological conditions. In pathological states, a-syn can misfold and aggregate, forming insoluble fibrils that are the main component of Lewy bodies, a hallmark of neurodegenerative diseases such as Parkinson’s disease and dementia with Lewy bodies.

[0284] The term “Parkinson's disease” refers to a neurodegenerative disorder characterized by the progressive loss of dopaminergic neurons in the substantia nigra pars compacta and the pathological accumulation of alpha-synuclein (a-syn) in the form of Lewy bodies. Parkinson’s disease is associated with motor symptoms such as bradykinesia, rigidity, resting tremor, and postural instability, as well as non-motor symptoms, including but not limited to autonomic dysfunction, cognitive impairment, and mood disorders.

[0285] The term “dementia with Lewy bodies (DLB)” refers to a progressive neurodegenerative disorder characterized by cognitive decline that interferes with daily functioning and is accompanied by fluctuating attention, visual hallucinations, and motor symptoms similar to Parkinson’s disease. Pathologically, DLB is defined by the presence of Lewy bodies — intracellular aggregates primarily composed of misfolded alpha-synuclein (a- syn) — in cortical and subcortical regions of the brain. It is considered one of the most common causes of dementia, distinct from Alzheimer’s disease by its clinical and pathological features.

[0286] The term “multiple system atrophy (MSA)” refers to a rare, progressive neurodegenerative disorder characterized by autonomic dysfunction, parkinsonism, and cerebellar ataxia. Pathologically, MSA is defined by the accumulation of misfolded alpha- synuclein (a-syn) in oligodendrocytes, forming glial cytoplasmic inclusions (GCIs). The disease affects multiple systems, including motor and autonomic pathways, and is subdivided into two main types based on the predominant symptoms: MSA-P (parkinsonian subtype) and MSA-C (cerebellar subtype).

[0287] The term “ AA amyloidosis” refers to a systemic disorder caused by the extracellular deposition of amyloid fibrils composed of misfolded serum amyloid A (SAA) protein. It is often secondary to chronic inflammatory conditions or infections and primarily affects organs such as the kidneys, liver, and spleen, leading to progressive organ dysfunction.

[0288] The term “Alzheimer’s disease” refers to a progressive neurodegenerative disorder characterized by cognitive decline, memory impairment, and behavioral changes. Pathologically, it is defined by the accumulation of extracellular amyloid-beta plaques and intracellular tau neurofibrillary tangles in the brain, along with neuronal loss and synaptic dysfunction.

[0289] The term “monoclonal immunoglobulin light-chain amyloidosis” (AL amyloidosis) refers to a disorder caused by the deposition of misfolded monoclonal light chains derived from clonal plasma cells. These misfolded proteins form insoluble amyloid fibrils that accumulate in various tissues, leading to progressive organ damage, particularly in the heart, kidneys, liver, and peripheral nerves.

[0290] The term “Huntington’s disease” refers to a genetic neurodegenerative disorder caused by an expanded CAG trinucleotide repeat in the HTT gene, leading to the production of mutant huntingtin protein. The disease is characterized by progressive motor dysfunction, psychiatric symptoms, and cognitive decline, with pathological hallmarks including neuronal loss and intracellular inclusions of aggregated huntingtin protein.

[0291] The term “Parkinson's disease” refers to a neurodegenerative disorder characterized by the progressive loss of dopaminergic neurons in the substantia nigra pars compacta and the pathological accumulation of alpha-synuclein (a-syn) in the form of Lewy bodies. Parkinson’s disease is associated with motor symptoms such as bradykinesia, rigidity, resting tremor, and postural instability, as well as non-motor symptoms, including but not limited to autonomic dysfunction, cognitive impairment, and mood disorders.

[0292] The term “Creutzfeldt-Jakob disease” (CID) refers to a rare, fatal neurodegenerative disorder caused by the accumulation of misfolded prion protein (PrPASc), which induces the aggregation of normal prion protein (PrPAC) in the brain. CJD is characterized by rapidly progressive dementia, motor dysfunction, and myoclonus, with spongiform changes observed in the brain tissue upon histological examination.

[0293] The term “prion disorders” refers to a group of fatal neurodegenerative diseases caused by the misfolding and aggregation of prion protein (PrP), leading to the propagation of the pathological isoform (PrPASc). These disorders include Creutzfeldt-Jakob disease, fatal familial insomnia, Gerstmann-Straussler-Scheinker syndrome, and kuru, all of which are characterized by spongiform changes, neuronal loss, and gliosis.

[0294] The term “amyotrophic lateral sclerosis” (ALS) refers to a progressive neurodegenerative disorder characterized by the loss of motor neurons in the brain and spinal cord, leading to muscle weakness, atrophy, and eventual paralysis. Pathological features include the aggregation of proteins such as TDP-43 and, in some cases, SOD1 or FUS, which contribute to motor neuron degeneration.

[0295] The term “type 2 diabetes” refers to a metabolic disorder characterized by chronic hyperglycemia due to insulin resistance and relative insulin deficiency. Pathophysiological features include impaired glucose uptake, pancreatic beta-cell dysfunction, and amyloid deposits composed of islet amyloid polypeptide (JAPP) in the pancreatic islets.

[0296] The term “transthyretin amyloidosis” (ATTR) refers to a systemic disorder caused by the misfolding and aggregation of transthyretin (TTR) protein into amyloid fibrils. ATTR can manifest in hereditary or wild-type forms, with clinical symptoms including polyneuropathy, cardiomyopathy, and progressive organ dysfunction due to amyloid deposition.

[0297] The term “anti -aggregation compound” refers to a substance or molecule that inhibits or prevents the aggregation of proteins, peptides, or other biological macromolecules. Such compounds are designed to interfere with the processes that lead to the misfolding, clustering, or accumulation of these molecules, which are often associated with pathological conditions, including neurodegenerative diseases like Parkinson’s disease, Alzheimer’s disease, and multiple system atrophy. Anti-aggregation compounds may act by stabilizing native protein conformations, disrupting aggregation-prone intermediates, or disassembling existing aggregates. The disclosure also relates to the following numbered Embodiments presented in no particular order of importance:

[0298] 1. A compound of the formula la, lb, or Ic: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein Yi is:

[0299] Z1is N or CH;

[0300] Z2is C, CH or N;

[0301] Z3is N or NRa;

[0302] Z1is O, S, or NRb; provided that Y1is not unsubstituted indolyl when Y2is: one of R'-R4is fluoro, chloro, I-NO2, C1-C2 alkyl or methoxy (OCH3); Raand Rbare each independently H or optionally substituted C1-C20 alkyl; and

[0303] R'-R8and R1-R3are each independently -H, halogen, -OH, -NH2, -NO2, -CN, -CF3, -COOH, -N3, -SO3H, -PO3H2, C1-C20 alkyl, C1-C20 hydroxyalkyl, C1-C20 alkylamino, C1-C20 dialkylamino, C1-20 alkoxy, C(0)0-(C1-C20) alkyl, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, or 3- to 15-membered heterocyclyl.

[0304] 2. The compound of Embodiment 1, wherein the compound is of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0305] Z1is N or CH;

[0306] Z3is O, S, or NRa; provided that Z1is not CH when Z3is NRa;

[0307] Rais H or optionally substituted C1-20 alkyl;

[0308] R'-R3and R5-R8are each independently -H, halogen, -CN, -CF3, -NH2, -NO2, optionally substituted C1-20 alkyl or optionally substituted C1-20 alkoxy.

[0309] 3. The compound of Embodiment 1, wherein the compound is of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0310] Z1is N or CH;

[0311] Z3is O, S, or NRa; provided that Z1is not CH when Z3is NRa;

[0312] Rais H or optionally substituted alkyl;

[0313] Zris O, S, or NRb;

[0314] Raand Rbare each independently H or optionally substituted C1-20 alkyl; and

[0315] R1-R3are each independently H, halogen, -OH, -NBL, -NO2, -CN, -COOH, -Ns, -SO3H, - PO3H2 C1-C20 alkyl, alkoxy, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, C3-C12 cycloalkenyl, or 3- to 15-membered heterocyclyl. 4. The compound of Embodiment 1, wherein the compound is of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0316] Z1is N or CH;

[0317] Z3is O, S, or NRa;

[0318] Rais H or optionally substituted C1-20 alkyl; and

[0319] RkR8are each independently H, halogen, -CF3,-CN, -NO2, or alkoxy.

[0320] 5. The compound of Embodiment 1, wherein the compound is of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0321] Z1is N or CH;

[0322] Z3is O, S, or NRa;

[0323] Rais H or optionally substituted alkyl;

[0324] Z1is O, S, or NRb;

[0325] Raand Rbare each independently H or optionally substituted C1-20 alkyl; and

[0326] R1-R3are each independently H, halogen, -OH, -NH2, -NO2, -CF3, -CN, -COOH, -Ns, -

[0327] SOsH, -PO3H2, C1-C20 alkyl, alkoxy, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, C3-

[0328] C12 cycloalkenyl, or 3- to 15-membered heterocyclyl.

[0329] 6. The compound of Embodiment 1, wherein the compound is of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0330] Yi is 4-amino-indole, 5-amino-indole, 6-amino-indole, or 7-amino-indole; Ri is -H, halogen, -OH, C1-C6 alkyl, C1-C6 alkyl-OH, C1-C6 alkoxy, or -NO2;

[0331] R2is -H, halogen, -OH, C1-C6 alkyl-OH, C1-C6 alkoxy, or -NO2; and

[0332] R3is -H, halogen, -OH, C1-C6 alkyl-OH, C1-C6 alkoxy, or -NO2.

[0333] 7. The compound of Embodiment 6, wherein Y1is optionally substituted with a C1-C6 alkyl or a halogen.

[0334] 8. The compound of Embodiment 6 or 7, wherein the halogen is I.

[0335] 9. The compound of Embodiment 6, wherein:

[0336] Y1is 4-amino-indole;

[0337] R1is -H, and

[0338] R3is -NO2.

[0339] 10. The compound of Embodiment 6, wherein

[0340] Y1is 6-amino-indole,

[0341] R1is -H, and

[0342] R3is -NO2.

[0343] 11. The compound of Embodiment 1, wherein the compound is of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0344] Z1is N or CH

[0345] Z3is O, S, or NRa

[0346] Rais H or optionally substituted C1-20 alkyl; and

[0347] RkR8are each independently H, halogen, -CF3, -CN, -NO2, or alkoxy.

[0348] 12. The compound of Embodiment 1, wherein the compound is of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0349] Z1is N or CH;

[0350] Z3is O, S, or NRa; Z1is O, S, or NRb;

[0351] Raand Rbare each independently H or optionally substituted C1-20 alkyl; and

[0352] R1-R3is H, halogen, -OH, -NIL, -NO2, -CF3, -CN, -COOH, -N3, -SO3H, -PO3H2, C1-C20 alkyl, C1-C20 alkoxy, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, C3-C12 cycloalkenyl, or 3- to 15-membered heterocyclyl.

[0353] 13. The compound of Embodiment 1, wherein the compound is of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein:

[0354] R2and R3are each independently -H, -CN or -NO2;

[0355] R4is C1-C6 hydroxyalkyl, -OH, or C(0)0-(C1-C20) alkyl, and

[0356] R5-R8are each independently -H, -NO2, -NH2, C1-C6 alkylamino, or C1-C6 dialkylamino,

[0357] 14. The compound of Embodiment 1, wherein the compound is of the formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), wherein

[0358] R2and R3are each independently -H, -CN or -NO2;

[0359] R4is C1-C6 hydroxyalkyl, -OH, or C(0)0-(C1-C20) alkyl, and

[0360] R6-R8are each independently -H, -NO2, -NH2, C1-C6 alkylamino, or C1-C6 dialkylamino.

[0361] 15. The compound of Embodiment 14, wherein:

[0362] R2or R3is -CN,

[0363] R4is -OCH3, and

[0364] R5is -H.

[0365] 16. The compound of Embodiment 14, wherein;

[0366] R2or R3is -NO2,

[0367] R4is -OCH3, and

[0368] R5is -H. 17. A compound of the formula:

[0369] or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester).

[0370] 18. A compound of formula: or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester) wherein: the indole group is optionally substitued; and

[0371] Rlb-R3bare each independently alkyl, amino, halo, alkoxy or OH.

[0372] 19. The compound of Embodiment 18, or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), having the formula: The compound of Embodiment 18 or 19, a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester), having the formula: mono- and di-alkylated versions of the amino (NH2) group. A pharmaceutical composition comprising at least one compound of any one of Embodiments 1-20 or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester) and a pharmaceutically acceptable carrier. A method of inhibiting a-synuclein (a-syn) protein aggregation, tau isoform 2N4R protein aggregation, or both in a subject having, or at risk for, said protein aggregation, comprising administering to the subject at least one compound of any one of Embodiments 1-20 or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester) or a pharmaceutical composition of Embodiment 21 in an amount effective to inhibit a-syn protein aggregation, tau isoform 2N4R protein aggregation, or both, whereupon said protein aggregation is inhibited in the subject having, or at risk for, said protein aggregation. The method of Embodiment 22, wherein the subject has, or is at risk for, Alzheimer’s disease. The method of Embodiment 22, wherein the subject has, or is at risk for, Parkinson’s disease or dementia with Lewy bodies (DLB). The method of Embodiment 22, wherein the subject has neuroblastoma and formation of a-syn inclusions is inhibited. 26. A method of inhibiting fibril formation in a subject having, or at risk for, fibril formation, wherein the method comprises administering to the subject at least one compound of any one of Embodiments 1-20 or a pharmaceutically acceptable salt or ester thereof (e.g., methyl or ethyl ester) or a pharmaceutical composition of Embodiment 21 in an amount effective to inhibit fibril formation, whereupon fibril formation is inhibited in the subject having, or at risk for, said fibril formation.

[0373] EXAMPLES

[0374] The following examples serve to illustrate the present disclosure. The examples are not intended to limit the scope of the claims in any way.

[0375] Experimental procedures

[0376] 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 sodium sulfate. The solvents were evaporated on a Biichi rotavapor R-100 equipped with a Biichi V-100 vacuum controller. The nuclear magnetic resonance (NMR) spectra were recorded on a Bruker 500 MHz spectrometer. Proton chemical shifts are reported in parts per million (ppm) with the solvent reference relative to tetramethyl silane (TMS) employed as the internal standard (CDC13, 5 7.26; DMSO-d6 5 2.54). The multiplicities of NMR signals are designated as s (singlet), d (doublet), dd (double doublet), t (triplet), q (quartet), br (broad), 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 (mp) were scored using an electrothermal apparatus (Barnstead International, Dubuque, Iowa, USA).

[0377] Chemical and protein source

[0378] Thioflavin-T (ThT) were purchased from Alfa Aesar (Ward Hill, MA) for the a-syn ThT assays. Heparin sodium salt was purchased from Millipore-Sigma. Procuration of recombinant a-syn and tau 2N4R was from rPeptide (WatKinsville, GA). Concerning the preparation of tau 0N4R, a bacterial expression plasmid consisting of the vector pET30a carrying a cDNA encoding the human Tau 0N4R isoform was a kind gift of Dr. Benjamin Wolozin (Boston University). E. coli stock (Rosetta BL21 Ecoli (CamR) containing pET30a[0N4R tau wt] (KanR)) were grown in LB media supplemented with kanamycin (50 pg / mL) and chloramphenicol (50 pg / mL). Protein over-expression was induced by the addition of 1 mM IPTG for ~18 h at 37°C, and cells were pelleted by centrifugation at 6,000 g for 15 min at 4°C. The cells were resuspended in lysis buffer (10 mM Hepes pH 7.4, 50 mM NaCl, 1 mM MgCh, 1 mM PMSF, IX PIC, 0.5 mM DTT) and lysed by sonication at 30 sec On 1 min Off at ~30- 45% power for ~3-5 minutes. Lysate was centrifuged at 10000 rpm for 10 minutes at 4°C and supernatant was transferred with 7.8 ml (total) of 3M NaCl. The lysate was incubated for 10 min in 80°C water bath, then cooled for 10 min in an ice bath. The lysate was centrifuge at 10000 rpm, 4°C for 10 minutes and supernatant was transferred to new tubes. The supernatant was dialyzed overnight against cation exchange buffer (50 mM MES, IM NaCl, 1 mM DTT, pH 6.0). The dialysate was loaded onto a HiPrep SP HP column, and proteins were eluted with a linear gradient ranging from 50 mM to 1 M NaCl. Fractions containing tau isoform 0N4R were pooled, and the resulting protein solution was dialyzed against PBS (pH 7.4) and stored at -80°C.

[0379] Thioflavin T (ThT) fluorescence assays

[0380] A technique commonly used to track the kinetics of a-syn fibril formation in response to various drug candidate treatments is the ThT fluorescence test.36,37The final concentration of the tested compounds was 100 pM while ThT was applied at a final concentration of 20 pM. The recombinant a-syn obtained from rPeptide was been validated with proper quality control to confirm the monomeric state of the protein. a-Syn was dissolved in 20 mM Tris-HCl (pH 7.4) supplemented with 100 mM NaCl to a stock solution of 280 pM prior to resuspension in ThT buffer to obtain a final concentration of 2 pM (in each well). Compounds and ThT were first added to the wells. The kinetics of fibril formation begin when the a-syn is solubilized in the ThT buffer (10 mM PBS buffer (pH 7.4), supplemented with 0.5 mM SDS and 300 mM NaCl) and added to a non-treated black 96 well microplate with a transparent flat bottom. Each well was filled with a maximum volume of 150 pL buffer with one 3 mm borosilicate bead.38The background fluorescence signal consisted of ThT in buffer and 0.25% DMSO without a- syn. The excitation and emission wavelengths consisted of 440 and 485 nm, respectively. Measurements were acquired with Synergy HT multi-mode microplate reader (BioTek, Winooski, VT) and taken at 37°C every 20 min with 10 sec shaking prior to reading the plate. Kinetics were of 40 to 70 hours duration. Samples were measured in three replicates. The experiments were repeated at least two to three additional times using different a-syn stock solutions and a final concentration of 2 pM. For each time point, arbitrary units of fluorescence were calculated from the mean values normalized against the maximum value. The percentage of fluorescence intensity at the plateau phase in Table 1-3 were expressed as mean ± SEM. Concerning the dose response curve depicted in Fig. 4, the data were plotted using GraphPad Prism.

[0381] Photo-induced cross-linking of unmodified proteins (PICUP) assay a-syn, 2N4R, and 0N4R tau isoforms were diluted in 10 mM phosphate buffer to achieve final concentrations of 60 pM for u-syn and 10 pM for the tau isoforms.392 pL of Ru(bpy) (2 pL, 300 pM final concentration) and ammonium persulfate (2 pL, 6 mM final concentration) were added to start the cross-linking process. To verify the progressive impact of our compounds on the suppression of protein oligomerization, controls that were not exposed to light and contained no Ru (bpy) were used (i.e., 0.125% DMSO). The samples underwent quick irradiation using a 53 W (120 V) incandescent bulb in a homemade dark box, and light exposures of 1 sec and 60 sec for the a-syn and tau isoforms, respectively. The ultimate capacity of each tube was 20 pL. Laemmli loading buffer (8.3 pL) containing 15% 2- mercaptoethanol was added to the solution right away after light exposure, and the mixture was then incubated at 95 °C for 10 minutes. On a 16% SDS-PAGE gel, the cross-linked samples were separated, and Coomassie blue staining was used for visualization.

[0382] Transmission Electron Microscopy

[0383] Tau isoform 2N4R (6 pM) mixed with DMSO (0.25%) was incubated in a 10 mM PBS buffer (pH 7.4) for 72 hours at 37 °C. a-syn samples (at 2 pM) recovered from kinetic investigations of ThT fibril formation were also examined. A volume of 10 pL of each sample was put to a 400-mesh Formvar-carbon-coated copper grid (Electron Microscopy Sciences, Hatfield, PA) to prepare the grids for TEM analysis. The sample was incubated with the grids for one minute before being rinsed three times with distilled water. A new solution of 1% uranyl acetate was then applied for one minute after being air-dried. Filter paper was used to absorb the solution, then grids were dried in the air. A transmission electron microscope (JEOL 1400 Flash, Japan) was used to analyze the grids, and micrographs were taken at a magnification of 40k and an acceleration voltage of 100 kV. a-Synuclein inclusion-forming neuroblastoma cell experiment

[0384] M17D-TR / aS-3K::YFP neuroblastoma cells that are doxycycline(dox)-induced have been previously used for this assay.40Compounds were introduced 24 hours after the cells had been plated in 96-well plates at a density of 30,000 cells per well. aS-3K::YFP transgene expression was stimulated 24 hours later by adding culture medium dox at a final concentration of 1 pg per mL. The Incucyte Zoom 2000 platform (Essen Biosciences) was used to incubate the cells, while taking continuous green bright field pictures. After 48 hours of induction (96 hours after plating), inclusion formation or growth endpoint analysis was carried out. The Incucyte processing definition “Inclusions” was developed as follows: parameters, fixed threshold, threshold (GCU) 50; edge split on; edge sensitivity 100; cleanup; hole fill (m2): 10; adjust size (pixels); filters; area (m2): max 50; mean intensity: min 60; integrated intensity: min 2000. The following processing definition of "Cells" was used to measure cell confluence: parameters, segmentation adjustment 0.7; Cleanup, all parameters are set to 0; filters, area (m2): min 345.00. A polyclonal antibody to GAPDH (Sigma-Aldrich, St. Louis, MO, G9545; 1 :5000) and an S-specific monoclonal antibody 4B12 (Thermofisher, Waltham, MA; 1 : 1000) were used, as previously described, for the evaluation of protein expression by SDS-PAGE and Western blotting in the LiCor system.41

[0385] Example 1

[0386] Design of Compounds

[0387] Structural hybridization of two biologically active pharmacophores has been widely used as a common strategy to build up new hybrid scaffolds capable of binding to multitarget proteins.32 35The methoxy benzothiazole pharmacophore from N744 dye was selected to guarantee high affinity and inhibitory activity toward the / Lsheet structures of 2N4R tau aggregates. On the other hand, structural analysis of Anlel38b showed that the phenyl ring substituted with the electron-withdrawing group (EWG) bromine atom has the highest activity against a-syn aggregates.25Therefore, compounds were designed by integrating the 4-methoxy benzothiazole moiety and various EWG-phenyl rings into one hybrid molecule. The two pharmacophores were separated by three different linkers: urea, amide, and sulfonamide. The most promising candidates were further optimized by replacing the benzothiazole ring with 4-

[0388] 5-, and 6-substituted indole scaffolds as shown in Fig. 1.

[0389] Example 2 Synthesis

[0390] The designed compounds were synthesized using the commercially available 2-amino- 4-methoxy benzothiazole (4-MBT). To prepare compounds with the urea linker, a quantitative amount of the appropriate phenyl isocyanate was used in dichloromethane (DCM). The amide or the sulfonamide counterparts were synthesized using the corresponding benzoyl chloride or benzene sulfonyl chloride, respectively, in the presence of anhydrous potassium carbonate and pyridine (Fig. 2). In addition, the unsubstituted, electron-releasing, and 2-thiophenyl- containing counterparts were prepared (Table 3). Then, the best candidates, with the 2-iodo and the 4-nitro sulfonamides and their urea analogs, were further modified by replacing the 4- MBT with 4-, 5-, or 6-amino indole (Al) using the same reaction conditions (Fig. 3). All products were obtained with moderate to excellent yields (61-93%).

[0391] General procedure for the synthesis of compounds with the urea linker (1-14, 43-48)

[0392] 4-Methoxy-2-amino benzothiazole, 4-, 5-, or 6-amino indole (100 mg, 1.0 eq), and the appropriate phenyl isocyanate (1.0 eq) were dissolved in di chloromethane (DCM, 10 mL) and stirred for 4-12 h at room temperature. The precipitate was then filtered and washed with hexane (10 mL), DCM (2 x 10 mL) and diethyl ether (10 mL). The desired products were obtained in excellent to quantitative yields. l-(2-Fluorophenyl)-3-(4-methoxybenzo[J|thiazol-2-yl)urea (1). Starting with 4- methoxy-2-amino benzothiazole and 2-fluoro phenyl isocyanate. 92%, white solid;1H NMR (500 MHz, DMSO) δ 11.01 (s, 1H), 9.13 (s, 1H), 8.12 - 8.08 (m, 1H), 7.47 (dd, J= 7.9, 1.0 Hz, 1H), 7.30 - 7.25 (m, 1H), 7.23 - 7.15 (m, 2H), 7.14 - 7.05 (m, 1H), 6.95 (dd, J= 8.0, 1.0 Hz, 1H).13C NMR (126 MHz, DMSO) δ 157.9, 153.9, 151.9, 133.3, 126.8, 126.7, 125.2, 125.2,

[0393] 124.5, 121.8, 115.8, 115.7, 113.9, 108.3, 56.2. Melting Point: 209.5 - 211.3°C. l-(4-Fluorophenyl)-3-(4-methoxybenzo[J|thiazol-2-yl)urea (2). Starting with 4- methoxy-2-amino benzothiazole and 4-fluoro phenyl isocyanate. 87%, white solid;1H NMR (500 MHz, DMSO) δ 10.86 (s, 1H), 9.16 (s, 1H), 7.55 - 7.48 (m, 2H), 7.48 - 7.42 (m, 1H), 7.21 - 7.12 (m, 3H), 6.95 (dd, J= 8.1, 1.0 Hz, 1H), 3.89 (s, 3H).13C NMR (126 MHz, DMSO) δ 159.4, 157.5, 151.6, 135.3, 133.1, 124.3, 121.4, 121.4, 116.0, 115.9, 113.9, 108.3, 56.2. Melting Point: 201.0 - 202.3 °C. l-(2,4-Difluorophenyl)-3-(4-methoxybenzo[J]thiazol-2-yl)urea (3). Starting with 4- methoxy-2-amino benzothiazole and 2,4-difluoro phenyl isocyanate. 94%, white solid; 'H NMR (500 MHz, DMSO) δ 11.01 (s, 1H), 9.08 (s, 1H), 8.09 - 7.95 (m, 1H), 7.46 (dd, J= 8.0, 0.9 Hz, 1H), 7.38 - 7.32 (m, 1H), 7.18 (t, J= 8.0 Hz, 1H), 7.13 - 7.03 (m, 1H), 6.95 (d, J= 7.2 Hz, 1H), 3.89 (s, 3H).13C NMR (126 MHz, DMSO) δ 157.3, 154.3, 151.8, 133.2, 124.5, 123.5, 123.2, 113.9, 111.8, 111.7, 108.3, 104.7, 104.5, 104.3, 56.2. Melting Point: 220.8 - 225.0 °C. l-(4-Chlorophenyl)-3-(4-methoxybenzo[J|thiazol-2-yl)urea (4). Starting with 4- methoxy-2-amino benzothiazole and 4-chloro phenyl isocyanate. 82%, white solid;1H NMR (500 MHz, DMSO) δ 10.94 (s, 1H), 9.27 (s, 1H), 7.53 (d, J= 8.8 Hz, 2H), 7.45 (dd, J= 7.9, 0.9 Hz, 1H), 7.36 (d, J= 8.8 Hz, 2H), 7.18 (t, J= 8.0 Hz, 1H), 6.95 (dd, J= 8.1, 1.0 Hz, 1H), 3.89 (s, 3H).13C NMR (126 MHz, DMSO) δ 158.9, 158.5, 152.5, 151.5, 138.0, 133.0, 129.2, 127.0, 124.4, 121.8, 121.0, 113.9, 108.3, 56.2. Melting Point: 238.9 - 240.5°C. l-(3,5-Dichlorophenyl)-3-(4-methoxybenzo[J|thiazol-2-yl)urea (5). Starting with 4- methoxy-2-amino benzothiazole and 2,4-dichloro phenyl isocyanate. 91%, white solid; 'H NMR (500 MHz, DMSO) δ 11.38 (s, 1H), 9.47 (s, 1H), 7.64 - 7.57 (m, 2H), 7.44 (d, J = 8.0 Hz, 1H), 7.31 - 7.12 (m, 2H), 6.96 (d, J = 8.0 Hz, 1H), 3.90 (s, 3H).13C NMR (126 MHz, DMSO) δ 152.5, 151.1, 142.3, 141.8, 134.6, 132.6, 124.5, 122.4, 117.6, 117.2, 114.0, 108.4, 56.2. Melting Point: 246.5 - 247.8°C. l-(4-Bromophenyl)-3-(4-methoxybenzo[J|thiazol-2-yl)urea (6). Starting with 4- methoxy-2-amino benzothiazole and 4-bromo phenyl isocyanate. 87%, white solid;1H NMR (500 MHz, DMSO) δ 10.95 (s, 1H), 9.27 (s, 1H), 7.48 (s, 4H), 7.45 (dd, J= 8.0, 1.0 Hz, 1H), 7.18 (t, J = 8.0 Hz, 1H), 6.95 (dd, J= 8.1, 1.0 Hz, 1H), 3.89 (s, 3H).13C NMR (126 MHz, DMSO) δ 158.4, 152.5, 151.5, 138.5, 133.0, 132.1, 124.4, 121.5, 121.4, 115.0, 113.9, 108.3, 56.2. Melting Point: 276.8 - 279.1°C. l-(2-Iodophenyl)-3-(4-methoxybenzo[J|thiazol-2-yl)urea (7). Starting with 4- methoxy-2-amino benzothiazole and 2-iodo phenyl isocyanate. 71%, white solid;1H NMR (500 MHz, DMSO) δ 11.55 (s, 1H), 8.47 (s, 1H), 7.87 (dd, J = 7.9, 1.5 Hz, 1H), 7.82 (dd, J = 8.2, 1.5 Hz, 1H), 7.49 - 7.34 (m, 2H), 7.19 (t, J= 8.0 Hz, 1H), 6.99 - 6.87 (m, 2H), 3.90 (s, 3H).13C NMR (126 MHz, DMSO) δ 158.1, 152.2, 151.9, 139.6, 139.3, 133.3, 129.3, 126.7, 124.4, 124.2, 113.9, 108.4, 92.8, 56.3. Melting Point: 224.0 - 225.2°C. l-(4-Iodophenyl)-3-(4-methoxybenzo[J|thiazol-2-yl)urea (8). Starting with 4- methoxy-2-amino benzothiazole and 4-iodo phenyl isocyanate. 85%, white solid;1H NMR (500 MHz, DMSO) δ 10.93 (s, 1H), 9.24 (s, 1H), 7.64 (d, J= 8.7 Hz, 2H), 7.49 - 7.41 (m, 1H), 7.35 (d, J= 8.7 Hz, 2H), 7.18 (t, J = 8.0 Hz, 1H), 6.95 (d, J= 8.0 Hz, 1H), 3.89 (s, 3H).13C NMR (126 MHz, DMSO) δ 158.4, 152.4, 151.5, 138.9, 138.0, 133.0, 127.2, 124.4, 121.6, 113.9, 108.3, 86.7, 56.2. Melting Point: 215.2 - 217.0°C. l-(4-Methoxybenzo[J]thiazol-2-yl)-3-(3-(trifluoromethyl)phenyl)urea (9). Starting with 4-methoxy-2-amino benzothiazole and 3 -trifluoromethyl phenyl isocyanate. 91%, white solid;1H NMR (500 MHz, DMSO) δ 11.12 (s, 1H), 9.51 (s, 1H), 8.01 (s, 1H), 7.69 (d, J= 8.2 Hz, 1H), 7.55 (t, J= 8.0 Hz, 1H), 7.45 (d, J= 7.9 Hz, 1H), 7.38 (d, J= 7.7 Hz, 1H), 7.19 (t, J = 8.0 Hz, 1H), 6.96 (d, J= 7.9 Hz, 1H), 3.89 (s, 3H).13C NMR (126 MHz, DMSO) δ 166.7, 151.7, 140.0, 130.6, 130.2, 129.9, 125.7, 124.4, 123.5, 123.1, 119.7, 115.4, 113.9, 108.3, 56.2. l-(4-Methoxybenzo[J|thiazol-2-yl)-3-(4-(trifluoromethyl)phenyl)urea (10).

[0394] Starting with 4-methoxy-2-amino benzothiazole and 4-trifluoromethyl phenyl isocyanate. 97%, white solid;1H NMR (500 MHz, DMSO) δ 11.03 (s, 1H), 9.53 (s, 1H), 7.72 (d, J = 8.4 Hz, 2H), 7.67 (d, J= 8.4 Hz, 2H), 7.46 (d, J= 7.9 Hz, 1H), 7.19 (t, J= 8.0 Hz, 1H), 6.96 (d, J = 7.9 Hz, 1H), 3.90 (s, 3H).13C NMR (126 MHz, DMSO) δ 142.8, 140.6, 140.0, 128.1, 126.7,

[0395] 126.0, 124.5, 123.4, 121.7, 119.2, 113.9, 108.3, 56.2. l-(4-Cyanophenyl)-3-(4-methoxybenzo[J|thiazol-2-yl)urea (11). Starting with 4- methoxy-2-amino benzothiazole and 4-cyano phenyl isocyanate. 88%, white solid;1H NMR

[0396] (500 MHz, DMSO) δ 11.18 (s, 1H), 9.62 (s, 1H), 7.76 (d, J= 8.8 Hz, 2H), 7.70 (d, J= 8.8 Hz,

[0397] 2H), 7.49 - 7.36 (m, 1H), 7.19 (t, J= 8.0 Hz, 1H), 6.96 (dd, J= 8.0, 0.9 Hz, 1H), 3.90 (s, 3H).13C NMR (126 MHz, DMSO) δ 151.3, 143.6, 133.8, 132.7, 126.8, 124.5, 119.6, 119.4, 119.3, 114.0, 108.3, 104.9, 56.2. Melting Point: 252.4 - 252.9°C. l-(4-Methoxybenzo[J|thiazol-2-yl)-3-(2-nitrophenyl)urea (12). Starting with 4-methoxy-2- amino benzothiazole and 2-nitro phenyl isocyanate. 99%, white solid;1H NMR (500 MHz, DMSO) δ 12.17 (s, 1H), 9.92 (s, 1H), 8.28 (d, J= 8.4 Hz, 1H), 8.12 (d, J= 6.7 Hz, 1H), 7.79

[0398] - 7.69 (m, 1H), 7.47 (d, J= 7.9 Hz, 1H), 7.28 (t, J= 8.0 Hz, 1H), 7.20 (t, J= 8.0 Hz, 1H), 6.96 (d, J= 8.0 Hz, 1H), 3.90 (s, 3H).13C NMR (126 MHz, DMSO) δ13C NMR (126 MHz, DMSO) δ 138.9, 135.6, 134.0, 128.9, 126.0, 124.6, 124.0, 123.6, 118.9, 115.1, 113.9, 108.4, 56.3. l-(4-Methoxybenzo[J|thiazol-2-yl)-3-(3-nitrophenyl)urea (13). Starting with 4- methoxy-2-amino benzothiazole and 3-nitro phenyl isocyanate. 92%, white solid;1H NMR (500 MHz, DMSO) δ 11.28 (s, 1H), 9.65 (s, 1H), 8.56 (s, 1H), 7.87 - 7.81 (m, 2H), 7.58 (t, J=

[0399] 8.2 Hz, 1H), 7.44 (d, J= 7.9 Hz, 1H), 7.18 (t, J = 8.0 Hz, 1H), 6.95 (d, J = 8.0 Hz, 1H), 3.90 (s, 3H).13C NMR (126 MHz, DMSO) δ 159.0, 153.2, 151.3, 148.6, 140.5, 132.7, 130.6, 125.5, 124.5, 117.7, 113.9, 113.4, 110.3, 108.3, 56.2. Melting Point: 266.0 - 266.7°C. l-(4-Methoxybenzo[J|thiazol-2-yl)-3-(4-nitrophenyl)urea (14). Starting with 4- methoxy-2-amino benzothiazole and 4-nitro phenyl isocyanate. 100%, off-white solid; 'H NMR (500 MHz, DMSO) δ 11.33 (s, 1H), 9.84 (s, 1H), 8.20 (d, J= 8.7 Hz, 2H), 7.76 (d, J = 8.7 Hz, 2H), 7.45 (d, J = 8.0 Hz, 1H), 7.20 (t, J = 8.0 Hz, 1H), 6.97 (d, J= 8.1 Hz, 1H), 3.90 (s, 3H).13C NMR (126 MHz, DMSO) δ 151.2, 145.8, 142.2, 132.7, 125.5, 124.6, 121.9, 118.8,

[0400] 114.0, 113.8, 108.6, 108.4, 56.2. Melting Point: 279.1 - 280.4°C. l-(lH-Indol-4-yl)-3-(2-iodophenyl)urea (43). Starting with 4-amino indole and 2- iodo phenyl isocyanate. 95%, white solid; *H NMR (500 MHz, DMSO) δ 11.11 (s, 1H), 9.08 (s, 1H), 8.21 (s, 1H), 7.84 (d, J = 8.2 Hz, 1H), 7.80 (d, J = 7.4 Hz, 1H), 7.61 (d, J= 7.6 Hz, 1H), 7.37 - 7.32 (m, 1H), 7.30 (t, J= 2.8 Hz, 1H), 7.06 (d, J= 8.0 Hz, 1H), 6.99 (t, J= 7.9 Hz, 1H), 6.89 - 6.78 (m, 1H), 6.62 (s, 1H).13C NMR (126 MHz, DMSO) δ 153.1, 140.5, 139.4, 137.0, 131.8, 129.0, 125.6, 124.5, 124.2, 122.0, 119.7, 108.6, 106.6, 98.6, 92.1. Melting Point:

[0401] 207.0 - 208.4°C. l-(lH-Indol-5-yl)-3-(2-iodophenyl)urea (44). Starting with 5-amino indole and 2- iodo phenyl isocyanate. 92%, white solid;1(H50 N0M MRHz, DMSO) δ 10.93 (s, 1H), 9.18 (s, 1H), 7.87 (d, J= 8.2 Hz, 1H), 7.84 - 7.54 (m, 3H), 7.43 - 7.17 (m, 3H), 7.07 (d, J= 8.8 Hz, 1H), 6.87 - 6.69 (m, 1H), 6.34 (s, 1H).13C NMR (126 MHz, DMSO) δ 153.2, 140.7, 139.4, 132.8, 131.7, 129.0, 128.2, 126.3, 125.0, 123.1, 115.1, 111.8, 110.5, 101.4, 91.2. Melting Point:

[0402] 206.1 - 207.9°C. l-(lH-Indol-6-yl)-3-(2-iodophenyl)urea (45). Starting with 6-amino indole and 2- iodo phenyl isocyanate. 100%, white solid; *HNMR (500 MHz, DMSO) δ 10.93 (s, 1H), 9.31 (s, 1H), 7.88 (d, J= 8.2 Hz, 1H), 7.81 (t, J= 6.7 Hz, 3H), 7.42 (d, J = 8.4 Hz, 1H), 7.33 (s, 1H), 7.21 (t, .7= 2.8 Hz, 1H), 6.87 (dd, J= 8.4, 1.9 Hz, 1H), 6.81 (d, J = 7.5 Hz, 1H), 6.33 (t, J= 2.6 Hz, 1H).13C NMR (126 MHz, DMSO) δ 153.0, 140.6, 139.4, 136.7, 134.1, 129.0, 125.2, 125.0, 123.7, 123.2, 120.5, 112.3, 101.7, 101.4, 91.4. Melting Point: 213.1 - 215.3°C. l-(lH-Indol-4-yl)-3-(4-nitrophenyl)urea (46). Starting with 4-amino indole and 4- nitro phenyl isocyanate. 89%, yellow solid;1H NMR (500 MHz, DMSO) δ 11.16 (s, 1H), 9.60 (s, 1H), 8.71 (s, 1H), 8.19 (d, J= 9.2 Hz, 2H), 7.72 (d, J = 92 Hz, 2H), 7.67 - 7.60 (m, 1H), 7.36 - 7.29 (m, 1H), 7.11 - 7.09 (m, 1H), 7.02 (t, J= 7.9 Hz, 1H), 6.55 (s, 1H).13C NMR (126 MHz, DMSO) δ 152.4, 146.9, 141.4, 137.0, 131.2, 125.7, 124.8, 122.0, 119.7, 117.7, 108.5, 107.1, 98.2. HRMS m / z: calcd for CI5HI2N4O2, 296.0909; found, 296.0988, M + H+. Melting Point: 223.5 - 224.1 °C. l-(lH-Indol-5-yl)-3-(4-nitrophenyl)urea (47). Starting with 5-amino indole and 4- nitro phenyl isocyanate. 86%, white solid; (15H00 N MMHRz, DMSO) δ 10.98 (s, 1H), 9.34 (s, 1H), 8.65 (s, 1H), 8.17 (d, J= 8.1 Hz, 2H), 7.86 - 7.54 (m, 3H), 7.45 - 7.18 (m, 2H), 7.11 (d, J= 8.6 Hz, 1H), 6.37 (s, 1H).13C NMR (126 MHz, DMSO) δ 152.8, 147.4, 141.2, 133.1, 131.1, 128.2, 126.4, 125.6, 117.7, 115.5, 111.8, 111.1, 101.5. HRMS m / z: calcd for CI5HI2N4O2, 296.0909; found, 296.0992, M + H+. Melting Point: 248.5 - 250.1°C. l-(lH-Indol-6-yl)-3-(4-nitrophenyl)urea (48). Starting with 6-amino indole and 4- nitro phenyl isocyanate. 79%, canary yellow solid; *HNMR (500 MHz, DMSO) δ 10.96 (s, 1H), 9.36 (s, 1H), 8.80 (s, 1H), 8.18 (d, J= 92 Hz, 2H), 7.80 (s, 1H), 7.69 (d, J= 92 Hz, 2H), 7.43 (d, J= 8.4 Hz, 1H), 7.23 (t, J= 2.7 Hz, 1H), 6.90 (dd, J= 8.4, 2.0 Hz, 1H), 6.34 (s, 1H).13C NMR (126 MHz, DMSO) δ 152.6, 147.2, 141.2, 136.6, 133.4, 125.6, 125.2, 124.1, 120.5, 117.7, 112.7, 102.1, 101.4. HRMS m / z: calcd for CI5HI2N4O2, 296.0909; found, 296.0993, M + H+. Melting Point: 247.3 - 248.9°C.

[0403] General procedure for the synthesis of compounds with the urea linker (55-58)

[0404] 4-Methoxy-2-amino benzothiazole (100 mg, 1.0 eq), and the un / substituted phenyl isocyanate or 2-thienyl isocyanate (1.0 eq) were dissolved in dichloromethane (DCM, 10 mL) and stirred for 4-12 h at room temperature. The precipitate was then filtered and washed with hexane (10 mL), DCM (2 x 10 mL) and diethyl ether (10 mL). The desired products were obtained in excellent to quantitative yields. l-(4-Methoxybenzo[J|thiazol-2-yl)-3-phenylurea (55). Starting with 4-methoxy-2- amino benzothiazole and phenyl isocyanate, 81%, white solid; (500 M1HHz N,M DMR SO) δ 10.85 (s, 1H), 9.18 (s, 1H), 7.53 - 7.47 (m, 2H), 7.45 (d, J= 7.9 Hz, 1H), 7.35 - 7.27 (m, 2H), 7.18 (t, J= 8.0 Hz, 1H), 7.04 (t, J= 7.46Hz, 1H), 6.95 (d, J= 7.9 Hz, 1H), 3.89 (s, 3H).13C NMR (126 MHz, DMSO) δ 158.3, 151.7, 142.2, 138.9, 133.3, 129.4, 127.5, 124.3, 123.4, 119.4, 113.9, 108.3, 56.2. Melting Point: 207.7- 208.9 °C. l-(4-Methoxybenzo[J|thiazol-2-yl)-3-(2-methoxyphenyl)urea (56). Starting with 4- methoxy-2-amino benzothiazole and 2-methoxy phenyl isocyanate. 86%, white solid;1H NMR (500 MHz, DMSO) δ 11.24 (s, 1H), 8.95 (s, 1H), 8.11 (dd, J = 8.0, 1.6 Hz, 1H), 7.46 (dd, J = 8.0, 1.0 Hz, 1H), 7.18 (t, J= 7.9 Hz, 1H), 7.08 - 6.98 (m, 2H), 6.98 - 6.89 (m, 2H), 3.90 (s, 3H), 3.88 (s, 3H).13C NMR (126 MHz, DMSO) δ 158.1, 151.8, 148.6, 139.27, 139.26, 133.3, 127.9, 124.4, 123.6, 121.1, 119.3, 113.9, 111.5, 108.4, 56.33, 56.28. Melting Point: 200.1 -

[0405] 201.5 °C. l-(4-Methoxybenzo[J|thiazol-2-yl)-3-(2-methylphenyl)urea (57). Starting with 4- methoxy-2-amino benzothiazole and 2-methyl phenyl isocyanate. 84%, white solid; 'H NMR (500 MHz, DMSO) δ 11.12 (s, 1H), 8.85 (s, 1H), 7.85 (d, J= 8.0 Hz, 1H), 7.46 (dd, J= 8.0, 0.9 Hz, 1H), 7.24 - 7.15 (m, 3H), 7.01 (t, J= 7.1 Hz, 1H), 6.95 (d, J= 7.1 Hz, 1H), 3.89 (s, 3H), 2.28 (s, 3H).13C NMR (126 MHz, DMSO) δ 158.6, 152.0, 151.8, 139.3, 136.8, 133.0,

[0406] 130.8, 128.7, 126.8, 124.4, 124.2, 121.7, 113.9, 108.4, 56.3, 18.2. l-(4-Methoxybenzo[J|thiazol-2-yl)-3-(thiophen-2-yl)urea (58). Starting with 4- methoxy-2-amino benzothiazole and 2-thienyl isocyanate. 89%, off-white solid;JH NMR (500 MHz, DMSO) δ 11.19 (s, 1H), 10.16 (s, 1H), 7.49 - 7.41 (m, 1H), 7.19 (t, J= 8.0 Hz, 1H), 7.00 - 6.91 (m, 2H), 6.84 (dd, J= 5.5, 3.7 Hz, 1H), 6.70 (dd, J= 3.7, 1.5 Hz, 1H), 3.89 (s, 3H).13C NMR (126 MHz, DMSO) δ 152.3, 151.3, 151.2, 140.4, 132.6, 124.9, 124.4, 117.5, 114.0, 111.8, 108.3, 56.2. Melting Point: 175.9 - 177.4°C

[0407] General procedure for the synthesis of compounds with the amide or sulfonamide linker (15-42, 49-54)

[0408] The appropriate benzoyl chloride or its corresponding benzene sulfonyl chloride (1.0 eq) was added slowly to a stirred solution of 4-methoxy-2-amino benzothiazole, 4-, 5-, or 6- amino indole (100 mg, 1.0 eq) in pyridine (3 mL) at 0°C. Potassium carbonate (1.5 eq) was added, and the reaction was allowed to stir overnight at room temperature. After completion, an aqueous solution of IN hydrochloric acid (7 mL) and dichloromethane (10 mL) was added to the reaction mixture and extracted. The organic layers were then washed with saturated solutions of ammonium chloride (7 mL), sodium bicarbonate (7 mL), and brine (7 mL), respectively. Organic layers were collected, filtered over anhydrous magnesium sulfate, and concentrated in-vacuo. The crude was purified using column chromatography to obtain the desired products with moderate to good yields.

[0409] 2-Fluoro-7V-(4-methoxybenzo[J|thiazol-2-yl)benzamide (15). Starting with 4- methoxy-2-amino benzothiazole and 2-fluoro benzoyl chloride. 58%, white solid;1H NMR

[0410] (500 MHz, CDCI3) δ 8.32 - 8.11 (m, 1H), 7.63 - 7.56 (m, 1H), 7.44 (dd, J= 8.0, 0.9 Hz, 1H),

[0411] 7.35 (d, J= 7.8, 1H), 7.30 (t, J= 8.0 Hz, 1H), 7.43 - 7.21 (m, 1H), 6.92 (dd, J= 8.0, 0.9 Hz, 1H), 4.04 (s, 3H).13C NMR (126 MHz, CDCI3) δ 161.3, 156.7, 152.1, 138.2, 135.2, 133.6, 132.3, 125.3, 125.3, 125.1, 116.7, 116.5, 113.5, 106.8, 55.9. Melting Point: 178.4 - 180.8°C.

[0412] 4-Fluoro-7V-(4-methoxybenzo[J|thiazol-2-yl)benzamide (16). Starting with 4- methoxy-2-amino benzothiazole and 4-fluoro benzoyl chloride. 67%, white solid;1H NMR (500 MHz, DMSO) δ 8.29 - 8.16 (m, 2H), 7.54 (dd, J= 8.0, 0.9 Hz, 1H), 7.39 (t, J= 8.8 Hz, 2H), 7.27 (t, J= 8.0 Hz, 1H), 7.00 (dd, J= 8.0, 0.9 Hz, 1H), 3.92 (s, 3H).13C NMR (126 MHz, DMSO) δ 172.5, 165.1, 157.7, 152.3, 133.3, 131.7, 131.6, 125.2, 116.3, 116.1, 113.9, 108.0, 56.1.

[0413] 2,4-Difluoro-7V-(4-methoxybenzo[J|thiazol-2-yl)benzamide (17). Starting with 4- methoxy-2-amino benzothiazole and 2,4-difluoro benzoyl chloride. 75%, white solid;1H NMR (500 MHz, DMSO) δ 7.88 (td, J= 8.5, 6.5 Hz, 1H), 7.55 (dd, J= 8.0, 0.9 Hz, 1H), 7.50 - 7.39 (m, 1H), 7.32 - 7.21 (m, 2H), 7.01 (dd, J= 8.1, 1.0 Hz, 1H), 3.91 (s, 3H).13C NMR (126 MHz, DMSO) δ 163.0, 159.8, 156.8, 152.4, 138.8, 133.3, 132.8, 125.4, 119.3, 119.2, 114.0, 112.4, 108.1, 105.5, 56.2. Melting Point: 147.6 - 149.1°C.

[0414] 4-Chloro-N -(4-methoxybenzo[J|thiazol-2-yl)benzamide (18). Starting with 4- methoxy-2-amino benzothiazole and 4-chloro benzoyl chloride. 81%, white solid;1H NMR (500 MHz, DMSO) δ 8.15 (d, J = 8.6 Hz, 2H), 7.62 (d, J= 8.6 Hz, 2H), 7.54 (dd, J= 8.0, 0.9 Hz, 1H), 7.27 (t, J = 8.0 Hz, 1H), 7.01 (dd, J= 8.1, 1.0 Hz, 1H), 3.92 (s, 3H).13C NMR (126 MHz, DMSO) δ 165.2, 152.4, 138.3, 133.4, 131.2, 131.1, 130.7, 129.3, 125.2, 120.8, 113.9,

[0415] 108.0, 56.1.

[0416] 3,5-Dichloro-N -(4-methoxybenzo[J|thiazol-2-yl)benzamide (19). Starting with 4- methoxy-2-amino benzothiazole and 3.5-di chloro benzoyl chloride 85%, white solid; *HNMR (500 MHz, DMSO) δ 8.15 (s, 2H), 7.90 (s, 1H), 7.54 (dd, J= 8.0, 0.9 Hz, 1H), 7.28 (t, J= 8.0 Hz, 1H), 7.01 (d, J= 8.1 Hz, 1H), 3.92 (s, 3H).13C NMR (126 MHz, DMSO) δ 165.4, 157.7,

[0417] 152.3, 135.7, 135.0, 133.2, 132.5, 128.3, 127.6, 125.4, 114.0, 108.1, 56.2.

[0418] 4-Bromo-7V-(4-methoxybenzo[J|thiazol-2-yl)benzamide (20). Starting with 4- methoxy-2-amino benzothiazole and 4-bromo benzoyl chloride. 71%, white solid;1H NMR (500 MHz, DMSO) δ 8.07 (d, J= 8.6 Hz, 2H), 7.75 (d, J= 8.6 Hz, 2H), 7.54 (dd, J= 7.9, 1.0 Hz, 1H), 7.27 (t, J= 8.0 Hz, 1H), 7.00 (dd, J= 8.1, 1.0 Hz, 1H), 3.91 (s, 3H).13C NMR (126 MHz, DMSO) δ 165.4, 157.6, 152.4, 133.4, 132.2, 131.5, 130.8, 129.3, 127.3, 125.2, 113.9, 108.0, 56.1. Melting Point: 225.5 - 226.9°C.

[0419] 2-Iodo-7V-(4-methoxybenzo[J]thiazol-2-yl)benzamide (21). Starting with 4- methoxy-2-amino benzothiazole and 2-iodo benzoyl chloride. 75%, white solid; (5001H NMR MHz, DMSO) δ 7.95 (dd, J= 7.9, 1.1 Hz, 1H), 7.60 - 7.46 (m, 3H), 7.31 - 7.22 (m, 2H), 7.00 (dd, J= 8.1, 1.0 Hz, 1H), 3.90 (s, 3H).13C NMR (126 MHz, DMSO) δ 168.4, 156.8, 152.5, 140.6, 139.8, 133.4, 132.4, 129.2, 128.6, 128.6, 125.3, 114.0, 108.1, 94.2, 56.2. Melting Point: 159.0 - 161.9°C.

[0420] 4-lodo- \-(4-methoxybenzo|7|thi:izol-2-yl)benzamide (22). Starting with 4- methoxy-2-amino benzothiazole and 4-iodo benzoyl chloride. 88%, white solid;1H NMR (500 MHz, DMSO) δ 7.94 - 7.91 (m, 2H), 7.91 - 7.88 (m, 2H), 7.67 (d, J= 8.4 Hz, 1H), 7.54 (dd, J = 7.9, 0.9 Hz, 1H), 7.27 (t, J= 8.0 Hz, 1H), 7.00 (dd, J= 8.0, 0.9 Hz, 1H), 3.91 (s, 3H).13C NMR (126 MHz, DMSO) δ 167.4, 165.7, 157.6, 152.3, 138.1, 133.3, 131.5, 130.5, 125.2, 113.9, 108.0, 101.7, 56.1. Melting Point: 210.2 - 211.0°C.

[0421] 3-Trifluoromethyl-7V-(4-methoxybenzo[J|thiazol-2-yl)benzamide (23). Starting with 4-methoxy-2-amino benzothiazole and 3 -trifluoromethyl benzoyl chloride. 67%, white solid;1H NMR (500 MHz, CDCI3) δ 8.23 (s, 1H), 8.09 (d, J= 8.1 Hz, 1H), 7.74 (dd, J= 7.8, 1.7 Hz, 1H), 7.51 - 7.39 (m, 2H), 7.27 (d, J= 8.0 Hz, 1H), 6.69 (d, J= 8.0 Hz, 1H).13C NMR (126 MHz, CDCI3) δ 164.8, 159.0, 151.4, 137.4, 133.1, 131.5, 131.1, 129.2, 125.4, 125.1, 124.4, 122.3, 113.5, 106.6, 55.0. Melting Point: 166.3 - 168.0°C.

[0422] 4-Trifluoromethyl-7V-(4-methoxybenzo[J|thiazol-2-yl)benzamide (24). Starting with 4-methoxy-2-amino benzothiazole and 4-trifluoromethyl benzoyl chloride. 73%, white solid;1H NMR (500 MHz, DMSO) δ 8.31 (d, J = 8.0 Hz, 2H), 7.92 (d, J= 7.9 Hz, 2H), 7.56 (dd, J= 8.0, 0.9 Hz, 1H), 7.29 (t, J= 8.0 Hz, 1H), 7.02 (dd, J= 8.1, 1.0 Hz, 1H), 3.92 (s, 3H).13C NMR (126 MHz, DMSO) δ 172.5, 165.2, 152.4, 136.2, 133.3, 132.9, 129.7, 126.1, 125.4, 123.2, 114.0, 108.0, 56.2.

[0423] 4-Cyano-7V-(4-methoxybenzo[J]thiazol-2-yl)benzamide (25). Starting with 4- methoxy-2-amino benzothiazole and 4-cyano benzoyl chloride. 72%, canary yellow solid;1H

[0424] NMR (500 MHz, DMSO) δ 8.26 (d, J= 8.0 Hz, 2H), 8.02 (d, J= 8.0 Hz, 2H), 7.55 (d, J= 7.9 Hz, 1H), 7.28 (t, J= 8.0 Hz, 1H), 7.01 (d, J= 8.0 Hz, 1H), 3.92 (s, 3H).13C NMR (126 MHz, DMSO) δ 165.0, 157.7, 152.3, 136.5, 133.3, 133.1, 130.4, 129.5, 125.3, 118.6, 115.4, 114.0, 108.0, 56.2. Melting Point: 223.3 - 224.1°C.

[0425] 2-Nitro-7V-(4-methoxybenzo[J]thiazol-2-yl)benzamide (26). Starting with 4- methoxy-2-amino benzothiazole and 2-nitro benzoyl chloride. 61%, white solid; 'HNMR (500 MHz, DMSO) δ 8.19 (dd, J= 8.2, 1.2 Hz, 1H), 7.90 - 7.78 (m, 3H), 7.57 (dd, J= 8.0, 1.0 Hz, 1H), 7.29 (t, .7= 8.0 Hz, 1H), 7.01 (dd, J= 8.1, 1.0 Hz, 1H), 3.90 (s, 3H).13C NMR (126 MHz, DMSO) δ 165.5, 156.7, 152.5, 146.9, 134.8, 133.4, 132.4, 130.6, 130.2, 125.4, 125.0, 125.0, 114.1, 108.3, 56.3. Melting Point: 233.7- 235.3°C.

[0426] 3-Nitro-7V-(4-methoxybenzo[J]thiazol-2-yl)benzamide (27). Starting with 4- methoxy-2-amino benzothiazole and 3-nitro benzoyl chloride. 64%, white solid;1H NMR (500 MHz, DMSO) δ 8.97 (s, 1H), 8.57 - 8.41 (m, 2H), 7.82 (t, J= 7.9 Hz, 1H), 7.53 (d, J= 7.9 Hz, 1H), 7.27 (t, J = 7.9 Hz, 1H), 7.00 (d, J = 8.0 Hz, 1H), 3.92 (s, 3H).13C NMR (126 MHz, DMSO) δ 164.5, 157.9, 152.2, 148.3, 135.0, 134.0, 133.3, 130.8, 127.6, 125.3, 123.7, 114.0, 110.1, 108.1, 56.2. Melting Point: 244.3 - 246.0°C. 4-Nitro-7V-(4-methoxybenzo[J]thiazol-2-yl)benzamide (28). Starting with 4- methoxy-2-amino benzothiazole and 4-nitro benzoyl chloride. 78%, yellow solid;1H NMR (500 MHz, DMSO) δ 8.33 (s, 4H), 7.55 (d, J= 8.0 Hz, 1H), 7.28 (t, J= 7.8 Hz, 1H), 7.01 (d, J = 7.8 Hz, 1H), 3.92 (s, 3H).13C NMR (126 MHz, DMSO) δ 164.8, 152.3, 150.2, 138.0, 133.3, 130.3, 129.3, 125.4, 124.1, 118.6, 114.0, 108.1, 56.2. Melting Point: 243.5 - 244.3°C.

[0427] 2-Fluoro-7V-(4-methoxybenzo[J]thiazol-2-yl)benzenesulfonamide (29). Starting with 4-methoxy-2-amino benzothiazole and 2-fluorophenyl sulfonyl chloride. 63%, white solid; ‘H NMR (500 MHz, CDCI3) δ 9.76 (s, 1H), 8.04 - 8.02 (m, 1H), 7.55 - 7.48 (m, 1H), 7.27 - 7.25 (m, 1H), 7.21 (t, J= 8.1 Hz, 1H), 7.17 - 7.09 (m, 2H), 6.87 (dd, J= 8.2, 0.9 Hz, 1H), 3.94 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.9, 160.5, 158.4, 146.0, 134.5, 129.5, 125.1, 124.1, 117.0, 116.8, 114.0, 108.3, 56.0. Melting Point: 195.3 - 197.1°C.

[0428] 4-Fluoro-7V-(4-methoxybenzo[J|thiazol-2-yl)benzenesulfonamide (30). Starting with 4-methoxy-2-amino benzothiazole and 4-fluoro phenyl sulfonyl chloride. 74%, beige colored solid;1H NM (5R00 MHz, CDCI3) δ 8.03 - 7.94 (m, 2H), 7.18 (t, J= 8.1 Hz, 1H), 7.14 - 7.05 (m, 3H), 6.84 (d, J = 8.1 Hz, 1H), 3.91 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.1, 166.0, 163.9, 146.0, 137.9, 129.2, 129.2, 125.3, 125.1, 116.0, 115.8, 114.0, 108.4, 56.0. Melting Point: 166.1 - 167.4°C.

[0429] 2,4-Difluoro-7V-(4-methoxybenzo[J|thiazol-2-yl)benzenesulfonamide (31). Starting with 4-methoxy-2-amino benzothiazole and 2,4-difluorophenyl sulfonyl chloride. 51%, white solid;1H NMR (500 MHz, CDCI3) δ 9.77 (s, 1H), 8.09 - 7.99 (m, 1H), 7.21 (t, J= 8.1 Hz, 1H), 7.12 (dd, J= 8.1, 0.9 Hz, 1H), 6.96 (m, 1H), 6.91 - 6.83 (m, 2H), 3.94 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.9, 166.5, 164.5, 161.1, 159.2, 146.0, 131.2, 125.2, 125.1, 114.0, 108.3,

[0430] 105.7, 105.5, 56.0. Melting Point: 188.2 - 190.2°C.

[0431] 4-Chloro-7V-(4-methoxybenzo[J|thiazol-2-yl)benzenesulfonamide (32). Starting with 4-methoxy-2-amino benzothiazole and 4-chlorophenyl sulfonyl chloride. 64%, white solid;1H NMR (500 MHz, DMSO) δ 7.83 (d, J= 8.7 Hz, 2H), 7.60 (d, J= 8.7 Hz, 2H), 7.34 (d, J= 8.0 Hz, 1H), 7.20 (t, J= 8.1 Hz, 1H), 7.02 (d, J= 8.1 Hz, 1H), 3.86 (s, 3H).13C NMR (126 MHz, DMSO) δ 167.67, 146.91, 141.53, 137.51, 129.67, 128.23, 126.44, 125.07, 114.81,

[0432] 109.60, 56.56. Melting Point: 153.9 - 155.3°C.

[0433] 3,5-Dichloro-7V-(4-methoxybenzo[J|thiazol-2-yl)benzenesulfonamide (33).

[0434] Starting with 4-methoxy-2-amino benzothiazole and 3.5-di chlorophenyl sulfonyl chloride. 75%, white solid; ‘H NMR (500 MHz, CDCI3) δ 9.94 (s, 1H), 7.81 (d, J= 1.9 Hz, 2H), 7.43 (t,

[0435] J= 1.9 Hz, 1H), 7.19 (d, J= 8.1 Hz, 1H), 7.10 (dd, J = 8.1, 0.9 Hz, 1H), 6.85 (d, J= 8.2 Hz, 1H), 3.92 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.6, 146.1, 144.6, 135.6, 132.2, 125.4,

[0436] 125.2, 125.2, 124.9, 114.0, 108.5, 56.1. Melting Point: 160.7 - 162.3°C.

[0437] 4-Bromo-7V-(4-methoxybenzo[J|thiazol-2-yl)benzenesulfonamide (34). Starting with 4-methoxy-2-amino benzothiazole and 4-bromophenyl sulfonyl chloride. 79%. pale yellow solid;1H NM (R500 MHz, CDCI3) δ 7.83 (d, J= 8.6 Hz, 2H), 7.58 (d, J= 8.6 Hz, 2H), 7.19 (t, J= 8.1 Hz, 1H), 7.10 (dd, J= 8.1, 0.9 Hz, 1H), 6.85 (dd, J= 8.2, 0.9 Hz, 1H), 3.92 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.2, 146.0, 140.8, 132.0, 128.1, 127.2, 125.3, 125.2, 125.1, 114.0, 108.4, 56.0. Melting Point: 175.6 - 177.1°C.

[0438] 2-Iodo-7V-(4-methoxybenzo[J]thiazol-2-yl)benzenesulfonamide (35). Starting with 4-methoxy-2-amino benzothiazole and 4-iodo phenyl sulfonyl chloride. 75%, white solid; 'H NMR (500 MHz, CDCI3) δ 9.68 (s, 1H), 8.31 (dd, J= 7.9, 1.7 Hz, 1H), 8.04 (dd, J= 7.8, 1.2 Hz, 1H), 7.48 (td, J= 7.6, 1.2 Hz, 1H), 7.22 - 7.11 (m, 3H), 6.87 (dd, J= 8.1, 0.9 Hz, 1H), 3.95 (s, 3H).13C NMR (126 MHz, CDCI3) δ 168.0, 145.9, 143.8, 142.3, 134.6, 133.0, 130.1, 128.1, 125.5, 125.1, 114.0, 108.3, 92.8, 56.0. HRMS m / z: calcd for Ci4HnIN2O3S2, 445.9256; found, 445.9351, M + H+. Melting Point: 224.9.0 - 226.5°C.

[0439] 4-Iodo-7V-(4-methoxybenzo[J]thiazol-2-yl)benzenesulfonamide (36). Starting with 4-methoxy-2-amino benzothiazole and 4-iodophenyl sulfonyl chloride. 71%, beige colored solid; ‘HNMR (500 MHz, CDCI3) δ 7.81 (d, J= 8.6 Hz, 2H), 7.69 (d, J= 8.6 Hz, 2H), 7.19 (d, J= 8.1 Hz, 1H), 7.11 (dd, J= 8.1, 0.9 Hz, 1H), 6.86 (dd, J= 8.2, 0.9 Hz, 1H), 3.93 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.2, 145.9, 141.4, 138.0, 128.0, 125.3, 125.2, 125.1, 114.1, 108.4, 99.6, 56.0. Melting Point: 206.4 - 208.0°C.

[0440] 3-Trifluoromethyl-7V-(4-methoxybenzo[J|thiazol-2-yl)benzenesulfonamide (37). Starting with 4-methoxy-2-amino benzothiazole and 3 -trifluoromethylphenyl sulfonyl chloride. 68%, beige-colored solid;1H ( N50M0R MHz, CDCI3) δ 9.86 (s, 1H), 8.23 (d, J = 1.7 Hz, 1H), 8.17 (d, J= 7.9, 1H), 7.76 (dd, J= 7.7, 1.7 Hz, 1H), 7.59 (t, J= 7.9 Hz, 1H), 7.19 (t, J= 8.1 Hz, 1H), 7.10 (dd, J = 8.0, 0.9 Hz, 1H), 6.85 (d, J= 8.1 Hz, 1H), 3.91 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.5, 146.1, 142.9, 129.6, 129.0, 128.9, 125.3, 125.2, 125.2, 123.6, 123.6, 114.0, 108.5, 56.0. Melting Point: 156.3 - 158.0°C.

[0441] 4-Trifluoromethyl-7V-(4-methoxybenzo[J|thiazol-2-yl)benzenesulfonamide (38). Starting with 4-methoxy-2-amino benzothiazole and 4-trifluoromethylphenyl sulfonyl chloride. 64%, beige-colored solid,JHNMR (500 MHz, CDC13) δ 9.75 (s, 1H), 8.10 (d, J= 8.1 Hz, 2H), 7.71 (d, J= 8.1, 2H), 7.21 (d, J= 8.1 Hz, 1H), 7.12 (dd, J= 8.1, 0.9 Hz, 1H), 6.86 (dd, J = 8.2, 0.9 Hz, 1H), 3.93 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.5, 146.1, 145.2, 134.1, 133.9, 127.0, 126.0, 125.9, 125.3, 114.1, 108.4, 56.0. Melting Point: 154.2 - 156.4°C.

[0442] 4-Cyano-7V-(4-methoxybenzo[J|thiazol-2-yl)benzenesulfonamide (39). 69%, yellow solid. Starting with 4-methoxy-2-amino benzothiazole and 4-cyanophenyl sulfonyl chloride. *HNMR (500 MHz, DMSO) δ 8.12 - 7.92 (m, 4H), 7.35 (d, J= 7.9 Hz, 1H), 7.20 (t, J= 8.1 Hz, 1H), 7.02 (d, J = 8.2 Hz, 1H), 3.86 (s, 3H).13C NMR (126 MHz, DMSO) δ 168.01, 147.11, 146.74, 133.79, 127.04, 125.15, 118.31, 115.05, 114.80, 110.96, 109.61, 56.57. Melting Point: 184.5 - 185.6°C.

[0443] 2-Nitro-7V-(4-methoxybenzo[J]thiazol-2-yl)benzenesulfonamide (40). Starting with 4-methoxy-2-amino benzothiazole and 2-nitrophenyl sulfonyl chloride.71%; white solid; 'H NMR (500 MHz, CDCI3) δ 9.88 (brs, 1H), 8.32 - 8.23 (m, 1H), 7.73 - 7.63 (m, 3H), 7.22 (t, J = 8.1 Hz, 1H), 7.15 - 7.08 (m, 1H), 6.87 (dd, J= 8.1, 1.0 Hz, 1H), 3.94 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.9, 147.6, 146.1, 135.3, 133.2, 132.1, 130.3, 125.5, 125.4, 125.0, 124.3, 114.0, 108.4, 56.0. Melting Point: 201.6 - 203.1°C.

[0444] 3-\itro- \-(4-methoxybenzo|7|thi:izol-2-yl)benzenesulfonamide (41). Starting with 4-methoxy-2-amino benzothiazole and 3-nitrophenyl sulfonyl chloride.59%, white solid; 'H NMR (500 MHz, CDCI3) δ 9.79 (s, 1H), 8.78 (t, J= 2.0 Hz, 1H), 8.38 - 8.33 (m, 1H), 8.32 - 8.29 (m, 1H), 7.68 (t, J = 8.0 Hz, 1H), 7.22 (t, J = 8.1 Hz, 1H), 7.13 (dd, J= 8.1, 0.9 Hz, 1H), 6.87 (dd, .7= 8.1, 0.9 Hz, 1H), 3.94 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.7, 148.1, 146.1, 143.8, 132.2, 130.2, 126.8, 125.5, 125.2, 125.1, 121.8, 114.1, 108.5, 56.1. Melting Point: 166.3 - 167.8°C.

[0445] 4-Nitro-7V-(4-methoxybenzo[J]thiazol-2-yl)benzenesulfonamide (42). Starting with 4-methoxy-2-amino benzothiazole and 4-nitrophenyl sulfonyl chloride.67%, white solid; 'H NMR (500 MHz, CDCI3) δ 8.28 (d, J= 8.9 Hz, 2H), 8.15 (d, J = 8.9 Hz, 2H), 7.22 (t, J= 8.1 Hz, 1H), 7.13 (dd, J= 8.0, 0.9 Hz, 1H), 6.87 (dd, J= 8.2, 0.9 Hz, 1H), 3.93 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.7, 149.8, 147.4, 146.2, 127.8, 125.5, 125.3, 125.2, 124.1, 114.1, 108.5, 56.1. HRMS m / z: calcd for Ci4HnN3O5S2, 365.0140; found, 365.0229, M + H+. Melting Point: 170.4 - 172.1°C.

[0446] 7V-(lH-Indol-4-yl)-2-iodobenzene sulfonamide (49). Starting with 4-amino indole and 2-iodophenyl sulfonyl chloride. 86%, white solid; 73%, grey solid;1H NMR (500 MHz, DMSO) δ 11.09 (s, 1H), 10.25 (s, 1H), 8.06 (d, J= 7.7 Hz, 1H), 7.95 (d, J= 8.2 Hz, 1H), 7.45

[0447] (t, J= 7.5 Hz, 1H), 7.29 - 7.15 (m, 2H), 7.09 (d, J= 8.0 Hz, 1H), 6.88 (t, J= 7.8 Hz, 1H), 6.77 (d, J= 7.7 Hz, 1H), 6.74 (t, J = 2.6 Hz, 1H).13C NMR (126 MHz, DMSO) δ 143.1, 142.7, 137.3, 134.1, 131.1, 129.1, 128.8, 125.0, 122.2, 121.5, 111.2, 108.9, 100.2, 93.8. Melting Point: 158.6 - 160.4°C.

[0448] 7V-(lH-Indol-5-yl)-2-iodobenzene sulfonamide (50). Starting with 5-amino indole and 2-iodophenyl sulfonyl chloride. 86%, white solid; (510H0 N MMHRz, CDCI3) δ 8.16 (s, 1H), 8.05 (dd, J = 7.9, 1.2 Hz, 1H), 7.93 (dd, J = 7.9, 1.6 Hz, 1H), 7.41 (d, J= 2.1 Hz, 1H), 7.32 - 7.27 (m, 1H), 7.26 - 7.19 (m, 2H), 7.18 (dd, J= 3.2, 2.5 Hz, 1H), 7.14 - 7.09 (m, 1H), 6.98 (dd, J = 8.6, 2.1 Hz, 1H), 6.48 - 6.43 (m, 1H).13C NMR (126 MHz, CDCI3) δ 142.0, 141.2, 134.4, 133.5, 132.0, 128.5, 128.1, 127.7, 125.5, 119.3, 116.7, 111.5, 102.9, 92.4. Melting Point: 163.2 - 164.4°C.

[0449] 7V-(lH-Indol-6-yl)-2-iodobenzene sulfonamide (51). Starting with 6-amino indole and 2-iodophenyl sulfonyl chloride. 78%, red oily liquid; (5010H M NHMz,R DMSO) δ 10.98 (s, 1H), 10.23 (s, 1H), 8.07 (dd, J= 7.8, 1.2 Hz, 1H), 7.97 (dd, J = 7.9, 1.6 Hz, 1H), 7.53 - 7.44 (m, 1H), 7.36 (d, J= 8.4 Hz, 1H), 7.24 - 7.17 (m, 3H), 6.82 (dd, J= 8.5, 2.0 Hz, 1H), 6.30 (s, 1H).13C NMR (126 MHz, DMSO) δ 143.0, 142.4, 136.3, 134.2, 131.2, 131.2, 128.9, 125.9,

[0450] 125.2, 120.8, 114.2, 104.3, 101.4, 93.7.

[0451] 7V-(lH-Indol-4-yl)-4-nitrobenzene sulfonamide (52). Starting with 4-amino indole and 4-nitrophenyl sulfonyl chloride. 83%, yellow solid; (501H0 M NMHzR, DMSO) δ 11.11

[0452] (s, 1H), 10.35 (s, 1H), 8.29 (d, J= 8.8 Hz, 2H), 7.95 (d, J= 8.8 Hz, 2H), 7.21 - 7.13 (m, 2H), 6.95 (t, J= 7.9 Hz, 1H), 6.82 (dd, J= 7.6, 0.9 Hz, 1H), 6.49 - 6.43 (m, 1H).13C NMR (126 MHz, DMSO) δ 150.1, 146.4, 137.3, 128.8, 128.4, 125.5, 124.8, 123.1, 121.6, 113.4, 109.8, 99.6. Melting Point: 185.0 - 186.3°C.

[0453] 7V-(lH-Indol-5-yl)-4-nitrobenzene sulfonamide (53). Starting with 5-amino indole and 4-nitrophenyl sulfonyl chloride. 77%, yellow solid; (501H0 M NMHzR, DMSO) δ 11.07 (s, 1H), 10.15 (s, 1H), 8.31 (d, J = 8.9 Hz, 2H), 7.90 (d, J = 8.9 Hz, 2H), 7.30 (t, J= 2.8 Hz, 1H), 7.25 (dd, J= 5.4, 3.2 Hz, 2H), 6.80 (dd, J= 8.7, 2.0 Hz, 1H), 6.37 - 6.30 (m, 1H).13C NMR (126 MHz, DMSO) δ 150.1, 145.7, 134.4, 128.8, 128.5, 128.2, 126.9, 124.9, 117.9,

[0454] 115.0, 112.3, 101.7. Melting Point: 142.1 - 144.5°C.

[0455] 7V-(lH-Indol-6-yl)-4-nitrobenzene sulfonamide (54). Starting with 6-amino indole and 4-nitrophenyl sulfonyl chloride. 76%, yellow solid; (510H0 M NMHzR, DMSO) δ 11.01 (s, 1H), 10.27 (s, 1H), 8.32 (d, J= 8.9 Hz, 2H), 7.92 (d, J= 8.9 Hz, 2H), 7.36 (d, J= 8.4 Hz, 1H), 7.26 (t, J= 2.7 Hz, 1H), 7.19 - 7.13 (m, 1H), 6.72 (dd, J= 8.4, 1.9 Hz, 1H), 6.34 - 6.28 (m, 1H).13C NMR (126 MHz, DMSO) δ 150.1, 145.6, 136.2, 130.7, 128.8, 126.3, 125.9, 124.9, 120.9, 115.1, 105.7, 101.4. Melting Point: 193.7 - 195.2°C.

[0456] General procedure for the synthesis of compounds with the amide linker (59-63)

[0457] The un / substituted benzoyl chloride or 2-thiophene carbonyl chloride (1.0 eq) was added slowly to a stirred solution of 4-methoxy-2-amino benzothiazole (100 mg, 1.0 eq) in pyridine (3 mL) at 0°C. Potassium carbonate (1.5 eq) was added, and the reaction was allowed to stir overnight at room temperature. After completion, an aqueous solution of IN hydrochloric acid (7 mL) and di chloromethane (10 mL) was added to the reaction mixture and extracted. The organic layers were then washed with saturated solutions of ammonium chloride (7 mL), sodium bicarbonate (7 mL), and brine (7 mL), respectively. Organic layers were collected, filtered over anhydrous magnesium sulfate, and concentrated in-vacuo. The crude was purified using column chromatography to obtain the desired products with moderate to good yields.

[0458] \-(4-Methoxybenzo|7|thi:izol-2-yl)benzamide (59). Starting with 4-methoxy-2- amino benzothiazole and benzoyl chloride. 89%, white solid;JH NMR (500 MHz, DMSO) δ 8.14 (d, J= 7.8 Hz, 2H), 7.64 (t, J= 7.4 Hz, 1H), 7.60 - 7.49 (m, 3H), 7.27 (t, J= 8.0 Hz, 1H), 7.00 (d, J = 7.9 Hz, 1H), 3.92 (s, 3H).13C NMR (126 MHz, DMSO) δ 166.1, 157.7, 152.4, 133.4, 132.3, 130.1, 129.2, 128.8, 125.2, 125,1, 113.9, 108.0, 56.1.

[0459] 2-Methoxy-7V-(4-methoxybenzo[J|thiazol-2-yl)benzamide (60). Starting with 4- methoxy-2-amino benzothiazole and 2-methoxy benzoyl chloride. 82%, white solid;1H NMR (500 MHz, DMSO) δ 12.07 (s, 1H), 7.76 - 7.72 (m, 1H), 7.60 - 7.52 (m, 2H), 7.26 (t, J= 8.0 Hz, 1H), 7.21 (d, J= 8.4 Hz, 1H), 7.11 - 7.08 (m, 1H), 7.00 (dd, J= 8.0, 1.0 Hz, 1H), 3.93 (s, 3H), 3.90 (s, 3H).13C NMR (126 MHz, DMSO) δ 165.2, 157.7, 156.6, 152.3, 138.9, 134.1, 133.4, 130.9, 125.1, 122.0, 121.1, 114.0, 112.8, 108.0, 56.6, 56.1. Melting Point: 193.7 - 194.9°C.

[0460] 2-Methyl-\-(4-methoxybenzo|7|thiazol-2-yl)benzamide (61). Starting with 4- methoxy-2-amino benzothiazole and 2-methyl benzoyl chloride. 67%, white solid;1H NMR (500 MHz, CDCI3) δ 7.56 (dd, J= 7.6, 1.4 Hz, 1H), 7.43 (dd, J= 8.0, 0.9 Hz, 1H), 7.36 (dd, J = 7.6, 1.4 Hz, 1H), 7.30 - 7.27 (m, 2H), 7.19 - 7.12 (m, 1H), 6.83 (dd, J = 8.0, 0.9 Hz, 1H), 3.89 (s, 3H), 2.60 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.3, 158.0, 151.9, 138.4, 137.8, 133.1, 132.5, 131.8, 131.6, 127.5, 126.0, 125.0, 113.4, 106.7, 55.7, 20.5. Melting Point: 137.2 - 138.2°C.

[0461] 2-Chloro- \-(4-inet hoxy benzo |7|thiazol-2-yl (benzamide (62). Starting with 4- methoxy-2-amino benzothiazole and 2-chloro benzoyl chloride. 76%, white solid;1H NMR (500 MHz, DMSO) δ 7.67 (dd, J= 7.6, 1.6 Hz, 1H), 7.62 - 7.51 (m, 3H), 7.48 - 7.45 (m, 1H), 7.28 (t, J = 8.0 Hz, 1H), 7.01 (dd, J = 8.1, 1.0 Hz, 1H), 3.91 (s, 3H).13C NMR (126 MHz, DMSO) δ 165.9, 156.6, 152.5, 138.8, 134.5, 133.4, 132.6, 130.9, 130.4, 130.1, 127.8, 125.4, 114.0, 108.2, 56.2. 7V-(4-methoxybenzo[J]thiazol-2-yl)thiophene-2-carboxamide (63). Starting with 4- methoxy-2-amino benzothiazole and 2-thiophene carbonyl chloride. 65%, white solid; 'H NMR (500 MHz, DMSO) δ 8.31 (s, 1H), 8.03 - 7.92 (m, 1H), 7.58 - 7.48 (m, 1H), 7.30 - 7.22 (m, 2H), 7.00 (d, J= 8.5 Hz, 1H), 3.91 (s, 3H).13C NMR (126 MHz, DMSO) δ 160.7, 157.4, 152.3, 138.8, 137.5, 134.6, 133.4, 131.8, 129.2, 125.2, 113.9, 108.0, 56.2.

[0462] General procedure for the synthesis of compounds with the sulfonamide linker (64-68)

[0463] Following the general procedure for the synthesis of amide-containing compounds replacing the aryl carbonyl chlorides with un / substituted benzene sulfonyl chlorides or 2- thiophene sulfonyl chloride. Flash column chromatography was used to elute the purified compound using different hexane and ethyl acetate ratios.

[0464] \-(4-Methoxybenzo|7|thi:izol-2-yl)benzenesulfonamide (64). Starting with 4- methoxy-2-amino benzothiazole and benzene sulfonyl chloride. 65%, white solid;1H NMR (500 MHz, CDCI3) δ 9.65 (s, 1H), 7.98 (dd, J= 7.3, 1.7 Hz, 2H), 7.52 (d, J= 6.9 Hz, 1H), 7.48 - 7.43 (m, 2H), 7.21 - 7.14 (m, 1H), 7.09 (dd, J= 8.1, 1.0 Hz, 1H), 6.84 (d, J= 8.1 Hz, 1H), 3.92 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.1, 145.9, 141.7, 132.4, 128.8, 126.5, 125.4, 125.1, 125.0, 114.0, 108.3, 56.0. Melting Point: 205.7 - 207.1°C.

[0465] 2-Methoxy-7V-(4-methoxybenzo[J|thiazol-2-yl)benzenesulfonamide (65). Starting with 4-methoxy-2-amino benzothiazole and 2-methoxyphenyl sulfonyl chloride. 61%, white solid;1H NMR (500 MHz, CDC13) δ 8.04 (dd, J= 7.9, 1.7 Hz, 1H), 7.51 - 7.43 (m, 1H), 7.19 (t, J= 8.1 Hz, 1H), 7.10 (dd, J= 8.1, 0.9 Hz, 1H), 7.04 - 7.01 (m, 1H), 6.94 (dd, J = 8.1, 1.0 Hz, 1H), 6.86 (dd, J= 8.1, 0.9 Hz, 1H), 3.93 (s, 3H), 3.75 (s, 3H).13C NMR (126 MHz, CDCI3) 5 168.0, 157.3, 145.9, 134.1, 129.6, 129.4, 125.9, 125.0, 124.8, 120.2, 114.0, 112.1, 108.2, 56.1, 56.0. Melting Point: 188.1 - 189.8°C.

[0466] 2-Methyl-7V-(4-methoxybenzo[J|thiazol-2-yl)benzenesulfonamide (66). Starting with 4-methoxy-2-amino benzothiazole and 2-m ethylphenyl sulfonyl chloride. 71%, white solid; ‘H NMR (500 MHz, CDCI3) δ 9.80 (s, 1H), 8.12 - 7.78 (m, 1H), 7.43 - 7.32 (m, 1H), 7.31 - 7.20 (m, 2H), 7.16 (t, J= 8.1 Hz, 1H), 7.07 (dd, J= 8.1, 0.8 Hz, 1H), 6.83 (d, J= 8.1 Hz, 1H), 3.92 (s, 3H), 2.69 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.0, 145.9, 139.6, 137.6, 133.2, 132.4, 132.3, 128.3, 125.7, 124.9, 123.6, 114.0, 108.2, 56.0, 20.6. Melting Point: 188.5 - 190.3°C.

[0467] 2-Chloro-7V-(4-methoxybenzo[J|thiazol-2-yl)benzenesulfonamide (67). Starting with 4-methoxy-2-amino benzothiazole and 2-chlorophenyl sulfonyl chloride. 59%, white solid; ‘H NMR (500 MHz, CDCI3) δ 9.76 (s, 1H), 8.27 - 8.18 (m, 1H), 7.50 - 7.42 (m, 2H),

[0468] 7.41 - 7.37 (m, 1H), 7.21 (t, J = 8.1 Hz, 1H), 7.12 (dd, J= 8.1, 0.9 Hz, 1H), 6.87 (dd, J= 8.1, 0.9 Hz, 1H), 3.94 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.9, 146.0, 139.0, 133.2, 132.7,

[0469] 131.6, 130.3, 126.8, 125.44, 125.1, 114.0, 108.3, 56.0. Melting Point: 187.1 - 188.6°C.

[0470] 7V-(4-Methoxybenzo[d]thiazol-2-yl)thiophene-2-sulfonamide (68). 61%, white solid; ‘H NMR (500 MHz, CDCI3) δ 9.55 (s, 1H), 7.70 (dd, J= 3.8, 1.4 Hz, 1H), 7.51 (dd, J= 5.0, 1.4 Hz, 1H), 7.20 (t, J = 8.1 Hz, 1H), 7.12 (dd, J= 8.0, 0.9 Hz, 1H), 7.04 (dd, J= 5.0, 3.8 Hz, 1H), 6.86 (dd, J= 8.1, 0.9 Hz, 1H), 3.94 (s, 3H).13C NMR (126 MHz, CDCI3) δ 167.3, 146.0, 142.7, 141.7, 131.2, 127.0, 125.4, 125.2, 125.1, 114.1, 108.3, 56.0. Melting Point: 231.6 - 233.4°C.

[0471] Example 3 Thioflavin-T (ThT) Fluorescence Assay

[0472] ThT is a fluorescent benzothiazole dye, which, upon binding to the cross P-sheet structures present in numerous prone-to-aggregate proteins, results in a significant fluorescent quantum yield increase with a red shift in its excitation and emission spectra. The ThT assay was used not only to detect the presence of fibrils but also to measure the kinetics of the latestage protein fibrillation process. Dense fibril formation occurs at the plateau phase of the kinetic curve, where equilibrium between disaggregation and aggregation occurs. Thus, the ThT assay was applied as a first-line screening step to monitor the anti-fibrillar activity of the newly synthesized 4-MBT compounds on u-syn protein. The 2-iodophenyl sulfonamide derivative 35 and the 4-nitrophenyl counterpart 42 demonstrated the highest anti-fibrillar activity as depicted by the decreasing ThT fluorescence to 5.0 and 9.5%, respectively, compared to the 100% control condition (Table 1).

[0473] Table 1. The effect of the synthesized compounds containing the methoxy benzothiazole moiety on a-syn fibrils using ThT fluorescence assay.

[0474] To identify the structural component that is crucial for biological activity, the benzothiazole moieties in 35 and 42 compounds were replaced by 4-, 5-, and 6-aminoindoles with sulfonamide or urea linkers to optimize further the anti-fibrillar activity (Table 2).

[0475] Table 2. The effect of the synthesized compounds containing different indole moieties on u- syn fibrils and 2N4R tau using ThT fluorescence assay. Ureas

[0476] Compound 46, resulting from these structural modifications, induced comparable or slightly higher ThT fluorescence percentages compared to 35 and 42 (Fig. 4A). Compound 46 displayed superior activity against the earlier stage u-syn oligomer formation (discussed below). Importantly, compound 46 showed a dose-response relationship with the u-syn antifibrillation assessment (Fig. 4B). Gradually increasing the compound concentration from 3.125 to 25 pM largely reduced the ThT fluorescence. Anti-fibrillary effects of best compounds (46, 47, 48) were evaluated on tau 2N4R and demonstrated substantial reduction of fibril formation (Fig. 5).

[0477] Table 3. The effect of the synthesized compounds on a-syn fibrils tau using ThT fluorescence assay.

[0478]

[0479] Example 4

[0480] Photo-Induced Cross-linking of Unmodified Protein (PICUP) assay

[0481] Suppression of oligomer formation is a crucial end goal to validate the effectiveness of candidates targeting protein misfolding. The PICUP assay is a rapid and efficient photochemical technique that was used to study the early events of protein oligomerization and protofibril formation thanks to its potent ability to cross-link covalently unmodified protein assemblies. Compounds that showed the best anti-fibrillary activity, as monitored by ThT assay, were subjected to the u-syn PICUP assay (Figs. 6 and 10). Unlike compounds 35 and 42, compound 46 showed promising anti-oligomer u-syn activity at a compound to protein ratio of 1 : 1.2. The compounds were further evaluated in a dose-dependent manner at varying concentrations (100 pM and 200 pM) while the concentration of the protein remained at 60 pM. However, compounds 46, 47, and 48 showed no effect on inhibiting 2N4R tau or 0N4R tau oligomers induced with the PICUP assay (Figs. 7A and 7B, respectively). That could be explained by the large variation in the core structures between u- syn and tau isoforms. The ThT assay and PICUP assay results definitively confirmed the ability of compound 46 to inhibit the oligomers and fibrils of u-syn protein and mature fibrils of 2N4R tau. Example 5

[0482] In vitro Transmission Electron Microscope (TEM) analysis

[0483] The top a-syn anti-fibrillar compounds were further tested using a-syn and 2N4R tau isoform using similar conditions (data not shown) to assess further the anti-aggregation ability using a direct method of visualization of fibrils, i.e., TEM. Samples were preincubated for a period of 68 hours (a-syn) and 72 hours (tau) prior to TEM analyses. Control with vehicle (DMSO 0.25%) resulted in dense mature fibrils of a-syn and 2N4R tau. TEM imaging of a-syn revealed that all three compounds (46, 47 and 48) significantly reduced protein fibrillation (Fig. 8, upper row). In contrast, short, needle-like and rounded-structured fibrils can be clearly visualized after treating 2N4R tau with compounds 46 and 48, respectively (Fig. 8, lower row).

[0484] Example 6 a-Synuclein Inclusion-Forming Neuroblastoma Cell Experiment

[0485] To examine the effects of the three compounds (46, 47, and 48) on cell viability and formation of inclusions, M17D neuroblastoma cells that express the fusion protein S3K::YFP were used (Figs. 9A-9D). In addition, compounds 35 and 42 were also examined using similar assay conditions (Figs. 11A-11E). M17D cells are normally regulated by doxycycline induction at 48 hours, and the effects of the compounds on inclusion formation were tested in a dose-dependent manner. Compound 48 had the highest effect in the reduction of inclusions as the concentration increased. A similar trend was observed for its effect on cell confluence at a concentration range of 10 pM to 40 pM. While compounds 46 and 47 demonstrated reasonable reducing effect of inclusions at varying concentrations, neither compound had an effect on cell confluence.

[0486] Benzothiazole- and indole-based scaffold compounds were rationally designed and synthesized for dual inhibition of 2N4R tau and a-syn aggregates. The elaborated compounds were prepared by hybridizing the active pharmacophore moieties of a strong 2N4R tau aggregation inhibitor, N744 dye, and a powerful a-syn anti-aggregation compound, Anlel38b. ThT fluorescence assay revealed the benzothiazoles 35 and 42 significantly decreased the ThT fluorescence to 5.0 and 9.5% towards a-syn fibrils. In addition, the ThT fluorescence was decreased to lower than 20% by the indole-based urea compounds 46 and 48 towards 2N4R tau and a-syn fibrils. Compound 46 showed a noticeable inhibition of a- syn oligomer formation as assessed by the PICUP assay. The TEM confirmed the striking anti-aggregation ability of 46 and 48 towards the dense mature fibrils of 2N4R tau and a-syn fibrils. Importantly, the u-syn inclusions were remarkably decreased after treatment with compound 46.

[0487] Example 7

[0488] Soluble Epoxide Hydrolase (sEH) Inhibition Experiment

[0489] Compound 46, was tested for inhibition of the soluble epoxide hydrolase (sEH) using the

[0490] Cayman kit 10011671 at low micromolar concentration as shown in Table 4:

[0491] Table 4. Compound 46 is an inhibitor of soluble epoxide hydrolase (sEH).

[0492] This result supports the dual activity of compound 46 at inhibiting the aggregation and soluble epoxide hydrolase which will reduce neuroinflammation. REFERENCES

[0493] "2010 Alzheimer's disease facts and figures." Alzheimer's and Dementia. 2010;6(2): 158-194. doi: 10.1016 / j .jalz.2010.01.009.

[0494] Aarsland D, Pahlhagen S, Ballard CG, Ehrt U, Svenningsson P. "Depression in Parkinson disease - Epidemiology, mechanisms and management." Nat Rev Neurol. 2012;S(l):35-47. doi : 10.1038 / nrneurol .2011.189.

[0495] Bassil F, Meymand ES, Brown HJ, et al. "a-Synuclein modulates tau spreading in mouse brains." Journal of Experimental Medicine . 2021;27S(l). doi: 10.1084 / JEM.20192193.

[0496] Berube G. "An overview of molecular hybrids in drug discovery." Expert Opin Drug Discov. 2016;77(3):281-305. doi:10.1517 / 17460441.2016.1135125.

[0497] Bhasne K, Sebastian S, Jain N, Mukhopadhyay S. "Synergistic Amyloid Switch Triggered by Early Heterotypic Oligomerization of Intrinsically Disordered a-Synuclein and Tau." J Mol Biol. 2018;430(16):2508-2520. doi: 10.1016 / j.jmb.2018.04.020.

[0498] Braak H, Del Tredici K, Bratzke H, Hamm-Clement J, Sandmann-Keil D, Rub U. "Staging of the intracerebral inclusion body pathology associated with idiopathic Parkinson's disease (preclinical and clinical stages)." Journal of Neurology, Supplement. 2002,249(3). doi : 10.1007 / s00415-002-1301 -4.

[0499] Braak H, Tredici K Del, Rub U, De Vos RAI, Jansen Steur ENH, Braak E. "Staging of Brain Pathology Related to Sporadic Parkinson's Disease." Vol 24.; 2003.

[0500] Breydo L, Wu JW, Uversky VN. "a-Synuclein misfolding and Parkinson's disease." Biochim Biophys Acta Mol Basis Dis. 2012, 1822(2)1261-285. doi: 10.1016 / j bbadis.2011.10.002.

[0501] Chirita C, Necula M, Kuret J. "Ligand-Dependent Inhibition and Reversal of Tau Filament Formation." Biochemistry. 2004;43(10):2879-2887. doi: 10.1021 / bi036094h.

[0502] Colom-Cadena M, Gelpi E, Charif S, et al. "Confluence of a-Synuclein, Tau, and A-Amyloid Pathologies in Dementia With Lewy Bodies." 2013. https: / / academic.oup.com / jnen / article / 72 / 12 / 1203 / 2917632.

[0503] Dasari AKR, Kayed R, Wi S, Lim KH. "Tau Interacts with the C-Terminal Region of a- Synuclein, Promoting Formation of Toxic Aggregates with Distinct Molecular Conformations." Biochemistry. 2019 ;5S(25):2814-2821. doi: 10.1021 / acs.biochem.9b00215.

[0504] Deeg AA, Reiner AM, Schmidt F, et al. "Anlel38b and related compounds are aggregation specific fluorescence markers and reveal high affinity binding to a-synuclein aggregates." Biochim Biophys Acta Gen Subj. 2015;7S50(9): 1884-1890. doi: 10.1016 / j .bbagen.2015.05.021.

[0505] Elbatrawy AA, Hyeon SJ, Yue N, et al. "'turn-On' Quinoline-Based Fluorescent Probe for Selective Imaging of Tau Aggregates in Alzheimer's Disease: Rational Design, Synthesis, and Molecular Docking." ACS Sens. 2021;6(6):2281-2289. doi: 10.1021 / acssensors.lc00338. Elkouzi A, Vedam-Mai V, Eisinger RS, Okun MS. "Emerging therapies in Parkinson disease — repurposed drugs and new approaches." Nat Rev Neurol. 2019;75(4):204-223. doi : 10.1038 / s41582-019-0155-7.

[0506] Fil A, Cano-de-la-Cuerda R, Munoz-Hellm E, Vela L, Ramiro-Gonzalez M, Fernandez-de-las- Penas C. "Pain in Parkinson disease: A review of the literature." Parkinsonism Relat Disord. 20J3;J9(3):285-294. doi: 10.1016 / j.parkreldis.2012.11.009.

[0507] Fortin JS, Shimanaka K, Saraswati AP, et al. "Anti-fibrillization effects of sulfonamide derivatives on a-synuclein and hyperphosphorylated tau isoform 1N4R." J Mol Struct. 2022; 1267. doi: 10.1016 / j.molstruc.2022.133574.

[0508] Gao Y, Tan L, Yu JT, Tan L. "Tau in Alzheimer's Disease: Mechanisms and Therapeutic Strategies." Curr Alzheimer Res. 2018;75(3):283-300. doi: 10.2174 / 1567205014666170417111859.

[0509] Giacobini E, Gold G. "Alzheimer disease therapy - Moving from amyloid-P to tau." Nat Rev Neurol. 2013;9(12):677-686. doi: 10.1038 / nrneurol.2013.223.

[0510] Gontijo VS, Viegas FPD, Ortiz CJC, et al. "Molecular Hybridization as a Tool in the Design of Multi -target Directed Drug Candidates for Neurodegenerative Diseases." Curr NeuropharmacoL 2020; 7S(5):348-407. doi: 10.2174 / 1385272823666191021124443.

[0511] Hardy J, Selkoe DJ. "The Amyloid Hypothesis of Alzheimer's Disease: Progress and Problems on the Road to Therapeutics." https: / / www.science.org.

[0512] Henderson MX, Trojanowski JQ, Lee VMY. "a-Synuclein pathology in Parkinson's disease and related a-synucleinopathies." Neurosci Lett. 2019;709. doi: 10.1016 / j.neulet.2019.134316.

[0513] Imberdis T, Negri J, Ramalingam N, et al. "Cell models of lipid-rich a-synuclein aggregation validate known modifiers of a-synuclein biology and identify stearoyl-CoA desaturase." Proc Natl Acad Sci USA. 2019; 776(41):20760-20769. doi: 10.1073 / pnas.1903216116.

[0514] Ivasiv V, Albertini C, Goncalves AE, Rossi M, Bolognesi ML. "Molecular Hybridization as a Tool for Designing Multitarget Drug Candidates for Complex Diseases." Curr Top Med Chem. 2019;79(19): 1694-1711. doi: 10.2174 / 1568026619666190619115735.

[0515] Langley CH, Charlesworth B. "Nature © Macmillan Publishers Ltd 1997 Expansion of Xenotransplantation." 1997.

[0516] Maity S, Shimanaka K, Rivet LN, et al. "In vitro characterization of urea derivatives to inhibit alpha-synuclein early-stage aggregation." J Mol Struct. 2022; 1249. doi: 10.1016 / j.molstruc.2021.131569.

[0517] Medeiros R, Baglietto-Vargas D, Laferla FM. "The Role of Tau in Alzheimer's Disease and Related Disorders." CNS Neurosci Ther. 2011; 77(5): 514-524. doi: 10.1111 / j .1755- 5949.2010.00177. Mok SA, Condello C, Freilich R, et al. "Mapping interactions with the chaperone network reveals factors that protect against tau aggregation." Nat Struct Mol Biol. 2018; 25(5):384-393. doi : 10.1038 / s41594-018-0057- 1.

[0518] Musiek ES, Holtzman DM. "Three dimensions of the amyloid hypothesis: Time, space and 'wingmen.'" Nat Neurosci. 2015; 7S(6):800-806. doi: 10.1038 / nn.4018.

[0519] Necula M, Chirita CN, Kuret J. "Cyanine dye N744 inhibits tau fibrillization by blocking filament extension: Implications for the treatment of tauopathic neurodegenerative diseases." Biochemistry. 2005;44(30): 10227-10237. doi:10.1021 / bi050387o.

[0520] Ramirez E, Ganegamage SK, Elbatrawy AA, et al. "5-Nitro-l,2-benzothiazol-3-amine and N- Ethyl-l-[(ethylcarbamoyl)(5-nitro-l,2-benzothiazol-3-yl)amino]formamide Modulate a- Synuclein and Tau Aggregation." ACS Omega. 2023;S(22):20102-20115. doi : 10.1021 / acsomega.3 c02668.

[0521] Saez-Atienzar S, Masliah E. "Cellular senescence and Alzheimer disease: the egg and the chicken scenario." Nat Rev Neurosci. 2020; 27(8):433-444. doi: 10.1038 / s41583-020-0325-z.

[0522] Sampath Kumar HM, Herrmann L, Tsogoeva SB. "Structural hybridization as a facile approach to new drug candidates." Bioorg Med Chem Lett. 2020; 30(23). doi: 10.1016 / j.bmcl.2020.127514.

[0523] Selkoe DJ, Hardy J. "The amyloid hypothesis of Alzheimer's disease at 25 years." EMBO Mol Med. 2016; S(6):595-608. doi : 10.15252 / emmm.201606210.

[0524] SenguptaU, Guerrero-Munoz MJ, Castillo-Carranza DL, et al. "Pathological Interface between Oligomeric Alpha-Synuclein and Tau in Synucleinopathies." Biol Psychiatry. 2015; 7S(10):672-683. doi: 10.1016 / j.biopsych.2014.12.019.

[0525] Seppi K, Ray Chaudhuri K, Coelho M, et al. "Update on treatments for nonmotor symptoms of Parkinson's disease — an evidence-based medicine review." Movement Disorders. 2019;34(2): 180-198. doi: 10.1002 / mds.27602.

[0526] Sipe JD, Cohen AS. "Review: History of the amyloid fibril." J Struct Biol. 2000; 730(2-3):88- 98. doi: 10.1006 / jsbi.2000.4221.

[0527] Srivastava S, Ahmad R, Khare SK. "Alzheimer's disease and its treatment by different approaches: A review ." Eur J Med Chem. 2021; 216. doi: 10.1016 / j.ejmech.2021.113320.

[0528] Terry-Kantor E, Tripathi A, Imberdis T, et al. "Rapid alpha-synuclein toxicity in a neural cell model and its rescue by a stearoyl-coa desaturase inhibitor." IntJMol Sci. 2020; 27(15): 1-14. doi: 10.3390 / ijms21155193.

[0529] Wagner J, Ryazanov S, Leonov A, et al. "Anlel38b: A novel oligomer modulator for diseasemodifying therapy of neurodegenerative diseases such as prion and Parkinson's disease." Acta Neuropathol. 2013; 725(6):795-813. doi: 10.1007 / s00401-013-1114-9.

[0530] Waheed Z, Choudhary J, Jatala FH, et al. "The Role of Tau Proteoforms in Health and Disease. " Mol Neurobiol. 2023 (online). doi: 10.1007 / sl2035-023-03387-8. Zeuner KE, Schaffer E, Hopfner F, Bruggemann N, Berg D. "Progress of Pharmacological Approaches in Parkinson's Disease." Clin Pharmacol Ther. 2019; 705(5): 1106-1120. doi: 10.1002 / cpt.1374.

[0531] 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. In the event of inconsistent usages between this document and those documents so incorporated by reference, the usage in the incorporated reference should be considered supplementary to that of this document; for irreconcilable inconsistencies, the usage in this document controls.

[0532] 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, for 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. The term "or" is used to refer to a nonexclusive "or" unless otherwise indicated.

[0533] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art. The following terms and phrases shall have the meaning indicated.

[0534] 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, such as within 10%, within 5%, or within 1% of a stated value or stated limit of a range) 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%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more of a stated value or of a stated limit of a range. In addition, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Any use of section headings is intended to aid reading of the document and is not to be interpreted as limiting. Further, information that is relevant to a section heading may occur within or outside of that particular section.

Claims

WHAT IS CLAIMED IS:

1. A compound of the formula la, lb, or Ic:la lb Ic or a pharmaceutically acceptable salt or ester thereof, wherein Yi is:and Y2iswherein:Z1is N or CH;Z2is C, CH or N;Z3is N or NRa;Zris O, S, or NRb; provided that Y1is not unsubstituted indolyl when Y2is:one of R'-R4is fluoro, chloro, I-NO2, C1-C2 alkyl or methoxy (OCH3);Raand Rbare each independently H or optionally substituted C1-C20 alkyl; andR'-R8and R1-R3are each independently -H, halogen, -OH, -NH2, -NO2, -CN, -CF3, -COOH, -N3, -SO3H, -PO3H2, C1-C20 alkyl, C1-C20 hydroxyalkyl, C1-C20 alkylamino, C1-C20 dialkylamino, C1-20 alkoxy, C(0)0-(C1-C20) alkyl, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, or 3- to 15-membered heterocyclyl.

2. The compound of claim 1, wherein the compound is of the formula:or a pharmaceutically acceptable salt or ester thereof, wherein:Z1is N or CH;Z3is O, S, or NRa; provided that Z1is not CH when Z3is NRa;Rais H or optionally substituted C1-20 alkyl;R'-R3and R5-R8are each independently -H, halogen, -CN, -CF3, -NH2, -NO2, optionally substituted C1-20 alkyl or optionally substituted C1-20 alkoxy.

3. The compound of claim 1, wherein the compound is of the formula:or a pharmaceutically acceptable salt or ester thereof, wherein:Z1is N or CH;Z3is O, S, or NRa; provided that Z1is not CH when Z3is NRa;Rais H or optionally substituted alkyl;Zris O, S, or NRb;Raand Rbare each independently H or optionally substituted C1-20 alkyl; andR1-R3are each independently H, halogen, -OH, -NIL, -NO2, -CN, -COOH, -Ns, -SO3H, - PO3H2 C1-C20 alkyl, alkoxy, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, C3-C12 cycloalkenyl, or 3- to 15-membered heterocyclyl.

4. The compound of claim 1, wherein the compound is of the formula:or a pharmaceutically acceptable salt or ester thereof, wherein:Z1is N or CH;Z3is O, S, or NRa;Rais H or optionally substituted C1-20 alkyl; andRkR8are each independently H, halogen, -CF3,-CN, -NO2, or alkoxy.

5. The compound of claim 1, wherein the compound is of the formula:or a pharmaceutically acceptable salt or ester thereof, wherein:Z1is N or CH;Z3is O, S, or NRa;Rais H or optionally substituted alkyl;Zris O, S, or NRb;Raand Rbare each independently H or optionally substituted C1-20 alkyl; andR1-R3are each independently H, halogen, -OH, -NH2, -NO2, -CF3, -CN, -COOH, -Ns, - SO3H, -PO3H2, C1-C20 alkyl, alkoxy, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, C3- C12 cycloalkenyl, or 3- to 15-membered heterocyclyl.

6. The compound of claim 1, wherein the compound is of the formula:or a pharmaceutically acceptable salt or ester thereof, wherein:Yi is 4-amino-indole, 5-amino-indole, 6-amino-indole, or 7-amino-indole;Ri is -H, halogen, -OH, C1-C6 alkyl, C1-C6 alkyl-OH, C1-C6 alkoxy, or -NO2;R2is -H, halogen, -OH, C1-C6 alkyl-OH, C1-C6 alkoxy, or -NO2; and R3is -H, halogen, -OH, C1-C6 alkyl-OH, C1-C6 alkoxy, or -NO2.

7. The compound of claim 6, wherein Y1is optionally substituted with a C1-C6 alkyl or a halogen.

8. The compound of claim 6 or 7, wherein the halogen is I.

9. The compound of claim 6, wherein:Y1is 4-amino-indole;R1is -H, andR3is -NO2.

10. The compound of claim 6, whereinY1is 6-amino-indole,R1is -H, andR3is -NO2.

11. The compound of claim 1, wherein the compound is of the formula:or a pharmaceutically acceptable salt or ester thereof, wherein:Z1is N or CHZ3is O, S, or NRaRais H or optionally substituted C1-20 alkyl; andR'-R8are each independently H, halogen, -CF3, -CN, -NO2, or alkoxy.

12. The compound of claim 1, wherein the compound is of the formula:or a pharmaceutically acceptable salt or ester thereof, wherein:Z1is N or CH;Z3is O, S, or NRa;Z1is O, S, or NRb;Raand Rbare each independently H or optionally substituted C1-20 alkyl; andR1-R3is H, halogen, -OH, -NH2, -NO2, -CF3, -CN, -COOH, -N3, -SO3H, -PO3H2, C1-C20 alkyl, C1-C20 alkoxy, C2-C20 alkenyl, C2-C20 alkynyl, C3-C12 cycloalkyl, C3-C12 cycloalkenyl, or 3- to 15-membered heterocyclyl.

13. The compound of claim 1, wherein the compound is of the formula:or a pharmaceutically acceptable salt or ester thereof, wherein:R2and R3are each independently -H, -CN or -NO2;R4is C1-C6 hydroxyalkyl, -OH, or C(0)0-(C1-C20) alkyl, andR5-R8are each independently -H, -NO2, -NH2, C1-C6 alkylamino, or C1-C6 dialkylamino,14. The compound of claim 1, wherein the compound is of the formula:or a pharmaceutically acceptable salt or ester thereof, whereinR2and R3are each independently -H, -CN or -NO2;R4is C1-C6 hydroxyalkyl, -OH, or C(0)0-(C1-C20) alkyl, andR6-R8are each independently -H, -NO2, -NH2, C1-C6 alkylamino, or C1-C6 dialkylamino.

15. The compound of claim 14, wherein:R2or R3is -CN,R4is -OCH3, andR5is -H.

16. The compound of claim 14, wherein;R2or R3is -NO2,R4is -OCH3, andR5is -H.

17. A compound of the formula:or a pharmaceutically acceptable salt or ester thereof.

18. A compound of formula:or a pharmaceutically acceptable salt or ester thereof wherein: the indole group is optionally substitued; andRlb-R3bare each independently alkyl, amino, halo, alkoxy or OH.

19. The compound of claim 18, or a pharmaceutically acceptable salt or ester thereof, having the formula:

20. The compound of claim 18 or 19, a pharmaceutically acceptable salt or ester thereof, having the formula:mono- and di-alkylated versions of the amino (NH2) group.

21. A pharmaceutical composition comprising at least one compound of claim 1 or 17 or a pharmaceutically acceptable salt or ester thereof and a pharmaceutically acceptable carrier.

22. A method of inhibiting a-synuclein (a-syn) protein aggregation, tau isoform 2N4R protein aggregation, or both in a subject having, or at risk for, said protein aggregation, comprising administering to the subject at least one compound of claim 1 or 17 or a pharmaceutically acceptable salt or ester thereof in an amount effective to inhibit a-syn protein aggregation, tau isoform 2N4R protein aggregation, or both, whereupon said protein aggregation is inhibited in the subject having, or at risk for, said protein aggregation.

23. The method of claim 22, wherein the subject has, or is at risk for, Alzheimer’s disease.

24. The method of claim 22, wherein the subject has, or is at risk for, Parkinson’s disease or dementia with Lewy bodies (DLB).

25. The method of claim 22, wherein the subject has neuroblastoma and formation of a- syn inclusions is inhibited.

26. A method of inhibiting fibril formation in a subject having, or at risk for, fibril formation, wherein the method comprises administering to the subject at least one compound of claim 1 or 17 or a pharmaceutically acceptable salt or ester thereof in an amount effective to inhibit fibril formation, whereupon fibril formation is inhibited in the subject having, or at risk for, said fibril formation.

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