Supramolecular Polymer Therapeutics and Diagnostics

Abstract

A method for inhibiting the propagation of protein misfolding associated with neurological disease is carried out by contacting an environment populated with propagating amyloid conformations of a protein (prion) associated with neurological disease with a molecule that binds to multiple adjacent sites on the protein aggregate, allowing the molecule to bind to multiple sites on the protein aggregate, thereby preventing the propagation of disease-associated conformations of the protein within the environment. Drug / prion complexes are formed, and the use of the drug in the detection and treatment of neurodegenerative diseases is disclosed.

Description

[Technical Field]

[0001] government support This invention was made with government support under Grant No. 66721 awarded by the Henry M. Jackson Foundation for the Uniformed Services University of the Health Services (USU) and Grant No. P01AG002132 awarded by the National Institutes of Health. The government has certain rights in this invention.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U.S.C. § 119(e) to the filing date of U.S. Provisional Patent Application No. 63 / 407,106, filed September 15, 2022, the disclosure of which is incorporated herein by reference.

[0003] Electronic sequence list reference The Sequence Listing is provided herewith as Sequence Listing xml "UCSF-691WO_Seq_List.xml" having a size of 11,371 bytes, created on September 14, 2023. The contents of the Sequence Listing xml are incorporated herein by reference in their entirety.

[0004] The present invention relates generally to the field of neurodegenerative diseases, and more particularly to drug / amyloid fibril complexes and methods for treating and detecting amyloid fibril diseases, and linking certain transmissible beta-sheet rich protein constructs (prions) to specific types of neurodegenerative diseases. [Background technology]

[0005] Over 200 years ago, James Parkinson described the disease that bears his name (2). Little progress had been made in deciphering the cause of Parkinson's disease (PD) for nearly a century before Fritz Lewy discovered the brain inclusions that were named after him (3). In his first manuscript, he described these inclusions as eosinophilic and insoluble in alcohol, chloroform, and benzene, consistent with the presence of a major protein component. Two years later, Konstantin Tretiakoff described the abundance of these inclusions in the substantia nigra in PD and named them Lewy bodies (4).

[0006] Drugs generally form a distinct molecular association with a target site that results in a therapeutic interaction. This occurs when drugs bind to one or more biological targets based on complementary shapes, characteristics, and / or reactivities of their surfaces. In some cases, the result is a binary drug-target complex that alters the conformation, activity, or fate of the target. In still others, multiple (identical or different) small molecules bind cooperatively at multiple distinct locations within the target complex with a net affinity or effect greater than the sum of the individual binding events (e.g., GPCRs: Lu, et al. Structural basis for the cooperative allosteric activation of the free fatty acid receptor GPR40 (2017) Nat Struct Mol Biol 24, 570-577; type III kinase inhibitors Martinez, et al. (2020). Avoiding or Co-Opting ATP Inhibition: Overview of Type III, IV, V, and VI Kinase Inhibitors.: In: Shapiro, P. (eds) Next Generation Kinase Inhibitors. Springer, Cham.).

[0007] In rare cases, two or three molecules of the same ligand have been observed to bind within a single site in the target complex while engaging in productive interactions with each other (supramolecular dimers of small molecule ligands: Shokat, K.A. drug-drug interaction crystallizes a new entry point into the UPR. Mol. Cell 38, 161-163 (2010); supramolecular trimers of ligands: Stornaiuolo, M., De Kloe, G., Rucktooa, P. et al. Assembly of a π-π stack of ligands in the binding site of an acetylcholine-binding protein. Nat Commun 4, 1875 (2013); dimers leveraged for drug discovery: Allen, et al. bioRxiv 2022.05.23.493001, https: / / doi.org / 10.1101 / 2022.05.23.493001.) Reversible intermolecular interactions between monomers in these supramolecular dimeric or trimeric complexes counteract the entropic penalty for simultaneously binding two different molecules at the same site.

[0008] Supramolecular polymers spontaneously assemble from appropriately arranged monomers and maintain their polymeric properties in solution (de Greef, T., Meijer, E. Supramolecular polymers. Nature 453, 171-173 (2008)). They are distinguished from supramolecular dimers and trimers by the increased number of self-associating monomer subunits.

[0009] We unexpectedly observed supramolecular polymeric assemblies of an α-synuclein prion inhibitor bound to α-synuclein amyloid fibrils (Figure 1). Based on this observation, we propose that supramolecular polymers of small molecule monomers are uniquely suited to interact with supramolecular protein assemblies characteristic of degenerative diseases of the central nervous system (CNS). Due to their helical symmetry, fibrillar oligomers and protein polymers characteristic of neurodegenerative diseases (e.g., α-synuclein, tau, amyloid beta, etc.) typically lack sufficiently concave, distinct binding pockets that can engage in strong binding interactions with small molecule ligands. Instead of such druggable binding pockets, these fibrillar protein assemblies display channels created by repeated display of surfaces and voids along the long axis of the fibrils. Drugs and diagnostic ligands large enough to span relatively long distances along the binding channel would be able to generate sufficient affinity to produce pharmacologically relevant changes in the activity of fibrillating proteins, but such large molecules are typically not sufficiently permeable to cross the blood-brain barrier. We identified supramolecular polymers composed of low-molecular-weight monomers that can cross the blood-brain barrier and assemble in the presence of ordered α-synuclein aggregates characteristic of multiple system atrophy (MSA). We demonstrated the generality of our approach to other proteins characteristic of neurodegenerative diseases by establishing the binding of "polyGTP-1" to the paired helical filaments of tau from another supramolecular polymer: Alzheimer's disease (Figure 2). Thus, we identified a general class of supramolecular polymers as therapeutic and diagnostic agents for treating diseases such as Alzheimer's disease, Parkinson's disease, and MSA, which are characterized by the ordered accumulation of misfolded proteins.

[0010] General information Before the present methods and uses are described, it is to be understood that this invention is not limited to the particular steps, apparatus, and compounds described, as such may, of course, vary. The scope of the present invention will be limited only by the appended claims, and therefore it is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0011] Where a range of values ​​is provided, unless the context clearly dictates otherwise, it is understood that each intervening value, to the tenth of the unit of the lower limit, between the upper and lower limits of that range is also specifically disclosed. Each smaller range between any stated or intervening value in a stated range and any other stated or intervening value within that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded, and each range in which either, neither, or both limits are included in the smaller range is also encompassed within the invention, subject to any specifically excluded limits in the stated range. When a stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0012] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs.Any method and material similar or equivalent to the methods and materials described herein can be used to implement or test this invention, but some possible preferred methods and materials are described here.All publications mentioned herein are incorporated by reference to disclose and describe the relevant method and / or material that the publication is cited for.It is understood that this disclosure supersedes any disclosure of the incorporated publication, if there is any discrepancy.

[0013] It should be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a "scan" includes a plurality of such scans, reference to a "biomarker of response" includes reference to one or more such biomarkers of response and equivalents thereof known to those skilled in the art, and so forth.

[0014] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein should be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the publication dates provided may be different from the actual publication dates, which may need to be independently confirmed.

[0015] All patents and publications, including all sequences disclosed within such patents and publications, referred to herein are expressly incorporated by reference. [Prior art documents] [Non-patent literature]

[0016] [Non-Patent Document 1] GPCR: Lu, et al. “Structural basis for the cooperative allosteric activation of the free fatty acid receptor GPR40(2017)” Nat Struct Mol Biol 24,570-577 [Non-patent document 2] type III kinase inhibitors Martinez, et al. (2020) "Avoiding or Co-Opting ATP Inhibition: Overview of Type III, IV, V, and VI Kinase Inhibitors.": Shapiro, P. (ed.) Next Generation Kinase Inhibitors. Springer, Cham. [Non-patent document 3] supramolecular dimer of small molecule ligands: Shokat, KM, “A drug-drug interaction crystallizes a new entry point into the UPR.” Mol.Cell 38, 161-163 (2010) [Non-patent document 4] supramolecular trimer of ligands: Stornaiuolo, M., De Kloe, G., Rucktooa, P. et al. “Assembly of a π-π stack of ligands in the binding site of an acetylcholine-binding protein.” Nat Commun 4, 1875 (2013) [Non-Patent Document 5] dimers leveraged for drug discovery: Allen, et al. bioRxiv 2022.05.23.493001, https: / / doi.org / 10.1101 / 2022.05.23.493001 [Non-patent document 6] de Greef, T., Meijer, E. "Supramolecular polymers." Nature 453, 171-173 (2008) Summary of the Invention [Means for solving the problem]

[0017] A method for inhibiting the propagation of protein misfolding associated with neurological disease is carried out by contacting an environment populated with propagating amyloid conformations of a protein (prion) associated with neurological disease with a molecule that binds to multiple adjacent sites on the protein aggregate, allowing the molecule to bind to multiple sites on the protein aggregate, thereby preventing the propagation of disease-associated conformations of the protein within the environment. Drug / prion complexes are formed, and the use of the drug in the detection and treatment of neurodegenerative diseases is disclosed.

[0018] A method for disrupting the propagation of stacked proteins associated with neurological diseases is carried out by contacting an environment densely populated with stacked proteins associated with neurological diseases with a molecule that binds to multiple sites on the stacked proteins, allowing the molecule to bind to multiple sites on the stacked proteins, thereby preventing the propagation of the stacked proteins in the environment. Drug / prion complexes are formed, and the use of the drug in the detection and treatment of neurodegenerative diseases is disclosed.

[0019] A method for preventing the progressive templated misfolding of a protein associated with a neurodegenerative disease includes administering a molecule to a biological environment containing both the propagating amyloid conformation of the protein and the native cellular form of the same protein, allowing the formation of a complex between a supramolecular polymeric assembly of the molecule and the supramolecular assembly of the protein, thereby preventing further sequestration of the native cellular protein and its conversion to the propagating amyloid form. The biological environment may be selected from the group consisting of a cell lysate, an active cell culture, and a mammalian brain, and may be an animal model brain or a human brain.

[0020] Drug / prion complexes are formed, and the use of drugs in the detection and treatment of neurodegenerative diseases is disclosed. The drug can be a supramolecular polymer that simultaneously binds to multiple stacked prion sites, or multiple small molecules that are provided so that each small molecule binds to a different site in the stacked prion complex, thereby forming a drug / prion complex. By tagging the supramolecular polymer or small molecule that binds to the stacked prion complex, detection can be achieved, and by administering a therapeutically effective amount, therapy can be provided in that the drug-target complex changes the conformation, activity, or fate of the target, preventing replication.

[0021] Also provided are compounds for use in the methods disclosed herein.

[0022] The invention will be best understood from the following detailed description when read in conjunction with the accompanying drawings. It is emphasized that, according to common practice, the various features of the drawings are not to scale. Conversely, the dimensions of the various features are arbitrarily increased or reduced for clarity. The drawings include the following figures: [Brief explanation of the drawings]

[0023] [Figure 1A] Figure 1 shows the observed electron density in the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (monomer = compound 79) bound to fibrils isolated from an MSA patient sample, as revealed by cryo-electron microscopy. [Figure 1B] FIG. 1 shows the observed electron density in the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (monomer = compound 83) bound to fibrils isolated from MSA patient samples, as revealed by cryo-EM. [Figure 1C] FIG. 1 shows the observed electron density in the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (monomer = compound 125) bound to fibrils isolated from MSA patient samples, as revealed by cryo-electron microscopy. [Figure 2A]Two images showing the observed electron density in the co-structure of a supramolecular polymeric ligand of tau (GTP-1) and its paired helical filament (tau) from an AD patient sample. [Figure 2B] FIG. 1 is a two-image diagram showing the observed electron density in the co-structure of a supramolecular polymeric ligand of tau (GTP-1 polymer) and its paired helical filament from an AD patient sample. [Figure 3] FIG. 4 is a conceptual diagram of the planar core of the binding moiety of a molecule that binds to a target site on the stacked protein of FIG. 3, showing the interplanar distance between the cores at 3.3-3.5 angstroms. [Figure 4] FIG. 5 shows the planar cores of two molecules as shown in FIG. 4, and also shows the distance between equivalent atoms in adjacent molecules at 4.8 Angstroms, and the angle between the line defined by two equivalent atoms on adjacent molecules and a line perpendicular to the planar core at 44°. [Figure 5] 4 and 5, showing the interplanar distance between cores at 3.3-3.5 Å, the distance between equivalent atoms in adjacent molecules at 4.8 Å, the angle between a line defined by two equivalent atoms on adjacent molecules and a line perpendicular to the planar core at 44°, and the in-plane displacement of the equivalent atoms. [Figure 6] A diagram of 821 structures from a list of chemical structures numbered 1-821 along with the IUPAC name for each of the structures shown. These compounds are molecules that bind to multiple sites on the stacked prion protein. [Figure 7] FIG. 11 is a diagram of compounds with EC50 in a cellular assay for the propagation of synuclein fibrils from MSA. [Figure 8] FIG. 1 is a diagram of compounds that extend symptom-free survival in a mouse model of MSA. [Figure 9]FIG. 11 is a two-image diagram showing the observed electron density of α-synuclein fibrils from an MSA patient and fibrils bound to an inhibitor compound (compound 22; round-bottom image bound to residues 86-99 of α-synuclein of SEQ ID NO: 9). [Figure 10A] 1A-1C show electron density observed in the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (monomer = compound 22) bound to fibrils isolated from an MSA patient sample, as resolved by cryo-EM. 1D discloses the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (monomer = compound 22; arrow) bound to fibrils isolated from an MSA patient sample, as resolved by cryo-EM. [Figure 10B] Electron density observed in the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (10B monomer = compound 83) bound to fibrils isolated from an MSA patient sample, as resolved by cryo-EM. Figure 1 discloses the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (monomer = compound 83; arrow) bound to fibrils isolated from an MSA patient sample, as resolved by cryo-EM. [Figure 10C] Figure 1 shows the electron density observed in the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (10C monomer = compound 149) bound to fibrils isolated from an MSA patient sample, as resolved by cryo-EM. Figure 2 discloses the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (monomer = compound 149; arrow) bound to fibrils isolated from an MSA patient sample, as resolved by cryo-EM. [Figure 10D] Figure 1 shows the electron density observed in the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (10D monomer = compound 293) bound to fibrils isolated from an MSA patient sample, as resolved by cryo-EM. Figure 2 discloses the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (monomer = compound 293; arrow) bound to fibrils isolated from an MSA patient sample, as resolved by cryo-EM. [Figure 10E] Figure 1 shows the electron density observed in the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (10E monomer = compound 294) bound to fibrils isolated from an MSA patient sample, as resolved by cryo-EM. Figure 2 discloses the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (monomer = compound 294; arrow) bound to fibrils isolated from an MSA patient sample, as resolved by cryo-EM. [Figure 10F] Figure 1 shows the electron density observed in the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (10F monomer = compound 427) bound to fibrils isolated from an MSA patient sample, as resolved by cryo-EM. Figure 2 discloses the co-structure of a supramolecular polymeric inhibitor of α-synuclein aggregate propagation (monomer = compound 427; arrow) bound to fibrils isolated from an MSA patient sample, as resolved by cryo-EM. [Figure 11] FIG. 1 discloses autoradiography of Compound 294 in the brain of mice inoculated with MSA prion. DETAILED DESCRIPTION OF THE INVENTION

[0024] definition As used herein, the term "alkyl," by itself or as part of another substituent, refers to a saturated branched or straight-chain monovalent hydrocarbon radical derived by removing one hydrogen atom from a single carbon atom of a parent alkane. Typical alkyl groups include, but are not limited to, methyl; ethyl; propyl, e.g., propan-1-yl or propan-2-yl; and butyl, e.g., butan-1-yl, butan-2-yl, 2-methyl-propan-1-yl, or 2-methyl-propan-2-yl. In some embodiments, an alkyl group contains 1 to 20 carbon atoms. In other embodiments, an alkyl group contains 1 to 10 carbon atoms. In still other embodiments, an alkyl group contains 1 to 6 carbon atoms, e.g., 1 to 4 carbon atoms.

[0025] "Alkanyl" by itself or as part of another substituent refers to a saturated branched, straight-chain, or cyclic alkyl group derived by removing one hydrogen atom from a single carbon atom of an alkane. Typical alkanyl groups include, but are not limited to, methanyl; ethanyl; propanyl, such as propan-1-yl, propan-2-yl (isopropyl), cyclopropan-1-yl, and the like; butanyl, such as butan-1-yl, butan-2-yl (sec-butyl), 2-methyl-propan-1-yl (isobutyl), 2-methyl-propan-2-yl (t-butyl), cyclobutan-1-yl, and the like.

[0026] "Alkylene" refers to a branched or unbranched saturated hydrocarbon chain typically having from 1 to 40 carbon atoms, more typically 1 to 10 carbon atoms, and even more typically 1 to 6 carbon atoms. This term is exemplified by groups such as methylene (-CH-), ethylene (-CHCH-), propylene isomers (e.g., -CHCHCH- and -CH(CH)CH-), and the like.

[0027] "Alkenyl," by itself or as part of another substituent, refers to an unsaturated branched, straight-chain, or cyclic alkyl group having at least one carbon-carbon double bond derived by the removal of a hydrogen atom from a single carbon atom of an alkene. The group may be in either the cis or trans conformation about the double bond. Typical alkenyl groups include, but are not limited to, ethenyl; propenyl, e.g., prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl (allyl), prop-2-en-2-yl, cycloprop-1-en-1-yl; cycloprop-2-en-1-yl; butenyl, e.g., but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl, cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl, and the like.

[0028] "Alkynyl," by itself or as part of another substituent, refers to an unsaturated branched, straight-chain, or cyclic alkyl group having at least one carbon-carbon triple bond derived by removing one hydrogen atom from a single carbon atom of an alkyne. Typical alkynyl groups include, but are not limited to, ethynyl; propynyl, such as prop-1-yn-1-yl, prop-2-yn-1-yl, etc.; butynyl, such as but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.

[0029] "Acyl" by itself or as part of another substituent refers to the radical -C(O)R 30 refers to R 30 is hydrogen, alkyl, cycloalkyl, cycloheteroalkyl, aryl, arylalkyl, heteroalkyl, heteroaryl, heteroarylalkyl, and substituted versions thereof, as defined herein. Representative examples include, but are not limited to, formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl, benzylcarbonyl, piperonyl, propionyl, succinyl, and malonyl, and the like.

[0030] The term "aminoacyl" refers to the group -C(O)NR 21 R 22 In the formula, R 21 and R 22 is independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic; R 21 and R 22is optionally joined together with the nitrogen bound thereto to form a heterocyclic or substituted heterocyclic group, wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic are as defined herein.

[0031] "Alkoxy" by itself or as part of another substituent means the radical -OR 31 refers to R 31 represents an alkyl or cycloalkyl group, as defined herein. Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclohexyloxy, and the like.

[0032] "Alkoxycarbonyl" by itself or as part of another substituent refers to the radical -C(O)OR 31 refers to R 31 represents an alkyl or cycloalkyl group as defined herein. Representative examples include, but are not limited to, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl, cyclohexyloxycarbonyl, and the like.

[0033] "Aryl" by itself or as part of another substituent refers to a monovalent aromatic hydrocarbon radical derived by removing one hydrogen atom from a single carbon atom of an aromatic ring system. Typical aryl groups include, but are not limited to, groups derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene, fluoranthene, fluorene, hexacene, hexaphene, hexalene, as-indacene, s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene, ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene, rubicene, triphenylene, trinaphthalene, and the like. In certain embodiments, aryl groups contain 6 to 20 carbon atoms. In certain embodiments, aryl groups contain 6 to 12 carbon atoms. Examples of aryl groups are phenyl and naphthyl.

[0034] "Arylalkyl" by itself or as part of another substituent refers to an alkyl group having a carbon atom, typically a terminal or sp 3 It refers to an acyclic alkyl radical in which one of the hydrogen atoms bonded to a carbon atom is replaced with an aryl group. Typical arylalkyl groups include, but are not limited to, benzyl, 2-phenylethan-1-yl, 2-phenylethen-1-yl, naphthylmethyl, 2-naphthylethan-1-yl, 2-naphthylethene-1-yl, naphthobenzyl, 2-naphthophenylethan-1-yl, and the like. When specific alkyl moieties are intended, the nomenclature arylalkanyl, arylalkenyl, and / or arylalkynyl is used. In certain embodiments, arylalkyl groups are (C7-C 30 ) arylalkyl, e.g., the alkanyl, alkenyl, or alkynyl portion of the arylalkyl group is (C-C 10 ), and the aryl moiety is (C6-C 20 In certain embodiments, the arylalkyl group is (C7-C 20) arylalkyl, e.g., the alkanyl, alkenyl, or alkynyl portion of the arylalkyl group is (C1-C8) and the aryl portion is (C6-C 12 )

[0035] "Arylaryl," by itself or as part of another substituent, refers to a monovalent hydrocarbon radical derived by removing one hydrogen atom from a single carbon atom of a ring system in which two or more identical or non-identical aromatic ring systems are directly joined together by single bonds, the number of such direct ring bonds being one less than the number of aromatic ring systems involved. Typical arylaryl groups include, but are not limited to, biphenyl, triphenyl, phenyl-naphthyl, binaphthyl, biphenyl-naphthyl, and the like. When the number of carbon atoms in an arylaryl group is specified, that number refers to the carbon atoms comprising each aromatic ring. For example, (C5-C 14 ) Arylaryl is an arylaryl group in which each aromatic ring contains 5 to 14 carbons, such as biphenyl, triphenyl, binaphthyl, phenylnaphthyl, etc. In certain embodiments, each aromatic ring system of the arylaryl group is independently (C5-C 14 In certain embodiments, each aromatic ring system of the arylaryl group is independently (C-C 10 ) aromatic. In certain embodiments, each aromatic ring system is identical, e.g., biphenyl, triphenyl, binaphthyl, trinaphthyl, etc.

[0036] "Cycloalkyl" by itself or as part of another substituent refers to a saturated or unsaturated cyclic alkyl group. Where a specific level of saturation is intended, the nomenclature "cycloalkanyl" or "cycloalkenyl" is used. Typical cycloalkyl groups include, but are not limited to, groups derived from cyclopropane, cyclobutane, cyclopentane, cyclohexane, and the like. In certain embodiments, cycloalkyl groups are selected from the group consisting of (C3-C 10 )cycloalkyl. In certain embodiments, the cycloalkyl group is (C3-C7)cycloalkyl.

[0037] "Cycloheteroalkyl" or "heterocyclyl," by itself or as part of another substituent, refers to a saturated or unsaturated cyclic alkyl group in which one or more carbon atoms (and any associated hydrogen atoms) are independently replaced with the same or different heteroatoms. Typical heteroatoms replacing carbon atoms include, but are not limited to, N, P, O, S, Si, and the like. Where a specific level of saturation is intended, the nomenclature "cycloheteroalkanyl" or "cycloheteroalkenyl" is used. Typical cycloheteroalkyl groups include, but are not limited to, groups derived from epoxides, azirines, thiiranes, imidazolidines, morpholines, piperazines, piperidines, pyrazolidines, pyrrolidines, quinuclidines, and the like.

[0038] "Heteroalkyl, heteroalkanyl, heteroalkenyl, and heteroalkynyl," by themselves or as part of another substituent, refer to alkyl, alkanyl, alkenyl, and alkynyl groups, respectively, in which one or more of the carbon atoms (and any associated hydrogen atoms) are independently replaced with the same or different heteroatom groups. Exemplary heteroatom groups that may be included in these groups include -O-, -S-, -SS-, -OS-, -NR 37 R 38 -, .=NN=, -N=N-, -N=N-NR 39 R 40 , -PR 41 -, -P(O)2-, -POR 42 -, -OP(O)2-, -SO-, -S-(O)-, -SO2-, -SnR 43 R 44 Examples include, but are not limited to, R 37 , R 38 , R 39 , R 40 , R 41 , R 42 , R 43 , and R 44is independently hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, heteroalkyl, substituted heteroaryl, substituted heteroaryl, heteroarylalkyl, or substituted heteroarylalkyl.

[0039] "Heteroaryl" by itself or as part of another substituent refers to a monovalent heteroaromatic radical derived by removing one hydrogen atom from a single atom of a heteroaromatic ring system. Typical heteroaryl groups include, but are not limited to, acridine, arsindole, carbazole, β-carboline, chromane, chromene, cinnoline, furan, imidazole, indazole, indole, indoline, indolizine, isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole, oxazole, perimidine, phenanthridine, phenanthroline, phenazine, phthalazine, pteridine, purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolidine, quinazoline, quinoline, quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole, thiophene, triazole, xanthene, benzodioxole, and the like. In certain embodiments, heteroaryl groups are from 5- to 20-membered heteroaryls. In certain embodiments, heteroaryl groups are from 5- to 10-membered heteroaryls. In certain embodiments, heteroaryl groups are derived from thiophene, pyrrole, benzothiophene, benzofuran, indole, pyridine, quinoline, imidazole, oxazole, and pyrazine.

[0040] A "heteroarylalkyl" by itself or as part of another substituent refers to a heteroaryl group having a carbon atom, typically a terminal or sp 3"Heteroaryl" refers to an acyclic alkyl radical in which one of the hydrogen atoms bonded to a carbon atom is replaced by a heteroaryl group. When specific alkyl moieties are intended, the nomenclature heteroarylalkanyl, heteroarylalkenyl, and / or heterorylalkynyl is used. In certain embodiments, a heteroarylalkyl group is a 6-30-membered heteroarylalkyl, e.g., the alkanyl, alkenyl, or alkynyl portion of the heteroarylalkyl is 1-10-membered, and the heteroaryl portion is a 5-20-membered heteroaryl. In certain embodiments, a heteroarylalkyl group is a 6-20-membered heteroarylalkyl, e.g., the alkanyl, alkenyl, or alkynyl portion of the heteroarylalkyl is 1-8-membered, and the heteroaryl portion is a 5-12-membered heteroaryl.

[0041] "Aromatic ring system," by itself or as part of another substituent, refers to an unsaturated cyclic or polycyclic ring system having a conjugated π-electron system. Specifically included within the definition of "aromatic ring system" are fused ring systems in which one or more of the rings is aromatic and one or more of the rings is saturated or unsaturated, such as, for example, fluorene, indane, indene, phenalene, etc. Exemplary aromatic ring systems include, but are not limited to, aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene, fluoranthene, fluorene, hexacene, hexaphene, hexalene, as-indacene, s-indacene, indane, indene, naphthalene, octacene, octaphene, octalene, ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene, picene, pleiadene, pyrene, pyranthrene, rubicene, triphenylene, trinaphthalene, and the like.

[0042] "Heteroaromatic ring system," by itself or as part of another substituent, refers to an aromatic ring system in which one or more carbon atoms (and any associated hydrogen atoms) are independently replaced with the same or different heteroatoms. Typical heteroatoms replacing carbon atoms include, but are not limited to, N, P, O, S, Si, and the like. Specifically included within the definition of "heteroaromatic ring system" are fused ring systems in which one or more of the rings is aromatic and one or more of the rings is saturated or unsaturated, such as, for example, arsindole, benzodioxane, benzofuran, chromane, chromene, indole, indoline, xanthene, and the like. Exemplary heteroaromatic ring systems include, but are not limited to, arsindole, carbazole, β-carboline, chroman, chromene, cinnoline, furan, imidazole, indazole, indole, indoline, indolizine, isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole, oxazole, perimidine, phenanthridine, phenanthroline, phenazine, phthalazine, pteridine, purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolidine, quinazoline, quinoline, quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole, thiophene, triazole, xanthene, and the like.

[0043] "Substituted" refers to a group in which one or more hydrogen atoms are independently replaced with the same or different substituents. Exemplary substituents include alkylenedioxy (e.g., methylenedioxy), -M, -R 60 , -O - , =O, -OR 60 , -SR 60 , -S - , =S, -NR 60 R 61 , =NR 60 , -CF3, -CN, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)2O - , -S(O)2OH, -S(O)2R 60 , -OS(O)2O -, -OS(O)2R 60 , -P(O)(O - )2, -P(O)(OR 60 )(O - ), -OP(O)(OR 60 )(OR 61 ), -C(O)R 60 , -C(S)R 60 , -C(O)OR 60 , -C(O)NR 60 R 61 , -C(O)O - , -C(S)OR 60 , -NR 62 C(O)NR 60 R 61 , -NR 62 C(S)NR 60 R 61 , -NR 62 C(NR 63 )NR 60 R 61 , and -C(NR 62 )NR 60 R 61 (wherein M is a halogen and R 60 , R 61 , R 62 , and R 63 are independently hydrogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, aryl, substituted aryl, heteroaryl, or substituted heteroaryl, or optionally R 60 and R 61 together with the nitrogen atom to which they are attached form a cycloheteroalkyl or substituted cycloheteroalkyl ring, and R 64 and R 65 are independently hydrogen, alkyl, substituted alkyl, aryl, cycloalkyl, substituted cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, aryl, substituted aryl, heteroaryl, or substituted heteroaryl, or optionally R 64 and R 65and, taken together with the nitrogen atom to which they are attached, form a cycloheteroalkyl or substituted cycloheteroalkyl ring. In certain embodiments, the substituents are -M, -R 60 , =O, -OR 60 , -SR 60 , -S - , =S, -NR 60 R 61 , =R 60 , -CF3, -CN, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)2R 60 , -OS(O)2O - , -OS(O)2R 60 , -P(O)(O - )2, -P(O)(OR 60 )(O - ), -OP(O)(OR 60 )(OR 61 ), -C(O)R 60 , -C(S)R 60 , -C(O)OR 60 , -C(O)NR 60 R 61 , -C(O)O - , -NR 62 C(O)NR 60 R 61 In certain embodiments, the substituents include -M, -R 60 , =O, -OR 60 , -SR 60 , -NR 60 R 61 , -CF3, -CN, -NO2, -S(O)2R 60 , -P(O)(OR 60 )(O - ), -OP(O)(OR 60 )(OR 61 ), -C(O)R 60 , -C(O)OR 60 , -C(O)NR 60 R 61 , -C(O)O - In certain embodiments, the substituents include -M, -R 60 , =O, -OR 60 , -SR 60 , -NR 60 R61 , -CF3, -CN, -NO2, -S(O)2R 60 , -OP(O)(OR 60 )(OR 61 ), -C(O)R 60 , -C(O)OR 60 , -C(O)O - Contains R 60 , R 61 , and R 62 is as defined above. For example, the substituent may have a methylenedioxy substituent, or 1, 2, or 3 substituents selected from a halogen atom, a (1-4C) alkyl group, and a (1-4C) alkoxy group.

[0044] "Amino" is the group -NR X R Y refers to R X and R Y are each independently H or a non-hydrogen substituent. Exemplary non-hydrogen substituents include alkyl groups (e.g., methyl, ethyl, and isopropyl).

[0045] "Ether" refers to a diradical group of formula -O-. For example, when the ether group is attached to an alkyl group, the overall group is an alkoxy group (e.g., OCH or methoxy). When the ether group is attached to a carbonyl group, the overall group is an ester group of formula -OC(O)-.

[0046] "Halo" and "halogen" refer to chloro, bromo, fluoro, and iodo groups.

[0047] "Nitro" refers to a group of the formula -NO2.

[0048] Unless otherwise specified, a reference to an atom is meant to include all isotopes of that atom. For example, a reference to H is 1 H, 2 H (i.e., D or deuterium), and 3 H (i.e., tritium), and references to C include 12 C and all other isotopes of carbon (e.g.,13 C). Unless otherwise specified, a group includes all possible stereoisomers.

[0049] With respect to any of the groups disclosed herein that contain one or more substituents, it is of course understood that such groups do not contain any substitutions or substitution patterns that are sterically impractical and / or synthetically infeasible. In addition, the subject compounds include all stereochemical isomers arising from the substitution of these compounds.

[0050] In certain embodiments, substituents may contribute to the optical isomerism and / or stereoisomerism of the compound.Salts, solvates, hydrates, and prodrug forms of the compound are also of interest.All such forms are encompassed by the present disclosure.Thus, the compounds described herein include their salts, solvates, hydrates, prodrugs, and isomeric forms, including their pharmaceutically acceptable salts, solvates, hydrates, prodrugs, and isomers.In certain embodiments, the compounds can be metabolized into pharmaceutically active derivatives.

[0051] Neurodegenerative diseases The present disclosure provides methods, drugs, molecules, labeled molecular compounds, and α-synuclein prion inhibitors for detecting and treating neurodegenerative diseases. The terms "neurodegenerative disease" and "neurological disease" may be used interchangeably. Neurodegenerative diseases include transmissible spongiform encephalopathies, such as Creutzfeldt-Jakob disease (CJD), multiple system atrophy (MSA), Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), amyotrophic lateral sclerosis / Parkinson's disease dementia complex, anti-IgLON5-associated tauopathy, Caribbean Parkinson's disease, chronic traumatic encephalopathy, diffuse neurofibrillary tangles with calcifications, Down's syndrome, familial British dementia, familial Danish dementia, Niemann-Pick disease type C, and non-guanine motor tangles. Any neurological disease associated with stacked proteins, including, but not limited to, neuronal diseases, postencephalitic Parkinson's disease, primary age-related tauopathies, progressive ataxia and palatal tremor, neurofibrillary senile dementia, familial frontotemporal dementia and Parkinson's disease, Pick's disease, argyrophilic grain dementia, corticobasal degeneration, Guadeloupe Parkinson's disease, glomerular tauopathy, Huntington's disease, progressive supranuclear palsy, SLC9a-associated Parkinson's disease, tau astrogliopathy, etc.

[0052] Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by memory loss, spatial disorientation, and a gradual decline in intellectual ability. Numerous pathological changes have been described in the postmortem brains of AD patients, including synaptic and neuronal loss, oxidative damage, activated inflammatory cells, amyloid plaques composed primarily of beta-amyloid peptide (Aβ), and neurofibrillary tangles (NFTs) composed of hyperphosphorylated and / or acetylated aggregates of the microtubule-associated protein tau; the latter two are considered pathological hallmarks. For several reasons, research on the involvement of Aβ in AD has progressed more rapidly than research on tau. The description of the "amyloid cascade hypothesis," based on the discovery of genetic mutations causing autosomal familial AD, was centered on research on Aβ. Furthermore, biochemical studies of amyloid precursor protein (APP) and presenilin have greatly advanced our understanding of the molecular pathways leading to Aβ generation. These studies have supported the systematic development of disease-modifying therapies based on the Aβ pathway.

[0053] Tau is a soluble protein that normally binds to microtubules and regulates their dynamic growth and shortening behavior. In Alzheimer's disease (AD) and other neurodegenerative diseases, tau dissociates from microtubules and self-associates to form abnormal fibrillar aggregates. The distribution of these abnormal tau structures correlates highly with neuronal cell death and the clinical progression of neurodegenerative diseases, suggesting a close link between abnormal tau structures and neurodegeneration / dementia. Recent efforts to identify neurotoxic species of tau have shifted their focus from mature fibrillar aggregates to smaller oligomeric tau species. Studies using antibodies that selectively recognize tau oligomers have demonstrated that these species are elevated in AD brains. Cell-to-cell transmission of oligomeric tau and other abnormal pre-fibrillar species may underlie the spread of tau pathology, making these species promising therapeutic targets. A structural understanding of early tau aggregates may lead to a deeper understanding of their neurotoxic mechanisms and the rational design of therapeutic agents.

[0054] As a result of alternative RNA splicing, six distinct tau isoforms exist, differing from one another depending on the presence or absence of three inserts encoded by exons 2, 3, and 10 of the tau gene. Each tau isoform contains two key domains: the N-terminal projection domain, which determines microtubule spacing between bundled microtubules and also mediates the interaction of microtubules with the plasma membrane. Exons 2 and 3 each encode a 29-residue acidic insert located in the N-terminal projection domain. In contrast, the microtubule-binding pseudo-repeat (MTBR) domain contains either three or four imperfect repeats (depending on the presence or absence of exon 10 coding sequences) and serves to directly bind to microtubules and regulate their dynamics. This same region of the protein constitutes the core of fibrillar tau aggregates, which undergo a regional transition from random coil to beta-sheet structure. Fibrillar tau aggregates possess a cross-beta structure typical of amyloid fibrils. Finally, the six tau isoforms differ not only structurally but also in their relative expression levels and the rate and extent of their fibrillation. Genetic evidence clearly demonstrates that functional differences must exist between the six different tau isoforms.

[0055] The amino acid sequences of the six human tau isoforms include:

[0056] Isoform 1 (SEQ ID NO: 1) MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPK SPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL.

[0057] Isoform 2 (SEQ ID NO: 2): MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPT REPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL.

[0058] Isoform 3 (SEQ ID NO: 3): MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEDVTAPLVDEGAPGKQAAAQPHTEIPEGTTTAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGT PGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL

[0059] Isoform 4 (SEQ ID NO: 4): MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKAEEAGIGDTPSLEDEAAGHVTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRLQTAPVPM PDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL.

[0060] Isoform 5 (SEQ ID NO: 5): MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEAEEAGIGDTPSLEDEAAGHVTQARMVSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKS PSSAKSRLQTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL

[0061] Isoform 6 (SEQ ID NO: 6): MAEPRQEFEVMEDHAGTYGLGDRKDQGGYTMHQDQEGDTDAGLKESPLQTPTEDGSEEPGSETSDAKSTPTAEDVTAPLVDEGAPGKQAAAQPHTEIPEGTTTAEEAGIGDTPSLEDEAAGH VTQARMVSKSKDGTGSDDKKAKGADGKTKIATPRGAAPPGQKGQANATRIPAKTPPAPKTPPSSGEPPKSGDRSGYSSPGSPGTPGSRSRTPSLPTPPTREPKKVAVVRTPPKSPSSAKSRL QTAPVPMPDLKNVKSKIGSTENLKHQPGGGKVQIINKKLDLSNVQSKCGSKDNIKHVPGGGSVQIVYKPVDLSKVTSKCGSLGNIHHKPGGGQVEVKSEKLDFKDRVQSKIGSLDNITHVPGGGNKKIETHKLTFRENAKAKTDHGAEIVYKSPVVSGDTSPRHLSNVSSTGSIDMVDSPQLATLADEVSASLAKQGL. Where reference is made to a specific amino acid number, the numbering refers to SEQ ID NO: 6 unless otherwise stated.

[0062] The classically described function of tau is as a neuronal microtubule-associated protein found primarily in axons. Under physiological conditions, tau exists as a highly soluble, natively unfolded protein that interacts with tubulin and promotes its assembly into microtubules, helping to stabilize its structure. Recent evidence points to additional functions for tau. For example, tau phosphorylation allows neurons to escape acute apoptotic death by stabilizing β-catenin. Tau also plays an essential role in balancing microtubule-dependent axonal transport of organelles and biomolecules by regulating kinesin-driven anterograde and dynein-driven retrograde transport.

[0063] Soluble oligomeric species of amyloid-β (Aβ) are thought to be another important mediator of cognitive impairment in Alzheimer's disease (AD) (M. Sheng, et al. (2012) Cold Spring Harb Perspect Biol 4; J. J. Palop et al. (2010) Nat Neurosci 13, 812). Neuritic plaques, a hallmark of AD, are accumulations of aggregated or oligomerized amyloid beta (Aβ) peptides, including Aβ1-40 (Aβ40) and Aβ1-42 (Aβ42), derived from the processing of amyloid precursor protein (APP) by beta- and gamma-secretases. The majority of mutations associated with autosomal familial AD (FAD) are associated with elevated levels of Aβ1-42. Transgenic mice expressing high levels of human Aβ experience memory loss and synaptic retraction (M. Faizi et al., (2012) Brain Behav 2, 142; C. Perez-Cruz et al., (2011) J Neurosci 31, 3926; S. Knafo et al., (2009) Cereb Cortex 19, 586; M. Cisse et al., (2011) Nature 469, 47). Aβ production is thought to be activity-dependent (F. Kamenetz et al., (2003) Neuron 37, 925; J. Wu et al., (2011) Cell 147, 615), and even in wild-type mice, the addition of soluble Aβ oligomers to hippocampal slices or cultures induces a loss of long-term potentiation (LTP), increases long-term depression (LTD), and reduces dendritic spine density (G.M. Shankar et al., (2007) J Neurosci 27, 2866; G.M. Shankar et al., (2008) Nat Med 14, 837; H. Hsieh et al., (2006) Neuron 52, 831). Currently, there are no effective therapies to halt or reverse the cognitive impairments that characterize AD.

[0064] Aβ oligomer levels also increase by approximately 200–300% throughout life in patients with Down syndrome (DS) (reviewed in Head and Lott (2004) Curr Opin Neurol 17(2):95–100). The use of γ-secretase inhibitors to reduce β-amyloid levels in young mice modeling DS corrected the learning deficits characteristic of these mice, suggesting that therapies that disrupt Aβ oligomers may improve cognitive function even in young DS patients (Netzer WJ, et al. (2010) PLoS One 5:e10943).

[0065] Aβ, or "amyloid beta," or "amyloid β," refers to a 36-43 amino acid peptide derived from processing of the amyloid precursor protein (APP) by β- and γ-secretase. "Aβ oligomer," "amyloid β oligomer," or "amyloid beta oligomer" refers to aggregates of Aβ peptides. Aβ is the major component of deposits called amyloid plaques found in the brains of patients with Alzheimer's disease (AD) and cerebral amyloid angiopathy (CAA), and it has also been associated with retinal ganglion cells in patients with glaucoma. Two major variants of Aβ 1-40 ("Aβ40") (DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVV) (SEQ ID NO: 7) and Aβ 1-42 ("Aβ42") (DAEFRHDSGYEVHHQKLVFAAEDVGSNKGAIIGLMVGGVVIA) (SEQ ID NO: 8) is produced by selective carboxy-terminal truncation of APP (Selkoe et al. (1988) Proc. Natl. Acad. Sci. USA 85:7341-7345; Selkoe, (1993) Trends Neurosci 16:403-409). 1-42 Aβ is the more fibrillar and more abundant of the two peptides in amyloid deposits of both AD and CAA. Derivatives of the above peptides can be synthesized using naturally occurring substituents, e.g., Aβ. 1-42 H13R, Aβ 1-42 V18A, Aβ 1-42F19P, Aβ 1-42 E22D, Aβ 1-42 E22V, Aβ 1-42 E22A, Aβ 1-42 D23A, Aβ 1-42 G25A, Aβ 1-42 N27A, Aβ 1-42 K28A, Aβ 1-42 G29A, Aβ 1-42 I31A, Aβ 1-42 These include G37A, the British mutation, the Iowa mutation, the Tottori-Japan mutation, the Flemish mutation, the Arctic mutation, and the Italian mutation. For example, in addition to amyloid deposits that can occur in CNS tissue, amyloid deposits can occur in the walls of blood vessels (Hardy (1997); Haan et al. (1990); Vinters (1987); Itoh et al. (1993); Yamada et al. (1993); Greenberg et al. (1993); Levy et al. (1990)). These vascular lesions are characteristic of CAA, which can exist in the absence of AD.

[0066] Aβ oligomers are known in the art to have several effects on cells. These include, for example, reduced cell viability, reduced synaptic plasticity, and promoted synapse loss in neurons. "Synapse" refers to the structure on a neuron that allows the neuron to pass electrical or chemical signals to another cell. "Synaptic plasticity" refers to the ability of a synapse to change strength, i.e., become stronger or weaker, in response to either use or disuse of transmission through that synaptic pathway. Such changes in strength are typically manifested by one or more of the following structural changes: changes in the number of presynaptic vesicles, changes in the amount of neurotransmitter loaded per vesicle, changes in the number of dendritic spines, and / or changes in the number of neurotransmitter receptors located on the postsynaptic neuron. A decrease or increase in synaptic plasticity can be observed by assessing the ability of the postsynaptic neuron to induce long-term potentiation ("long-term potentiation," LTP) or long-term depression (LTD) in the activity of the presynaptic neuron, and / or by assaying for subsequent changes in synaptic strength, e.g., by detecting one or more of the above-mentioned structural changes. "Improved synaptic plasticity" refers to greater synaptic strengthening (LTP), more stable synapses, and the inability to eliminate synapses and the spines that carry them. "Decreased synaptic plasticity" refers to increased synaptic weakening (LTD), less stable synapses, and fewer spines and synapses. "Synapse loss" refers to a decrease in the number of synapses, for example, the loss of a connection between two neurons, or, if multiple synapses exist between two neurons, the loss of one or more of these synapses. As is well known in the art, changes in synaptic activity and synaptic strength and number are central to almost all neurobiological processes, including learning, memory, and neuronal development. In further describing embodiments of the present invention, the following description will focus on the effects of Aβ oligomers on neurons. However, the subject methods and compositions can also be used to inhibit the effects of Aβ oligomers on other types of cells, such as microglia.

[0067] Parkinson's disease (PD) is a major neurodegenerative disorder that primarily affects the motor system but can also cause cognitive and behavioral problems. PD brains exhibit widespread neuronal degeneration, affecting up to 70% of dopaminergic neurons in the substantia nigra (SN) by the time of death. Neuropathological hallmarks of PD include Lewy bodies (LBs) in the SN, brainstem, and rostral and forebrain regions, as well as selective loss of dopaminergic neurons in the SN. Cell death-induced damage in the SN may be the cause of patients' motor impairments. While the cause of this cell death is generally unknown, researchers have observed enrichment of alpha-synuclein in neuronal Lewy bodies. Tau aggregates can also be observed in PD, for example, in patients with LKKR2 mutations. Tau has also been associated with increased alpha-synuclein deposits. Immunohistochemistry using anti-tau antibodies demonstrated high levels of NFTs in the substantia nigra from postmortem human brain tissue. The researchers also reported that tauopathy in PD and PD with dementia (PDD) was observed only in DA neurons in the nigrostriatal region, in contrast to the widespread brain-wide expression pattern of tau in AD.

[0068] In Parkinson's disease, pigmented neurons in the substantia nigra, locus ceruleus, and other brainstem dopaminergic cell groups degenerate. Loss of substantia nigra neurons leads to dopamine depletion in the dorsal aspect of the putamen (part of the basal ganglia), causing many of the motor symptoms of Parkinson's disease.

[0069] Genetic predisposition is likely in at least some cases of Parkinson's disease. Genetic associations with polymorphisms surrounding the tau gene have been found in Parkinson's disease and Alzheimer's disease. Approximately 10% of PD patients have a family history of Parkinson's disease. Several abnormal genes have been identified. Inheritance is autosomal dominant for some genes and autosomal recessive for others. Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common mutations in sporadic cases of Parkinson's disease in patients and the most common autosomal dominant mutation among inherited forms of this disease. Recent data indicate that tau is particularly important in PD patients with LRRK2 mutations, and therefore the therapy proposed here may be particularly effective.

[0070] The diagnosis of Parkinson's disease is clinical. Parkinson's disease is suspected in patients with characteristic unilateral resting tremor, hypokinesia, or rigidity. During finger-nose coordination testing, tremor disappears (or attenuates) in the limb being tested. During neurological examination, patients are unable to successfully perform rapidly alternating or rapidly successive movements. Sensation and strength are usually normal. Reflexes are normal but may be difficult to elicit due to significant tremor or rigidity. Slowing and decreased movement due to Parkinson's disease must be distinguished from decreased movement and spasticity due to corticospinal tract lesions. To help distinguish Parkinson's disease from secondary or atypical parkinsonism, clinicians often test the patient's response to levodopa. A large and sustained response strongly supports Parkinson's disease.

[0071] Amyotrophic lateral sclerosis (ALS) is a group of rare neurological disorders primarily involving nerve cells (neurons) involved in controlling voluntary muscle movement. It is characterized by steady, unrelenting, and progressive degeneration of the corticospinal tract, anterior horn cells, bulbar motor nuclei, or a combination thereof. Symptoms vary in severity and can include muscle weakness and atrophy, fasciculations, emotional lability, and respiratory muscle weakness. Diagnosis involves nerve conduction studies, electromyography, and the exclusion of other disorders through MRI and laboratory testing. Current treatment is symptomatic. The majority of ALS cases (over 90 percent) are considered sporadic.

[0072] Most patients with ALS present with random, asymmetric symptoms consisting of cramps, weakness, and muscle atrophy in the hands (most commonly) or feet. Weakness progresses to the forearms, shoulders, and lower limbs. Fasciculations, spasticity, hyperactive deep tendon reflexes, extensor plantar reflexes, clumsiness, rigidity of movement, weight loss, fatigue, and difficulty controlling facial expressions and tongue movements soon follow. Other symptoms include hoarseness, difficulty swallowing, and slurred speech because swallowing is difficult, salivation appears to increase, and patients tend to choke on liquids. In the later stages of the disorder, emotional dysregulation occurs, accompanied by inappropriate, involuntary, uncontrollable, excessive laughter or crying. Sensory systems, consciousness, cognition, voluntary eye movements, sexual function, and urinary and anal sphincters are usually spared. Death is usually caused by failure of the respiratory muscles; 50% of patients die within 3 years of onset, 20% survive for 5 years, and 10% survive for 10 years. Survival >30 years is rare.

[0073] MSA is an adult-onset sporadic synucleinopathy characterized by symptoms of Parkinsonism, cerebellar ataxia, and autonomic dysfunction. MSA cases are classified into MSA-P, which represents predominant Parkinsonism caused by striatonigral degeneration, and MSA-C, which represents cerebellar ataxia associated with olivopontocerebellar atrophy. Autonomic dysfunction is common in both subtypes. Neuropathologically, MSA is defined by focal neuronal loss and the presence of abundant filamentous α-synuclein inclusions in oligodendrocytes: glial cytoplasmic inclusions known as Papp-Lantos bodies. Smaller numbers of α-synuclein inclusions are also present in neurons. The average duration of the disease is 6–10 years, although survival times of 18–20 years have been reported. The later appearance of autonomic dysfunction correlates with longer survival.

[0074] Neuropathies are pathologically defined by the presence of α-Syn (α-synuclein or alpha synuclein) aggregates. Lewy body diseases consist of synucleinopathies with pathological Lewy bodies and Lewy neurites, which clinically manifest as PD, Parkinson's disease dementia (PDD), or dementia with Lewy bodies (DLB). α-Syn aggregates in the form of glial cytoplasmic inclusions (GCIs) are a hallmark of MSA. It has previously been shown that α-Syn fibrils are present in PD and are expected to be present in other synucleinopathies. α-Syn fibrils formed in vitro also induce α-Syn inclusions when injected into animal models.

[0075] α-Syn is a 140-amino acid protein encoded by the gene synuclein alpha (SNCA), whose normal function is thought to be related to synaptic vesicle transmission. Residues 1–60 constitute a lysine-rich N-terminal region containing KTK lipid-binding repeats for vesicle binding. Familial SNCA gene mutations, including A30P, A30G, E46K, G51D, A53E, A53V, and A53T, are found in this N-terminal region. Residues 61–95 contain the non-amyloid b-component (NAC) region, which has been shown to be essential for aggregation. Small peptides derived from this region were able to readily aggregate. Furthermore, b-synuclein, which is similar to α-Syn but lacks amino acids 71–82 in the NAC, has not been found to aggregate like α-Syn. Removal of this region from α-Syn also prevents aggregation in vitro, further supporting the importance of this region in aggregation. The amino acid sequence of α-Syn is MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVHGVATVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNEEGAPQEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA (SEQ ID NO: 9).

[0076] Clinically, PD is the second most common neurodegenerative disorder, affecting 2%–3% of individuals over the age of 65, while DLB is believed to be the underlying cause of 10%–15% of all dementia cases. PD is characterized by neuronal loss in the substantia nigra, which causes striatal dopamine deficiency and leads to bradykinesia and other motor symptoms. However, PD can progress to PDD, and DLB can progress to motor dysfunction similar to that seen in PD. PD, PDD, and DLB are generally considered to be on the same disease spectrum, sharing clinical and pathological similarities, including α-Syn aggregates in the form of Lewy bodies and Lewy neurites. MSA differs clinically and pathologically from PD and DLB and has a low prevalence in the general population. MSA is sporadic in onset and characterized by Parkinsonism, cerebellar ataxia, and / or autonomic failure. Similar to PD and DLB, MSA has neuronal asyn inclusions and neuronal cell loss, but α-Syn inclusions are more prevalent in oligodendrocytes as GCIs.

[0077] As seen with tau fibrils, the structure of α-Syn fibrils varies depending on the disease state. GCIs from MSA have also been shown to be 103 times more agglutinating seeds than fibrils derived from PD Lewy bodies. Recent characterization of amplified patient-derived fibrils suggests that PD and MSA fibrils share many characteristics, but MSA fibrils are more potent inducers of movement disorders, neurodegeneration, and α-Syn pathology. Although DLB and PD are considered to be on the same disease spectrum, DLB fibrils do not result in the significant neuropathology seen with PD fibrils. Although more recent, these results may suggest that PD fibrils contained in Lewy bodies and DLB fibrils may not be as similar as previously thought.

[0078] Familial SNCA mutations disrupt known stabilizing interactions in the in vitro folding conformation of wild-type α-Syn. Under the same conditions, the A53T, A53E, G51D, and E46K mutants of α-Syn have been shown to form fibrils with morphologies distinct from those of wild-type fibrils. E46K mutant α-Syn is toxic to neuronal cells. These fibrils were less stable and fragmented more readily, but also seeded better than wild-type fibrils, potentially indicating that toxicity is related to seeding rather than fibril stability. These in vitro data suggest that mutations may affect fibril structure, stability, and their impact on disease progression.

[0079] The G51D and A53E mutations can cause mixed MSA and PD pathology in patients. G51 is located at the protofilament interface of ex vivo MSA fibrils, near K43, K45, and H50. Structurally, there may be room for a glycine-to-aspartate switch; however, this switch reduces the positive charge of the central cavity. It is difficult to predict how changes in charge or amino acid side chains will affect the central cavity and the entire in vivo fibril. However, this could alter nonproteinaceous compounds within the central cavity, potentially disrupting fibril stability or improving fibril stability, making fibrils more stable and more pathogenic than wild-type α-Syn fibrils, leading to mixed MSA and PD pathology.

[0080] Some of the imaged in vitro fibrils can explain how some of the familial mutations assemble in vivo, and the ex vivo structures solved can accommodate some of the familial SNCA mutations. However, to confirm the actual structure, further ex vivo α-Syn structures of mutant fibrils need to be solved. The structures of fibrils from patients with familial mutations are important for understanding why those mutations specifically cause disease and may also lead to breakthroughs in understanding the onset and progression of sporadic disease.

[0081] compound The present disclosure provides novel compounds.The novel compounds and pharmaceutical preparations containing such compounds or combinations of these compounds with at least one additional therapeutic agent can be used to treat diseases described herein, such as, inter alia, multiple system atrophy (MSA).

[0082] symbol [ka] indicates the point at which the displayed moiety is attached to the remainder of the compound, whether utilized as a bond or displayed perpendicular to a bond.

[0083] In one aspect, the present invention provides a compound of the present invention. In an exemplary embodiment, the present invention is a compound described herein. In an exemplary embodiment, the present invention is a compound according to a formula described herein. In some embodiments, the present invention is any of the compounds disclosed in Figure 6. In some embodiments, the present invention is any of the compounds disclosed in Figure 6, or a pharmaceutically acceptable salt, hydrate, or solvate thereof.

[0084] In one aspect, the present invention provides a compound having a structure according to formula (I), or a salt or hydrate or solvate thereof: [ka] (In the formula, T is a substituted or unsubstituted naphthyridinone or a substituted or unsubstituted dihydronaphthyridinone; X is a substituted or unsubstituted aryl or a substituted or unsubstituted heteroaryl, X is monocyclic; Z is substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted ethenyl.

[0085] Group T In an exemplary embodiment, the compound is according to Formula (I), where X and Z are as described herein and T is: [ka] where R a , R b , R c , and R d is H, halogen, substituted or unsubstituted C 1-3 Alkyl, C2-C4 alkenyl, substituted or unsubstituted C 1-3 alkoxy, and when the connection between C* and C** is a single bond, R a and R b can optionally be attached to C* or C** to form a substituted or unsubstituted cyclopropyl.

[0086] In an exemplary embodiment, the compound is according to Formula (I), where X and Z are as described herein and T is: [ka] where R a , R b , R c , and R d are each independently H, halogen, or unsubstituted C 1-3 Alkyl, C substituted with one or more halogens 1-3 Alkyl, C2-C4 alkenyl, unsubstituted C 1-3 Alkoxy and C substituted with one or more halogens 1-3 In an exemplary embodiment, the compound is according to Formula (I), wherein X and Z are as described herein and T is selected from the group consisting of: [ka] where R a , R b , R c , and R dare independently selected from the group consisting of H, F, Cl, Br, methyl, ethenyl, isopropyl, difluoromethyl, trifluoromethyl, 1,1-difluoroethyl, methoxy, trifluoromethoxy, difluoromethoxy, and 1,1-difluoroethoxy. In an exemplary embodiment, the compound is according to Formula (I), wherein X and Z are as described herein and T is [ka] where R a , R b , R c , and R d are independently selected from the group consisting of H, F, Cl, Br, methyl and methoxy.

[0087] In an exemplary embodiment, the compound is according to Formula (I), where X and Z are as described herein and T is: [ka] or [ka] and In the formula, R a , R b , R c , and R d is as described herein.

[0088] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, is according to Formula (I), wherein X and Z are as described herein, and T is [ka] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, is according to Formula (I), wherein X and Z are as described herein, and T is [ka] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, is according to Formula (I), wherein X and Z are as described herein, and T is [ka] ,or [ka] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, is according to Formula (I), wherein X and Z are as described herein, and T is [ka] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, is according to Formula (I), wherein X and Z are as described herein, and T is [ka] is.

[0089] Group X In an exemplary embodiment, the compound is according to Formula (I), where T and Z are as described herein, and X is substituted or unsubstituted phenyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted pyrimidinyl, or substituted or unsubstituted thienyl.

[0090] In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is substituted or unsubstituted phenyl. In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is unsubstituted phenyl, or halogen, C2-C4 alkenyl, unsubstituted C 1-3 Alkyl, C substituted with one or more halogens 1-3 Alkyl, unsubstituted C 1-3 Alkoxy and C substituted with one or more halogens 1-3In an exemplary embodiment, the compound is according to formula (I), wherein T and Z are as described herein, and X is unsubstituted phenyl or phenyl substituted with one or more members selected from the group consisting of F, Cl, Br, methyl, ethenyl, isopropyl, difluoromethyl, trifluoromethyl, 1,1-difluoroethyl, methoxy, trifluoromethoxy, difluoromethoxy, and 1,1-difluoroethoxy. In an exemplary embodiment, the compound is according to formula (I), wherein T and Z are as described herein, and X is unsubstituted phenyl. In an exemplary embodiment, the compound is according to formula (I), wherein T and Z are as described herein, and X is [ka] wherein R e , R f , and R g are each independently H, halogen, C2-C4 alkenyl, unsubstituted C 1-3 Alkyl, C substituted with one or more halogens 1-3 Alkyl, unsubstituted C 1-3 Alkoxy and C substituted with one or more halogens 1-3 alkoxy; R e , R f , and R g and at least one of is not H. In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is [ka] wherein R e , R f , and R gare each independently selected from the group consisting of H, F, Cl, Br, methyl, ethenyl, isopropyl, difluoromethyl, trifluoromethyl, 1,1-difluoroethyl, methoxy, trifluoromethoxy, difluoromethoxy, and 1,1-difluoroethoxy; R e , R f , and R g and at least one of is not H. In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is [ka] wherein R e , R f , and R g are each independently selected from the group consisting of H, F, Cl, methyl, isopropyl, trifluoromethyl, trifluoromethoxy, and difluoromethoxy; R e , R f , and R g At least one of them is not H.

[0091] In an exemplary embodiment, the compound is according to Formula (I), where T and Z are as described herein, and X is substituted or unsubstituted pyridinyl. In an exemplary embodiment, the compound is according to Formula (I), where T and Z are as described herein, and X is unsubstituted pyridinyl or halogen, C2-C4 alkenyl, unsubstituted C 1-3 Alkyl, C substituted with one or more halogens 1-3 Alkyl, unsubstituted C 1-3 Alkoxy and C substituted with one or more halogens 1-3In an exemplary embodiment, the compound is according to formula (I), wherein T and Z are as described herein, and X is unsubstituted pyridinyl or pyridinyl substituted with one or more members selected from the group consisting of F, Cl, Br, methyl, ethenyl, isopropyl, difluoromethyl, trifluoromethyl, 1,1-difluoroethyl, methoxy, trifluoromethoxy, difluoromethoxy, and 1,1-difluoroethoxy. In an exemplary embodiment, the compound is according to formula (I), wherein T and Z are as described herein, and X is unsubstituted pyridinyl. In an exemplary embodiment, the compound is according to formula (I), wherein T and Z are as described herein, and X is [ka] wherein R e and R f are each independently H, halogen, C2-C4 alkenyl, unsubstituted C 1-3 Alkyl, C substituted with one or more halogens 1-3 Alkyl, unsubstituted C 1-3 Alkoxy and C substituted with one or more halogens 1-3 alkoxy; R e and R f and at least one of is not H. In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is [ka] wherein R e and R f are each independently selected from the group consisting of H, F, Cl, Br, methyl, ethenyl, isopropyl, difluoromethyl, trifluoromethyl, 1,1-difluoroethyl, methoxy, trifluoromethoxy, difluoromethoxy, and 1,1-difluoroethoxy; R e and R fand at least one of is not H. In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is [ka] wherein R e and R f are each independently selected from the group consisting of H, F, methyl, ethenyl, isopropyl, trifluoromethyl, and 1,1-difluoroethyl; R e and R f At least one of them is not H.

[0092] In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is: [ka] wherein R e , R f , and R g are each independently H, halogen, C2-C4 alkenyl, unsubstituted C 1-3 Alkyl, C substituted with one or more halogens 1-3 Alkyl, unsubstituted C 1-3 Alkoxy and C substituted with one or more halogens 1-3 alkoxy; R e , R f , and R g and at least one of is not H. In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is [ka] wherein R e , R f , and R gare each independently selected from the group consisting of H, F, Cl, Br, methyl, ethenyl, isopropyl, difluoromethyl, trifluoromethyl, 1,1-difluoroethyl, methoxy, trifluoromethoxy, difluoromethoxy, and 1,1-difluoroethoxy; R e , R f , and R g and at least one of is not H. In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is [ka] wherein R e , R f , and R g are each independently selected from the group consisting of H, Cl, and trifluoromethyl; R e , R f , and R g At least one of them is not H.

[0093] In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is substituted or unsubstituted pyrimidinyl. In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is unsubstituted pyrimidinyl or halogen, C-C alkenyl, unsubstituted C 1-3 Alkyl, C substituted with one or more halogens 1-3 Alkyl, unsubstituted C 1-3 Alkoxy and C substituted with one or more halogens 1-3In an exemplary embodiment, the compound is according to formula (I), wherein T and Z are as described herein, and X is unsubstituted pyrimidinyl or pyrimidinyl substituted with one or more members selected from the group consisting of F, Cl, Br, methyl, ethenyl, isopropyl, difluoromethyl, trifluoromethyl, 1,1-difluoroethyl, methoxy, trifluoromethoxy, difluoromethoxy, and 1,1-difluoroethoxy. In an exemplary embodiment, the compound is according to formula (I), wherein T and Z are as described herein, and X is unsubstituted pyrimidinyl. In an exemplary embodiment, the compound is according to formula (I), wherein T and Z are as described herein, and X is [ka] wherein R e is H, halogen, C2-C4 alkenyl, unsubstituted C 1-3 Alkyl, C substituted with one or more halogens 1-3 Alkyl, unsubstituted C 1-3 Alkoxy or C substituted with one or more halogens 1-3 In an exemplary embodiment, the compound is according to Formula (I), where T and Z are as described herein and X is: [ka] wherein R e is F, Cl, Br, methyl, ethenyl, isopropyl, difluoromethyl, trifluoromethyl, 1,1-difluoroethyl, methoxy, trifluoromethoxy, difluoromethoxy, or 1,1-difluoroethoxy. In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein and X is [ka] wherein Re is trifluoromethyl.

[0094] In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is substituted or unsubstituted thienyl. In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein, and X is unsubstituted thienyl or halogen, C-C alkenyl, unsubstituted C 1-3 Alkyl, C substituted with one or more halogens 1-3 Alkyl, unsubstituted C 1-3 Alkoxy and C substituted with one or more halogens 1-3 In an exemplary embodiment, the compound is according to formula (I), wherein T and Z are as described herein, and X is unsubstituted thienyl or thienyl substituted with one or more members selected from the group consisting of F, Cl, Br, methyl, ethenyl, isopropyl, difluoromethyl, trifluoromethyl, 1,1-difluoroethyl, methoxy, trifluoromethoxy, difluoromethoxy, and 1,1-difluoroethoxy. In an exemplary embodiment, the compound is according to formula (I), wherein T and Z are as described herein, and X is unsubstituted thienyl. In an exemplary embodiment, the compound is according to formula (I), wherein T and Z are as described herein, and X is [ka] wherein R e is halogen, C2-C4 alkenyl, unsubstituted C 1-3 Alkyl, C substituted with one or more halogens 1-3 Alkyl, unsubstituted C 1-3 Alkoxy or C substituted with one or more halogens 1-3 In an exemplary embodiment, the compound is according to Formula (I), where T and Z are as described herein and X is: [ka] wherein R e is F, Cl, Br, methyl, ethenyl, isopropyl, difluoromethyl, trifluoromethyl, 1,1-difluoroethyl, methoxy, trifluoromethoxy, difluoromethoxy, or 1,1-difluoroethoxy. In an exemplary embodiment, the compound is according to Formula (I), wherein T and Z are as described herein and X is [ka] wherein R e is trifluoromethyl.

[0095] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and Z are as described herein, and X is 4-(trifluoromethyl)phenyl, 4-fluorophenyl, 2-(trifluoromethyl)pyridin-5-yl, or 2-(1,1-difluoroethyl)pyridin-5-yl.

[0096] Group Z: In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted ethenyl.

[0097] part a In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is substituted or unsubstituted phenyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted furan, substituted or unsubstituted thienyl, substituted or unsubstituted pyrimidinyl, substituted or unsubstituted oxazolyl, or substituted or unsubstituted ethenyl.

[0098] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is phenyl, pyridinyl, furan, thienyl, pyrimidinyl, or oxazolyl.

[0099] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is pyridinyl or ethenyl substituted with phenyl.

[0100] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is substituted or unsubstituted 4-pyrimidinyl, substituted or unsubstituted 2-pyrimidinyl, substituted or unsubstituted 6-isoquinolyl, substituted or unsubstituted 3-quinolyl, substituted or unsubstituted 6-quinolyl, substituted or unsubstituted 1-pyrrolidinyl, substituted or unsubstituted 2,3-dihydro-1-benzofuran-5-yl, substituted or unsubstituted xylyl, substituted or unsubstituted tolyl. , substituted or unsubstituted 4-mesylphenyl, substituted or unsubstituted 2-pyridinyl, substituted or unsubstituted 3-pyridinyl, substituted or unsubstituted 4-pyridinyl, substituted or unsubstituted 2,4-diaza-2-indanyl, substituted or unsubstituted 1,6-diaza-2-naphthyl, substituted or unsubstituted 1,5-diaza-2-naphthyl, substituted or unsubstituted 2,3-dihydro-1,4-dioxa-5-aza-7-naphthyl, substituted or unsubstituted 2,3-dihydro-1,4-benzodioxin-6-yl, substituted or unsubstituted 1-oxa-4-aza-2- Indenyl, substituted or unsubstituted 6-quinoxalinyl, substituted or unsubstituted 2-quinoxalinyl, substituted or unsubstituted 2-furanyl, substituted or unsubstituted 1-oxa-6-aza-2-indenyl, substituted or unsubstituted 1,7a-diaza-2-indenyl, substituted or unsubstituted 1,7a-diaza-2-indenyl, substituted or unsubstituted 1,7a-diaza-2-indenyl, substituted or unsubstituted 1,3a-diaza-2-indenyl, substituted or unsubstituted 1-oxa-3,4-diaza-2-indenyl, substituted or unsubstituted 1,3a,6-diaza- riaza-2-indenyl, substituted or unsubstituted 1,3,3a-triaza-2-indenyl, substituted or unsubstituted 1,3a-diaza-2-indenyl, substituted or unsubstituted 1-oxa-4-aza-2-indenyl, substituted or unsubstituted 1,3a-diaza-3-indenyl, substituted or unsubstituted 1,3-oxazol-2-yl, substituted or unsubstituted 1,3-benzoxazol-2-yl, substituted or unsubstituted 5-benzothiophenyl, substituted or unsubstituted 1-pyrrolyl, substituted or unsubstituted 1-pyrazolyl, substituted or unsubstituted 1H-1,3-Benzimidazol-1-yl, substituted or unsubstituted 2H-indazol-2-yl, substituted or unsubstituted 2H-indazol-5-yl, substituted or unsubstituted 1H-indazol-5-yl, substituted or unsubstituted indazol-5-yl, substituted or unsubstituted 2H-indazol-6-yl, substituted or unsubstituted 2H-1,2,7-triazainden-5-yl, substituted or unsubstituted 2H-1,2,4-triazainden-5-yl, substituted or unsubstituted 2H-1,2,6-triazainden-5-yl, substituted or unsubstituted 1-benzofuran-2-yl, substituted or unsubstituted 2,3-dihydro-1-benzofuran-4-yl, substituted or unsubstituted 1-benzofuran-4-yl, substituted or unsubstituted 1-benzofuran-6-yl, substituted or unsubstituted 2,3-dihydro 1-benzofuran-7-yl, substituted or unsubstituted 1-benzofuran-7-yl, substituted or unsubstituted 2,4-diaza-2-indanyl, substituted or unsubstituted 2,5-diaza-2-indanyl, substituted or unsubstituted 2,4,7-triaza-2-indanyl, substituted or unsubstituted 3,4-dihydro-2H-1,4-benzoxazin-6-yl, substituted or unsubstituted 3,4-dihydro-2H-1,4-benzoxazin-7-yl, substituted or unsubstituted 1,3-thiazol-2-yl, substituted or unsubstituted 2-thienyl, substituted or unsubstituted 1,3-benzothiazol-2-yl, substituted or unsubstituted imidazolidinone, substituted or unsubstituted 2-isoindolinyl, substituted or unsubstituted 1-oxo-2-isoindolinyl, or substituted or unsubstituted 6-indolyl.

[0101] In an exemplary embodiment, the compound, or a salt or hydrate or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is substituted or unsubstituted 2-(pyridinyl)ethenyl or substituted or unsubstituted 2-(phenyl)ethenyl. In an exemplary embodiment, the compound, or a salt or hydrate or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is substituted or unsubstituted 2-(4-pyridinyl)ethenyl, substituted or unsubstituted 2-(3-pyridinyl)ethenyl, or substituted or unsubstituted 2-(phenyl)ethenyl. In an exemplary embodiment, the compound, or a salt or hydrate or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is 2-(4-pyridinyl)ethenyl, 2-(3-pyridinyl)ethenyl, or 2-(phenyl)ethenyl. In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), wherein T and X are as described herein, and Z is a halogen or C 1-3 2-(4-pyridinyl)ethenyl substituted with alkyl, halogen or C 1-3 2-(3-pyridinyl)ethenyl substituted with alkyl, or halogen or C 1-3 It is 2-(phenyl)ethenyl substituted with alkyl.

[0102] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), wherein T and X are as described herein, and Z is: [ka] [ka] ,or [ka] is.

[0103] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is: [ka] [ka] ,or [ka] where R y and R z are each independently selected from H, halogen, cyano, cyclopropyl, C1-C3 alkyl, C1-C3 alkyl substituted with cyclopropyl, C1-C3 alkyl substituted with C1-C3 alkoxy, C1-C3 alkyl substituted with hydroxy, C1-C3 alkyl substituted with one or more halogens, C1-C3 alkyl substituted with hydroxy and one or more halogens, C2-C4 alkenyl, C1-C3 alkoxy, C1-C3 alkoxy substituted with cyclopropyl, C1-C3 alkoxy substituted with one or more halogens, C1-C3 alkoxy substituted with one or more halogens, C1-C3 alkoxy substituted with hydroxy and one or more halogens, C1-C3 alkylamino, C1-C3 dialkylamino, pyridinyl, pyridinyl substituted with C1-C3 alkyl, 1H-1,2,4-triazol-3-yl, and 1-methyl-1H-1,2,4-triazol-3-yl substituted with C1-C3 alkyl; R y and R z At least one of them is not H.

[0104] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is: [ka] [ka] ,or [ka] where R y and R z are each independently H, F, Cl, cyano, cyclopropyl, methyl, cyclopropylmethyl, ethenyl, isopropyl, methoxy, 2-ethoxy, difluoromethyl, trifluoromethyl, trifluoromethoxy, difluoromethoxy, 3,3,3-trifluoro-2-hydroxypropyl, methoxymethyl, 2,2-difluoroethyl, dimethylamino, 2,2,2-trifluoroethyl, 2-hydroxyethyl, 2-methyl-4-pyridyl, 1-methyl-1H-1,2,4-triazol-3-yl, and R y and R z At least one of them is not H.

[0105] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is 1-methyl-6-isoquinolyl, 1-methoxy-6-isoquinolyl, 3-chloro-6-isoquinolyl, m-tolyl, 2-fluoro-3-tolyl, 4-fluoro-3-tolyl, 2-ethoxy-4-pyrimidinyl, 4,5-dimethyl-2-pyridyl, p-(trimethylsilyl)-2-pyridyl, 2-methyl-4-pyrimidin ... fluoromethyl)phenyl, 2,3-dihydro-1-benzofuran-5-yl, (3S,4S)-3,4-dimethyl-1-pyrrolidinyl, 3-fluoro-4-mesylphenyl, 5,6-dimethyl-3-pyridyl, p-(1-methyl-1H-1,2,4-triazol-3-yl)phenyl, 4-cyanophenyl, 3-chloro-4-methoxyphenyl, phenyl, 3,5-dichlorophenyl, 3-chloro-4-fluorophenyl, 5-chloro-2-fluorophenyl, m-chlorophenyl, p-chlorophenyl, p-fluorophenyl, 3-chloro-2-fluorophenyl, 2-fluoro-4-cyanophenyl, p-difluoromethoxyphenyl, 2-fluoro-3-methoxyphenyl, p-methoxyphenyl, p-(dimethylamino)phenyl, 5-methyl-2,4-diaza-2-indanyl, p-mesylphenyl, 2-naphthyl, 1,5-diaza-2-na phthalic anhydride, 1,6-diaza-2-naphthyl, 1,8-diaza-2-naphthyl, 2,3-dihydro-1,4-dioxa-5-aza-7-naphthyl, 3-methoxy-6-quinolyl, 1-methyl-1,2,3,4-tetrahydro-6-quinolyl, 1-methyl-4,4-dimethyl-2-oxo-3,4-dihydro-6-quinolyl, 6-methyl-3-quinolyl, 6-fluoro-3-quinolyl, 7-(trifluoromethyl)-3-quinolyl, 2,6-dimethyl-4-pyridyl, 6-methoxy-5-methyl-3-pyridyl, 5-cyclopropyl-3-pyridinyl, 5-chloro-6-methyl-3-pyridinyl, 5-chloro-6-(methoxymethyl)-3-pyridinyl, 5-chloro-6-methoxy-3-pyridinyl, 6-(dimethylamino)-3-pyridinyl, 3-pyridinyl, 6-cyano-2-pyridinyl, 4-ethynyl-2-pyridinyl, 2-pyridinyl, 6-chloro-2-pyridinyl, 6-(trifluoromethyl)-2-pyridyl, 6-methyl-2-pyridinyl, 5- Chloro-2-furanyl, 5-(trifluoromethyl)-2-furanyl, 4,5-dimethyl-2-furanyl, 6-quinoxalinyl, 2-quinoxalinyl, 7-methoxy-1-oxa-6-aza-2-indenyl, 6-chloro-1,7a-diaza-2-indenyl, 7-methyl-1,7a-diaza-2-indenyl, 5-methyl-1,7a-diaza-2-indenyl, 7-methyl-1,3a-diaza-2-indenyl, 5-methyl-1-oxa-3,4-diaza-2-indenyl, 1,3a,6-triaza-2-indenyl, 1 ,3,3a-triaza-2-indenyl, 1,3a-diaza-2-indenyl, 1-oxa-4-aza-2-indenyl, 7-methoxy-1,3a-diaza-3-indenyl, 5-cyclopropyl-4-methyl-1,3-oxazol-2-yl, 5-methoxy-1,3-benzoxazol-2-yl, 5-fluoro-1,3-benzoxazol-2-yl, 1,3-benzoxazol-2-yl, 5-cyano-benzoxazol-2-yl, 1-benzothiophen-5-yl, 3-bromo-4-methyl-1-pyro lyl, 3-(trifluoromethyl)-1-pyrazolyl, 5-cyclopropyl-1-methyl-3-pyrazolyl, 1-(2,2-difluoroethyl)-5-methyl-3-pyrazolyl, 1-isopropyl-5-methyl-3-pyrazolyl, 4-methoxy-1H-1,3-benzimidazol-1-yl, 1H-1,3-benzimidazol-2-yl, 1-methyl-1H-1,3-benzimidazol-2-yl, 2H-indazol-2-yl, 3-(difluoromethyl)-2-methyl-2H-indazol-5-yl, 1-(2,2-Difluoroethyl)-1H-indazol-5-yl, 1-(difluoromethyl)-1H-indazol-5-yl, 1-ethyl-3-methyl-1H-indazol-5-yl, 2-(3,3,3-trifluoro-2-hydroxypropyl)-2H-indazol-5-yl, 2-(2,2,2-trifluoroethyl)-2H-indazol-5-yl, 2-methylcyano-indazol-5-yl, 2-ethyl-3-methyl-2H-indazol-5-yl, 2-cyclobutyl-2H-indazol-5-yl 2-isopropyl-2H-indazol-5-yl, 2-(cyclopropylmethyl)-2H-indazol-5-yl, 2-methyl-2H-indazol-6-yl, 2-methyl-3-methyl-2H-1,2,7-triazainden-5-yl, 2-methyl-2H-1,2,4-triazainden-5-yl, 2-methyl-3-methyl-2H-1,2,6-triazainden-5-yl, 4-fluoro-1-benzofuran-2-yl, 7-fluoro-1-benzofuran-2-yl, 7-methoxy-1-benzofuran-2-yl, 2-indanyl, 3-methyl-1-benzofuran-2-yl, 1-benzofuran-2-yl, 2,3-dihydro-1-benzofuran-4-yl, 2-methyl-1-benzofuran-6-yl, 2,3-dihydro-1-benzofuran-7-yl, 2,4-diaza-2-indanyl, 2,5-diaza-2-indanyl, 2,4,7-triaza-2-indanyl, 4-methyl-3,4-dihydro-2H-1,4-benzoxazin-6-yl, 4-methyl-3,4-dihydro-2H-1,4-benzoxazin-7-yl, 5 -cyclopropyl-4-methyl-1,3-thiazol-2-yl, 4,5-dimethyl-1,3-thiazol-2-yl, 2-pyrimidinyl, 4-pyrimidinyl, 4,5-dimethyl-2-thienyl, 1,3-benzothiazol-2-yl, p-phenyl-2-imidazolidinone, 2-isoindolinyl, 7-fluoro-1-oxo-2-isoindolinyl, 1-oxo-2-isoindolinyl, 4-methoxy-2-isoindolinyl, 5-methoxy-2-isoindolinyl, or 1-methyl-6-indolyl.

[0106] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is (E)-2-(3-fluoro-4-pyridyl)ethenyl, (E)-2-(2-fluoro-4-pyridyl)ethenyl, (E)-2-phenylethenyl, (E)-2-(2,6-dimethyl-4-pyridyl)ethenyl, (E)-2-(4-pyridyl)ethenyl, (E)-2-(2-methyl-4-pyridyl)ethenyl, or (E)-2-(3-pyridyl)ethenyl.

[0107] part b In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is substituted or unsubstituted phenyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted furan, substituted or unsubstituted thienyl, substituted or unsubstituted pyrimidinyl, and substituted or unsubstituted oxazolyl.

[0108] In an exemplary embodiment, the compound, or salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is substituted or unsubstituted pyrimidinyl, substituted or unsubstituted isoquinolyl, substituted or unsubstituted quinolyl, substituted or unsubstituted xylyl, substituted or unsubstituted pyrrolyl, substituted or unsubstituted cyclopropyl, substituted or unsubstituted oxazolyl, substituted or unsubstituted triazaindenyl, substituted or unsubstituted diazaindenyl, substituted or unsubstituted phenyl, substituted or unsubstituted naphthyl, substituted or unsubstituted benzofuranyl, substituted or unsubstituted indazolyl, substituted or unsubstituted benzothiazolyl, substituted or unsubstituted benzoxazolyl, substituted or unsubstituted indolyl, substituted or unsubstituted 2,3-dihydro-1,4-benzodioxin-6-yl, substituted or unsubstituted thienyl, substituted or unsubstituted isoindoinyl, or substituted or unsubstituted 1-oxa-4-aza-2-indenyl.

[0109] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), wherein T and X are as described herein, and Z is: [ka] ,or [ka] where R y and R z are each independently selected from H, halogen, cyano, cyclopropyl, C1-C3 alkyl, C1-C3 alkyl substituted with cyclopropyl, C1-C3 alkyl substituted with C1-C3 alkoxy, C1-C3 alkyl substituted with hydroxy, C1-C3 alkyl substituted with one or more halogens, C1-C3 alkyl substituted with hydroxy and one or more halogens, C2-C4 alkenyl, C1-C3 alkoxy, C1-C3 alkoxy substituted with cyclopropyl, C1-C3 alkoxy substituted with one or more halogens, C1-C3 alkoxy substituted with one or more halogens, C1-C3 alkoxy substituted with hydroxy and one or more halogens, C1-C3 alkylamino, C1-C3 dialkylamino, pyridinyl, pyridinyl substituted with C1-C3 alkyl, 1H-1,2,4-triazol-3-yl, and 1-methyl-1H-1,2,4-triazol-3-yl substituted with C1-C3 alkyl; R y and R z At least one of them is not H.

[0110] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), wherein T and X are as described herein, and Z is: [ka] ,or [ka] where Ry and R z are each independently selected from H, fluoro, chloro, cyclopropyl, methyl, isopropyl, methoxy, 2-hydroxyethyl, and 2-methyl-4-pyridyl; R y and R z At least one of them is not H.

[0111] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is 2-cyclopropyl-4-pyrimidinyl, 1-methyl-6-isoquinolyl, 1-(2-hydroxyethyl)-6-isoquinolyl, 7-isoquinolyl, 8-fluoro-3-quinolyl, 1-methoxy-6-isoquinolyl, 3,4-xylyl, 1-isopropyl-6-isoquinolyl, 8-fluoro-7-methyl-3-quinol ...methyl-6-isoquinolyl, 3,4-xylyl, 1-isopropyl-6-isoquinolyl, 8-fluoro-7-methyl-3-quinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-methyl-6-iso Noryl, 3-(2-methoxy-3-pyridyl)-1-pyrrolyl, 2-cyclopropyl, 2-(2-methyl-4-pyridyl)-1,3-oxazol-5-yl, 2-methyl-6-quinolyl, 2-methyl-2H-1,2,7-triazainden-5-yl, p-phenyl-1,3-oxazolidin-2-one, 5-fluoro-1,3-benzoxazol-2-yl, 4-chloro-5-methoxy-1-benzofuran-2-yl, 6-isoquinolyl, 6-fluoro-1-methyl-1H-in 5-naphthyl, 2-methyl-1,3-benzoxazol-5-yl, 5-methoxy-1,3-benzothiazol-2-yl, 6-fluoro-1,3-benzoxazol-2-yl, 3-thia-1,4-diaza-2-indenyl, 2-indole-5-carbonitrile, 2,3-dihydro-1,4-benzodioxin-6-yl, 2-methyl-1,3-benzoxazol-6-yl, 2-fluoro-4-methoxyphenyl, 5-methoxy-1,3-benzoxazol- thiazol-2-yl, 5-methoxy-2-isoindolinyl, 6-methoxy-1,3-benzothiazol-2-yl, 2-methyl-2H-indazol-5-yl, 1-oxa-4-aza-2-indenyl, 4-methoxy-2-isoindolinyl, 4-fluoro-5-methoxy-2-isoindolinyl, phenyl, p-(dimethylamino)phenyl, 1-benzofuran-5-yl, m-methoxyphenyl, 5-methyl-2-thienyl, p-methoxyphenyl, and p-chlorophenyl.

[0112] part c In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is substituted or unsubstituted 6-isoquinolyl, substituted or unsubstituted 2-quinolyl, substituted or unsubstituted 3-quinolyl, substituted or unsubstituted 4-quinolyl, substituted or unsubstituted 6-quinolyl, substituted or unsubstituted 7-quinolyl, substituted or unsubstituted 7-quinazolinyl, substituted or unsubstituted 1H-indazole. substituted or unsubstituted 2H-indazol-5-yl, substituted or unsubstituted 1-benzofuran-6-yl, substituted or unsubstituted 1-benzofuran-2-yl, substituted or unsubstituted 2,3-dihydro-1-benzofuran-6-yl, substituted or unsubstituted 1,2,3-benzotriazol-5-yl, substituted or unsubstituted benzothiazol-2-yl, substituted or unsubstituted 1,3a-diaza-2-indenyl, substituted or unsubstituted 1,7a-diaza-2-indenyl substituted or unsubstituted 1,3a-diaza-5-indenyl, substituted or unsubstituted 3,4-diaza-2-indenyl, substituted or unsubstituted 1-thia-4-aza-2-indenyl, substituted or unsubstituted indazol-5-yl, substituted or unsubstituted 1H-indazol-5-yl, substituted or unsubstituted 2H-indazol-5-yl, substituted or unsubstituted 1,2,6-triazainden-5-yl, substituted or unsubstituted 1H-1,2,6-triazainden-5-yl, substituted or unsubstituted 2H-1,2,6-triazainden-5-yl, substituted or unsubstituted phenyl, substituted or unsubstituted 1-pyrrolyl, substituted or unsubstituted 2-indolyl, substituted or unsubstituted 5-indolinyl, substituted or unsubstituted 2-oxo-5-indolyl, substituted or unsubstituted 2-isoindolinyl, substituted or unsubstituted 3-oxo-5-isoindolinyl, substituted or unsubstituted 1-oxo-5-isoindolinyl, or substituted or unsubstituted 2-thienyl.

[0113] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is: [ka] [ka] [ka] ,or [ka] where R y and R z are each independently H, halogen, cyano, cyclopropyl, cyclopropyl substituted with C1-C3 alkyl, cyclopropyl substituted with hydroxy-substituted C1-C3 alkyl, C1-C3 alkyl, C1-C3 alkyl substituted with cyclopropyl, C1-C3 alkyl substituted with an amino group, C1-C3 alkyl substituted with a C1-C3 alkoxy group, C1-C3 alkyl substituted with a hydroxy group, C1-C3 alkyl substituted with one or more halogens, C1-C3 alkyl substituted with hydroxy and one or more halogens, C1-C3 alkyl substituted with cyano, C1-C3 alkyl substituted with -S(O)2CH3 is selected from 1-C3 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C1-C3 alkoxy, C1-C3 alkoxy substituted with cyclopropyl, C1-C3 alkoxy substituted with one or more hydroxy, C1-C3 alkoxy substituted with one or more halogens, C1-C3 alkoxy substituted with hydroxy and one or more halogens, C1-C3 alkylamino, C1-C3 dialkylamino, pyridinyl, pyridinyl substituted with C1-C3 alkyl, 1H-1,2,4-triazol-3-yl, and 1-methyl-1H-1,2,4-triazol-3-yl substituted with C1-C3 alkyl; R y and R z At least one of them is not H.

[0114] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is: [ka] [ka] or [ka] where R y and R z are each independently selected from H, F, Cl, cyano, (1S,2R)-2-(methoxymethyl)cyclopropyl, [(1R,2R)-2-(hydroxymethyl)cyclopropyl, (1S,2S)-2-(hydroxymethyl)cyclopropyl, (1R,2R)-2-methylcyclopropyl, cyclopropyl, 2-hydroxypropyl, methoxy, ethoxy, 2-hydroxyethoxy, methyl, difluoromethyl, trifluoromethyl, ethyl, 2-methoxyethyl, 2-aminoethyl, 2-hydroxyethyl, 2-fluoroethyl, 1,1-difluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, propyl, isopropyl, 2-propynyl, methylamino, dimethylamino, cyanomethyl, and 2-mesylethyl; R y and R z At least one of them is not H.

[0115] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is [(1S,2R)-2-(methoxymethyl)cyclopropyl]-6-isoquinolyl, (2-methoxyethyl)-6-isoquinolyl, [(1R,2R)-2-(hydroxymethyl)cyclopropyl]-6-isoquinolyl, [(1S,2S)-2-(hydroxymethyl)cyclopropyl]-6-isoquinolyl, (1R,2R)-2-methylcyclopropyl]-6-isoquinolyl. , (2-hydroxypropyl)-6-isoquinolyl, 1-methoxy-6-isoquinolyl, (2-hydroxyethoxy)-6-isoquinolyl, 1-methyl-6-isoquinolyl, 1-(methylamino)-6-isoquinolyl, 7-fluoro-1-methoxy-6-isoquinolyl, 2-methyl-1-oxo-3,4-dihydro-6-isoquinolyl, 1,3-dimethyl-6-isoquinolyl, 1-ethyl-6-isoquinolyl, 2-methyl-1-oxo-6-isoquinolyl, 5-fluoro-1-methoxy-6-isoquinolyl, 1-ethoxy-6-isoquinolyl, 1- (Dimethylamino)-6-isoquinolyl, 3-methyl-6-isoquinolyl, 3-fluoro-6-isoquinolyl, 6-isoquinolyl, 4-methyl-2-quinolyl, 7-fluoro-3-quinolyl, 8-methyl-3-quinolyl, 8-methoxy-3-quinolyl, 7-chloro-3-quinolyl, 8-chloro-3-quinolyl, 3-quinolyl, 7-methyl-3-quinolyl, 8-methoxy-4-quinolyl, 8-methoxy-2-methyl-4-quinolyl, 2-ethoxy-6-quinolyl, 1-methyl-3-methyl-2-oxo-3,4-dihydro-6-quinolyl, 3-methyl quinolyl, 2-methoxy-6-quinolyl, 2-methyl-6-quinolyl, 1-methyl-2-oxo-3,4-dihydro-6-quinolyl, 1-methyl-2-oxo-6-quinolyl, 6-quinolyl, 8-fluoro-7-quinolyl, 7-quinolyl, 2-methyl-7-quinazolinyl, 7-fluoro-2,3-dihydro-1-benzofuran-6-yl, 2,3-dihydro-1-benzofuran-6-yl, 3-chloro-1-benzofuran-2-yl, 4-methoxy-1-benzofuran-2-yl, 5,6-dimethoxy-1-benzofuran-2-yl, 4,6-Dimethoxy-1-benzofuran-2-yl, 4-chloro-5-methoxy-1-benzofuran-2-yl, 5-methoxy-1-benzofuran-2-yl, 4-fluoro-1-benzofuran-2-yl, 1-benzofuran-2-yl, (2-aminoethyl)-1H-indazol-5-yl, (2-propynyl)-1H-indazol-5-yl, (2-propynyl)-2H-indazol-5-yl, 6-methoxy-1,3a-diaza-2-indenyl, 5-methyl 1,7a-diaza-2-indenyl, 6-methoxy-1,7a-diaza-2-indenyl, 4-chloro-1,7a-diaza-2-indenyl, 6-methyl-1,7a-diaza-2-indenyl, 2-methyl-2H-1,2,3-benzotriazol-5-yl, 5-(1,1-difluoroethyl)-1,3-benzoxazol-2-yl, 5-(2,2,2 trifluoroethyl)-1,3-benzoxazol-2-yl, 5-isopropyl-1,3- Benzoxazol-2-yl, 4-methoxy-1,3-benzoxazol-2-yl, 5-ethyl-1,3-benzoxazol-2-yl, 4-fluoro-1,3-benzoxazol-2-yl, 5-methoxy-1,3-benzoxazol-2-yl, 1,3-benzoxazol-2-yl, 2-methyl-1,3-benzoxazol-6-yl, 3-benzothiazol-2-yl, 4-methoxy-1,3-benzothiazol-2-yl, o-fluoro brophenyl, 2-fluoro-3-methoxyphenyl, m-tolyl, 1-thia-4-aza-2-indenyl, 1,7a-diaza-2-indenyl, 2-methyl-1,3a-diaza-5-indenyl, 5-cyclopropyl-1-oxa-3,4-diaza-2-indenyl, 1-(2-hydroxyethyl)-1H-indazol-5-yl, 1-(2-mesylethyl)-1H-indazol-5-yl, 1-cyanomethyl-1H-indazol-5-yl, 1-(2,2,2-trifluoroethyl)-1H-indazol-5-yl, 1-methyl-1H-indazol-5-yl, 1-cyclopropyl-1H-indazol-5-yl, 1-ethyl-1H-indazol-5-yl, 7-chloro-1-methyl-1H-indazol-5-yl, 3-chloro-2-methyl-2H-indazol-5-yl, 2-methyl-6-methyl-2H-indazol-5-yl, 2-methyl-7-methyl-2 2H-indazol-5-yl, (2-fluoroethyl)-2H-indazol-5-yl, 2-methyl-3-methyl-2H-indazol-5-yl, 2-(2,2-difluoroethyl)-2H-indazol-5-yl, 2-(2-hydroxyethyl)-2H-indazol-5-yl, 2-propyl-2H-indazol-5-yl, 2-(difluoromethyl)-2H-indazol-5-yl, 7-chloro-2-methyl 2-methyl-2H-indazol-5-yl, 7-fluoro-2-methyl-2H-indazol-5-yl, 2-ethyl-2H-indazol-5-yl, 2-methyl-2H-indazol-6-yl, 6-fluoro-2-methyl-2H-indazol-5-yl, 2-methyl-2H-indazol-5-yl, 2-methyl-2H-1,2,6-triazainden-5-yl, 1-methyl-1H-1,2,6-triazainden- 5-yl, 2-ethyl-2H-1,2,6-triazainden-5-yl, 3,4-dimethyl-1-pyrrolyl, 5-cyano-2-indolyl, 1-methyl-2-oxo-5-indolinyl, 2-methyl-3-oxo-5-isoindolinyl, 2-methyl-1-oxo-5-isoindolinyl, 5-cyano-isoindolin-2-yl, 4-fluoro-2-isoindolinyl, or 4,5-dimethyl-2-thienyl.

[0116] part d In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is substituted or unsubstituted phenyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted furan, substituted or unsubstituted thienyl, substituted or unsubstituted pyrimidinyl, and substituted or unsubstituted oxazolyl.

[0117] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is substituted or unsubstituted isoquinolyl, substituted or unsubstituted quinolyl, substituted or unsubstituted indenyl, substituted or unsubstituted indazolyl, substituted or unsubstituted benzoxazolyl, substituted or unsubstituted pyrimidinyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted benzofuranyl, or substituted or unsubstituted phenyl.

[0118] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is isoquinolyl, quinolyl, indenyl, indazolyl, benzoxazolyl, pyrimidinyl, pyridinyl, benzofuranyl, or phenyl, each of which is selected from the group consisting of halogen, C-C alkenyl, unsubstituted C 1-3 C substituted with alkyl, one or more halogen and / or hydroxy 1-3 Alkyl, unsubstituted cyclopropyl, unsubstituted C 1-3 Alkoxy and C substituted with one or more halogens 1-3 It may be substituted with one or more members selected from the group consisting of alkoxy.

[0119] In an exemplary embodiment, the compound, or salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is isoquinolyl, quinolyl, indenyl, indazolyl, benzoxazolyl, pyrimidinyl, pyridinyl, benzofuranyl, or phenyl, each of which can be substituted with one or more members selected from the group consisting of F, methyl, trifluoromethyl, (2-hydroxy)ethyl, (2-fluoro)ethyl, and cyclopropyl.

[0120] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is: [ka] ,or [ka] is.

[0121] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is: [ka] ,or [ka] where R z is halogen, C2-C4 alkenyl, unsubstituted C 1-3 Alkyl, C substituted with one or more halogens 1-3 Alkyl and / or hydroxy, cyclopropyl, unsubstituted C 1-3 Alkoxy or C substituted with one or more halogens 1-3 It is an alkoxy.

[0122] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is: [ka] ,or [ka] where R z is F, methyl, trifluoromethyl, (2-hydroxy)ethyl, (2-fluoro)ethyl, or cyclopropyl.

[0123] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has a structure according to Formula (I), where T and X are as described herein, and Z is 1-methylisoquinolin-6-yl, 2-(trifluoromethyl)pyrimidin-4-yl, 2-methylpyrimidin-4-yl, 1-methyl-6-isoquinolyl, 1-(2-hydroxyethyl)-6-isoquinolyl, 3-isoquinolyl, 6-quinolyl, 8- fluoro-3-quinolyl, 8-fluoro-7-quinolyl, 4-methyl-1,7a-diaza-2-indenyl, 1-thia-5-aza-2-indenyl, 1-(2-fluoroethyl)-1H-indazol-5-yl, 3-quinolyl, 2-cyclopropyl-2H-indazol-5-yl, 6-fluoro-1,3-benzoxazol-2-yl, 5-fluoro-2-pyridyl, 1-benzofuran-2-yl, or phenyl.

[0124] Groups T and X: In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has the formula (II): [ka] wherein Z is as defined herein.

[0125] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has the formula (III): [ka] wherein Z is as defined herein.

[0126] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has the formula (IV): [ka] wherein Z is as defined herein.

[0127] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has the formula (V): [ka] wherein Z is as defined herein.

[0128] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has the formula (VI): [ka] wherein Z is as defined herein.

[0129] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has Formula (VII): [ka] wherein Z is as defined herein.

[0130] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has Formula (VIII): [ka] wherein Z is as defined herein.

[0131] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has Formula (IX): [ka] wherein Z is as defined herein.

[0132] Groups X and Z: In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has the formula (X): [ka] wherein T is as defined herein and Z is 1-methylisoquinolin-6-yl, 2-(trifluoromethyl)pyrimidin-4-yl, 2-methylpyrimidin-4-yl, 1-methyl-6-isoquinolyl, 1-(2-hydroxyethyl)-6-isoquinolyl, 3-isoquinolyl, 6-quinolyl, 8-fluoro-3-quinolyl, 8-fluoro-7-quinolyl, 4-methyl-1,7a-diaza-2-indenyl, 1-thia-5-aza-2-indenyl, 1-(2-fluoroethyl)-1H-indazol-5-yl, 3-quinolyl, 2-cyclopropyl-2H-indazol-5-yl, 6-fluoro-1,3-benzoxazol-2-yl, 5-fluoro-2-pyridyl, 1-benzofuran-2-yl, or phenyl.

[0133] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has the formula (XI): [ka] wherein T is as defined herein and Z is 1-methylisoquinolin-6-yl, 2-(trifluoromethyl)pyrimidin-4-yl, 2-methylpyrimidin-4-yl, 1-methyl-6-isoquinolyl, 1-(2-hydroxyethyl)-6-isoquinolyl, 3-isoquinolyl, 6-quinolyl, 8-fluoro-3-quinolyl, 8-fluoro-7-quinolyl, 4-methyl-1,7a-diaza-2-indenyl, 1-thia-5-aza-2-indenyl, 1-(2-fluoroethyl)-1H-indazol-5-yl, 3-quinolyl, 2-cyclopropyl-2H-indazol-5-yl, 6-fluoro-1,3-benzoxazol-2-yl, 5-fluoro-2-pyridyl, 1-benzofuran-2-yl, or phenyl.

[0134] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has the formula (XII): [ka] wherein T is as defined herein and Z is 1-methylisoquinolin-6-yl, 2-(trifluoromethyl)pyrimidin-4-yl, 2-methylpyrimidin-4-yl, 1-methyl-6-isoquinolyl, 1-(2-hydroxyethyl)-6-isoquinolyl, 3-isoquinolyl, 6-quinolyl, 8-fluoro-3-quinolyl, 8-fluoro-7-quinolyl, 4-methyl-1,7a-diaza-2-indenyl, 1-thia-5-aza-2-indenyl, 1-(2-fluoroethyl)-1H-indazol-5-yl, 3-quinolyl, 2-cyclopropyl-2H-indazol-5-yl, 6-fluoro-1,3-benzoxazol-2-yl, 5-fluoro-2-pyridyl, 1-benzofuran-2-yl, or phenyl.

[0135] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has Formula (XIII): [ka] wherein T is as defined herein and Z is 1-methylisoquinolin-6-yl, 2-(trifluoromethyl)pyrimidin-4-yl, 2-methylpyrimidin-4-yl, 1-methyl-6-isoquinolyl, 1-(2-hydroxyethyl)-6-isoquinolyl, 3-isoquinolyl, 6-quinolyl, 8-fluoro-3-quinolyl, 8-fluoro-7-quinolyl, 4-methyl-1,7a-diaza-2-indenyl, 1-thia-5-aza-2-indenyl, 1-(2-fluoroethyl)-1H-indazol-5-yl, 3-quinolyl, 2-cyclopropyl-2H-indazol-5-yl, 6-fluoro-1,3-benzoxazol-2-yl, 5-fluoro-2-pyridyl, 1-benzofuran-2-yl, or phenyl.

[0136] Groups T and Z: In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has the formula (XIV): [ka] wherein X is as defined herein and Z is 1-methylisoquinolin-6-yl, 2-(trifluoromethyl)pyrimidin-4-yl, 2-methylpyrimidin-4-yl, 1-methyl-6-isoquinolyl, 1-(2-hydroxyethyl)-6-isoquinolyl, 3-isoquinolyl, 6-quinolyl, 8-fluoro-3-quinolyl, 8-fluoro-7-quinolyl, 4-methyl-1,7a-diaza-2-indenyl, 1-thia-5-aza-2-indenyl, 1-(2-fluoroethyl)-1H-indazol-5-yl, 3-quinolyl, 2-cyclopropyl-2H-indazol-5-yl, 6-fluoro-1,3-benzoxazol-2-yl, 5-fluoro-2-pyridyl, 1-benzofuran-2-yl, or phenyl.

[0137] In an exemplary embodiment, the compound, or a salt, hydrate, or solvate thereof, has the formula (XV): [ka] wherein X is as defined herein and Z is 1-methylisoquinolin-6-yl, 2-(trifluoromethyl)pyrimidin-4-yl, 2-methylpyrimidin-4-yl, 1-methyl-6-isoquinolyl, 1-(2-hydroxyethyl)-6-isoquinolyl, 3-isoquinolyl, 6-quinolyl, 8-fluoro-3-quinolyl, 8-fluoro-7-quinolyl, 4-methyl-1,7a-diaza-2-indenyl, 1-thia-5-aza-2-indenyl, 1-(2-fluoroethyl)-1H-indazol-5-yl, 3-quinolyl, 2-cyclopropyl-2H-indazol-5-yl, 6-fluoro-1,3-benzoxazol-2-yl, 5-fluoro-2-pyridyl, 1-benzofuran-2-yl, or phenyl.

[0138] In an exemplary embodiment, the compound, or salt, hydrate, or solvate thereof, is 7-{5-[6-(1,1-difluoroethyl)-3-pyridyl]-2-(1-methyl-6-isoquinolyl)-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-[1-(2-hydroxyethyl)-6-isoquinolyl]-5-[6-(trifluoromethyl)-3-pyridyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-[1-(2- hydroxyethyl)-6-isoquinolyl]-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-[5-(p-fluorophenyl)-2-(3-isoquinolyl)-1,3-oxazol-4-yl]-1,7-diaza-8(7H)-naphthalenone, 7-[5-(p-fluorophenyl)-2-(6-quinolyl)-1,3-oxazol-4-yl]-1,7-diaza-8(7H)-naphthalenone, 7-{2-(8-fluoro-3-quinolyl)-1,7-diaza-8(7H)-naphthalenone 7-[5-(p-fluorophenyl)-2-(8-fluoro-7-quinolyl)-1,3-oxazol-4-yl]-1,7-diaza-8(7H)-naphthalenone, 7-{2-(4-methyl-1,7a-diaza-2-indenyl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-(4-methyl-1,7a-diaza-2-indenyl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2 -(1-Thia-5-aza-2-indenyl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-[1-(2-fluoroethyl)-1H-indazol-5-yl]-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-(3-quinolyl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-Diaza-8(7H)-naphthalenone, 7-{2-(2-cyclopropyl-2H-indazol-5-yl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-(6-fluoro-1,3-benzoxazol-2-yl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{5-[p-(trifluoromethyl)phenyl]-2-[2-(trifluoromethyl)-4-pyrimidinyl]-1,3-oxazol-4 7-{2-(5-fluoro-2-pyridyl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-[2-(1-benzofuran-2-yl)-5-(p-fluorophenyl)-1,3-oxazol-4-yl]-1,7-diaza-8(7H)-naphthalenone, or 7-{2-phenyl-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone. In an exemplary embodiment, the compound, or a salt or hydrate or solvate thereof, is 7-(2-(2-methylpyrimidin-4-yl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)-6,7-dihydro-1,7-naphthyridin-8(5H)-one. In an exemplary embodiment, the compound, or a salt or hydrate or solvate thereof, is 7-(5-(4-(trifluoromethyl)phenyl)-2-(2-(trifluoromethyl)pyrimidin-4-yl)oxazol-4-yl)-1,7-naphthyridin-8(7H)-one. In an exemplary embodiment, the compound, or a salt or hydrate or solvate thereof, is 7-(2-(1-methylisoquinolin-6-yl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)-1,7-naphthyridin-8(7H)-one.

[0139] In an exemplary embodiment, the salts of the compounds in this section are pharmaceutically acceptable salts. In an exemplary embodiment, the salts of the compounds described herein are pharmaceutically acceptable salts. In an exemplary embodiment, the salts of the compounds of the invention are pharmaceutically acceptable salts.

[0140] In an exemplary embodiment, the present invention provides a compound described herein, or a salt, hydrate, or solvate thereof, or a combination thereof. In an exemplary embodiment, the present invention provides a compound described herein, or a salt, hydrate, or solvate thereof. In an exemplary embodiment, the present invention provides a compound described herein, or a salt thereof. In an exemplary embodiment, the salt is a pharmaceutically acceptable salt. In an exemplary embodiment, the present invention provides a compound described herein, or a hydrate thereof. In an exemplary embodiment, the present invention provides a compound described herein, or a solvate thereof. In an exemplary embodiment, the present invention provides a salt of a compound described herein. In an exemplary embodiment, the present invention provides a pharmaceutically acceptable salt of a compound described herein. In an exemplary embodiment, the present invention provides a hydrate of a compound described herein. In an exemplary embodiment, the present invention provides a solvate of a compound described herein. In an exemplary embodiment, the present invention provides any of the compounds disclosed herein, wherein one or more atoms of the disclosed compound are replaced with [2H], [3H], [11C], [18F], or [13N]. The compounds disclosed above and throughout bind to 4 to 8 amino acid residues 86 to 99 of SEQ ID NO:9.

[0141] Embodiments of Formula (XVI) In addition to the formulae disclosed above, the present disclosure also provides compounds of formula (XVI): [ka] (In the formula, R1, R5, and R6 are each independently an aryl, a substituted aryl, a heteroaryl, a substituted heteroaryl, an alkyl, a substituted alkyl, -CH2R 20 , -CHMeR20 , -CH(OH)R 20 , cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, alkenyl, alkynyl, acyl, nitro, halo, amino, substituted amine, ether, thioether, and H; Ar is selected from aryl, substituted aryl, heteroaryl, and substituted heteroaryl; A is selected from N, CH, C(halo); X is selected from O and N-R2; R2 and R 20 are each independently selected from aryl, substituted aryl, heteroaryl, substituted heteroaryl, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclic, substituted heterocyclic, alkenyl, alkynyl, acyl, and H; Ring B is a 5-membered heteroaryl ring. Embodiments in which X is NR2

[0142] In some embodiments, X is NR2, also referred to herein as "N-R2." In some such cases, A is N. In other cases, A is C(halo), such as CF, CCl, or CBr. In other cases, A can be CH. Further, in some embodiments, R5 is H, R6 is H, or both R5 and R6 are H. In some embodiments, ring B is oxazole, the structure and atomic numbering of which is shown below. [ka]

[0143] In some cases, ring B is oxazole and R is C 2 Ar is bonded to the oxazole at C 5 The oxazole is bonded to ring B at C 4 to the remainder of the molecule at This bonding arrangement is shown in the drawing below and is also referred to herein as the embodiment in which ring B has the structure (M). [ka]

[0144] Thus, in some embodiments of formula (XVI), X is NR2, A is N, R5 and R6 are both H, and ring B has the structure (M). [ka]

[0145] In some embodiments, R is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclic, and substituted heterocyclic. 20 , -CHMeR 20 or -CH(OH)R 20 In some embodiments, R 20 is H, alkyl, or substituted alkyl.

[0146] Embodiments in which X is O In other embodiments of Formula (XVI), X is O. In some such cases, A is N. In other cases, A is C(halo), such as CF, CCl, or CBr. In other cases, A can be CH. Further, in some embodiments, R5 is H, R6 is H, or both R5 and R6 are H. In some embodiments, ring B is an oxazole, for example, an oxazole having the bonding configuration shown in the following diagram: [ka] .

[0147] Thus, in some embodiments of formula (XVI), X is O, A is N, R5 and R6 are both H, and ring B is [ka] is.

[0148] In some embodiments, R is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclic, and substituted heterocyclic. 20 , -CHMeR 20 or -CH(OH)R 20 In some embodiments, R 20 is H, alkyl, or substituted alkyl. In some embodiments, R is aryl, substituted aryl, heteroaryl, or substituted heteroaryl.

[0149] Embodiments of Formula (XVII) In some embodiments of Formula (XVI), the compound has Formula (XVII), shown below: Formula (XVII) represents an embodiment of Formula (XVI) wherein Ring B is oxazole and R is C 2 Ar is bonded to the oxazole at C 5 The oxazole is bonded to ring B at C 4 It is attached to the rest of the molecule. [ka]

[0150] The embodiment of formula (XVII) can have each of the variations described above with respect to formula (XVII). For example, A can be N or C (halo), such as CF. Furthermore, X can be O or NR2. One or both of R5 and R6 can be H.

[0151] In some embodiments, R is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclic, and substituted heterocyclic. 20 , -CHMeR 20 or -CH(OH)R20 In some embodiments, R 20 is H, alkyl, or substituted alkyl. In some embodiments, R is aryl, substituted aryl, heteroaryl, or substituted heteroaryl.

[0152] Embodiments of Formula (XIII) In some embodiments of Formula (XII), the compound can have Formula (XIII). Formula (XIII) is a compound of Formula (XII) where the Ar group is a phenyl ring having meta substituents at R7 and R9 along with a para substituent at R8. Such R7-R9 groups are selected from halogen, H, alkyl, substituted alkyl, alkoxy, and substituted alkoxyl. Thus, in some cases, each of such groups is H and the phenyl ring is unsubstituted. [ka]

[0153] In some cases, the compound has formula (XIII)(a), (XIII)(b), (XIII)(c), (XIII)(d), (XIII)(e), or (XIII)(f): [ka] [ka] [ka] It has.

[0154] In embodiments of Formula (XIII), optionally A is N. Optionally, X is NR. Optionally, X is O, and optionally, both R5 and R6 are H. Optionally, ring B has structure (M), as described above.

[0155] In some embodiments, R is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclic, and substituted heterocyclic. 20 , -CHMeR 20 or -CH(OH)R 20 In some embodiments, R 20 is H, alkyl, or substituted alkyl. In some embodiments, R is aryl, substituted aryl, heteroaryl, or substituted heteroaryl.

[0156] Embodiments of Formula (XIV) In some embodiments of Formula (XII), the compound has Formula (XIV)(a) or (XIV)(b): [ka] (In the formula, Z9-Z 13 are each independently N and CR Z is selected from Z 14 -Z 18 are each independently N, O, S, and CR Z is selected from Each R Z are independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted alkyl, heterocycloalkyl, and substituted heterocycloalkyl.

[0157] In some embodiments of Formula (XIV)(a), Z 10 -Z 12 One or more of the Z and each R Z is a non-hydrogen group.

[0158] The embodiments of formula (XIV)(a) and (XIV)(b) can have each of the variations described above with respect to formula (XVI). For example, A can be N or C (halo), such as CF. Furthermore, X can be O or NR2. One or both of R5 and R6 can be H.

[0159] In an exemplary embodiment, the invention provides any of the compounds disclosed herein, wherein one or more atoms of the disclosed compounds are replaced with [2H], [3H], [11C], [18F], or [13N]. The compounds disclosed above and throughout bind to 4 to 8 amino acid residues 86-99 of SEQ ID NO:9.

[0160] The compounds disclosed above form π-π interactions with each other, thereby stacking with each other and with stacked proteins. In some cases, π-π interactions are achieved by having a planar core composed of one or two rings, possibly heterocyclic, and accessible molecular conformations that allow three or more of the molecules to self-associate in parallel, displaced stacks that repeat along the stacked protein, where the distance between adjacent atoms in the planar core is 3.3-3.5 Å, the distance between equivalent atoms on two adjacent molecules is 4.8 Å, the angle (θ) between the line defined by two equivalent atoms on adjacent molecules and a line perpendicular to the plane of the planar core is 44°, and the minimum distance between any atom in the plane of the core and an equivalent atom in an adjacent molecule bound to a stacked protein is 3.2-3.6 Å. The compounds bind to the same site on each stacked protein, thereby forming stacked compounds that can interrupt the propagation of stacked proteins. Any compounds with the above characteristics related to π-π interactions can form π-π interactions among themselves and with the stacked proteins to which they bind. In some cases, the compounds of the present disclosure exist as low-molecular-weight monomers, which allow for greater passive cell permeability and easier crossing of the blood-brain barrier, making them uniquely suited to interacting with stacked proteins associated with neurodegenerative diseases of the central nervous system (CNS). The monomers can then form oligomers upon binding to the stacked proteins of the present disclosure. The oligomers can then detect the presence of stacked proteins, prions, amyloid fibrils, and template misfolded proteins. The oligomers can also disrupt the propagation of misfolded proteins. The oligomers can also disrupt the propagation of amyloid conformations of proteins by preventing the sequestration of native cellular proteins and their conversion into propagating amyloid forms.

[0161] Methods for detecting neurological disorders The present disclosure provides a method for detecting a neurological disease, comprising isolating brain tissue from its natural environment, contacting the brain tissue with a labeled molecule that binds to multiple sites of a stacked protein associated with a neurodegenerative disease, and determining binding of the labeled molecule, thereby determining the neurological disease associated with the brain tissue.

[0162] The present disclosure also provides a method for detecting a neurological disease, the method comprising contacting brain tissue with a labeled molecule that binds to a plurality of sites of a stacked protein associated with the neurological disease, determining binding of the labeled molecule, and thereby determining the neurological disease associated with the brain tissue.

[0163] The contacting can be any form of contacting that results in the stacked protein being bound by the labeled molecule. In some embodiments, the contacting is carried out by administering the labeled molecule to an individual who has or is predicted to have a neurological disease. The administration can be any form of administration that results in the stacked protein being bound by the molecule. In some embodiments, the administration is carried out by administering the molecule to an individual who has or is predicted to have a neurological disease. In some embodiments, the administration is intravenous administration. In some embodiments, the contacting involves direct administration of the labeled molecule to isolated brain tissue.

[0164] The natural environment can be any environment in which stacked proteins associated with neurodegenerative diseases are found. In some embodiments, the natural environment is brain tissue. In some embodiments, the brain tissue is mammalian brain tissue. In some embodiments, the brain tissue is human brain tissue.

[0165] The brain tissue of the present disclosure can be any brain tissue that is deemed useful. In some embodiments, the brain tissue has or is suspected of having a neurological disease. In some embodiments, the brain tissue is human brain tissue that has or is suspected of having a neurological disease. Neurological diseases include transmissible spongiform encephalopathies, such as Creutzfeldt-Jakob disease (CJD), multiple system atrophy (MSA), Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), amyotrophic lateral sclerosis / Parkinson's disease dementia complex, anti-IgLON5-associated tauopathy, Caribbean Parkinson's disease, chronic traumatic encephalopathy, diffuse neurofibrillary tangles with calcifications, Down's syndrome, familial British dementia, familial Danish dementia, Niemann-Pick disease type C, non-guanine motor neuron with neurofibrillary tangles, and the like. The condition may be any neurological disease associated with stacked proteins, including, but not limited to, Neuronal disease, postencephalitic Parkinson's disease, primary age-related tauopathy, progressive ataxia and palatal tremor, neurofibrillary senile dementia, familial frontotemporal dementia and Parkinson's disease, Pick's disease, argyrophilic grain dementia, corticobasal degeneration, Guadeloupe Parkinson's disease, glomerular tauopathy, Huntington's disease, progressive supranuclear palsy, SLC9a-associated Parkinson's disease, tauastrogliopathy, etc. In some embodiments, the condition is multiple system atrophy (MSA). In some embodiments, the condition is Parkinson's disease. In some embodiments, the condition is Alzheimer's disease.

[0166] The stacked protein of the present disclosure can be any stacked protein associated with a neurological disease. Stacked proteins associated with neurological diseases include, but are not limited to, prions associated with transmissible spongiform encephalopathies, such as α-synuclein associated with Creutzfeldt-Jakob disease (CJD), multiple system atrophy (MSA), and Parkinson's disease, amyloid beta associated with Alzheimer's disease and Parkinson's disease, and tau associated with Alzheimer's disease and Parkinson's disease. In some embodiments, the stacked protein is α-synuclein. In some embodiments, the stacked protein is amyloid beta. In some embodiments, the stacked protein is tau.

[0167] The detection of neurological diseases of the present disclosure involves contacting brain tissue with a labeling molecule. The labeling molecule can be any molecule that can bind to multiple sites of the stacked protein of a neurodegenerative disease and has a detectable label. The detectable label can be any detectable label that can be detected using positron emission tomography (PET). Detectable labels include, but are not limited to, [2H], [3H], [11C], [18F], [13N], etc. Detectable labels that can be detected using PET are known in the art and are described, for example, by Sun et al. (Acc Chem Res. 2015 Feb 17;48(2):286-94), which is specifically incorporated herein by reference.

[0168] In some embodiments, the labeled molecule is a molecule according to Formula (I). In some embodiments, the labeled molecule is a molecule according to Formula (II). In some embodiments, the labeled molecule is a molecule according to Formula (III). In some embodiments, the labeled molecule is a molecule according to Formula (IV). In some embodiments, the labeled molecule is a molecule according to Formula (V). In some embodiments, the labeled molecule is a molecule according to Formula (VI). In some embodiments, the labeled molecule is a molecule according to Formula (VII). In some embodiments, the labeled molecule is a molecule according to Formula (VIII). In some embodiments, the labeled molecule is a molecule according to Formula (IX). In some embodiments, the labeled molecule is a molecule according to Formula (X). In some embodiments, the labeled molecule is a molecule according to Formula (XI). In some embodiments, the labeled molecule is a molecule according to Formula (XII). In some embodiments, the labeled molecule is a molecule according to Formula (XIII)(a). In some embodiments, the labeled molecule is a molecule according to Formula (XIII)(b). In some embodiments, the labeled molecule is a molecule according to Formula (XIII)(c). In some embodiments, the labeled molecule is a molecule according to Formula (XIII)(d). In some embodiments, the labeled molecule is a molecule according to Formula (XIII)(e). In some embodiments, the labeled molecule is a molecule according to Formula (XIII)(f). In some embodiments, the labeled molecule is a molecule according to Formula (XIV)(a). In some embodiments, the labeled molecule is a molecule according to Formula (XIV)(b). In some embodiments, the labeled molecule is a molecule according to any one of compounds 1-821 in FIG. 6. In some embodiments, the labeled molecule is any of the detectably labeled compounds described above.

[0169] Determining the binding of the labeled molecule of the present disclosure involves detecting the presence of the labeled molecule in brain tissue. The determining can be any method capable of visualizing the presence of the labeled molecule in brain tissue. The determining can be performed by any method, including, but not limited to, PET, autoradiography, imaging mass spectrometry, magnetic resonance imaging, etc. In some embodiments, the determining is performed using PET. In some embodiments, the determining is performed using imaging mass spectrometry. In some embodiments, the determining is performed using magnetic resonance imaging. In some embodiments, the determining is performed using autoradiography. When a protein associated with a neurological disease is accumulated in brain tissue, the labeled molecule binds to the accumulated protein in the brain tissue, thereby determining the neurological disease in the brain tissue. In some embodiments, when a protein associated with a neurological disease is accumulated in brain tissue, the labeled molecule co-localizes with the accumulated protein in the brain tissue, thereby determining the neurological disease in the brain tissue.

[0170] In some embodiments, the binding moiety-labeled molecules of the present disclosure, i.e., the proteins of the labeled molecules that bind to stacked proteins, are characterized by a planar core composed of one or two rings, optionally heterocyclic, and an accessible molecular conformation that allows three or more of the molecules to self-associate in parallel, displaced stacks that repeat along the stacked protein. In some embodiments, the binding moiety of the labeled molecule is further characterized by the configuration shown in FIG. 3, in which the distance between adjacent atoms in the planar core is 3.3-3.5 Å. In some embodiments, the binding moiety of the labeled molecule is further characterized by the configuration shown in FIG. 4, in which the distance between equivalent atoms on two adjacent molecules is 4.8 Å. In some embodiments, the binding moiety of the labeled molecule is further characterized by the configuration shown in FIG. 5, in which the angle (θ) between the line defined by two equivalent atoms on adjacent molecules and a line perpendicular to the plane of the planar core is 44°. In some embodiments, the binding moiety of the labeled molecule is further characterized by the configuration shown in FIG. 4, in which the minimum distance between any atom in the plane of the core and an equivalent atom in an adjacent molecule bound to the stacked protein is 3.2-3.6 Å.

[0171] In some embodiments, the binding moiety of the labeling molecule is further characterized by a substituent that forms a non-covalent interaction with the stacked protein, wherein the interaction is selected from the group consisting of a hydrogen bond, a van der Waals contact, a π-π interaction, and a chalcogen bond. In some embodiments, the interaction is a hydrogen bond. In some embodiments, the interaction is a van der Waals contact. In some embodiments, the interaction is a hydrogen bond. In some embodiments, the interaction is a π-π interaction. In some embodiments, the interaction is a hydrogen bond. In some embodiments, the interaction is a chalcogen bond.

[0172] Supramolecular polymer assembly conjugates The present disclosure provides supramolecular polymer assembly conjugates isolated from their natural environment that contain α-synuclein prion stacked proteins associated with neurodegenerative diseases, and α-synuclein prion inhibitors that reversibly bind to α-synuclein prion amyloid fibrils. In some embodiments, the reversible binding is non-covalent.

[0173] The natural environment can be any environment in which alpha-synuclein prion-stacked proteins associated with neurodegenerative diseases are found. In some embodiments, the natural environment is brain tissue. In some embodiments, the brain tissue is human brain tissue.

[0174] The α-synuclein prion inhibitor can be any α-synuclein prion inhibitor that inhibits α-synuclein from forming stacked proteins, e.g., prions. In some embodiments, the α-synuclein prion inhibitor has a structure according to Formula (I). In some embodiments, the α-synuclein prion inhibitor has a structure according to Formula (II). In some embodiments, the α-synuclein prion inhibitor has a structure according to Formula (III). In some embodiments, the α-synuclein prion inhibitor has a structure according to Formula (IV). In some embodiments, the α-synuclein prion inhibitor has a structure according to Formula (V). In some embodiments, the α-synuclein prion inhibitor has a structure according to Formula (VI). In some embodiments, the α-synuclein prion inhibitor has a structure according to Formula (VII). In some embodiments, the α-synuclein prion inhibitor has a structure according to Formula (VIII). In some embodiments, the α-synuclein prion inhibitor has a structure according to Formula (IX). In some embodiments, an α-synuclein prion inhibitor has a structure according to Formula (X). In some embodiments, an α-synuclein prion inhibitor has a structure according to Formula (XI). In some embodiments, an α-synuclein prion inhibitor has a structure according to Formula (XII). In some embodiments, an α-synuclein prion inhibitor has a structure according to Formula (XIII)(a). In some embodiments, an α-synuclein prion inhibitor has a structure according to Formula (XIII)(b). In some embodiments, an α-synuclein prion inhibitor has a structure according to Formula (XIII)(c). In some embodiments, an α-synuclein prion inhibitor has a structure according to Formula (XIII)(d). In some embodiments, an α-synuclein prion inhibitor has a structure according to Formula (XIII)(e). In some embodiments, an α-synuclein prion inhibitor has a structure according to Formula (XIII)(f). In some embodiments, the α-synuclein prion inhibitor has a structure according to formula (XIV)(a): In some embodiments, the α-synuclein prion inhibitor has a structure according to formula (XIV)(b):In some embodiments, the α-synuclein prion inhibitor has a structure according to any one of compounds 1-821 in Figure 6. In some embodiments, the α-synuclein prion inhibitor is any of the compounds described above.

[0175] In some embodiments, the disclosed α-synuclein prion inhibitors are characterized by a planar core composed of one or two rings, optionally heterocyclic, and accessible molecular conformations that allow three or more of the molecules to self-associate in repeating parallel displaced stacks along the stacked protein. In some embodiments, the α-synuclein prion inhibitors are further characterized by the configuration shown in FIG. 3, in which the distance between adjacent atoms in the planar core is 3.3-3.5 Å. In some embodiments, the α-synuclein prion inhibitors are further characterized by the configuration shown in FIG. 4, in which the distance between equivalent atoms on two adjacent molecules is 4.8 Å. In some embodiments, the α-synuclein prion inhibitors are further characterized by the configuration shown in FIG. 5, in which the angle (θ) between the line defined by two equivalent atoms on adjacent molecules and a line perpendicular to the plane of the planar core is 44°. In some embodiments, the α-synuclein prion inhibitor is further characterized by a configuration as shown in Figure 4, in which the minimum distance between any atom in the plane of the core and an equivalent atom in an adjacent molecule bound to a stacked protein is 3.2-3.6 Å. In some embodiments, the α-synuclein prion inhibitor is further characterized as being uniquely suited to interact with stacked proteins associated with neurodegenerative diseases of the central nervous system (CNS).

[0176] In some cases, the disclosed α-synuclein prion inhibitors are uniquely suited to interact with stacked proteins associated with neurodegenerative diseases because the α-synuclein prion inhibitors form π-π interactions with each other, thereby stacking with each other and with the stacked proteins. In some cases, the π-π interactions are achieved by having a planar core composed of one or two rings, optionally heterocyclic, and accessible molecular conformations that allow three or more of the molecules to self-associate in parallel displaced stacks that repeat along the stacked protein, wherein the distance between adjacent atoms in the planar core is 3.3-3.5 Å, the distance between equivalent atoms on two adjacent molecules is 4.8 Å, the angle (θ) between a line defined by two equivalent atoms on adjacent molecules and a line perpendicular to the plane of the planar core is 44°, and the minimum distance between any atom in the plane of the core and an equivalent atom in an adjacent molecule bound to the stacked protein is 3.2-3.6 Å. In some cases, the alpha-synuclein prion inhibitors of the present disclosure are uniquely suited to interact with stacked proteins associated with neurodegenerative diseases of the central nervous system (CNS) because they exist as low molecular weight monomers that allow for greater passive cell permeability, allowing for easier crossing of the blood-brain barrier. The monomers can then form oligomers upon binding to the stacked proteins of the present disclosure.

[0177] In some embodiments, the alpha-synuclein prion inhibitor is detectably labeled, including but not limited to, [2H], [3H], [11C], [18F], [13N], etc.

[0178] Pharmaceutical preparations The compounds, α-synuclein prion inhibitors, labeled molecules, and molecules discussed herein can be formulated using any convenient excipients, reagents, and methods. The compositions are provided in formulations containing pharmaceutically acceptable excipients. A wide variety of pharmaceutically acceptable excipients are known in the art and need not be discussed in detail herein. Pharmaceutically acceptable excipients are, for example, those described in A. Gennaro (2000) "Remington: The Science and Practice of Pharmacy," 20th edition, Lippincott, Williams, & Wilkins; Pharmaceutical Dosage Forms and Drug Delivery Systems (1999) H.C. Cansel et al., eds., 7 th ed., Lippincott, Williams, & Wilkins; and Handbook of Pharmaceutical Excipients (2000) AHKibbe et al., eds., 3 rd It has been fully described in various publications, including ed. Amer. Pharmaceutical Assoc.

[0179] In an exemplary embodiment, the present invention provides a pharmaceutical formulation comprising: a) a compound described herein, or a pharmaceutically acceptable salt, hydrate, or solvate thereof; and b) a pharmaceutically acceptable excipient. The pharmaceutical formulation can be administered by any suitable means, for example, orally and / or via a parenteral depot. Oral administration in the form of a pill, capsule, elixir, syrup, lozenge, troche, or the like is particularly preferred. Dosage levels of about 5 mg to about 250 mg / kg body weight / day, more preferably about 25 mg to about 150 mg / kg body weight / day, are useful in treating the disorders described herein. Dosage frequency may also vary depending on the compound used and the specific disorder being treated. However, for the treatment of most disorders, a dosing regimen of four times daily or less is preferred. It will be understood, however, that the specific dose level for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, time of administration, route of administration and excretion rate, drug combination, and the severity of the specific disorder being treated. Preferred compounds of the present invention will have desirable pharmacological properties, including, but not limited to, oral bioavailability, low toxicity, low serum protein binding, and desirable in vitro and in vivo half-lives. Penetration of the blood-brain barrier is necessary for compounds used to treat CNS disorders, while low brain levels are often desirable for compounds used to treat peripheral disorders. In an exemplary embodiment, the present invention provides an oral pharmaceutical formulation comprising: a) a compound described herein, or a pharmaceutically acceptable salt, hydrate, or solvate thereof; and b) a pharmaceutically acceptable excipient suitable for oral administration. In an exemplary embodiment, the present invention provides a parenteral pharmaceutical formulation comprising: a) a compound described herein, or a pharmaceutically acceptable salt, hydrate, or solvate thereof; and b) a pharmaceutically acceptable excipient suitable for parenteral administration. In an exemplary embodiment, the present invention provides an intravenous pharmaceutical formulation comprising: a) a compound described herein, or a pharmaceutically acceptable salt, hydrate, or solvate thereof; and b) a pharmaceutically acceptable excipient suitable for intravenous administration. In some embodiments, the pharmaceutical formulation is an injectable formulation.In some embodiments, the pharmaceutical formulation is an oral formulation. In some embodiments, the pharmaceutical formulation is a parenteral formulation.

[0180] The subject compounds may be administered in unit dosage form and may be prepared by any method known in the art. Such methods include combining the subject compounds with pharmaceutically acceptable carriers or diluents, which constitute one or more accessory components. Pharmaceutically acceptable carriers are selected based on the selected route of administration and standard pharmaceutical practice. Each carrier must be "pharmaceutically acceptable" in the sense that it is compatible with the other components of the formulation and is not harmful to the subject. The carrier may be solid or liquid, and its type is generally selected based on the type of administration used.

[0181] The compounds of the present invention can also be used in combination with additional therapeutic agents.In an exemplary embodiment, the present invention provides a combination comprising: a) a compound described herein, or its pharmaceutically acceptable salt, hydrate, or solvate; and b) at least one additional therapeutic agent.In an exemplary embodiment, the additional therapeutic agent is useful for treating neurodegenerative diseases.

[0182] Any drug delivery device or system that provides the dosing regimen of the present disclosure can be used. A wide variety of delivery devices and systems are known to those skilled in the art.

[0183] In some embodiments, the compound, alpha-synuclein prion inhibitor, labeled molecule, and molecule in the formulation are detectably labeled, including, but not limited to, [2H], [3H], [11C], [18F], [13N], etc. A method to disrupt the propagation of stacked proteins

[0184] The present disclosure provides methods for disrupting the propagation of stacked proteins associated with a neurological disease, comprising contacting an environment densely populated with stacked proteins associated with a neurological disease with a molecule that binds to multiple sites on the stacked proteins, allowing the molecule to bind to the multiple sites on the stacked proteins, thereby preventing the propagation of the stacked proteins in the environment. In some embodiments, the methods for disrupting the propagation of stacked proteins associated with a neurological disease treat the neurological disease.

[0185] The environment can be any environment in which the stacked proteins are associated with a neurodegenerative disease, including, but not limited to, a cell lysate, a cell culture, mammalian brain tissue, etc. In some embodiments, the environment is mammalian brain tissue. In some embodiments, the mammalian brain tissue is human brain tissue. In some embodiments, the environment is a cell lysate. In some embodiments, the environment is a cell culture. In some embodiments, the cell culture is a mammalian cell culture. In some embodiments, the cell culture contains neurons. In some embodiments, the brain or brain tissue is within an individual having or suspected of having a neurological disease.

[0186] Neurological diseases include transmissible spongiform encephalopathies, e.g., Creutzfeldt-Jakob disease (CJD), multiple system atrophy (MSA), Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), amyotrophic lateral sclerosis / Parkinson's disease dementia complex, anti-IgLON5-associated tauopathy, Caribbean Parkinson's disease, chronic traumatic encephalopathy, diffuse neurofibrillary tangles with calcifications, Down's syndrome, familial British dementia, familial Danish dementia, Niemann-Pick disease type C, non-guanine motor neuropathies with neurofibrillary tangles, and The neurological disease may be any neurological disease associated with stacked proteins, including, but not limited to, Neuronal disease, postencephalitic Parkinson's disease, primary age-related tauopathy, progressive ataxia and palatal tremor, neurofibrillary senile dementia, familial frontotemporal dementia and Parkinson's disease, Pick's disease, argyrophilic grain dementia, corticobasal degeneration, Guadeloupe Parkinson's disease, glomerular tauopathy, Huntington's disease, progressive supranuclear palsy, SLC9a-associated Parkinson's disease, tauastrogliopathy, etc. In some embodiments, the neurological disease is multiple system atrophy (MSA). In some embodiments, the neurological disease is Parkinson's disease. In some embodiments, the neurological disease is Alzheimer's disease.

[0187] The stacked protein of the present disclosure can be any stacked protein associated with a neurological disease. Stacked proteins associated with neurological diseases include, but are not limited to, prions associated with transmissible spongiform encephalopathies, such as α-synuclein associated with Creutzfeldt-Jakob disease (CJD), multiple system atrophy (MSA), and Parkinson's disease, amyloid beta associated with Alzheimer's disease and Parkinson's disease, and tau associated with Alzheimer's disease and Parkinson's disease. In some embodiments, the stacked protein is α-synuclein. In some embodiments, the stacked protein is amyloid beta. In some embodiments, the stacked protein is tau.

[0188] The contacting can be any form of contacting that results in the stacked proteins being bound by the molecule. In some embodiments, the contacting is carried out by administering the molecule to an individual having or suspected of having a neurological disorder. In some embodiments, the administration is by oral administration. In some embodiments, the administration is by parenteral depot. In some embodiments, the administration is intravenous administration. In some embodiments, the contacting involves administering an effective dose of the molecule to an individual having or suspected of having a neurological disorder.

[0189] As used herein, an effective dose refers to a dose sufficient to alleviate symptoms associated with a neurological disease. The term effective dose also refers to an amount of a drug sufficient to produce beneficial or desired results. The effective dose varies depending on the subject and neurological disease being treated, the subject's weight and age, the severity of the disease state, the mode of administration, etc., and can be easily determined by those skilled in the art. This term also applies to a dose that provides an image for detection by any one of the detection methods described herein. The specific dose varies depending on the specific compound or molecule selected, the dosing regimen to be followed, whether it is administered in combination with other compounds, the timing of administration, the tissue to be imaged, and the physical delivery system in which it is carried. An effective dose is any dose that stops, reverses, or alleviates a neurological condition.

[0190] The cessation, reversal, or alleviation of a neurological condition can be determined by evaluating an individual having or suspected of having a neurological disease before and after contact. The cessation, reversal, or alleviation of a neurological condition can be determined by determining whether propagation of stacked proteins associated with a neurological disease is disrupted, for example, by using the detection methods described above or disclosed herein.

[0191] The molecule can be any molecule that inhibits the propagation of stacked proteins, e.g., prions. In some embodiments, the molecule is a molecule according to Formula (I). In some embodiments, the molecule is a molecule according to Formula (II). In some embodiments, the molecule is a molecule according to Formula (III). In some embodiments, the molecule is a molecule according to Formula (IV). In some embodiments, the molecule is a molecule according to Formula (V). In some embodiments, the molecule is a molecule according to Formula (VI). In some embodiments, the molecule is a molecule according to Formula (VII). In some embodiments, the molecule is a molecule according to Formula (VIII). In some embodiments, the molecule is a molecule according to Formula (IX). In some embodiments, the molecule is a molecule according to Formula (X). In some embodiments, the molecule is a molecule according to Formula (XI). In some embodiments, the molecule is a molecule according to Formula (XII). In some embodiments, the molecule is a molecule according to Formula (XIII)(a). In some embodiments, the molecule is a molecule according to Formula (XIII)(b). In some embodiments, the molecule is a molecule according to Formula (XIII)(c). In some embodiments, the molecule is a molecule according to Formula (XIII)(d). In some embodiments, the molecule is a molecule according to Formula (XIII)(e). In some embodiments, the molecule is a molecule according to Formula (XIII)(f). In some embodiments, the molecule is a molecule according to Formula (XIV)(a). In some embodiments, the molecule is a molecule according to Formula (XIV)(b). In some embodiments, the molecule is a molecule according to any one of compounds 1-821 in Figure 6. In some embodiments, the molecule is any of the compounds described above.

[0192] In some embodiments, the molecules of the present disclosure are characterized by a planar core composed of one or two rings, optionally heterocyclic, and accessible molecular conformations that allow three or more of the molecules to self-associate in parallel, displaced stacks that repeat along the stacked protein. In some embodiments, the molecules are further characterized by the configuration shown in FIG. 3, in which the distance between adjacent atoms in the planar core is 3.3-3.5 Å. In some embodiments, the molecules are further characterized by the configuration shown in FIG. 4, in which the distance between equivalent atoms on two adjacent molecules is 4.8 Å. In some embodiments, the molecules are further characterized by the configuration shown in FIG. 5, in which the angle (θ) between the line defined by two equivalent atoms on adjacent molecules and a line perpendicular to the plane of the planar core is 44°. In some embodiments, the molecules are further characterized by the configuration shown in FIG. 4, in which the minimum distance between any atom in the plane of the core and an equivalent atom in an adjacent molecule bound to the stacked protein is 3.2-3.6 Å.

[0193] In some embodiments, the molecule that binds to the stacked protein is further characterized by a substituent that forms a non-covalent interaction with the stacked protein, wherein the interaction is selected from the group consisting of a hydrogen bond, a van der Waals contact, a π-π interaction, and a chalcogen bond. In some embodiments, the interaction is a hydrogen bond. In some embodiments, the interaction is a van der Waals contact. In some embodiments, the interaction is a hydrogen bond. In some embodiments, the interaction is a π-π interaction. In some embodiments, the interaction is a hydrogen bond. In some embodiments, the interaction is a chalcogen bond.

[0194] In some cases, the molecules disclosed herein are uniquely suited to interacting with stacked proteins associated with neurological diseases because the molecules form π-π interactions with each other, thereby stacking with each other and with the stacked proteins. In some cases, the π-π interactions are achieved by having a planar core composed of one or two rings, possibly heterocyclic, and accessible molecular conformations that allow three or more of the molecules to self-associate in parallel, displaced stacks that repeat along the stacked protein, where the distance between adjacent atoms in the planar core is 3.3-3.5 Å, the distance between equivalent atoms on two adjacent molecules is 4.8 Å, the angle (θ) between the line defined by the two equivalent atoms on adjacent molecules and a line perpendicular to the plane of the planar core is 44°, and the minimum distance between any atom in the plane of the core and an equivalent atom in an adjacent molecule bound to a stacked protein is 3.2-3.6 Å. The molecules bind to the same site on each stacked protein, thereby forming stacked molecules that can interrupt the propagation of the stacked proteins. In some cases, the molecules of the present disclosure are uniquely suited to interact with stacked proteins associated with neurodegenerative diseases of the central nervous system (CNS) because they exist as low molecular weight monomers, which allow for greater passive cell permeability and easier crossing of the blood-brain barrier. The monomers can then form oligomers upon binding to the stacked proteins of the present disclosure.

[0195] Methods for preventing progressive template misfolding of proteins The present disclosure provides herein a method for preventing progressive templated misfolding of proteins associated with neurodegenerative diseases, comprising administering a molecule to a biological environment containing both a propagating amyloid conformation of the protein and a native cellular form of the same protein, allowing the formation of a complex between a supramolecular polymeric assembly of the molecule and the supramolecular assembly of the protein, thereby preventing further sequestration of the native cellular protein and its conversion to a propagating amyloid form.

[0196] The biological environment can be any biological environment in which protein misfolding is associated with neurodegenerative diseases, including, but not limited to, cell lysates, cell cultures, mammalian brain tissue, etc. In some embodiments, the biological environment is mammalian brain tissue. In some embodiments, the mammalian brain tissue is human brain tissue. In some embodiments, the biological environment is a cell lysate. In some embodiments, the biological environment is a cell culture. In some embodiments, the cell culture is a mammalian cell culture. In some embodiments, the cell culture contains neurons.

[0197] Neurodegenerative diseases include transmissible spongiform encephalopathies, e.g., Creutzfeldt-Jakob disease (CJD), multiple system atrophy (MSA), Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), amyotrophic lateral sclerosis / Parkinson's disease dementia complex, anti-IgLON5-associated tauopathy, Caribbean Parkinson's disease, chronic traumatic encephalopathy, diffuse neurofibrillary tangles with calcifications, Down's syndrome, familial British dementia, familial Danish dementia, Niemann-Pick disease type C, non-guanine motor neuropathies with neurofibrillary tangles, and others. The neurodegenerative disease may be any neurodegenerative disease associated with stacked proteins, including, but not limited to, Neuronal disease, postencephalitic Parkinson's disease, primary age-related tauopathy, progressive ataxia and palatal tremor, neurofibrillary senile dementia, familial frontotemporal dementia and Parkinson's disease, Pick's disease, argyrophilic grain dementia, corticobasal degeneration, Guadeloupe Parkinson's disease, glomerular tauopathy, Huntington's disease, progressive supranuclear palsy, SLC9a-associated Parkinson's disease, tauastrogliopathy, etc. In some embodiments, the neurodegenerative disease is multiple system atrophy (MSA). In some embodiments, the neurodegenerative disease is Parkinson's disease. In some embodiments, the neurodegenerative disease is Alzheimer's disease.

[0198] The template-misfolded protein of the present disclosure can be any template-misfolded protein associated with a neurological disease, including, but not limited to, prions associated with transmissible spongiform encephalopathies, such as a-synuclein associated with Creutzfeldt-Jakob disease (CJD) and multiple system atrophy (MSA), amyloid b associated with Alzheimer's disease and Parkinson's disease, and tau associated with Alzheimer's disease and Parkinson's disease.

[0199] The administering can be any form of administration that results in the misfolded template protein being bound by the molecule. In some embodiments, the administering is carried out by administering the molecule to an individual having or suspected of having a neurological disorder. In some embodiments, the administering is by oral administration. In some embodiments, the administering is by parenteral depot. In some embodiments, the administering is intravenous administration. In some embodiments, the contacting involves administering an effective dose of the molecule to an individual having or suspected of having a neurological disorder. In some embodiments, the administering involves administering an effective amount of the molecule to an individual having or suspected of having a neurological disorder.

[0200] As used herein, an effective dose refers to a dose sufficient to alleviate symptoms associated with a neurodegenerative disease. The term effective dose also refers to an amount of a molecule sufficient to produce beneficial or desired results. The effective dose varies depending on the subject and neurological disease being treated, the subject's weight and age, the severity of the disease state, the mode of administration, etc., and can be easily determined by those skilled in the art. This term also applies to the dose that provides an image for detection by any one of the detection methods described herein. The specific dose varies depending on the specific compound or molecule selected, the dosing regimen to be followed, whether it is administered in combination with other compounds, the timing of administration, the tissue to be imaged, and the physical delivery system in which it is carried. An effective dose is any dose that stops, reverses, or alleviates a neurological condition.

[0201] The halting, reversal or alleviation of neurological conditions can be determined by assessing individuals who have or suspect that they have neurodegenerative diseases before and after administration for cognitive impairment associated with neurodegenerative diseases.The halting, reversal or alleviation of neurodegenerative diseases can be determined by determining whether the progressive template misfolding of proteins associated with neurodegenerative diseases is prevented, for example, by using the detection method described above or disclosed herein.

[0202] The molecule can be any molecule that inhibits template misfolded proteins, e.g., prions, from capturing native cellular proteins and their propagating amyloid forms. In some embodiments, the molecule is a molecule according to Formula (I). In some embodiments, the molecule is a molecule according to Formula (II). In some embodiments, the molecule is a molecule according to Formula (III). In some embodiments, the molecule is a molecule according to Formula (IV). In some embodiments, the molecule is a molecule according to Formula (V). In some embodiments, the molecule is a molecule according to Formula (VI). In some embodiments, the molecule is a molecule according to Formula (VII). In some embodiments, the molecule is a molecule according to Formula (VIII). In some embodiments, the molecule is a molecule according to Formula (IX). In some embodiments, the molecule is a molecule according to Formula (X). In some embodiments, the molecule is a molecule according to Formula (XI). In some embodiments, the molecule is a molecule according to Formula (XII). In some embodiments, the molecule is a molecule according to Formula (XIII)(a). In some embodiments, the molecule is a molecule according to Formula (XIII)(b). In some embodiments, the molecule is a molecule according to Formula (XIII)(c). In some embodiments, the molecule is a molecule according to Formula (XIII)(d). In some embodiments, the molecule is a molecule according to Formula (XIII)(e). In some embodiments, the molecule is a molecule according to Formula (XIII)(f). In some embodiments, the molecule is a molecule according to Formula (XIV)(a). In some embodiments, the molecule is a molecule according to Formula (XIV)(b). In some embodiments, the molecule is a molecule according to any one of compounds 1-821 in Figure 6. In some embodiments, the molecule is any of the compounds described above.

[0203] In some embodiments, the molecules of the present disclosure are characterized by a planar core composed of one or two rings, optionally heterocyclic, and accessible molecular conformations that allow three or more of the molecules to self-associate in parallel, displaced stacks that repeat along the stacked protein. In some embodiments, the molecules are further characterized by the configuration shown in FIG. 3, in which the distance between adjacent atoms in the planar core is 3.3-3.5 Å. In some embodiments, the molecules are further characterized by the configuration shown in FIG. 4, in which the distance between equivalent atoms on two adjacent molecules is 4.8 Å. In some embodiments, the molecules are further characterized by the configuration shown in FIG. 5, in which the angle (θ) between the line defined by two equivalent atoms on adjacent molecules and a line perpendicular to the plane of the planar core is 44°. In some embodiments, the molecules are further characterized by the configuration shown in FIG. 4, in which the minimum distance between any atom in the plane of the core and an equivalent atom in an adjacent molecule bound to the stacked protein is 3.2-3.6 Å.

[0204] In some embodiments, the molecule that binds to the stacked protein is further characterized by a substituent that forms a non-covalent interaction with the stacked protein, wherein the interaction is selected from the group consisting of a hydrogen bond, a van der Waals contact, a π-π interaction, and a chalcogen bond. In some embodiments, the interaction is a hydrogen bond. In some embodiments, the interaction is a van der Waals contact. In some embodiments, the interaction is a hydrogen bond. In some embodiments, the interaction is a π-π interaction. In some embodiments, the interaction is a hydrogen bond. In some embodiments, the interaction is a chalcogen bond.

[0205] In some cases, the molecules disclosed herein are uniquely suited to interacting with templated misfolded proteins associated with neurodegenerative diseases because the molecules form π-π interactions with each other, thereby stacking with each other and with stacked proteins. In some cases, the π-π interactions are achieved by having a planar core composed of one or two rings, possibly heterocyclic, and accessible molecular conformations that allow three or more of the molecules to self-associate in parallel, displaced stacks that repeat along the stacked protein, where the distance between adjacent atoms in the planar core is 3.3-3.5 Å, the distance between equivalent atoms on two adjacent molecules is 4.8 Å, the angle (θ) between the line defined by two equivalent atoms on adjacent molecules and a line perpendicular to the plane of the planar core is 44°, and the minimum distance between any atom in the plane of the core and an equivalent atom in an adjacent molecule bound to a stacked protein is 3.2-3.6 Å. The molecules bind to the same site on each stacked protein, thereby forming stacked molecules that can interrupt the propagation of the stacked proteins.

[0206] Example The following examples are presented to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the following experiments are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be accounted for. Unless otherwise specified, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric.

[0207] Example 1 Many neurodegenerative diseases (NDs) are characterized by the formation of prions that propagate into amyloid filaments, which adopt disease-specific conformations in the brain. Small molecules have recently been developed that show promise as diagnostic and possibly therapeutic agents for NDs. The binding mechanism of many of these small molecules to amyloid filaments remains unclear. Here, we use cryo-electron microscopy (cryo-EM) to determine the 2.7 Å structure of paired helical filaments (PHFs) of patient-derived Alzheimer's disease tau incubated with a GTP-1 PET probe. The cryo-EM structure reveals a novel stacked arrangement of the GTP-1 PET ligand bound to the Alzheimer's disease tau filament.

[0208] GTP-1 is bound stoichiometrically along the exposed cleft of each protofilament in a stacked configuration that simultaneously satisfies the translational symmetry of amyloids and the π-π stacking of aromatic small molecules. Following modeling analysis, further calculations established that highly favorable ligand-ligand interactions facilitate this binding mode. Thus, this structure provides new insights into designing compounds for the diagnosis and treatment of specific NDs.

[0209] The accumulation of misfolded tau protein in the brain is a hallmark of a large subset of neurodegenerative diseases (NDs) known as tauopathies (1, 2), the most common and widely studied of which is Alzheimer's disease (AD) (3). The spread of tau deposits, known as neurofibrillary tangles (NFTs), in AD parallels neuronal loss and cognitive impairment (4, 5) and serves as a marker of disease progression (6). Furthermore, NFT accumulation has been shown to occur through the conversion of soluble tau into prions, a process that transforms them into ordered, stable amyloid filaments. Prions are self-propagating and transmit across neurons via synaptic junctions (7-10). Prions were first identified in PrPSc, which causes Creutzfeldt-Jakob disease (CJD), Gerstmann-Straussler-Scheinker syndrome (GSS), and other incurable diseases (11, 12). Structures determined by cryo-electron microscopy (cryo-EM) of tau filaments purified from patient brains revealed that the conformation of the microtubule-binding repeats, including the cross-β-sheet filament core, varies among different NDs. (13-18) This opens the possibility for small molecules to bind site-specifically to different tau prion conformers, and here we present an example of a mechanism for achieving site specificity.

[0210] Small molecules have been developed that can distinguish between amyloid proteins (19, 20) and even strains of the same prion (21, 22). However, the mechanism of this specificity remains unclear. Despite this limitation, several promising tau-selective PET ligands for AD have been developed and tested in vivo (23-27). Many such molecules contain heterocyclic aromatic moieties (28), including Flortaucipir, a first-generation tau PET ligand that is FDA-approved and clinically available (29). Second-generation PET tracers have been developed to reduce off-target binding and optimize pharmacokinetic properties (30, 31), but the mechanism of specificity remains unclear, limiting their rational design and the design of better diagnostics and more effective therapeutics to expand the array of NDs. Current models provided by the cryo-EM structure of the PET ligand APN-1607 at low resolution (32) and docking studies (33-36) indicate that small molecules bind end-to-end and parallel to the fibril axis. These models show heterogeneous binding sites within the tau prion fibril core, so conformational specificity is not explained. Further co-structures that identify site-specific ligand binding modes are needed to develop models of binding to the amyloid fold and advance the development of conformation-specific probes.

[0211] Using cryo-EM, we determined the structure of GTP-1 (Genentech Tau Probe 1), a high-affinity (11 nM Kd) second-generation tau PET tracer currently in clinical trials. Tau filament samples were purified from the frontal cortex of AD patients as previously described (13) and exhibit high infectivity in cell-based assays. Samples were incubated with 20 μM GTP-1 and then vitrified. A concentration well above the estimated Kd of 11 nM (23) was used to achieve site saturation of tau fibrils under cryo-EM conditions. Micrograph images and their 2D classification revealed well-resolved filaments primarily in the PHF conformation, with crossover distances ranging from 700 to 800 Å. A minor population of linear filaments (SFs) was also identified; however, due to their limited abundance, further structural characterization was not feasible. Using standard helical reconstruction methods (see Methods), we determined the structure of the PHF at an overall resolution of 2.7 Å, exhibiting clear β-strand separation. The PHF structure consists of two protofilaments related by two-fold symmetry with a 2.37 Å rise and a 179.45° twist, consistent with previously reported structures of PHFs prepared from AD brain (13, 15). The protofilament is composed of 3R and 4R tau domains (residues 306–378), which form the canonical C-shaped cross-β fold seen in AD, interacting laterally via an antiparallel PGGGQ motif (residues 332–336). This central region has the highest resolution at approximately 2.5 Å, while the periphery, at approximately 3.2 Å, exhibits high resolution across the β-sheet core with well-resolved side chain density.

[0212] Notably, the structure reveals strong additional density representing a GTP-1 small molecule bound to a solvent-exposed cleft (residues 351–360) adjacent to the three-stranded β-helix (β5–7). Notably, this density is identical in both protofilaments, indicating comparable binding. While other density is present around the filament core, these are poorly resolved in comparison and are similar to density present in previously reported tau filament structures. Importantly, difference map analysis comparing the GTP-1 co-structure with the previously determined PHF map (EMDB:0259) (15) identifies this density as uniquely present and without additional density in the difference map, indicating specific binding by GTP-1. Thus, in contrast to previous PET-ligand studies identifying multiple possible binding modes, only a single, well-defined binding site for GTP-1 within the structured core of tau was observed.

[0213] The density shows that GTP-1 binds tau with a 1:1 stoichiometry, with the compound stacked in a geometric repeat that precisely matches the geometric repeat of the protein monomers in the fibril. The ligand forms parallel, displaced stacks in which each GTP-1 spans three tau monomers. This arrangement is in stark contrast to previously described models that predict end-to-end binding parallel to the fibril axis (32-36). Notably, the ligand resolution is similar to that of adjacent filament structures (approximately 2.6 Å), and the density persists at a high sigma threshold, indicating near-complete occupancy. Taken together, our observations confirm that GTP-1 binds tau in a single conformation within the conserved binding site.

[0214] The atomic model of the tau PHF was achieved by docking and refinement of a previous 3.2 Å resolution structure of the PHF solved in the absence of exogenous ligand. The overall filament structure is nearly identical to the previous structure of the AD PHF (α-carbon RMSD = 0.5 Å). However, small differences are observed in the side chains of residues lining the binding pocket, namely Lys353, Asp358, and Ile360. It is unclear whether perturbations in the binding site are the result of improved resolution relative to the published apo form, or whether they result from ligand-induced perturbations to the cleft. To fit GTP-1 to a fully resolved ligand density, the best approach was found to result from a combination of molecular mechanics to generate conformers and density functional theory to perform constrained optimizations of dimers to capture small molecule-small molecule interactions. By using a crush filter (<2.5 Å) for small molecule-small molecule and small molecule-protein interactions compared to the electron density, it was possible to rapidly converge on a suitable starting conformation for final refinement using Phenix. (37) The final modeled conformer gave excellent map-model agreement and was energetically reasonable.

[0215] GTP-1 binds to the groove of the AD fibril with precise physicochemical and geometric complementarity. The binding site consists of strands β6 and β7, separated by a kink at Gly355, creating a concave cleft that complements the convex shape of the GTP-1 stack. Within the uniform, parallel-displaced stack, each GTP-1 molecule binds across three β-strands, making direct contact with Gln351 on strand 1, Gln351 and Lys353 on strand 2, Ile360 on strand 3, and the backbone between Gln351 and Lys353 on strands 1 and 2. Remarkably, a short section of GTP-1 is parallel to the filament and intercalates between the two β-strands. Although the binding site is primarily composed of polar residues, there is a precise match between the nonpolar portions of their side chains and those of small molecules. The aliphatic carbon of Ile360 contacts C7 of the phenyl ring, and the nonpolar carbon of the Gln353 side chain is aligned with the portion of the pocket occupied by the relatively nonpolar fluoroethyl tail. Specific hydrogen-bonding interactions also contribute significantly to GTP-1 binding. Lys353 lies at the bottom of the binding groove, where it forms a bifurcated hydrogen bond with the benzimidazole nitrogen (2.8 Å N-N distance) and the pyrimido nitrogen of GTP-1 (3.4 Å), satisfying the hydrogen-bonding potential of the buried polar atoms in the tricyclic aromatic ring. Lys353 also completes its hydrogen-bonding potential by forming a strong salt bridge with Asp358 in the same chain and a weaker hydrogen bond with Asp358 in the adjacent chain. The oxygen of the Gln351 side chain is well-positioned to form a noncanonical hydrogen bond with the C-H bond of the beta-carbon of the fluoroethyl tail, which points toward the interior of the fibril backbone. This tail orientation allows for close van der Waals contacts with the main chain atoms of the two strands and interactions with the side chain of Gln351.

[0216] A clear observation from the modeled GTP-1 ligand is that the stacked heterocycles are positioned at optimal distances for π-π stacking (3.3–3.5 Å). To assess the favorability of these π-π interactions, Hartree-Fock London dispersion calculations were performed (38). The aromatic and nonaromatic regions of GTP-1 (aromatic, pyrimido[1,2-a]benzimidazole, and nonaromatic, 2-fluoro-4-ethylpiperidine) make distinct contributions to the overall interactions. The major component (57%) is indeed from aromatic-aromatic interactions, while the smallest contribution comes from cross-interactions between nonaromatic and aromatic regions (19%), and the remainder comes from nonaromatic-nonaromatic interactions (24%). Considering that these subunits (aromatic and nonaromatic) have similar surface areas (340 Å and 315 Å), this implies an electronic favorability for stacking aromatic molecules as opposed to nonaromatic molecules, even before considering entropy, which favors more rigid aromatic molecules. The tilt of the aromatic region of GTP-1 relative to the amyloid backbone was hypothesized to be a result of the optimal π-π stacking distance and the geometry of the 4.77 Å repeat of amyloid (although the helical twist is negligible in shorter assemblies). Approximating the tilt as a simple cosine relationship between these two values ​​(44°) fits the observed data. Given the commonality of heterocyclic aromatic rings in small molecules that bind to amyloid and the constancy of amyloid buildup, the adoption of a tilted heterocycle relative to the amyloid backbone is a common motif in such systems, allowing significantly more favorable π-π interactions between small molecules while maintaining amyloid translational symmetry.

[0217] This structure represents a powerful strategy for the discovery and design of small molecules that bind to amyloid with high affinity in both a sequence- and conformation-specific manner. Filaments present a particular challenge for small molecule design because their accessible surfaces tend to be relatively flat. This limits the amount of surface area that can be lost upon binding of monomeric small molecules, and docking studies therefore tend to demonstrate face-on binding to amyloid. While GTP-1 forms several favorable contacts with amyloid, the surface area lost upon binding of a single monomer is negligible. However, when two GTP-1 molecules stack together, the overall loss of surface area increases to 85 Å, most of which is on the nonpolar face of GTP-1, generating a large driving force associated with the burial of hydrophobic groups. This effect is not observed when the two monomers are separated by a ligand-free binding site, suggesting that the system may be cooperative. To further investigate this cooperativity, we performed single-point DFT calculations for the binding of one, two, and three GTP-1 molecules to the five chains of our truncated model of tau (residues 351–360). While the accuracy of the calculations is inherently limited due to their static nature and lack of explicit solvation, trends can be gleaned. Notably, the binding energy of a single tracer to the five chains is the same at all three potential binding sites, suggesting that the protein-small molecule interaction is limited to the three chains crossed by GTP-1. For two tracers bound to adjacent sites, the energy is the sum of the protein-small molecule binding energy and the small molecule-small molecule dimerization energy, indicating positive cooperativity. The same trend continues with the minimal model of an extended stack, three tracers, suggesting that the calculations relate to the entire ensemble. In contrast, two tracers separated by an unliganded binding site (the minimal model of loose binding) do not exhibit the desired small molecule-small molecule binding energy.

[0218] The positive cooperativity observed in these models suggests that such assemblies of π-stacked small molecules interacting with amyloid are likely related to the behavior of similarly positioned ligands, even though this structure was obtained at a concentration of GTP-1 (20 μM) higher than the measured IC50 (22 nM) (23). Furthermore, this observed behavior, in which both protein-small molecule and small molecule-small molecule interactions are localized and the latter are positively cooperative, is similar to other well-studied biological systems. These systems, including the random coil-to-helix transition of polypeptides or the binding of dye molecules to DNA, have been well described by mathematical models (39-41), suggesting a path to better understanding the thermodynamic and kinetic behavior of small molecule-amyloid interactions under physiological conditions. Templated assembly and symmetry matching have also been observed in the assembly of similar aromatic molecules with globular proteins, although the limited size of the binding pocket limits the assembly size to a maximum of four molecules (42-45).

[0219] Rather than binding to a featureless surface along a uniform beta sheet, the strong geometric and physical complementarity between GTP-1 and this AD-specific cleft likely confers considerable specificity. The local structure of Gln351–Ile360, which comprises the GTP-1 binding site, is strikingly different in filament structures from other tauopathies. In many cases, key residues that form close contacts in the AD structure are not solvent-exposed or form convex surfaces, as opposed to the concave cleft favored for binding. While the CTE protofilament has a similar C-shaped structure in AD, this region of the CTE filament structure is defined by a much shallower angle formed by the kink at Gly355. This shifts Ile360 approximately 3 Å further from Gln351 than in the AD structure, resulting in the loss of a nonpolar interaction between Ile360 and C7 of the GTP-1 phenyl ring. This capping interaction, along with the close nonpolar contact between the fluoroethyl tail at the opposite end of GTP-1 and the AD amyloid filaments, may help prevent positional and conformational heterogeneity, thereby facilitating ligand stacking. Based on structural differences between known tau conformers, GTP-1 was predicted to bind specifically to AD filaments. It is possible that binding to other conformers may occur, possibly at different sequences in the tau filaments, and may involve alternative binding modes.

[0220] The symmetry agreement observed in the structure of GTP-1 bound to PHFs from AD patients may provide a powerful strategy for increasing the druggability of available binding sites in the filaments. Because small changes in the binding site are likely to have a significant impact on GTP-1 binding, it is feasible to design small molecule compounds with high specificity and affinity for a single site within the amyloid filament conformation. This analysis suggests that the development of future tools for diagnostics and potentially therapeutics should focus on heterocycles that stack favorably in the context of amyloid translational constraints, as well as achieving geometric and electrostatic synergy with targeted binding clefts. Understanding not only amyloid assemblies as supramolecular entities but also small molecules opens new routes for designing amyloid filament binders.

[0221] Drugs generally form distinct molecular associations that result in therapeutic interactions. This occurs when drugs bind to one or more biological targets based on complementary shapes, characteristics, and / or reactivities of their surfaces. In some cases, the result is a binary drug-target complex that alters the conformation, activity, or fate of the target. In still others, multiple (identical or different) small molecules bind cooperatively at multiple distinct locations within a target complex with a net affinity or efficacy greater than the sum of the individual binding events (e.g., GPCRs: Lu, et al. Structural basis for the cooperative allosteric activation of the free fatty acid receptor GPR40 (2017) Nat Struct Mol Biol 24, 570-577; type III kinase inhibitors Martinez, et al. (2020). Avoiding or Co-Opting ATP Inhibition: Overview of Type III, IV, V, and VI Kinase Inhibitors. In: Shapiro, P. (eds) Next Generation Kinase Inhibitors. Springer, Cham.). In rare cases, two or three molecules of the same ligand have been observed to bind within a single site in a target complex while engaging in productive interactions with each other (supramolecular dimers of small molecule ligands: Shokat, KMA). point into the UPR.Mol.Cell 38,161-163(2010);supramolecular trimer of ligands:Stornaiuolo,M.,De Kloe,G.,Rucktooa,P.et al.Assembly of aπ-πstack of ligands in the binding site of an acetylcholine-binding protein.Nat Commun 4,1875(2013).(Dimers leveraged for drug discovery: Allen, et al. bioRxiv 2022.05.23.493001, https: / / doi.org / 10.1101 / 2022.05.23.493001.) The reversible intermolecular interactions between monomers in these supramolecular dimeric or trimeric complexes counteract the entropic penalty for simultaneously binding two different molecules at the same site.

[0222] Supramolecular polymers spontaneously assemble from appropriately arranged monomers and maintain their polymeric properties in solution (de Greef, T., Meijer, E. Supramolecular polymers. Nature 453, 171-173 (2008)). They are distinguished from supramolecular dimers and trimers by the increased number of self-associating monomer subunits. Following the observation of unprecedented supramolecular polymer assemblies of an α-synuclein prion inhibitor bound to amyloid fibrils of α-synuclein (Figure 1), it was discovered that supramolecular polymers of small molecule monomers are uniquely suited to interact with supramolecular assemblies of proteins characteristic of degenerative diseases of the central nervous system (CNS). Due to their helical symmetry, fibrillar oligomers and polymers typically lack separate druggable binding pockets. Drugs and diagnostic ligands large enough to span the long binding channels can bind large enough to affect pharmacologically relevant changes in the activity of fibrillating proteins. While this approach is advantageous for generating affinity, such molecules are not sufficiently permeable to cross the blood-brain barrier. Supramolecular polymers composed of small monomers have been identified that can cross the blood-brain barrier and assemble in the presence of ordered α-synuclein aggregates characteristic of multiple system atrophy (MSA). By establishing the binding of GTP-1 to the paired helical filaments of tau from another supramolecular polymer, Alzheimer's disease (Figure 2), we demonstrated the generality of our approach to other proteins characteristic of neurodegenerative diseases. Thus, we identified a general class of supramolecular polymers as potential therapeutic and diagnostic agents for diseases characterized by the ordered accumulation of misfolded proteins, such as Alzheimer's disease, Parkinson's disease, and MSA.

[0223] Example 2 MSA Efficacy Testing Protocol Subjects: Forty-two M83 mice (21 males and 21 females) bred in the University of California, San Francisco, Institute facility were used in these experiments. M83 mice (B6 / C3 background) express human alpha-synuclein with the A53T mutation under the control of the mouse prion promoter. Mice were housed in an AAALAC-accredited facility maintained in a temperature-controlled environment with a 12-hour light:dark cycle. Food and water were provided ad libitum throughout the experimental period. All procedures were approved by the University of California, San Francisco, Institute of Clinical Investigation and Control (ICCUC).

[0224] Procedure: Mice were matched to experimental groups (18 control animals and 24 drug-treated animals) at 9 weeks of age, and all mice were manually inoculated into the thalamus with cell lysate (30 μl of 1 mg / ml MSA cell lysate) from HEK cells stably propagating the human MSA prion before 10 weeks of age. On the day of inoculation, mice in the drug-treated condition began receiving chow formulated with the compound under investigation, while control animals received drug-free chow. Where indicated (i.e., efficacy studies of Compound 3), 1-aminobenzotriazole was added to both the control and test chow, delivering approximately 50 mg / kg / day of a pan-cytochrome p450 inhibitor to all mice in the study. Animals underwent daily health checks throughout the experiment.

[0225] On day 14, additional animals were collected from each group so that 12 experimental animals remained in the control group and 12 experimental animals plus 6 PK animals remained in the drug treatment group. On day 49, serial blood sampling and final blood and brain collection were performed on three male and three female mice in the PK group to assess drug levels in the blood and brain. Starting on day 50, daily neurological assessments were added to the daily health checks. All assessments were performed by two individuals blinded to the animals' treatment conditions.

[0226] Neurological signs were observed if they developed (including proprioceptive deficits, ataxia, bradykinesia, loss of righting reflex, or loss of grasping ability), consistent with the presence of neurodegenerative disease. Mice were then sacrificed, and their brains were assessed for prion infectivity in a cellular assay to confirm that mice with neurological disease had been collected. The time to onset of neurological signs (days postinoculation) was the primary experimental endpoint. Survival curves for control versus treated mice were compared using the Mantel-Cox (log-rank) statistical test. Numerical survival benefit represents the median survival time to onset of neurological signs in the test group divided by the median survival time in the control group.

[0227] Example 3 Alpha-synuclein prion inoculum preparation Preparation of MSA-A inoculum Fragments of the basal ganglia (BG) from MSA patient PD080 were resuspended in 9 volumes of PBS (ml / g brain) and homogenized for 3 × 12 seconds using a probe homogenizer (Thomas Scientific) equipped with a disposable Omni Tip plastic homogenizing probe (Thomas Scientific catalog number 3409Y77) to generate a 10% brain homogenate (10% PD080-BH). Mice hemizygous for the full-length human A53T alpha-synuclein transgene (B6;C3-Tg(Prnp-SNCA*A53T)83Vle / J), commonly referred to as M83 mice (Glasson et al. Neuron 34(4)), were inoculated into the thalamus with 30 μL of 1% PD080BG-BH per mouse. Brains were harvested at the time of neurological symptoms. The latter were pooled and homogenized as described above to produce 10% BH. An aliquot of the latter BH was diluted to 1% and inoculated into hemizygous M83 mice as described above for the first inoculation. The brains of these mice were collected when the mice exhibited neurological symptoms. The brains were pooled and homogenized to produce 10% BH, designated SHO6. SHO6 10% BH was clarified of insoluble debris by centrifugation at 800 x g for 5 minutes. The resulting supernatant was collected, and the protein concentration of this clarified BH (SHO6-CBH) was determined by bicinchoninic acid assay (BCA) (Thermo Fisher catalog number 23227). HEK293 cells (S104) stably expressing C-terminally YFP-tagged full-length wild-type α-synuclein (1-140) were transfected with SHO6-CBH using lipofectamine 2000 (LF2K) (Thermo Fisher catalog number 11668500). Briefly, 0.1 μg of SHO6-CBH was mixed with 0.15 μl of LF2K in 2.5 μL of PBS per well of a 384-well plate and incubated at room temperature (RT) for 90 minutes. The transfection mixture was then diluted by adding 7.5 μL of Opti-MEM (Thermo Scientific catalog number 31985070), and the transfection complex was added to a single well of a 384-well plate containing 3,000 S104 cells.After 3 days, cells were harvested, counted, and seeded at a density of 1 cell / well in 96-well plates. Clonal cells containing YFP-positive synuclein aggregates were selected and designated A8 cells. Following expansion of these A8 cells, cell lysates were prepared by freeze-thawing. Cells from a confluent T175 flask were harvested by scraping in 1 ml of PBS containing 1x protease inhibitor cocktail. The cell suspension was subjected to four cycles of freezing in liquid nitrogen for 5 min followed by thawing at 37°C for 5 min. The resulting cell lysate was centrifuged at 2000 RPM for 10 min to remove cellular debris. The protein concentration of the resulting MSA-A lysate was determined by BCA assay, and aliquots were frozen for future use.

[0228] Preparation of MSA-B inoculum Fragments of substantia nigra (SN) from MSA patient 1720 were resuspended in 9 volumes of PBS (ml / g brain) and homogenized for 3 × 12 seconds using a probe homogenizer (Thomas Scientific) equipped with a disposable Omni Tip plastic homogenizing probe (Thomas Scientific catalog number 3409Y77). The resulting 10% brain homogenate (BH) was cleared of insoluble debris by centrifugation at 800 × g for 5 minutes. The protein concentration of the clarified BH (1720SN-CBH) was determined by bicinchoninic acid assay (BCA) (Thermo Fisher catalog number 23227). HEK293 cells (S208.4 cells) stably and constitutively expressing C-terminally YFP-tagged full-length α-synuclein (1-140) with the A53T mutation were transfected with 1720SN-CBH using lipofectamine 2000 (LF2K) (Thermo Fisher catalog number 11668500). Briefly, 0.57 μg / well of 1720SN-CBH was mixed with 0.86 μl / well of LF2K in 10 μL of PBS and incubated at room temperature (RT) for 90 minutes. The transfection mixture was diluted by adding 40 μL of Opti-MEM (Thermo Scientific catalog number 31985070) to a concentration of 1.7 × 10 4The cells were added to a single well of a 96-well plate containing 100 S208.4 cells. After 3 days, the cells were harvested, counted, and seeded at a density of 1 cell / well in a 96-well plate. Clonal cells containing YFP-positive synuclein aggregates were selected and designated M3 cells. Following expansion of these M3 cells, cell lysates were prepared by freeze-thawing. Cells from a confluent T175 flask were harvested by scraping in 1 ml of PBS containing 1x protease inhibitor cocktail (Thermo Scientific catalog no. A32953). The cell suspension was subjected to four cycles of freezing in liquid nitrogen for 5 minutes followed by thawing at 37°C for 5 minutes. The resulting cell lysate was centrifuged at 2000 RPM for 10 minutes to remove cell debris, and the protein concentration of the resulting M3 cells was determined by BCA assay. Subsequently, HEK293 cells (S501 cells) stably expressing a C-terminally YFP-tagged truncated form of α-synuclein (1-95) with the A53T mutation were transfected with M3 lysate using lipofectamine 2000 (LF2K). Briefly, 0.57 μg / well of M3 lysate was mixed with 0.86 μl / well of LF2K in 10 μL of PBS and incubated at room temperature for 90 minutes. The transfection mixture was diluted by adding 40 μL of Opti-MEM, resulting in a total of 1.7 × 10 4 The MSA-B aggregates were added to a single well of a 96-well plate containing 100 S501 cells. After 3 days, the cells were harvested, counted, and seeded at a density of 1 cell / well in a 96-well plate. Clonal cells containing YFP-positive synuclein aggregates were selected and designated M1 cells. Following expansion of these M1 cells, cell lysates were prepared by freeze-thawing. Cells from a confluent T175 flask were harvested by scraping in 1 ml of PBS containing 1x protease inhibitor cocktail. The cell suspension was subjected to four cycles of freezing in liquid nitrogen for 5 min followed by thawing at 37°C for 5 min. The resulting cell lysate was centrifuged at 2000 RPM for 10 min to remove cellular debris. The protein concentration of the resulting MSA-B lysate was determined by BCA assay, and aliquots were frozen for future use.

[0229] α-synuclein prion cell assay MSA-A dose-response assay in HEK293T cells HEK293 cells (DSS121 cells) stably expressing C-terminally YFP-tagged full-length α-synuclein (1–140) with the A53T mutation were incubated with 0.1 μM tetracycline and 0.1 μg / ml Hoechst Dye (Thermo Cells were seeded at a density of 3000 cells per well of a 384-well black-walled, clear-bottom plate (Greiner Catalog No. 5678-1091Q) in 60 μl of complete DMEM (DMEM (Corning Catalog No. 10-013-CV) containing 10% FBS (VWR Catalog No. 97068-085), 0.5% penicillin / streptomycin (Gibco Catalog No. 15140-122)) containing 0.24 μg of MSA-A lysate diluted in 5 μl of PBS per well to be transfected and 0.24 μl of Lipofectamine Pre-diluted MSA-A lysate was prepared by diluting LF2K (2000) (ThermoFisher Cat. No. 11668500) into a separate tube with 5 μl of Opti-MEM per well to be transfected. The pre-diluted MSA-A lysate and LF2K were then mixed and incubated at room temperature for 90 minutes. A 10 μL aliquot of the MSA-A transfection complex was added to each well of a 384-well plate containing DSS121 cells using a Bravo liquid handling platform. Serial dilutions of test compounds were performed using a Bravo liquid handling platform (Agilent). Briefly, 25 μL of each desired lysate was added to a separate tube. The compound was added to row A of a 96-well plate (Greiner catalog number 651261) and diluted 1:1 with DMSO. Subsequently, a 3-fold serial dilution in DMSO was performed for each compound from rows B through G of the 96-well plate. Compounds were further diluted by transferring 2 μL of the serially diluted DMSO stock to a 96-well plate containing 123 μL / well of complete DMEM to generate working stocks. After addition of the MSA-A transfection complex, 10 μL of working compound stock was transferred from a single well of the 96-well compound plate to each of four wells of a 384-well cell plate using a Bravo liquid handling platform.Cells were incubated at 37°C / 5% CO2 for 96 hours and then imaged using an InCell Analyzer 6000 (Cytiva GE). Images were captured using FITC (488 nm excitation / 525 + / - 20 nm emission) and DAPI (405 nm excitation / 455 + / - 50 nm emission) filter sets to image YFP-tagged α-σ inucrine and Hoechst-positive nuclei, respectively. FITC images were processed using InCell Developer image analysis software to determine the intensity of pixels within YFP-positive cell aggregates and the number of cells containing YFP-positive aggregates. Additionally, images captured with the DAPI filter set were processed using InCell Developer to determine the number of cells per image. Finally, processed data were plotted as a function of compound concentration and the EC50 and efficacy window values ​​calculated from these dose-response curves.

[0230] MSA-B dose-response assay in HEK293T cells HEK293 cells (DSS121 cells) stably expressing C-terminally YFP-tagged full-length α-synuclein (1–140) with the A53T mutation were incubated with 0.1 μM tetracycline and 0.1 μg / ml Hoechst Dye (Thermo Cells were seeded at a density of 3,000 cells per well of a 384-well black-walled, clear-bottom plate (Greiner catalog no. 5678-1091Q) in 60 μl of complete DMEM (DMEM (Corning catalog no. 10-013-CV), 10% FBS (VWR catalog no. 97068-085), 0.5% penicillin / streptomycin (Gibco catalog no. 15140-122) containing 1% FBS (Scientific catalog no. H3570). Transfection complexes were prepared by mixing 0.1 μg / well of MSA-B lysate with 0.15 μL / well of LF2K in PBS and incubated at room temperature for 90 minutes. The MSA-B transfection complex was then diluted with 4 volumes of Opti-MEM, and 10 μL of MSA-B transfection complex was added to each well of the 384-well plate containing DSS121 cells using a Bravo liquid handling platform. Serial dilutions of test compounds were performed. Dilutions were performed using a Bravo liquid handling platform (Agilent). Briefly, 25 μL of each compound of interest was added to row A of a 96-well plate (Greiner catalog number 651261) and diluted 1:1 with DMSO. Subsequently, a 3-fold serial dilution in DMSO was performed for each compound from rows B–G of the 96-well plate. Compounds were further diluted by transferring 2 μL of the serially diluted DMSO stock to a 96-well plate containing 123 μL / well of complete DMEM to generate working stocks. After addition of the MSA-B transfection complex, a Bravo liquid handling platform was used to transfer 10 μL of working compound stock from a single well of the 96-well compound plate to each of four wells of a 384-well cell plate (i.e., 96-well A1 was transferred to 384-wells A1, B1, A2, and B2). Cells were incubated at 37°C / 5% CO2 for 72 hours, followed by InCell™ fusion. Imaging was performed using an Analyzer 6000 (Cytiva GE).To image YFP-tagged α-synuclein and Hoechst-positive nuclei, respectively, images were captured using FITC (488 nm excitation / 525 ± 20 nm emission) and DAPI (405 nm excitation / 455 ± 50 nm emission) filter sets. FITC images were processed using InCell Developer image analysis software to determine the intensity of pixels within YFP-positive cell aggregates and the number of cells containing YFP-positive aggregates. Additionally, images captured with the DAPI filter set were processed using InCell Developer to determine the number of cells per image. Finally, processed data were plotted as a function of compound concentration and the EC50 and efficacy window values ​​calculated from these dose-response curves.

[0231] Example 4 Trisubstituted oxazoles Aminopyrimidone intermediate synthesis [ka]

[0232] 2-Chloro-N-ethyl-5-fluoropyrimidin-4-amine. To a solution of 2,4-dichloro-5-fluoropyrimidine (4.00 g, 1 eq, 24.0 mmol) and triethylamine (3.15 g, 4.34 mL, 1.3 eq, 31.1 mmol) in ethanol (60 mL) was added ethylamine (2 M in THF, 14.4 mL, 1.2 eq, 28.7 mmol) over 5 minutes at 0° C. under nitrogen. The reaction mixture was stirred at 0° C. for 10 minutes and at room temperature for 20 hours. The resulting mixture was concentrated under reduced pressure. The residue was diluted with aqueous NaCl. The organic material was extracted with ethyl acetate, dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography (silica gel, 10-35% EtOAc in heptane) to give 2-chloro-N-ethyl-5-fluoropyrimidin-4-amine (4.3 g, 24.2 mmol, 100%) as a white solid.

[0233] 1H NMR(400 MHz,CHLOROFORM-d)d ppm 1.30(t,J=7.34 Hz,3H),3.57(qd,J=7.25,5.62 Hz,2H),5.18(bs,1H),7.87(d,J=2.93 Hz,1H).

[0234] 4-(Ethylamino)-5-fluoropyrimidin-2(1H)-one. To a solution of 2-chloro-N-ethyl-5-fluoropyrimidin-4-amine (4.26 g, 1 eq., 24.2 mmol) in formic acid (16 g, 13 mL, 14 eq., 0.34 mol), water (1.3 g, 1.3 mL, 3.0 eq., 72 mmol) was added, and the reaction mixture was stirred at 90 °C for 10 hours. After cooling to room temperature, the mixture was concentrated under reduced pressure. The residue was mixed with ethanol, and the resulting precipitate was collected by vacuum filtration. The material was then washed with ethanol and ethyl acetate to give 4-(ethylamino)-5-fluoropyrimidin-2(1H)-one (3.47 g, 22.1 mmol, 91%) as a white solid.

[0235] 1 H NMR(400 MHz,DMSO-d6)d ppm 1.15(t,J=7.21 Hz,3H),3.45(quin,J=6.85 Hz,2H),8.03(d,J=6.11 Hz,1H),9.61(bs,1H),11.70(bs,1H).

[0236] LCMS: rt 0.14 min. [M+H] + 158.1 m / z. [ka]

[0237] 2-Chloro-5-fluoro-N-methylpyrimidin-4-amine. To a stirred solution of 2,4-dichloro-5-fluoro-pyrimidine (1.0 equiv.) in anhydrous THF (10.0 vol.) and triethylamine (1.2 equiv.) cooled in an ice bath, a 2 M solution of methylamine in THF (1.1 equiv.) was added. The mixture was stirred at room temperature for 16 h. Saturated aqueous NaHCO3 was added, and the mixture was extracted three times with ethyl acetate. The organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure to give 2-chloro-5-fluoro-N-methylpyrimidin-4-amine (80-100% yield).

[0238] 1 H NMR (300 MHz, DMSO-d6): d 8.13 (s, 1H), 8.04 (d, J = 3.5 Hz, 1H), 2.84 (d, J = 4.7 Hz, 3H).

[0239] 5-Fluoro-4-(methylamino)pyrimidin-2(1H)-one. 2-Chloro-5-fluoro-N-methylpyrimidin-4-amine (1.0 equiv.) was dissolved in formic acid (20.0 equiv.) and water (1.1 equiv.). The reaction mixture was stirred at 90°C for 24 hours. An additional portion of water (2.5 equiv.) and formic acid (9.2 equiv.) was then added, and the reaction was continued at 90°C for an additional 2 days. After cooling to ambient temperature, EtOAc was added. The resulting precipitate was collected by vacuum filtration, washed with EtOAc, and dried to give 5-fluoro-4-(methylamino)-1,2-dihydropyrimidin-2-one (75-95% yield).

[0240] 1 H NMR(300 MHz,DMSO-d6):d 9.92(s,1H),8.06(d,J=6.1 Hz,1H),2.97(d,J=3.5 Hz,3H).

[0241] LCMS: rt 0.12 min. [M+H] + 144.2 m / z. [ka]

[0242] 2-Chloro-5-fluoro-N-(methyl-d3)pyrimidin-4-amine 2-Chloro-5-fluoro-N-(methyl-d3)pyrimidin-4-amine was prepared similarly to the preparation of 2-chloro-N-ethyl-5-fluoropyrimidin-4-amine.

[0243] 1 H NMR(400 MHz,DMSO-d6)d ppm 7.28(bs,1H),7.20(dd,J=3.42,0.49 Hz,1H),1.66(bs,6H).

[0244] 19 F NMR(376 MHz,DMSO-d6)d ppm-158.11(s,1F).

[0245] 5-Fluoro-4-((methyl-d3)amino)pyrimidin-2(1H)-one. 5-Fluoro-4-((methyl-d3)amino)pyrimidin-2(1H)-one was prepared similarly to the preparation of 4-(ethylamino)-5-fluoropyrimidin-2(1H)-one.

[0246] LCMS: rt 0.13 min. [M+H] + 147.1 m / z. [ka]

[0247] 2-Chloro-N-(2,2-difluoroethyl)-5-fluoropyrimidin-4-amine 2-Chloro-N-(2,2-difluoroethyl)-5-fluoropyrimidin-4-amine was prepared similarly to the preparation of 2-chloro-N-ethyl-5-fluoropyrimidin-4-amine.

[0248] 1H NMR(400 MHz,CHLOROFORM-d)d ppm 3.96(tdd,J=14.61,6.24,4.16 Hz,2H),5.44(bs,1H),6.00(tt,J=55.75,3.91 Hz,1H),7.99(d,J=2.45 Hz,1H).

[0249] 4-((2,2-Difluoroethyl)amino)-5-fluoropyrimidin-2(1H)-one. 4-((2,2-Difluoroethyl)amino)-5-fluoropyrimidin-2(1H)-one was prepared similarly to the preparation of 4-(ethylamino)-5-fluoropyrimidin-2(1H)-one.

[0250] 1 H NMR(400 MHz,DMSO-d6)d ppm 3.78(t,J=14.92 Hz,2H),6.30(tt,J=55.51,3.91 Hz,1H),7.89(d,J=6.11 Hz,1H),8.82(bs,1H).

[0251] LCMS: rt 0.15 min. [M+H] + 194.1 m / z. [ka]

[0252] 2-Chloro-N-cyclopropyl-5-fluoropyrimidin-4-amine 2-Chloro-N-cyclopropyl-5-fluoropyrimidin-4-amine was prepared similarly to the preparation of 2-chloro-N-ethyl-5-fluoropyrimidin-4-amine.

[0253] LCMS: rt 1.34 min. [M+H] + 188.1 m / z.

[0254] 4-(Cyclopropylamino)-5-fluoropyrimidin-2(1H)-one. 4-(Cyclopropylamino)-5-fluoropyrimidin-2(1H)-one was prepared similarly to the preparation of 4-(ethylamino)-5-fluoropyrimidin-2(1H)-one.

[0255] LCMS: rt 0.12 min. [M+H] + 170.1 m / z.

[0256] Thionyloxazole chloride cyclization Synthesis of Compound 1 [ka]

[0257] 1-(4-Chloro-3-fluorophenyl)-2,2-dihydroxyethan-1-one. Selenium dioxide (25.4 g, 95 wt%, 1.5 equiv., 217 mmol) was placed in water (30.0 mL) and 1,4-dioxane (300 mL) in a 500 mL medium pressure flask and heated to 70 °C until all materials were completely dissolved. To the reaction mixture at 70 °C, 1-(4-chloro-3-fluorophenyl)ethan-1-one (25.0 g, 1 equiv., 145 mmol) was added. The flask was sealed and stirred at 100 °C for 21 h. The reaction was cooled to room temperature and filtered through a pad of Celite, further eluting with EtOAc. The filtrate was concentrated to a viscous brown oil, which was treated with HO (50 mL) and heated to 100 °C for 8 h under a reflux condenser. The heating bath was then removed, and the mixture was stirred at room temperature for 16 h. The resulting precipitate was collected by vacuum filtration, washed with cold water, and dried under vacuum to give 2-(4-chloro-3-fluorophenyl)-2-oxoacetaldehyde (24.1 g, 129 mmol, 89.2%) as a light brown solid.

[0258] LCMS: rt 1.31 min. [M+H] + 186.9 m / z.

[0259] 3-Chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide. To a solution of 2-(4-chloro-3-fluorophenyl)-2-oxoacetaldehyde (2.0 g, 1 equiv., 11 mmol) in dioxane (40 mL) was added 3-chlorobenzamide (1.8 g, 1.1 equiv., 12 mmol), and the reaction mixture was heated to 90° C. for 14 h. The reaction was then hot filtered through a pad of Celite and concentrated to a brown viscous oil. The crude material was recrystallized from EtOAc / heptane and dried under vacuum to give 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide (0.738 g, 2.16 mmol, 20%) as a beige solid.

[0260] 1 H NMR(400 MHz,DMSO-d6)d ppm 9.58(d,J=7.58 Hz,1H),7.91(t,J=4.89 Hz,2H),7.73-7.86(m,3H),7.62(d,J=8.07 Hz,1H),7.47-7.54(m,1H),6.80(d,J=6.60 Hz,1H),6.43(t,J=7.21 Hz,1H).LCMS:rt 2.14 min.[M+H] + 344.0 m / z

[0261] 3-Chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide. To a slurry of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide (0.738 g, 1 equiv., 2.16 mmol) in DCM (20 mL) was added PCl5 (496 mg, 95 wt%, 1.05 equiv., 2.26 mmol). The resulting cloudy reaction mixture was stirred at 50° C. for 2 h. The reaction was concentrated to a yellow solid and dried under vacuum to give the intermediate chloro adduct as a pale yellow solid. A solution of 5-fluoro-4-(methylamino)pyrimidin-2(1H)-one (401 mg, 1.3 equiv., 2.80 mmol) and triethylamine (655 mg, 902 μL, 3 equiv., 6.47 mmol) in DMF (20 mL) was stirred at room temperature for 15 minutes. The intermediate chloro adduct was added, and the reaction mixture was stirred at room temperature for 16 hours. The reaction was concentrated in vacuo and treated with water (10 mL). The resulting precipitate was collected by vacuum filtration, washed with cold water, and dried under vacuum to give 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide (276 mg, 591 μmol, 27.4%) as a pale yellow solid.

[0262] LCMS: rt 2.21 min. [M+H] + 467.0 m / z 1-(5-(4-chloro-3-fluorophenyl)-2-(3-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one. Compound 125

[0263] 3-Chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide (58.6 mg, 1 equivalent, 125 μmol) was treated with thionyl chloride (1.63 g, 1.00 mL, 109 equivalents, 13.7 mmol) and the reaction mixture was stirred at 80° C. for 3 hours. The reaction was cooled to room temperature and a precipitate formed. The solid was collected by vacuum filtration, washed with EtOAc, and dried under vacuum to give 1-(5-(4-chloro-3-fluorophenyl)-2-(3-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one (26.8 mg, 59.7 μmol, 47.6%) as a white solid.

[0264] 1 H NMR(400 MHz,DMSO-d6)d ppm 8.68(d,J=4.65 Hz,1H),8.25(d,J=0.98 Hz,1H),8.16(d,J=6.36 Hz,1H),8.10(d,J=6.60 Hz,1H),7.86(d,J=10.51 Hz,1H),7.74(t,J=7.95 Hz,1H),7.66-7.71(m,1H),7.60-7.66(m,1H),7.43(d,J=8.31 ​​Hz,1H),2.92(d,J=4.40 Hz,3H).

[0265] LCMS: rt 2.43 min. [M+H] + 449.0 m / z.

[0266] Synthesis of compound 83 [ka]

[0267] N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide. 3-Toluamide (2.0 g, 14.8 mmol, 1.0 equiv.), 1-(4-chloro-3-fluorophenyl)-2,2-dihydroxyethan-1-one (4.72 g, 22.2 mmol, 1.5 equiv.) were dissolved in dioxane (40.0 mL), and the resulting mixture was stirred at 100 °C for 2 h. The solvent was evaporated, and the resulting material was purified by silica gel column chromatography (0–10% MeOH in DCM) to give N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide (3.92 g, 12.2 mmol, 82%).

[0268] 1 H NMR(300 MHz,DMSO-d6)d 9.37(d,J=7.9 Hz,1H),7.91(dd,J=10.1,1.6 Hz,1H),7.85-7.73(m,2H),7.71-7.61(m,2H),7.35(dd,J=4.7,2.2 Hz,2H),6.64(s,1H),6.39(s,1H),2.34(s,3H).

[0269] N-(1-chloro-2-(4-chloro-3-fluorophenyl)-2-oxoethyl)-3-methylbenzamide. N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide (2.0 g, 6.22 mmol, 1.0 equiv) was dissolved in DCM (40.0 mL) and phosphorus pentachloride (1.36 g, 6.53 mmol, 1.05 equiv) was added. The resulting mixture was stirred at ambient temperature for 16 hours. The solvent was removed under reduced pressure. The resulting material was suspended in hexane and collected by vacuum filtration to give N-[1-chloro-2-(4-chloro-3-fluorophenyl)-2-oxoethyl]-3-methylbenzamide (0.605 g, 1.78 mmol, 29%).

[0270] 1H NMR(300 MHz,DMSO-d6):d 9.38(d,J=7.9 Hz,1H),7.91(dt,J=10.2,3.0 Hz,1H),7.83-7.76(m,2H),7.71-7.59(m,2H),7.46-7.32(m,2H),6.39(d,J=7.9 Hz,1H),2.34(d,J=4.3 Hz,3H).

[0271] N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide. N-(1-chloro-2-(4-chloro-3-fluorophenyl)-2-oxoethyl)-3-methylbenzamide (0.605 g, 1.78 mmol, 1.0 equiv.) was added to a mixture of 5-fluoro-4-(methylamino)pyrimidin-2(1H)-one (0.229 g, 1.6 mmol, 0.9 equiv.) and sodium bicarbonate (0.747 g, 8.89 mmol, 5.0 equiv.) in DMF (4.6 mL) at 0° C. The reaction mixture was then stirred at room temperature for 30 minutes and allowed to warm to ambient temperature with stirring for 48 hours. The solvent was evaporated, and the resulting solid was taken up in water and collected by vacuum filtration. The solid was then dissolved in CHCl and washed three times with brine. The organic layer was collected and dried over sodium sulfate. The solvent was removed to give N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxo-1,2-dihydropyrimidin-1-yl)-2-oxoethyl)-3-methylbenzamide (0.439 g, 0.982 mmol, 35%).

[0272] LCMS(ESI+):m / z 446.90,[M+H] + . 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one. Compound 83

[0273] N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxo-1,2-dihydropyrimidin-1-yl)-2-oxoethyl)-3-methylbenzamide (0.438 g, 0.618 mmol) was dissolved in thionyl chloride (1.58 mL, 21.6 mmol, 35 equiv.) and stirred at 60 °C for 2 h. The solvent was removed under vacuum, and the product was precipitated with methanol and ethyl ether. The solid precipitate was collected by vacuum filtration and purified by silica gel column chromatography (0–10% MeOH in DCM) to give 1-(5-(4-chloro-3-fluorophenyl)-2-(3-methylphenyl)-1,3-oxazol-4-yl)-5-fluoro-4-(methylamino)-1,2-dihydropyrimidin-2-one (0.233 g, 0.543 mmol, 87%).

[0274] LCMS(ESI+):m / z 428.86,[M+H] + .

[0275] 1 H NMR(300 MHz,DMSO-d6)δ 8.47(d,J=5.0 Hz,1H),8.13(d,J=6.7 Hz,1H),8.03-7.92(m,2H),7.81-7.70(m,2H),7.54-7.36(m,3H),2.91(d,J=4.6 Hz,3H),2.43(s,3H).

[0276] Synthesis of Compound 126 [ka] 4-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide

[0277] 4-Chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide.

[0278] 1 H NMR(400 MHz,DMSO-d6)d ppm 9.52(d,J=7.83 Hz,1H),7.85-7.98(m,3H),7.72-7.83(m,2H),7.54(d,J=8.31 ​​Hz,2H),6.76(d,J=6.85 Hz,1H),6.41(t,J=7.34 Hz,1H).

[0279] 19 F NMR(376 MHz,DMSO-d6)d ppm-115.16(s,1F).

[0280] LCMS: rt 2.23 min. [M+Na] + 363.9 m / z. 4-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide

[0281] 4-Chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide.

[0282] LCMS: rt 2.29 min. [M+H] + 467.0 m / z. Compound 126. 1-(5-(4-chloro-3-fluorophenyl)-2-(4-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one hydrochloride.

[0283] 1-(5-(4-chloro-3-fluorophenyl)-2-(4-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one hydrochloride was prepared similarly to the preparation of (5-(4-chloro-3-fluorophenyl)-2-(3-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one.

[0284] 1 H NMR(400 MHz,DMSO-d6)d ppm 8.54(d,J=5.40 Hz,1H),8.18(d,J=8.56 Hz,2H),8.10-8.14(m,1H),7.72-7.79(m,2H),7.67(d,J=8.80 Hz,2H),7.35-7.44(m,1H),2.91(d,J=4.65 Hz,3H).

[0285] LCMS: rt 2.55 min. [M+H] + 448.9 m / z.

[0286] Synthesis of compound 127 [ka] 4-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(4-(ethylamino)-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide.

[0287] 4-Chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(4-(ethylamino)-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide.

[0288] LCMS: rt 2.35 min. [M+H] + 481.0 m / z.

[0289] 1-(5-(4-chloro-3-fluorophenyl)-2-(4-chlorophenyl)oxazol-4-yl)-4-(ethylamino)-5-fluoropyrimidin-2(1H)-one. Compound 127 1-(5-(4-chloro-3-fluorophenyl)-2-(4-chlorophenyl)oxazol-4-yl)-4-(ethylamino)-5-fluoropyrimidin-2(1H)-one was prepared similarly to the preparation of (5-(4-chloro-3-fluorophenyl)-2-(3-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one.

[0290] LCMS: rt 2.63 min. [M+H] + 462.9 m / z.

[0291] Synthesis of compound 128 [ka]

[0292] N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide. N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide.

[0293] 1 H NMR(400 MHz,DMSO-d6)d ppm 9.39(d,J=7.82 Hz,1H),7.90(d,J=10.27 Hz,1H),7.74-7.83(m,2H),7.69(s,1H),7.65(d,J=6.36 Hz,1H),7.28-7.41(m,2H),6.66(d,J=6.85 Hz,1H),6.40(t,J=7.34 Hz,1H),2.33(s,3H).

[0294] 13 C NMR(101 MHz,DMSO-d6)d ppm 193.56,166.03,155.73,137.72,133.25,132.41,131.07,128.30,128.05,125.70,125.67,124.67,116.57,116.36,74.04,20.90.

[0295] LCMS: rt 2.14 min. [M+Na] + 344.0 m / z.

[0296] N-(2-(4-chloro-3-fluorophenyl)-1-(4-(ethylamino)-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide. N-(2-(4-chloro-3-fluorophenyl)-1-(4-(ethylamino)-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide.

[0297] LCMS:rt 2.32 min.[M+H]+461.0 m / z.

[0298] 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-4-(ethylamino)-5-fluoropyrimidin-2(1H)-one hydrochloride. Compound 128 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-4-(ethylamino)-5-fluoropyrimidin-2(1H)-one hydrochloride was prepared similarly to the preparation of (5-(4-chloro-3-fluorophenyl)-2-(3-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one.

[0299] LCMS: rt 2.60 min. [M+H]+ 443.0 m / z.

[0300] Synthesis of compound 129 [ka]

[0301] N-(2-(4-chloro-3-fluorophenyl)-1-(4-(cyclopropylamino)-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide. N-(2-(4-chloro-3-fluorophenyl)-1-(4-(cyclopropylamino)-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide.

[0302] LCMS: rt 2.32 min. [M+H] + 473.0 m / z.

[0303] 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-4-(cyclopropylamino)-5-fluoropyrimidin-2(1H)-one hydrochloride. Compound 129 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-4-(cyclopropylamino)-5-fluoropyrimidin-2(1H)-one hydrochloride was prepared similarly to the preparation of (5-(4-chloro-3-fluorophenyl)-2-(3-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one.

[0304] LCMS: rt 2.59 min. [M+H] + 455.0 m / z.

[0305] Synthesis of Compound 130 [ka]

[0306] N-(2-(4-chloro-3-fluorophenyl)-1-(4-((2,2-difluoroethyl)amino)-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide. N-(2-(4-chloro-3-fluorophenyl)-1-(4-((2,2-difluoroethyl)amino)-5-fluoro-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide.

[0307] LCMS: rt 2.35 min. [M+H] + 497.1 m / z.

[0308] Compound 130. 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-4-((2,2-difluoroethyl)amino)-5-fluoropyrimidin-2(1H)-one hydrochloride. 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-4-((2,2-difluoroethyl)amino)-5-fluoropyrimidin-2(1H)-one hydrochloride was prepared similarly to the preparation of (5-(4-chloro-3-fluorophenyl)-2-(3-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one.

[0309] LCMS: rt 2.62 min. [M+H] + 479.0 m / z.

[0310] Synthesis of compound 131. [ka]

[0311] N-(2-(5-bromothiophen-2-yl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide N-(2-(5-bromothiophen-2-yl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide.

[0312] 1 H NMR(400 MHz,DMSO-d6)d ppm 9.41(d,J=8.07 Hz,1H),7.74-7.77(m,1H),7.73(s,1H),7.69(d,J=6.36 Hz,1H),7.39(dd,J=4.16,0.73 Hz,1H),7.34-7.37(m,2H),6.87(d,J=6.60 Hz,1H),6.19(t,J=7.34 Hz,1H),2.35(s,3H).

[0313] LCMS: rt 2.50 min. [M+Na] + 353.0 m / z.

[0314] N-(2-(5-bromothiophen-2-yl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide N-(2-(5-Bromothiophen-2-yl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide.

[0315] 1H NMR(400 MHz,DMSO-d6)d ppm 9.77(d,J=8.56 Hz,1H),8.18(d,J=4.65 Hz,1H),7.90(d,J=6.85 Hz,1H),7.76(s,2H),7.72(d,J=7.34 Hz,1H),7.65(d,J=4.16 Hz,1H),7.42(d,J=3.42 Hz,2H),7.34(d,J=8.56 Hz,1H),2.81(d,J=4.65 Hz,3H),2.36(s,3H).

[0316] 19 F NMR(376 MHz,DMSO-d6)d ppm-169.62--168.03(m,1 F).

[0317] LCMS: rt 2.11 min. [M+H] + 479.0 m / z.

[0318] 1-(5-(5-Bromothiophen-2-yl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one. Compound 131 1-(5-(5-Bromothiophen-2-yl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one hydrochloride was prepared similarly to the preparation of (5-(4-chloro-3-fluorophenyl)-2-(3-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one.

[0319] 1 H NMR(400 MHz,DMSO-d6)d ppm 8.58(d,J=4.40 Hz,1H),8.13(d,J=6.60 Hz,1H),7.89(s,1H),7.86(d,J=7.82 Hz,1H),7.45-7.53(m,1H),7.39-7.44(m,1H),7.37(d,J=3.91 Hz,1H),7.30(d,J=3.91 Hz,1H),2.91(d,J=4.65 Hz,3H),2.42(s,3H).

[0320] 19 F NMR(376 MHz,DMSO-d6)d ppm-169.88--166.14(m,1 F).

[0321] LCMS: rt 2.38 min. [M+H] + 460.9 m / z.

[0322] Synthesis of compound 132. [ka]

[0323] N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)-5-methylthiophene-2-carboxamide N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)-5-methylthiophene-2-carboxamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide.

[0324] 1 H NMR(400 MHz,DMSO-d6)d ppm 9.31(d,J=8.07 Hz,1H),7.89(d,J=10.27 Hz,1H),7.73-7.83(m,2H),7.68(d,J=3.67 Hz,1H),6.83(d,J=3.67 Hz,1H),6.71(d,J=6.60 Hz,1H),6.38(t,J=7.34 Hz,1H),2.45(s,3H).

[0325] 19 F NMR(376 MHz,DMSO-d6)d ppm-115.13(s,1 F).

[0326] LCMS: rt 2.11 min. [M+Na] + 349.9 m / z.

[0327] N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-5-methylthiophene-2-carboxamide N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-5-methylthiophene-2-carboxamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide.

[0328] LCMS: rt 2.10 min. [M+H] + 453.0 m / z.

[0329] 1-(5-(4-chloro-3-fluorophenyl)-2-(5-methylthiophen-2-yl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one. Compound 132 1-(5-(4-chloro-3-fluorophenyl)-2-(5-methylthiophen-2-yl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one hydrochloride was prepared similarly to the preparation of (5-(4-chloro-3-fluorophenyl)-2-(3-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one.

[0330] 1 H NMR(400 MHz,DMSO-d6)d ppm 8.60(d,J=4.65 Hz,1H),8.12(d,J=6.60 Hz,1H),7.81(d,J=3.67 Hz,1H),7.73(t,J=8.19 Hz,1H),7.61(dd,J=10.27,1.71 Hz,1H),7.34(dd,J=8.44,1.83 Hz,1H),7.01(d,J=3.67 Hz,1H),2.90(d,J=4.65 Hz,3H),2.55(s,3H).

[0331] 19 F NMR(376 MHz,DMSO-d6)d ppm-116.98--111.63(m,1F),-169.61--166.81(m,1F).

[0332] LCMS: rt 2.33 min. [M+H] + 435.0 m / z.

[0333] Synthesis of compound 133 [ka]

[0334] N-(2-(3,4-difluorophenyl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide N-(2-(3,4-difluorophenyl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide.

[0335] LCMS: rt 2.02 min. [M+Na] + 328.0 m / z.

[0336] N-(2-(3,4-difluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide N-(2-(3,4-difluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide.

[0337] 1H NMR(400 MHz,DMSO-d6)d ppm 8.18(d,J=4.65 Hz,1H),7.88-7.98(m,3H),7.76-7.84(m,1H),7.73(s,1H),7.60-7.71(m,2H),7.44(d,J=8.31 Hz,1H),7.33-7.42(m,2H),2.80(d,J=4.40 Hz,3H),2.35(s,3H).

[0338] 19 F NMR(376 MHz,DMSO-d6)d ppm-129.86(d,J=22.40 Hz,1F),-136.65(d,J=23.00 Hz,1F),-169.14(s,1F).

[0339] LCMS: rt 2.06 min. [M+H] + 431.0 m / z.

[0340] 1-(5-(3,4-Difluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one hydrochloride. Compound 133 1-(5-(3,4-Difluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one hydrochloride was prepared similarly to the preparation of (5-(4-chloro-3-fluorophenyl)-2-(3-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one.

[0341] 1 H NMR(400 MHz,DMSO-d6)d ppm 8.83(d,J=4.65 Hz,1H),8.20(d,J=6.60 Hz,1H),7.99(s,1H),7.94(d,J=7.58 Hz,1H),7.76-7.86(m,1H),7.56-7.65(m,1H),7.45-7.52(m,1H),7.37-7.45(m,2H),2.93(d,J=4.40 Hz,3H),2.42(s,3H).

[0342] LCMS: rt 2.31 min. [M+H] + 413.0 m / z.

[0343] Synthesis of compound 134 [ka]

[0344] 2-(3-fluoro-4-methylphenyl)-2-oxoacetaldehyde 2-(3-Fluoro-4-methylphenyl)-2-oxoacetaldehyde was prepared similarly to the preparation of 1-(4-chloro-3-fluorophenyl)-2,2-dihydroxyethan-1-one.

[0345] LCMS: rt 1.17 min. [M+H] + 167.0 m / z.

[0346] N-(2-(3-fluoro-4-methylphenyl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide N-(2-(3-fluoro-4-methylphenyl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide.

[0347] LCMS: rt 2.27 min. [M+Na] + 324.0 m / z.

[0348] N-(1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-(3-fluoro-4-methylphenyl)-2-oxoethyl)-3-methylbenzamide N-(1-(5-Fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-(3-fluoro-4-methylphenyl)-2-oxoethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide.

[0349] LCMS: rt 2.23 min. [M+Na] + 449.1 m / z.

[0350] Compound 134. 5-Fluoro-1-(5-(3-fluoro-4-methylphenyl)-2-(m-tolyl)oxazol-4-yl)-4-(methylamino)pyrimidin-2(1H)-one. 5-Fluoro-1-(5-(4-fluoro-3-methylphenyl)-2-(m-tolyl)oxazol-4-yl)-4-(methylamino)pyrimidin-2(1H)-one was prepared similarly to the preparation of (5-(4-chloro-3-fluorophenyl)-2-(3-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one.

[0351] 1 H NMR:(400 MHz,DMSO-d6)d ppm 8.48(d,J=4.4 Hz,1H),8.15(d,J=6.4 Hz.1H),7.98(s,1H),7.93(d,J=7.6 Hz,1H),7.50-7.41(m,4H),7.31(d,J=8.0 Hz,1H),2.90(d,J=4.8 Hz,3H),2.43(s,3H),2.28(s,3H).

[0352] LCMS: rt 2.56 min. [M+Na] + 409.3 m / z.

[0353] Synthesis of compound 135. [ka]

[0354] 2-(4-chlorothiophen-2-yl)-2-oxoacetaldehyde hydrate 2-(4-Chlorothiophen-2-yl)-2-oxoacetaldehyde hydrate was prepared similarly to the preparation of 1-(4-chloro-3-fluorophenyl)-2,2-dihydroxyethan-1-one.

[0355] LCMS: rt 0.58 min. [M+Na] + 214.1 m / z.

[0356] N-(2-(4-chlorothiophen-2-yl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide N-(2-(4-chlorothiophen-2-yl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide.

[0357] LCMS: rt 2.18 min. [M+Na] + 232.0 m / z.

[0358] N-(2-(4-chlorothiophen-2-yl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide N-(2-(4-chlorothiophen-2-yl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide.

[0359] LCMS: rt 2.18 min. [M+Na] + 457 m / z.

[0360] 1-(5-(4-chlorothiophen-2-yl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one. Compound 135 1-(5-(4-Chlorothiophen-2-yl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one was prepared similarly to the preparation of (5-(4-chloro-3-fluorophenyl)-2-(3-chlorophenyl)oxazol-4-yl)-5-fluoro-4-(methylamino)pyrimidin-2(1H)-one.

[0361] 1 H NMR(400 MHz,DMSO-d6)d ppm 8.54(d,J=4.40 Hz,1H),8.14(d,J=6.60 Hz,1H),7.93(s,1H),7.88(d,J=7.58 Hz,1H),7.76(d,J=1.47 Hz,1H),7.50(d,J=1.47 Hz,1H),7.47(d,J=7.58 Hz,1H),7.38-7.44(m,1H),2.90(d,J=4.65 Hz,3H),2.42(s,3H).

[0362] LCMS: rt 2.53 min. [M+H] + 417.0 m / z.

[0363] Additional compounds prepared in a similar manner to Compound 135:

[0364] [Table 1]

[0365] Synthesis of 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-((methyl-d3)amino)pyrimidin-2(1H)-one. [ka]

[0366] N-(1-(3-benzoyl-5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-(4-chloro-3-fluorophenyl)-2-oxoethyl)-3-methylbenzamide To a slurry of N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)-3-methylbenzamide (4.0 g, 12 mmol) in DCM (50 mL) was added PCl5 (2.9 g, 95 wt%, 1.05 equiv, 13 mmol). The resulting cloudy reaction mixture was stirred at 50 °C for 2 h. The reaction was concentrated to a yellow solid and dried under vacuum to give the chloro adduct as a pale yellow solid, which was used directly. To a solution of 3-benzoyl-5-fluoropyrimidine-2,4(1H,3H)-dione (3.5 g, 1.2 equiv, 15 mmol) and triethylamine (3.8 g, 5.2 mL, 3 equiv, 37 mmol) in DMF (50 mL) at 0 °C, the chloro adduct was added, and the reaction mixture was stirred at room temperature for 16 h. The reaction was concentrated to a viscous oil and mixed with water. The precipitate that formed was collected by vacuum filtration and dried under vacuum to give N-(1-(3-benzoyl-5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-(4-chloro-3-fluorophenyl)-2-oxoethyl)-3-methylbenzamide (3.7 g, 6.9 mmol, 55%) as a brown solid. This material was used crude in the next reaction.

[0367] LCMS: rt 3.61 min. [M+Na] + 559.9 m / z.

[0368] 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoropyrimidine-2,4(1H,3H)-dione N-(1-(3-Benzoyl-5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-(4-chloro-3-fluorophenyl)-2-oxoethyl)-3-methylbenzamide (0.305 g, 1 equivalent, 567 μmol) was treated with thionyl chloride (4.89 g, 3.00 mL, 72.5 equivalents, 41.1 mmol) and stirred at 80° C. for 2.5 hours. The reaction was cooled to room temperature and concentrated to give a viscous brown oil. This material was further dried under vacuum to give crude 3-benzoyl-1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoropyrimidine-2,4(1H,3H)-dione as a brown foam.

[0369] LCMS: rt 2.88 min. [M+H] + 519.7 m / z.

[0370] The crude 3-benzoyl-1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoropyrimidine-2,4(1H,3H)-dione was dissolved in DCM (2 mL) and treated with TFA (1.48 g, 1.00 mL, 22.9 equiv., 13.0 mmol). The mixture was stirred at room temperature for 2 hours. The reaction was concentrated and dried under vacuum to give 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoropyrimidine-2,4(1H,3H)-dione (0.161 g, 387 μmol, 68.3%) as a brown foam.

[0371] LCMS: rt 2.56 min. [M+H] + 416.0 m / z.

[0372] 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one To a slurry of 1H-1,2,4-triazole (86 mg, 7 equiv., 1.2 mmol) in MeCN (2 mL) at 0° C. was added phosphoryl trichloride (68 mg, 41 μL, 2.5 equiv., 0.44 mmol) and triethylamine (0.13 g, 0.17 mL, 7 equiv., 1.2 mmol). The resulting yellow slurry was stirred at 0° C. for 30 minutes and then at 25° C. for 30 minutes. To the reaction was added a solution of 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoropyrimidine-2,4(1H,3H)-dione (0.074 g, 1 equiv., 0.18 mmol) in MeCN (2 mL), and the reaction was stirred at 25° C. for 16 hours and then at 80° C. for an additional hour. The mixture was cooled to room temperature, HO was added, and stirring was continued for 10 min. The mixture was then filtered, and the yellow residue was washed with water (3 × 3 mL) and dried under vacuum to give 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one (0.073 g, 0.16 mmol, 88%) as a yellow solid.

[0373] LCMS: rt 2.50 min. [M+H] + 467.0 m / z.

[0374] 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-((methyl-d3)amino)pyrimidin-2(1H)-one To a slurry of 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one (73.0 mg, 1 equiv., 156 μmol) in DMF (3 mL) was added triethylamine (111 mg, 153 μL, 7 equiv., 1.09 mmol), followed by d3-methylamine hydrochloride (33 mg, 3 equiv., 0.468 mmol). The resulting yellow slurry was stirred at 80° C. for 1 h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was treated with HO and stirred for 20 min. The resulting precipitate was collected by vacuum filtration, washed with water, and dried under vacuum at 50°C to give 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-((methyl-d3)amino)pyrimidin-2(1H)-one (65.7 mg, 152 µmol, 97.3%) as a yellow solid.

[0375] 1 H NMR(400 MHz,DMSO-d6)d ppm 8.43(s,1H),8.13(d,J=6.85 Hz,1H),8.00(s,1H),7.95(d,J=7.34 Hz,1H),7.68-7.80(m,2H),7.45-7.53(m,1H),7.35-7.44(m,2H),3.33(s,6H),2.43(s,3H).

[0376] 19 F NMR(376 MHz,DMSO-d6)d ppm-116.44--113.90(m,1F),-170.28--167.34(m,1F).

[0377] LCMS: rt 2.41 min. [M+H] + 432.0 m / z.

[0378] Synthesis of compound 149. [ka]

[0379] 7-Chloro-3-iodoquinolin-4-ol 7-Chloroquinolin-4(1H)-one (3 g, 16.7 mmol) was dissolved in DMF (30 ml) and treated with 1-iodopyrrolidine-2,5-dione (NIS, 1.5 equivalents, 5.62 g, 25 mmol) and stirred at room temperature overnight. The reaction was then diluted with water (30 ml) and the resulting precipitate was filtered off, washed with ethyl acetate and water, and then dried under vacuum to give 7-chloro-3-iodoquinolin-4-ol (5.00 g, 98.0%) as a white solid. tert-Butyl 2-(7-chloro-3-iodo-4-oxo-1,4-dihydroquinolin-1-yl)acetate

[0380] 7-Chloro-3-iodoquinolin-4-ol (2.3 g, 7.52 mmol) was dissolved in DMF (10 mL), followed by the addition of tert-butyl 2-bromoacetate (2.18 g, 11.2 mmol) and dipotassium carbonate (3.10 g, 22.5 mmol) and stirring for 16 hours at 50° C. The reaction was diluted with 10 ml of water, and the resulting precipitate was filtered off, washed with water and a mixture of ethyl acetate and heptane (1:3), and then dried to give tert-butyl 2-(7-chloro-3-iodo-4-oxo-1,4-dihydroquinolin-1-yl)acetate (3.10 g, 98.4%) as a pale yellow solid.

[0381] LCMS(ESI+):m / z 420.0[M+H]+

[0382] tert-Butyl 2-[7-chloro-3-(furan-3-yl)-4-oxo-1,4-dihydroquinolin-1-yl]acetate tert-Butyl 2-(7-chloro-3-iodo-4-oxo-1,4-dihydroquinolin-1-yl)acetate (3.02 g, 7.21 mmol) was dissolved in toluene (25 mL), followed by the addition of (furan-3-yl)boronic acid (1 g, 8.93 mmol), bis(cyclopentyldiphenylphosphane)dichloromethanedichloropalladium (567 mg, 686 μmol), dipotassium carbonate (2.84 g, 20.6 mmol), and water (2.5 mL). The reaction was stirred at 110° C. for 2 hours. The reaction was then diluted with ethyl acetate and brine, and the organic fraction was separated and dried over sodium sulfate. The solvent was removed under vacuum and the residue was purified by silica gel column chromatography eluting with (1:1) ethyl acetate and heptane to give tert-butyl 2-[7-chloro-3-(furan-3-yl)-4-oxo-1,4-dihydroquinolin-1-yl]acetate (1.65 g, 67.0%) as a brown solid.

[0383] LCMS(ESI+):m / z 360.0[M+H]+

[0384] 2-[3-(furan-3-yl)-4-oxo-7-(pyrrolidin-1-yl)-1,4-dihydroquinolin-1-yl]acetic acid tert-Butyl 2-[7-chloro-3-(furan-3-yl)-4-oxo-1,4-dihydroquinolin-1-yl]acetate (1.35 g, 3.75 mmol) was dissolved in DMF (20 mL). Pyrrolidine (1.33 g, 18.7 mmol), chloro(2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (292 mg, 375 μmol), and sodium (tert-butoxy) (1.10 g, 11.2 mmol) were then added, and the reaction was heated to 110°C for 4 hours. The resulting solid was filtered off and washed with water. The solid was then dissolved in 1N NaOH and filtered. The filtrate was then acidified with 1N HCl, and the solid was filtered. The solid was then washed with water, ethyl acetate, and then dried under vacuum to give 2-[3-(furan-3-yl)-4-oxo-7-(pyrrolidin-1-yl)-1,4-dihydroquinolin-1-yl]acetic acid (890 mg, 70.6%) as a yellow solid. 1 H NMR(400MHz,DMSO-d6,ppm)d 8.51(s,1H),8.33(s,1H),8.09-8.06(m,1H),7.68(s,1H),6.90(s,1H),6.76- 6.74(m,1H),6.13(s,1H),5.08(s,2H),3.40-3.30(m,4H),4.60-4.40(m,4H).

[0385] LCMS(ESI+):m / z 339.0[M+H]+

[0386] 2-[3-(furan-3-yl)-4-oxo-7-(pyrrolidin-1-yl)-1,4-dihydroquinolin-1-yl]-N-[2-(trifluoromethyl)phenyl]acetamide. Compound 149 2-[3-(furan-3-yl)-4-oxo-7-(pyrrolidin-1-yl)-1,4-dihydroquinolin-1-yl]acetic acid (890 mg, 2.63 mmol) was dissolved in toluene (10 mL), followed by triethylamine (1.32 g, 13.1 mmol), tripropyl-1,3,5,2λ 5 ,4λ 5 ,6λ5 -trioxatriphosphinane-2,4,6-trione (T3P, 5.02 g, 7.89 mmol) and 2-(trifluoromethyl)aniline (634 mg, 3.94 mmol) were added and stirred at 40° C. for 2 hours. The reaction was quenched with 1N HCl. The resulting solid was filtered, then washed with water, ethyl acetate, collected by filtration, and dried under vacuum to give 2-[3-(furan-3-yl)-4-oxo-7-(pyrrolidin-1-yl)-1,4-dihydroquinolin-1-yl]-N-[2-(trifluoromethyl)phenyl]acetamide (1 g, 79%) as a yellow solid. LCMS (ESI+): m / z 482.0 [M+H]+

[0387] Synthesis of compound 85 [ka]

[0388] 3-Benzoyl-5-bromopyrimidine-2,4(1H,3H)-dione A mixture of 5-bromo-1,2,3,4-tetrahydropyrimidine-2,4-dione (10 g, 52.3 mmol) and benzoyl chloride (18.2 g, 130 mmol) in a mixture of acetonitrile (100 mL) and pyridine (50 mL) was stirred at room temperature for 4 days. The reaction mixture was evaporated in vacuo. The residue was suspended in 1,4-dioxane (200 mL), treated with 0.25 M K2CO3 solution (100 mL), and stirred for 16 hours. To remove 1,4-dioxane, the mixture was evaporated, and then water (100 mL) was added to the mixture. The precipitate was collected on a filter paper. The resulting crude solid was suspended in ethanol and stirred at 50 °C for 3 hours. The precipitate was collected on a paper filter to give 3-benzoyl-5-bromopyrimidine-2,4(1H,3H)-dione (12.0 g, 78%) as a white solid.

[0389] 1H NMR(300 MHz,DMSO-d6)d 12.04(s,1H),8.19(s,1H),8.09-7.97(m,2H),7.86-7.70(m,1H),7.70-7.54(m,2H). N-(1-(3-benzoyl-5-bromo-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-(4-chloro-3-fluorophenyl)-2-oxoethyl)-3-methylbenzamide

[0390] 3-Benzoyl-5-bromopyrimidine-2,4(1H,3H)-dione (1.0 equiv.), (N-(1-chloro-2-(4-chloro-3-fluorophenyl)-2-oxoethyl)-3-methylbenzamide (1.0 equiv.), and triphenylphosphine (1.1 equiv.) were placed in a flask and suspended in dry toluene. The toluene was evaporated under high vacuum. This process was repeated three times. This mixture was then dissolved in anhydrous THF (10.0 vol.) and diluted with di-tert-butylazodicarboxylate. To the resulting mixture was added 1.3 equiv. of N-benzoyl-5-bromo-2,4-dioxo-1,2,3,4-tetrahydropyrimidin-1-yl)-2-(4-chloro-3-fluorophenyl)-2-oxoethyl]-3-methylbenzamide. The resulting mixture was stirred at room temperature for 16 hours. The solvent was removed under reduced pressure. The crude material was dissolved in methanol and placed in an ice bath. The resulting precipitate was collected by vacuum filtration, washed with methanol and hexane, and dried under vacuum to give N-[1-(3-benzoyl-5-bromo-2,4-dioxo-1,2,3,4-tetrahydropyrimidin-1-yl)-2-(4-chloro-3-fluorophenyl)-2-oxoethyl]-3-methylbenzamide.

[0391] 1 H NMR(300 MHz,DMSO-d6)d 9.98(d,J=8.4 Hz,1H),8.44(s,1H),8.02-7.93(m,2H),7.93-7.76(m,4H),7.76-7.53(m,5H),7.50-7.34(m,2H),2.36(s,3H).

[0392] 3-Benzoyl-5-bromo-1-[5-(4-chloro-3-fluorophenyl)-2-(3-methylphenyl)-1,3-oxazol-4-yl]-1,2,3,4-tetrahydropyrimidine-2,4-dione N-[1-(3-benzoyl-5-bromo-2,4-dioxo-1,2,3,4-tetrahydropyrimidin-1-yl)-2-(4-chloro-3-fluorophenyl)-2-oxoethyl]-3-methylbenzamide (1.0 equivalent), hexachloroethane (2.0 equivalents), and triphenylphosphine (2.0 equivalents) were dissolved in anhydrous MeCN (20.0 volumes). After stirring at room temperature for 10 minutes, pyridine (4.0 equivalents) was added, and the reaction mixture was stirred at 60° C. for 90 minutes. Upon cooling to room temperature, the formed precipitate was collected by vacuum filtration, washed with methanol, and dried under vacuum to give 3-benzoyl-5-bromo-1-[5-(4-chloro-3-fluorophenyl)-2-(3-methylphenyl)-1,3-oxazol-4-yl]-1,2,3,4-tetrahydropyrimidine-2,4-dione.

[0393] 1 H NMR(300MHz,DMSO-d6)d 8.71(s,1H),8.31-8.13(m,2H),8.06-7.91(m,3H),7.90-7.79(m,1H),7.79-7.58(m,4H),7.57-7.40(m,2H),2.43(s,3H).

[0394] 5-Bromo-1-[5-(4-chloro-3-fluorophenyl)-2-(3-methylphenyl)-1,3-oxazol-4-yl]-4-(1H-1,2,4-triazol-1-yl)-1,2-dihydropyrimidin-2-one 3-Benzoyl-5-bromo-1-[5-(4-chloro-3-fluorophenyl)-2-(3-methylphenyl)-1,3-oxazol-4-yl]-1,2,3,4-tetrahydropyrimidine-2,4-dione (1.0 equiv.) was suspended in a mixture of anhydrous MeCN (20.0 vol.) and anhydrous DCM (5.0 vol.). Triethylamine (12.0 equiv.) and 1,2,4-triazole (8.0 equiv.) were then added, followed by the dropwise addition of phosphorus(V) oxychloride (2.0 equiv.). The reaction mixture was stirred at room temperature for 16 hours. The precipitate that formed was collected by vacuum filtration, washed with MeCN, and dried under vacuum to give 5-bromo-1-[5-(4-chloro-3-fluorophenyl)-2-(3-methylphenyl)-1,3-oxazol-4-yl]-4-(1H-1,2,4-triazol-1-yl)-1,2-dihydropyrimidin-2-one.

[0395] 1 H NMR(300 MHz,DMSO-d6)d 9.42(s,1H),9.10(s,1H),8.49(s,1H),8.05(s,1H),8.00(d,J=7.5 Hz,1H),7.91(dd,J=10.4,2.1 Hz,1H),7.72(t,J=8.1 Hz,1H),7.59-7.43(m,3H),2.45(s,3H).

[0396] 5-Bromo-1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-4-(methylamino)pyrimidin-2(1H)-one. Compound 85 To a suspension of 5-bromo-1-[5-(4-chloro-3-fluorophenyl)-2-(3-methylphenyl)-1,3-oxazol-4-yl]-4-(1H-1,2,4-triazol-1-yl)-1,2-dihydropyrimidin-2-one (1.0 equiv.) in anhydrous MeOH (20.0 vol.) was added methylamine solution (2 M in THF, 1.5 equiv.). The reaction mixture was stirred at room temperature for 16 hours. The precipitate that formed was collected by vacuum filtration and washed with methanol and acetonitrile. The solid was dried under vacuum to give 5-bromo-1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-4-(methylamino)pyrimidin-2(1H)-one. [M+H] + 451.1 m / z.

[0397] Synthesis of Compound 150 [ka]

[0398] N-(1-hydroxy-2-oxo-2-(4-(trifluoromethyl)phenyl)ethyl)-3-methylbenzamide N-(1-hydroxy-2-oxo-2-(4-(trifluoromethyl)phenyl)ethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)benzamide.

[0399] LCMS: rt 2.28 min. [M+H] + 360.0 m / z.

[0400] N-(1-(3-benzoyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-oxo-2-(4-(trifluoromethyl)phenyl)ethyl)-3-methylbenzamide N-(1-(3-Benzoyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-oxo-2-(4-(trifluoromethyl)phenyl)ethyl)-3-methylbenzamide was prepared similarly to the preparation of 3-chloro-N-(2-(4-chloro-3-fluorophenyl)-1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-oxoethyl)benzamide.

[0401] LCMS: rt 2.26 min. [M+Na] + 558.0 m / z.

[0402] 1-(2-(m-tolyl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)pyrimidine-2,4(1H,3H)-dione To a solution of N-(1-(3-benzoyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-oxo-2-(4-(trifluoromethyl)phenyl)ethyl)-3-methylbenzamide (0.500 g, 1 eq, 934 μmol) in DCE (5.0 mL) was added 2,2,2-trifluoroacetic acid (2.66 g, 25 eq, 23.3 mmol) and the reaction mixture was heated to 60° C. for 1 h. The reaction was concentrated and dried under vacuum at 50° C. to give crude N-(1-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-oxo-2-(4-(trifluoromethyl)phenyl)ethyl)-3-methylbenzamide as a brown solid. LCMS: rt 2.31 min. [M+H] + 432 m / z. The crude residue was treated with thionyl chloride (4.9 g, 3.0 mL, 44 equiv, 41 mmol) and stirred at 60° C. for 1 hour. The reaction was then heated to 80° C. for 4 hours. The reaction was cooled to room temperature and concentrated. Ice water was added and a precipitate formed. The precipitate was collected by vacuum filtration and triturated with ether. The solid was collected by vacuum filtration and dried under vacuum to give 1-(2-(m-tolyl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)pyrimidine-2,4(1H,3H)-dione (95.4 mg, 231 μmol, 24.7%) as a white solid.

[0403] LCMS: rt 2.63 min. [M+H] + 414.0 m / z.

[0404] 1-(2-(m-tolyl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one 1-(2-(m-Tolyl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one was prepared similarly to the preparation of 1-(5-(4-chloro-3-fluorophenyl)-2-(m-tolyl)oxazol-4-yl)-5-fluoro-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one.

[0405] LCMS: rt 2.74 min. [M+H] + 465.0 m / z.

[0406] 4-(Azetidin-1-yl)-1-(2-(m-tolyl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)pyrimidin-2(1H)-one. Compound 150 To a slurry of 1-(2-(m-tolyl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one (30.0 mg, 1 equiv., 64.6 μmol) in DMF (1 mL) was added triethylamine (32.7 mg, 45.0 μL, 5 equiv., 323 μmol), followed by azetidine hydrochloride (21.8 mg, 97 wt%, 3.5 equiv., 226 μmol). The resulting brown slurry was stirred at 50° C. for 1.5 h. The reaction was cooled to room temperature and treated with HO. The resulting precipitate was collected by vacuum filtration, washed with water, and dried under vacuum to give 4-(azetidin-1-yl)-1-(2-(m-tolyl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)pyrimidin-2(1H)-one (23.4 mg, 51.7 μmol, 80.1%) as a light brown solid. 1 H NMR(400 MHz,DMSO-d6)d ppm 7.98(s,1H),7.93(d,J=7.82 Hz,1H),7.83-7.91(m,3H),7.79(d,J=8.07 Hz,2H),7.47-7.53(m,1H),7.41-7.46(m,1H),5.96(d,J=7.34 Hz,1H),4.23(t,J=7.58 Hz,2H),4.14(t,J=7.58 Hz,2H),2.43(s,3H),2.35-2.40(m,2H). 19 F NMR(376 MHz,DMSO-d6)d ppm-61.18(s,3F).LCMS:rt 2.56 min.[M+H] + 453.0 m / z.

[0407] Additional compounds prepared in a similar manner to Compound 150:

[0408] [Table 2]

[0409] Synthesis of 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one [ka]

[0410] 1-(4-chloro-3-fluorophenyl)-2,2-dihydroxyethan-1-one To a stirred solution of 1-(4-chloro-3-fluorophenyl)ethan-1-one (20.0 g, 1.0 equiv., 116 mmol) in DMSO (100 mL) was added HBr (58.6 g, 39.3 mL, 48 wt % in water, 3.0 equiv., 348 mmol), and the mixture was stirred at room temperature for 1 hour. The mixture was then stirred at 50° C. for 16 hours. The reaction mixture was poured into ice water, and a precipitate formed. The solid was collected by vacuum filtration and washed with hexane to give 1-(4-chloro-3-fluorophenyl)-2,2-dihydroxyethan-1-one (14.0 g, 68.4 mmol, 59.1%) as an off-white solid.

[0411] 1 H NMR(400 MHz,DMSO-d6)δ ppm 8.00(dd,1H,J=1.8,10.1 Hz),7.93(td,1H,J=1.0,8.4 Hz),7.78(dd,1H,J=7.6,8.2 Hz),6.98(d,2H,J=5.5 Hz),5.61(bs,1H).

[0412] N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)formamide To a stirred solution of 1-(4-chloro-3-fluorophenyl)-2,2-dihydroxyethan-1-one (5.0 g, 1 equivalent, 24 mmol) in dioxane (50.0 mL) was added formamide (5.5 g, 4.9 mL, 5.0 equivalents, 0.12 mol) at room temperature under an argon atmosphere. The resulting reaction mixture was stirred at 90 °C for 3 hours. After cooling to room temperature, the solvent was evaporated under reduced pressure to give the crude compound as a pale yellow liquid. The crude compound was triturated with MTBE and hexane. The precipitated solid was collected by vacuum filtration and dried under vacuum to give N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)formamide (3.0 g, 13 mmol, 53%) as an off-white solid.

[0413] 1 H NMR(400 MHz,DMSO-d6)δ ppm 9.00(d,J=8.4 Hz,1H),8.09(dd,J=0.7,1.6 Hz,1H),7.92(dd,J=1.4,10.1 Hz,1H),7.83-7.77(m,2H),6.86(d,J=7.2 Hz,1H),6.31(t,J=8.0 Hz,1H).

[0414] N-(1-(3-benzoyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-(4-chloro-3-fluorophenyl)-2-oxoethyl)formamide To a stirred solution of N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)formamide (12.0 g, 1 equiv., 51.8 mmol) in DCM (120 mL) was added PCl5 (12.9 g, 1.2 equiv., 62.2 mmol) in portions at room temperature. The resulting reaction mixture was stirred for 2 h. The reaction mixture was concentrated under reduced pressure to give the crude chloro compound. This material was then triturated with hexanes (100 mL), collected by vacuum filtration, and dried under vacuum to give the chloro intermediate, which was used directly. In a separate three-neck round-bottom flask under a nitrogen atmosphere, 3-benzoylpyrimidine-2,4(1H,3i)-dione (11.2 g, 1.0 equiv., 51.8 mmol), DMF (100 mL), and TEA (15.7 g, 21.7 mL, 3.0 equiv., 155 mmol) were combined. The above chloro intermediate was then added dropwise as a solution in DMF (100 mL). The resulting reaction mixture was stirred at room temperature for 4 hours. The reaction was concentrated in vacuo and diluted with ice water. The resulting precipitate was collected by vacuum filtration, washed with saturated sodium bicarbonate, and dried in vacuo to give the crude compound. The crude material was adsorbed onto silica gel and purified by silica gel column chromatography (40-65% EtOAc in petroleum ether) to give N-(1-(3-benzoyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-(4-chloro-3-fluorophenyl)-2-oxoethyl)formamide (14.0 g, 32.4 mmol, 62.5%) as a yellow sticky solid.

[0415] 1 H NMR(400 MHz,DMSO-d6)δ ppm 9.64(d,1H,J=7.6 Hz),8.26(s,1H),8.06(d,1H,J=8.2 Hz),7.8-7.9(m,3H),7.69(dd,1H,J=1.6,8.4 Hz),7.61(d,2H,J=2.1 Hz),7.5-7.5(m,2H),7.29(d,1H,J=8.0 Hz),6.05(d,1H,J=8.2 Hz).

[0416] 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)pyrimidine-2,4(1H,3H)-dione A stirred solution of N-(1-(3-benzoyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-(4-chloro-3-fluorophenyl)-2-oxoethyl)formamide (14 g, 1 equiv., 33 mmol) in thionyl chloride (22.8 g, 14.0 mL, 5.9 equiv., 192 mmol) was heated in a sealed tube at 65° C. for 32 h. Upon cooling to room temperature, the reaction mixture was concentrated in vacuo, diluted with saturated aqueous sodium bicarbonate, and extracted with 30% IPA in chloroform (3×15 ml). The combined organic layers were dried over sodium sulfate and concentrated in vacuo to give the crude compound. The crude compound was adsorbed onto silica gel and purified by silica gel column chromatography (4–10% methanol in EtOAc) to give still impure material. This material was then triturated with MTBE to give 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)pyrimidine-2,4(1H,3H)-dione (5.0 g, 16 mmol, 50%) as an off-white solid.

[0417] 1 H NMR(400 MHz,DMSO-d6)δ ppm 11.70(s,1H),8.69(s,1H),7.7-7.8(m,2H),7.58(dd,1H,J=2.0,10.2 Hz),7.36(ddd,1H,J=0.8,2.0,8.5 Hz),5.84(dd,1H,J=2.2,8.0 Hz)

[0418] 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one To a stirred solution of 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)pyrimidine-2,4(1H,3H)-dione (5.0 g, 1 eq, 16 mmol) in MeCN (100 mL) was added 1H-1,2,4-triazole (9.0 g, 8 eq, 0.13 mol) and triethylamine (20 g, 27 mL, 12 eq, 0.20 mol). Phosphoryl trichloride (5.0 g, 3.0 mL, 2.0 eq, 33 mmol) was then added dropwise under nitrogen at 0°C, and the reaction mixture was stirred at 0°C for 1 hour. A solid precipitate formed during the reaction, which was collected by vacuum filtration. This material was then washed with 50% aqueous MeCN (100 mL), water (100 mL), and MeCN (100 mL). The solid was then dried under vacuum. The material was then triturated with EtOAc and the solid was collected by vacuum filtration to give 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one (3.6 g, 9.2 mmol, 57%) as an off-white solid after drying under vacuum.

[0419] 1 H NMR(400 MHz,DMSO-d6)δ ppm 9.55(1H,s),8.77(1H,s),8.56(1H,d,J=7.2 Hz),8.49(1H,s),7.71(1H,t,J=8.1 Hz),7.60(1H,dd,J=10.2 Hz,2.0 Hz),7.36(1H,ddd,J=8.4 Hz,2.0 Hz,0.7 Hz),7.20(1H,d,J=7.2 Hz).

[0420] Synthesis of compound 155. [ka]

[0421] 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(5-azaspiro[2.3]hexan-5-yl)pyrimidin-2(1H)-one 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(5-azaspiro[2.3]hexan-5-yl)pyrimidin-2(1H)-one was prepared from 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one in a manner analogous to the preparation of 4-(azetidin-1-yl)-1-(2-(m-tolyl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)pyrimidin-2(1H)-one.

[0422] LCMS: rt 2.00 min. [M+H] + 373.1 m / z.

[0423] 1-(5-(4-chloro-3-fluorophenyl)-2-(4,6-dimethylpyridin-2-yl)oxazol-4-yl)-4-(5-azaspiro[2.3]hexan-5-yl)pyrimidin-2(1H)-one. Compound 155 1-(5-(4-Chloro-3-fluorophenyl)-2-(4,6-dimethylpyridin-2-yl)oxazol-4-yl)-4-(5-azaspiro[2.3]hexan-5-yl)pyrimidin-2(1H)-one was prepared similarly to the preparation of 1-(5-(4-chloro-3-fluorophenyl)-2-(4,6-dimethylpyridin-2-yl)oxazol-4-yl)-4-(3-fluoro-3-methylazetidin-1-yl)pyrimidin-2(1H)-one.

[0424] 1 H NMR(400 MHz,DMSO-d6)δ 7.94(s,1H),7.86(d,J=7.4 Hz,1H),7.79(t,J=8.1 Hz,1H),7.62-7.54(m,1H),7.40-7.35(m,1H),7.31(s,1H),5.96(d,J=7.4 Hz,1H),4.28(s,2H),4.18(s,2H),2.55(s,3H),2.41(s,3H),0.74(s,4H).

[0425] LCMS: rt 2.29 min. [M+H]+ 478.0 m / z.

[0426] Synthesis of compound 156. [ka]

[0427] 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(3-ethyl-3-fluoroazetidin-1-yl)pyrimidin-2(1H)-one 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(3-ethyl-3-fluoroazetidin-1-yl)pyrimidin-2(1H)-one was prepared from 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one in a manner analogous to the preparation of 4-(azetidin-1-yl)-1-(2-(m-tolyl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)pyrimidin-2(1H)-one.

[0428] LCMS: rt 2.61 min. [M+H] + 393.0 m / z.

[0429] 1-(5-(4-chloro-3-fluorophenyl)-2-(6-methylpyridin-2-yl)oxazol-4-yl)-4-(3-ethyl-3-fluoroazetidin-1-yl)pyrimidin-2(1H)-one. Compound 156 1-(5-(4-chloro-3-fluorophenyl)-2-(4,6-dimethylpyridin-2-yl)oxazol-4-yl)-4-(3-ethyl-3-fluoroazetidin-1-yl)pyrimidin-2(1H)-one was prepared similarly to the preparation of 1-(5-(4-chloro-3-fluorophenyl)-2-(4,6-dimethylpyridin-2-yl)oxazol-4-yl)-4-(3-fluoro-3-methylazetidin-1-yl)pyrimidin-2(1H)-one.

[0430] 1 H NMR(400 MHz,DMSO-d6)d ppm 7.88-7.97(m,2H),7.77(t,J=8.31 ​​Hz,1H),7.57(d,J=10.27 Hz,1H),7.38(d,J=8.31 Hz,1H),7.31(s,1H),6.03(d,J=7.34 Hz,1H),4.15-4.40(m,4H),2.54(s,3H),2.40(s,3H),1.89-2.04(m,2H),0.96(t,J=7.21 Hz,3H).

[0431] LCMS: rt 2.21 min. [M+H] + 498.2 m / z.

[0432] Synthesis of compound 157. [ka]

[0433] 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(3-fluoro-3-methylazetidin-1-yl)pyrimidin-2(1H)-one 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(3-fluoro-3-methylazetidin-1-yl)pyrimidin-2(1H)-one was prepared from 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one in a manner analogous to the preparation of 4-(azetidin-1-yl)-1-(2-(m-tolyl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)pyrimidin-2(1H)-one.

[0434] LCMS: rt 1.81 min. [M+H] + 379.0 m / z.

[0435] 1-(5-(4-chloro-3-fluorophenyl)-2-(4,6-dimethylpyridin-2-yl)oxazol-4-yl)-4-(3-fluoro-3-methylazetidin-1-yl)pyrimidin-2(1H)-one. Compound 157 1-(5-(4-chloro-3-fluorophenyl)oxazol-4-yl)-4-(3-fluoro-3-methylazetidin-1-yl)pyrimidin-2(1H)-one (75.0 mg, 1 equiv., 198 μmol), 2-bromo-4,6-dimethyl-pyridine (55.3 mg, 1.5 equiv., 297 μmol), cesium carbonate (194 mg, 3 equiv., 594 μmol), 1,1′-bis(diphenylphosphino)ferrocene-palladium(II) dichloride (21.7 mg, 0.15 equiv., 29.7 μmol), and copper(I) iodide (2.83 mg, 0.075 equiv., 14.9 μmol) were combined in DMF (3.0 mL). The reaction was then purged and refilled with argon three times and stirred at 90° C. for 16 h. The reaction was cooled to room temperature and filtered through a pad of Celite. The Celite was further eluted with DCM, and the combined organic solution was concentrated to dryness. The residue was purified by flash column chromatography (SiO, 40 g, 0–10% MeOH in DCM) to give 1-(5-(4-chloro-3-fluorophenyl)-2-(4,6-dimethylpyridin-2-yl)oxazol-4-yl)-4-(3-fluoro-3-methylazetidin-1-yl)pyrimidin-2(1H)-one (23 mg, 47.5 μmol, 24% yield).

[0436] 1 H NMR(400 MHz,DMSO-d6)d ppm 7.93(s,1H),7.89(d,J=7.34 Hz,1H),7.77(t,J=8.30 Hz,1H),7.55(d,J=10.50 Hz,1H),7.37(d,J=8.60 Hz,1H),7.31(s,1H),5.99(d,J=7.34 Hz,1H),4.28-4.41(m,2H),4.21(d,J=19.60 Hz,2H),2.54(s,3H),2.40(s,3H),1.67(d,J=22.30 Hz,3H).

[0437] 19 F NMR(376 MHz,DMSO-d6)d ppm-114.69(s,1F),-139.40--139.05(m,1F).

[0438] LCMS: rt 2.11 min. [M+H] + 484.1 m / z.

[0439] Synthesis of compound 175. [ka]

[0440] 2-(3-fluoro-4-methylphenyl)-2-oxoacetaldehyde Selenium dioxide (13.1 g, 1.8 equiv., 118 mmol) was combined with water (10.0 mL) and dioxane (100 mL) in a 200 mL medium pressure flask. The flask was heated to 70 °C until all materials were completely dissolved. To the reaction mixture at 70 °C, 4-acetyl-2-fluorotoluene (10.0 g, 1 equiv., 65.7 mmol) was added, and the reaction flask was sealed and stirred at 100 °C for 16 h. The reaction was cooled to room temperature and filtered through a pad of Celite, further eluting with EtOAc. The filtrate was concentrated to a viscous brown oil, which was treated with HO (50 mL) and heated to 100 °C under a reflux condenser for 6 h. Upon cooling to room temperature, the formed precipitate was collected by vacuum filtration, washed with cold water, and dried under vacuum to give 2-(3-fluoro-4-methylphenyl)-2-oxoacetaldehyde (10.5 g, 63.2 mmol, 96.2%) as a white solid, which was used directly in the next step without characterization.

[0441] N-(2-(4-chloro-3-fluorophenyl)-1-hydroxy-2-oxoethyl)formamide To a slurry of 2-(3-fluoro-4-methylphenyl)-2-oxoacetaldehyde (3.0 g, 1.1 equiv., 16.3 mmol) in dioxane (20 mL) was added formamide (2 g, 1.77 mL, 3.0 equiv., 48.9 mmol). The reaction mixture was heated to 90° C. for 16 hours. Upon cooling to room temperature, the reaction was concentrated and diluted with ethanol. The resulting solid was collected by vacuum filtration and dried in a vacuum oven to give N-(2-(3-fluoro-4-methylphenyl)-1-hydroxy-2-oxoethyl)formamide (1.52 g, 7.20 mmol, 48.6%). This material was used in the next step without purification or characterization.

[0442] N-(1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-(3-fluoro-4-methylphenyl)-2-oxoethyl)formamide To a slurry of N-(2-(3-fluoro-4-methylphenyl)-1-hydroxy-2-oxoethyl)formamide (2.34 g, 1 equiv., 10.4 mmol) in DCM (70 mL) was added phosphorus pentachloride (2.27 g, 1.05 equiv., 10.9 mmol). The resulting homogeneous solution was stirred at 65° C. for 2 h. After cooling to room temperature, the reaction was concentrated and dried under vacuum to provide the intermediate chloro adduct as a yellow solid. To a solution of 4-(methylamino)pyrimidin-2(1H)-one (1.43 g, 1.1 equiv., 11.4 mmol) in DMF (5 mL) was added triethylamine (3.15 g, 4.3 mL, 3 equiv., 31.2 mmol) and stirred at room temperature for 15 min. The reaction mixture was cooled to 0° C., and the intermediate chloro adduct was added portionwise. The cooling bath was removed, and the resulting reaction mixture was stirred at ambient temperature for 30 min. Water (40 mL) was then added and the resulting precipitate was collected by vacuum filtration, washed with cold water, and dried under vacuum at 50° C. to give N-(1-(5-fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-(3-fluoro-4-methylphenyl)-2-oxoethyl)formamide (1.83 g, 5.47 mmol, 52.7%).

[0443] 5-Fluoro-1-(5-(3-fluoro-4-methylphenyl)oxazol-4-yl)-4-(methylamino)pyrimidin-2(1H)-one N-(1-(5-Fluoro-4-(methylamino)-2-oxopyrimidin-1(2H)-yl)-2-(3-fluoro-4-methylphenyl)-2-oxoethyl)formamide (2.82 g, 1 equivalent, 8.00 mmol) was treated with thionyl chloride (30 equivalents, 17.5 mL) and heated to 65° C. for 3 hours. The reaction was cooled to room temperature and concentrated to give a dark brown viscous oil. The oil was then triturated with ether and dried under vacuum to give 5-fluoro-1-(5-(3-fluoro-4-methylphenyl)oxazol-4-yl)-4-(methylamino)pyrimidin-2(1H)-one (1.8 g, 5.7 mmol, 71%) as a beige solid.

[0444] LCMS(ESI+):m / z 319.1,[M+H] + .RT 1.83 min.

[0445] 5-Fluoro-1-(5-(3-fluoro-4-methylphenyl)-2-(3-fluoro-6-methylpyridin-2-yl)oxazol-4-yl)-4-(methylamino)pyrimidin-2(1H)-one. Compound 175 5-Fluoro-1-(5-(3-fluoro-4-methylphenyl)oxazol-4-yl)-4-(methylamino)pyrimidin-2(1H)-one (100 mg, 0.314 mmol, 1 equiv.), 2-bromo-3-fluoro-6-methylpyridine (3 equiv., 179 mg, 0.942 mmol), and cesium carbonate (307 mg, 3 equiv., 0.943 mmol) were combined in DMF (1 mL). Copper(I) iodide (4.49 mg, 0.075 equiv., 23.6 μmol) and Pd(dppf)Cl (34.5 mg, 0.15 equiv., 47.1 μmol) were then added. The reaction vial was evacuated, refilled with argon, and stirred at 90 °C for 24 h. Upon cooling to room temperature, the reaction was diluted with DCM and filtered through Celite. Water was added, and the organic fraction was separated, dried over magnesium sulfate, and concentrated. The residue was purified by silica gel column chromatography (0-20% MeOH in DCM containing 0.1% ammonium hydroxide). The collected product was then recrystallized in MeOH to give 5-fluoro-1-(5-(3-fluoro-4-methylphenyl)-2-(3-fluoro-6-methylpyridin-2-yl)oxazol-4-yl)-4-(methylamino)pyrimidin-2(1H)-one (5 mg, 0.01 mmol, 4%).

[0446] LCMS(ESI+):m / z 428.1,[M+H] + .RT 2.25 min.

[0447] 1 H NMR:(400 MHz,DMSO-d6)d 8.48-8.47(m,1H),8.13(d,J=6.4 Hz,1H),7.91(t,J=8.8 Hz,1H),7.57(dd,J=8.4 Hz,3.2 Hz,1H),7.48(t,J=8.0 Hz,1H),7.31(d,J=8.0 Hz,1H),7.25(d,J=10.8 Hz,1H),2.91(d,J=4.8 Hz,3H),2.59(s,3H),2.29(s,3H).

[0448] Additional compounds prepared in a similar manner to Compound 175:

[0449] [Table 3]

[0450] Synthesis of compound 77. [ka]

[0451] 3-Benzoyl-5-methylpyrimidine-2,4(1H,3H)-dione To a mixture of 5-methylpyrimidine-2,4(1H,3H)-dione (10.0 g, 79.2 mmol) in pyridine (50 mL) was added benzoyl chloride (15.7 mL, 24.4 g, 134 mmol) at 0°C. After stirring at room temperature for 20 hours, ice (75 g) was added at 0°C. The mixture was stirred for 1 hour and then at room temperature for 20 hours. The resulting mixture was diluted with water (200 mL) and extracted twice with CHCl. ​​The cloudy organic layer was washed three times with aqueous NaHCO (to obtain a nearly clear solution) and dried over NaSO. The mixture was concentrated under reduced pressure. The residue was triturated with CHCl, and then heptane was added to suspend the solid. The solid material obtained by filtration and washing with heptane was dried under vacuum at 45° C. for 2 hours to give 3-benzoyl-5-methylpyrimidine-2,4(1H,3H)-dione (11.4 g, 62.6%) as a colorless solid. 1 H NMR(400 MHz,CDCl3)d 9.50(s,1H),7.94(d,2H,J=8.4),7.67(t,1H,J=7.6),7.53-7.50(m,2H),7.05-7.03(m,1H),1.92(s,3H).

[0452] N-(1-(3-benzoyl-5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-oxo-2-phenylethyl)benzamide N-(1-chloro-2-oxo-2-phenylethyl)benzamide (1.30 g, 4.74 mmol) was added to a mixture of 3-benzoyl-5-methylpyrimidine-2,4(1H,3H)-dione (1.0 g, 4.34 mmol) and sodium bicarbonate (898 mg, 10.7 mmol) in N,N-dimethylformamide (25 mL) at 0 °C and stirred at the same temperature for 30 minutes. Then, it was stirred at room temperature for 18 hours. The mixture was diluted with EtOAc, washed twice with water and brine, dried over Na SO , and evaporated in vacuo. The residue was purified by column chromatography (silica gel, heptane:ethyl acetate=1:1) to give N-(1-(3-benzoyl-5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-oxo-2-phenylethyl)benzamide (1.77 g, 87.6%) as a white solid. 1 H NMR(400 MHz,CDCl3)d 8.13(d,1H,J=6.4 Hz),7.95-7.90(m,4H),7.75-7.70(m,2H),7.60-7.52(m,8H),7.32-7.27(m,2H),6.91(d,1H,J=6.8 Hz),2.03(s,3H).

[0453] 3-Benzoyl-1-(2,5-diphenyloxazol-4-yl)-5-methylpyrimidine-2,4(1H,3H)-dione N,N-Dimethylformamide (828 mg, 11.3 mmol) was added dropwise to phosphoryl trichloride (3.47 g, 22.6 mmol) at 0°C and stirred at the same temperature for 10 minutes. The mixture was warmed to room temperature, and a solution of N-(1-(3-benzoyl-5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-oxo-2-phenylethyl)benzamide (1.77 g, 3.78 mmol) in CHCl (2 mL) was added, followed by stirring at 110°C for 3 hours. A large amount of precipitate was formed, and the mixture could not be stirred. 1,4-Dioxane (20 mL) was added and the resulting precipitate was collected on a filter and washed with water to give 3-benzoyl-1-(2,5-diphenyloxazol-4-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (372 mg, 22.0%) as a white solid. 1 H NMR(400 MHz,CDCl3)d 8.11-8.10(m,2H),8.09-7.98(m,2H),7.70-7.62(m,3H),7.52-7.27(m,9H),2.05(s,3H).

[0454] 1-(2,5-diphenyloxazol-4-yl)-5-methylpyrimidine-2,4(1H,3H)-dione A suspension of 3-benzoyl-1-(2,5-diphenyloxazol-4-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (372 mg, 0.8276 mmol) in tetrahydrofuran (16 mL) and methanol (4 mL) was treated with 1 mL of 6N NaOH solution at room temperature for 1 hour. The precipitate was collected on a filter to give 1-(2,5-diphenyloxazol-4-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (46.0 mg, 16.1%) as a white solid.

[0455] 1 H NMR(400 MHz,CDCl3)d 8.17(d,2H,J=7.6 Hz),8.16-7.93(m,3H),7.72-7.70(m,1H),7.62-7.54(m,5 H),7.39(d,1H,J=9.2 Hz),1.56(s,3H).

[0456] 1-(2,5-diphenyloxazol-4-yl)-5-methyl-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one A mixture of 1-(2,5-diphenyloxazol-4-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (40 mg, 0.1158 mmol), triethylamine (138 mg, 1.37 mmol), and 1H-1,2,4-triazole (63.8 mg, 0.9251 mmol) in acetonitrile (1 mL) was treated with phosphoryl trichloride (35.4 mg, 0.2314 mmol) at room temperature for 12 hours. The mixture was diluted with AcOEt, washed with saturated NaHCO3 solution and brine, dried over Na2SO4, and evaporated in vacuo. The residue was suspended in AcOEt / heptane, and the resulting precipitate was collected on filter paper to give 1-(2,5-diphenyloxazol-4-yl)-5-methyl-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one (37.0 mg, 80.6%) as a white solid. 1 H NMR(400 MHz,DMSO-d6)d 9.48(s,1H),8.59(s,1H),8.47(s,1H),8.17-8.16(m,2H),7.67-7.64(m,5H),7.53-7.47(m,3),2.40(s,3H).

[0457] 1-(2,5-Diphenyl-1,3-oxazol-4-yl)-4-(2-hydroxyethoxy)-5-methyl-1,2-dihydropyrimidin-2-one. Compound 77 A mixture of 1-(2,5-diphenyloxazol-4-yl)-5-methyl-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-one (30 mg, 0.07568 mmol), 2,3,4,6,7,8,9,10 octahydropyrimido[1,2-a]azepine (0.02304 g, 0.1510 mmol), and ethane-1,2-diol (46.9 mg, 0.7555 mmol) in acetonitrile (1 mL) was stirred at 80° C. for 24 hours. The mixture was diluted with dichloromethane, washed with water and brine, dried over NaSO, and evaporated in vacuo. The residue was suspended in EtOAc and collected on a filter to give 1-(2,5-diphenyloxazol-4-yl)-4-(2-hydroxyethoxy)-5-methylpyrimidin-2(1H)-one (27.0 mg, 91.8%) as a white solid.

[0458] LCMS(ESI+):m / z 390.0,[M+H] + . 1 H NMR(400MHz,CDCl3)d 8.12-8.10(m,2H),7.62-7.60(m,2H),7.53-7.50(m,3H),7.46-7.41(m,4H),4.69-4.67(m,2H),4.06-4.02(m,2H),2.05(s,3H).

[0459] Synthesis of 2H,3H-furo[2,3-d]pyrimidin-2-one [ka]

[0460] 5-[(1E)-2-Bromoethenyl]-1,2,3,4-tetrahydropyrimidine-2,4-dione A solution of N-bromosuccinimide (0.513 g, 2.882 mmol, 1.05 equiv.) in a mixture of acetone (10.5 mL, 21.0 vol.) and water (10.5 mL, 21.0 vol.) was added dropwise over 90 minutes to a boiling solution of (2E)-3-(2,4-dioxo-1,2,3,4-tetrahydropyrimidin-5-yl)prop-2-enoic acid (0.5 g, 2.745 mmol, 1.0 equiv.) and potassium acetate (0.283 g, 2.884 mmol, 1.05 equiv.) in water (27.5 mL, 55.0 vol.). The reaction mixture was then concentrated under reduced pressure to half of its original volume and cooled to 4° C. for 16 hours. The precipitate so obtained was filtered off and washed with ice water to give 5-[(1E)-2-bromoethenyl]-1,2,3,4-tetrahydropyrimidine-2,4-dione (0.194 g, 0.894 mmol, 31%).

[0461] 1H NMR(300 MHz,DMSO-d6)δ 11.32(s,2H),7.67(s,1H),7.23(d,J=13.5 Hz,1H),6.84(d,J=13.5 Hz,1H).

[0462] 2H,3H-Furo[2,3-d]pyrimidin-2-one Under an inert atmosphere, 5-[(1E)-2-bromoethenyl]-1,2,3,4-tetrahydropyrimidine-2,4-dione (0.204 g, 0.893 mmol, 0.999 equiv.) and potassium tert-butoxide (1.304 g, 11.621 mmol, 13.0 equiv.) were suspended in anhydrous dimethylformamide (19.4 ml, 100.0 vol.), and the resulting mixture was stirred at 55°C for 3 hours. The solid material was filtered off, and the filtrate was neutralized with HCl in methanol. The volatiles were then removed under reduced pressure. The solid residue was washed with ice water and dried under reduced pressure to give 2H,3H-furo[2,3-d]pyrimidin-2-one (0.027 g, 0.198 mmol, 22%).

[0463] 1H NMR(300 MHz,DMSO-d6)δ 12.14(s,1H),8.34(s,1H),7.70(d,J=2.7 Hz,1H),6.74(d,J=2.7 Hz,1H).

[0464] Synthesis of compound 181. [ka]

[0465] N-[2-(4-chloro-3-fluorophenyl)-2-oxo-1-{2-oxo-2H,3H-furo[2,3-d]pyrimidin-3-yl}ethyl]-3-methylbenzamide N-[1-chloro-2-(4-chloro-3-fluorophenyl)-2-oxoethyl]-3-methylbenzamide (0.029 g, 0.213 mmol, 0.906 equiv) was added to a stirred solution of sodium bicarbonate (0.099 g, 1.178 mmol, 5.011 equiv) and 2H,3H-furo[2,3-d]pyrimidin-2-one (0.1 g, 0.235 mmol, 1.0 equiv) in dimethylformamide (0.8 ml, 10.0 vol) at 0°C. The resulting mixture was stirred at 0°C for 30 minutes and then at room temperature for 18 hours. Complete conversion, 80% DP in RM by UPLC analysis. Dichloromethane and water were added, and the layers were separated. The organic layer was washed with brine (3 times), dried over sodium sulfate, and concentrated under reduced pressure to give N-[2-(4-chloro-3-fluorophenyl)-2-oxo-1-{2-oxo-2H,3H-furo[2,3-d]pyrimidin-3-yl}ethyl]-3-methylbenzamide (0.095 g, 0.216 mmol, 85%)

[0466] LC-MS: [M+H] + =439.85.

[0467] 3-[5-(4-chloro-3-fluorophenyl)-2-(3-methylphenyl)-1,3-oxazol-4-yl]-2H,3H-furo[2,3-d]pyrimidin-2-one. Compound 181. Under an inert atmosphere, 4-chloro-N-(1-chloro-2-oxo-2-phenylethyl)benzamide (0.097 g, 0.205 mmol, 1.002 equivalents), hexachloroethane (0.097 g, 0.41 mmol, 2.003 equivalents), and triphenylphosphine (0.107 g, 0.408 mmol, 1.994 equivalents) were suspended in anhydrous acetonitrile (1.8 ml, 20.0 volumes). The resulting mixture was stirred for 10 minutes, and then pyridine (0.066 ml, 0.819 mmol, 4.004 equivalents) was added. The resulting mixture was then stirred at 60° C. for 2 hours. Complete conversion, 46% of the desired amount in the reaction mixture by UPLC analysis. Dichloromethane and brine were added, and the layers were separated. The organic layer was washed with brine (three times), dried over sodium sulfate, and concentrated under reduced pressure to give the crude product, which was triturated with ethyl acetate to give (4) 3-[5-(4-chloro-3-fluorophenyl)-2-(3-methylphenyl)-1,3-oxazol-4-yl]-2H,3H-furo[2,3-d]pyrimidin-2-one (0.013 g, 0.031 mmol, 14%).

[0468] 1H NMR(300 MHz,DMSO-d6)δ 8.81(s,1H),8.03(s,1H),7.98(d,J=7.5 Hz,1H),7.88(d,J=2.7 Hz,1H),7.80(dd,J=10.4,2.1 Hz,1H),7.69(t,J=8.1 Hz,1H),7.48(dt,J=14.0,7.6 Hz,2H),7.35(dd,J=8.6,2.0 Hz,1H),6.89(d,J=2.8 Hz,1H),2.44(s,3H).

[0469] LCMS: [M+H] + =422.06. [ka]

[0470] Methyl 4-(2-oxopropoxy)benzoate A mixture of methyl 4-hydroxybenzoate (2.0 g, 13.145 mmol, 1.0 equiv.), chloroacetone (2.432 g, 26.270 mmol, 2.0 equiv.), potassium carbonate (5.450 g, 39.435 mmol, 3.0 equiv.), and dimethylformamide (40.0 ml, 20.0 equiv.) was stirred at 50 °C for 16 h. The reaction mixture was cooled to room temperature, diluted with ethyl acetate, and washed with saturated sodium bicarbonate solution and brine. The organic layer was then dried over sodium sulfate and concentrated under reduced pressure to give the crude product. The crude product was purified via FCC (cHx / AcOEt 1 / 0 to 7 / 3 v / v) to give methyl 4-(2-oxopropoxy)benzoate (1.613 g, 13.15 mmol, 59%).

[0471] 1H NMR(300 MHz,DMSO-d6)δ 7.90(d,J=9.0 Hz,2H),7.02(d,J=9.0 Hz,2H),4.95(s,2H),3.82(s,3H),2.17(s,3H).

[0472] Methyl 4-(2-hydroxypropoxy)benzoate Methyl 4-(2-oxopropoxy)benzoate (1.613 g, 7.747 mmol, 1.0 equiv) was dissolved in ethanol (32.26 ml, 20.0 equiv). The mixture was cooled to -10 °C, and sodium borohydride (0.352 g, 9.296 mmol, 1.2 equiv) was added in portions. The reaction mixture was stirred at -10 °C for 1 h. TLC (hex / AcOEt 8 / 2 v / v) showed complete consumption of SM. UPLC analysis showed 89% DP in RM. Water (20 ml) was added, and the product was extracted with dichloromethane (3 times). The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated under reduced pressure to give methyl 4-(2-hydroxypropoxy)benzoate (1.514 g, 7.75 mmol, 93%).

[0473] 1H NMR(300 MHz,DMSO-d6)δ 7.91(d,J=9.0 Hz,2H),7.05(d,J=9.0 Hz,2H),4.93(d,J=4.7 Hz,1H),4.02-3.93(m,1H),3.92-3.85(m,2H),3.82(s,3H),1.16(d,J=6.2 Hz,3H).

[0474] Synthesis of compound 182. [ka]

[0475] Methyl 4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}benzoate To a stirred mixture of methyl 4-(2-hydroxypropoxy)benzoate (1.514 g, 7.202 mmol, 1.0 equiv.) and imidazole (1.961 g, 28.807 mmol, 4.0 equiv.) in dry dichloromethane (22.71 ml, 15.0 vol.) was added tert-butyldimethylsilyl chloride (2.171 g, 14.403 mmol, 2.0 equiv.) along with DMAP (0.880 g, 7.202 mmol, 1.0 equiv.) at room temperature. The resulting mixture was stirred at room temperature for 16 hours.

[0476] The reaction mixture was quenched by the addition of water (25 ml). The product was extracted with dichloromethane (3 times). The combined organic extracts were washed with water and brine (2 times), dried over sodium sulfate, and concentrated under reduced pressure to give methyl 4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}benzoate (2.304 g, 7.20 mmol, 94%).

[0477] 1H NMR(300 MHz,DMSO-d6)δ 7.91(d,J=8.9 Hz,2H),7.03(d,J=8.9 Hz,2H),4.23-4.11(m,1H),4.01-3.84(m,2H),3.82(s,3H),1.18(d,J=6.3 Hz,3H),0.86(s,9H),0.07(d,J=10.3 Hz,6H).

[0478] 4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}benzamide Sodium hydride (0.755 g, 18.884 mmol, 2.8 equiv.) was added to a solution of borane-ammonia complex (0.451 g, 16.186 mmol, 2.4 equiv.) in anhydrous tetrahydrofuran (46.07 ml, 20.0 vol.), and the resulting mixture was stirred at room temperature for 30 minutes. Methyl 4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}benzoate (1) (2.304 g, 6.744 mmol, 1.0 equiv.) was then added dropwise as a solution in THF (6 ml). The resulting mixture was then stirred at room temperature for 18 hours. Water, brine, and ethyl acetate were added to the reaction mixture, and the phases were separated. The aqueous layer was extracted with ethyl acetate (three times). The combined organic extracts were washed with brine, dried over sodium sulfate and concentrated under reduced pressure to give 4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}benzamide (2.052 g, 6.74 mmol, 93%).

[0479] 1H NMR(300 MHz,DMSO-d6)δ 7.90-7.79(m,3H),7.18(s,1H),6.96(d,J=8.9 Hz,2H),4.21-4.11(m,1H),3.97-3.82(m,2H),1.19(d,J=6.3 Hz,3H),0.87(s,9H),0.09(s,3H),0.06(s,3H).

[0480] 4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}-N-[2-(4-fluorophenyl)-1-hydroxy-2-oxoethyl]benzamide 4-Fluorophenylglyoxal hydrate (1.179 g, 6.929 mmol, 1.1 equiv.) and 4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}benzamide (2.052 g, 6.30 mmol, 1.0 equiv.) were dissolved in dioxane (30.78 ml, 15.0 vol.), and the resulting mixture was stirred at 100° C. for 5 h. Dioxane was removed under reduced pressure to give the crude product, which was purified by FCC (cHx / AcOEt 100 / 0 to 7 / 3 v / v) to give 4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}-N-[2-(4-fluorophenyl)-1-hydroxy-2-oxoethyl]benzamide (3) (1.047 g, 6.30 mmol, 34%).

[0481] 1H NMR(300 MHz,DMSO-d6)δ 9.22(d,J=7.9 Hz,1H),8.15-8.01(m,3H),7.86(d,J=8.9 Hz,2H),7.36(t,J=8.9 Hz,2H),6.97(d,J=8.9 Hz,2H),6.46(d,J=8.1 Hz,1H),4.21-4.12(m,1H),3.97-3.82(m,2H),1.19-1.17(m,3H),0.86(s,9H),0.07(d,J=10.1 Hz,6H). 4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}-N-[2-(4-fluorophenyl)-2-oxo-1-{2-oxo-2H,3H-furo[2,3-d]pyrimidin-3-yl}ethyl]benzamide

[0482] 4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}-N-[2-(4-fluorophenyl)-1-hydroxy-2-oxoethyl]benzamide (3), 2H,3H-furo[2,3-d]pyrimidin-2-one (0.421 g, 1.949 mmol, 1.0 equiv.), and triphenylphosphine (0.562 g, 2.144 mmol, 1.1 equiv.) were dried under vacuum with toluene (3 times). The dried starting material was then dissolved in anhydrous tetrahydrofuran (9.47 ml, 10.0 vol.). Di-tert-butyl azodicarboxylate (0.583 g, 2.534 mmol, 1.3 equiv.) was added in one portion, and the resulting mixture was stirred at room temperature for 18 hours. The solvent was removed under reduced pressure. The crude material was purified by FCC (DCM / AcOEt 100 / 0 to 8 / 2 v / v) to give 4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}-N-[2-(4-fluorophenyl)-2-oxo-1-{2-oxo-2H,3H-furo[2,3-d]pyrimidin-3-yl}ethyl]benzamide (0.194 g, 1.95 mmol, 17%).

[0483] 1H NMR(300 MHz,DMSO-d6)δ 9.98(d,J=8.4 Hz,1H),8.68(s,1H),8.08-8.02(m,2H),7.93(dd,J=8.9,1.2 Hz,2H),7.81(d,J=2.7 Hz,1H),7.76(d,J=8.1 Hz,1H),7.44(t,J=8.8 Hz,2H),7.03(d,J=8.9 Hz,2H),6.89(dd,J=2.7,0.6 Hz,1H),4.21-4.13(m,1H),3.99-3.83(m,2H),1.18(d,J=6.3 Hz,3H),0.85(s,9H),0.07(d,J=10.8 Hz,6H).

[0484] 3-[2-(4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}phenyl)-5-(4-fluorophenyl)-1,3-oxazol-4-yl]-2H,3H-furo[2,3-d]pyrimidin-2-one 4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}-N-[2-(4-fluorophenyl)-2-oxo-1-{2-oxo-2H,3H-furo[2,3-d]pyrimidin-3-yl}ethyl]benzamide (4) (0.194 g, 0.335 mmol, 1.0 equiv.) was suspended in anhydrous acetonitrile (3.88 ml, 20.0 vol.). Hexachloroethane (0.238 g, 1.004 mmol, 3.0 equiv.) and triphenylphosphine (0.263 g, 1.004 mmol, 3.0 equiv.) were added. The resulting mixture was stirred at room temperature for 10 minutes, and pyridine (0.162 ml, 2.008 mmol, 6.0 equiv.) was added. The reaction was carried out at 60 °C for 2 hours. The solvent was removed under reduced pressure. The crude 3-[2-(4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}phenyl)-5-(4-fluorophenyl)-1,3-oxazol-4-yl]-2H,3H-furo[2,3-d]pyrimidin-2-one (0.709 g, 0.33 mmol, 98%) was used directly in the next step without purification.

[0485] Synthesis of compound 182: 3-[5-(4-fluorophenyl)-2-[4-(2-hydroxypropoxy)phenyl]-1,3-oxazol-4-yl]-2H,3H-furo[2,3-d]pyrimidin-2-one. Trifluoroacetic acid (0.374 g, 3.282 mmol, 10.0 equiv.) was added to 3-[2-(4-{2-[(tert-butyldimethylsilyl)oxy]propoxy}phenyl)-5-(4-fluorophenyl)-1,3-oxazol-4-yl]-2H,3H-furo[2,3-d]pyrimidin-2-one (0.709 g, 0.328 mmol, 1.0 equiv.) in dichloromethane (21.27 ml, 30.0 vol.). The resulting mixture was stirred and sealed at room temperature for 18 hours. The solvent was removed under reduced pressure to give the crude product, which was triturated with DMSO. The solid was filtered off, and the filtrate was purified via preparative HPLC. The solid residue (70% pure by UPLC) was separately purified by FCC (DCM / MeOH 1 / 0 to 95 / 5 v / v). After purification, both fractions were combined to give 3-[5-(4-fluorophenyl)-2-[4-(2-hydroxypropoxy)phenyl]-1,3-oxazol-4-yl]-2H,3H-furo[2,3-d]pyrimidin-2-one (0.028 g, 0.33 mmol, 19%) as a white solid.

[0486] 1H NMR(400 MHz,DMSO-d6)δ 8.82(s,1H),8.07(d,J=8.9 Hz,2H),7.88(d,J=2.6 Hz,1H),7.64-7.59(m,2H),7.34(t,J=8.9 Hz,2H),7.16(d,J=8.9 Hz,2H),6.87(d,J=2.7 Hz,1H),4.94(d,J=4.7 Hz,1H),4.02-3.96(m,1H),3.95-3.90(m,2H),1.18(d,J=6.2 Hz,3H).

[0487] LCMS: [M+H] + =448.14

[0488] Additional compounds prepared in a similar manner to Compound 182.

[0489] [Table 4]

[0490] Synthesis of compound 185. [ka]

[0491] (Z)-1-((2-oxodihydro-2H-pyran-3(4H)-ylidene)methyl)urea To a stirred solution of tetrahydro-2H-pyran-2-one (5 g, 1 equiv., 0.05 mol) in THF (50 mL) at 0 °C, NaOMe (4 g, 1.3 equiv., 0.06 mol) was added dropwise, followed by methyl formate (4 g, 4 mL, 1.3 equiv., 0.06 mol). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was filtered, washed with THF, and dried under vacuum to give sodium (Z)-(2-oxodihydro-2H-pyran-3(4H)-ylidene)methanolate (5 g, 0.03 mol, 70%) (highly hygroscopic) as an off-white solid, which was used immediately in the next step.

[0492] To a stirred solution of urea (5 g, 2.5 equivalents, 0.08 mol) in 3N HCl (40 mL) was added sodium (Z)-(2-oxodihydro-2H-pyran-3(4H)-ylidene)methanolate (5 g, 1 equivalent, 0.03 mol) portionwise at 0° C. The reaction mixture was stirred at room temperature for 16 hours. The white solid that formed was filtered and dried under vacuum. The solid compound was recrystallized from water (5 g of crude material dissolved in water (140 mL), refluxed for 1 hour to give a clear solution, then cooled to room temperature; white crystals formed, filtered, and dried under vacuum to give (Z)-1-((2-oxodihydro-2H-pyran-3(4H)-ylidene)methyl)urea (2.5 g, 15 mmol, 40%) as a white solid.

[0493] LCMS: [MH] - = m / z 169.4.

[0494] 5-(3-hydroxypropyl)pyrimidine-2,4(1H,3H)-dione To a stirred solution of KOH (1.2 g, 1.5 equivalents, 22 mmol) in methanol (30 mL) at room temperature, (Z)-1-((2-oxodihydro-2H-pyran-3(4H)-ylidene)methyl)urea (2.5 g, 1 equivalent, 15 mmol) was added in one portion. The reaction mixture was stirred at 65° C. for 4 hours. The reaction mixture was filtered and washed with methanol. The solid compound was dissolved in water (10 mL) and acidified with 3N HCl (pH approx. 3) to form a solid ppt, which was filtered, washed with water, and dried under vacuum to give the compound. The compound was recrystallized from water (the white solid was diluted with water (20 ml) and heated to reflux for 1 h to give a clear solution. The solution was then cooled to room temperature and kept at room temperature for 16 h. The white crystals that formed were filtered and dried under vacuum to give pure 5-(3-hydroxypropyl)pyrimidine-2,4(1H,3H)-dione (1.1 g, 6.5 mmol, 44%) as a white solid.

[0495] 1H NMR(400 MHz,DMSO-d6)δ ppm 10.97(s,1 H),10.60(d,J=4.4 Hz,1H),7.17(d,J=5.6 Hz,1H),4.60-4.15(m,1H),3.36(t,J=6.4 Hz,2H),2.18(t,J=7.2 Hz,2H),1.57-1.50(m,2H).

[0496] 3,5,6,7-Tetrahydro-2H-pyrano[2,3-d]pyrimidin-2-one To a solution of 5-(3-hydroxypropyl)pyrimidine-2,4(1H,3H)-dione (2 g, 1 equiv., 0.01 mol) in pyridine (40 mL) stirred at -10 °C was added MsCl (2.01 g, 1.36 mL, 1 equiv., 17.6 mmol) dropwise. The reaction mixture was stirred at 0 °C for 2 h. The reaction mixture was monitored by TLC (5% MeOH / DCM SM rf: 0.2, product rf: 0.4), and a non-polar spot indicating SM completion was observed. Workup: The reaction mixture was poured into ice water (200 mL) and stirred for 10 min. The white solid ppt formed was filtered and dried under vacuum to give 3-(2,4-dioxo-1,2,3,4-tetrahydropyrimidin-5-yl)propyl methanesulfonate (2.2 g, 8.9 mmol, 80%) as a white solid.

[0497] To a stirred solution of 3-(2,4-dioxo-1,2,3,4-tetrahydropyrimidin-5-yl)propyl methanesulfonate (2.2 g, 1 eq, 8.9 mmol) in methanol (50 mL) was added sodium methoxide (0.48 g, 1 eq, 8.9 mmol) at 0 °C. The reaction mixture was heated to 65 °C for 4 h. The reaction mixture was monitored by TLC, and a SM completion polar spot was observed. Workup: The reaction mixture was diluted with DCM (50 mL) and filtered. The filtrate was concentrated to give the crude compound. Purification: The crude compound was triturated with IPA (10 mL), filtered, and dried under vacuum to give 3,5,6,7-tetrahydro-2Hpyrano[2,3-d]pyrimidin-2-one (1.2 g, 7.9 mmol, 89%) as an off-white solid.

[0498] N-(2-(4-fluorophenyl)-2-oxo-1-(2-oxo-6,7-dihydro-2H-pyrano[2,3-d]pyrimidin-3(5H)-yl)ethyl)formamide To a stirred solution of 3,5,6,7-tetrahydro-2H-pyrano[2,3-d]pyrimidin-2-one (1.9 g, 1.1 equiv., 12 mmol) in DMF (19 mL) at 0 °C was added TEA (3.4 g, 4.4 mL, 3 equiv., 33 mmol), followed by the dropwise addition of N-(1-chloro-2-(4-fluorophenyl)-2-oxoethyl)formamide (2.4 g, 1 equiv., 11 mmol) diluted with DMF (24 mL). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was monitored by TLC, and the sm-completed product was observed. Workup: The reaction mixture was concentrated to give the crude material, triturated with ethyl acetate, filtered, and dried under vacuum to give a solid which was dissolved in water, stirred for 30 minutes, filtered, and dried under vacuum to give N-(2-(4-fluorophenyl)-2-oxo-1-(2-oxo-6,7-dihydro-2H-pyrano[2,3-d]pyrimidin-3(5H)-yl)ethyl)formamide (2 g, 6 mmol, 50%, 95% purity) as a brown solid.

[0499] LCMS: [MH] - =330.1 m / z

[0500] 3-(5-(4-fluorophenyl)oxazol-4-yl)-3,5,6,7-tetrahydro-2H-pyrano[2,3-d]pyrimidin-2-one To a stirred solution of N-(2-(4-fluorophenyl)-2-oxo-1-(2-oxo-6,7-dihydro-2H-pyrano[2,3-d]pyrimidin-3(5H)-yl)ethyl)formamide (3.5 g, 1 equiv., 11 mmol) in a 100 mL single-neck RBF was added Eaton's reagent (21 mL). The reaction mixture was heated at 70° C. for 2 h. The reaction mixture was monitored by TLC, and the sm completion product was observed. Workup: The reaction mixture was poured onto crushed ice and basified with NaHCO3. The aqueous layer was extracted with 10% MeOH in DCM. The combined organic layers were concentrated to give the crude compound. Purification: The crude compound was purified by flash chromatography, eluting the product with 2% to 4% MeOH in DCM, and the pure fractions were concentrated to give pure 3-(5-(4-fluorophenyl)oxazol-4-yl)-3,5,6,7-tetrahydro-2H-pyrano[2,3-d]pyrimidin-2-one (1.6 g, 5.1 mmol, 48%) as an off-white solid.

[0501] LCMS: [M+H] + = m / z 314.1

[0502] 1-(5-(4-fluorophenyl)-2-phenyloxazol-4-yl)-5-(3-hydroxypropyl)pyrimidine-2,4(1H,3H)-dione To a stirred solution of 3-(5-(4-fluorophenyl)oxazol-4-yl)-3,5,6,7-tetrahydro-2H-pyrano[2,3-d]pyrimidin-2-one (200 mg, 1 equiv., 638 μmol) in DMF (4 mL) was added CsCO (624 mg, 3 equiv., 1.92 mmol), followed by bromobenzene (200 mg, 134 μL, 2 equiv., 1.28 mmol) and purged with argon for 15 minutes. Then CuI (12.2 mg, 0.1 equiv., 63.8 μmol) and Pd(dppf)Cl (46.7 mg, 0.1 equiv., 63.8 μmol) were added and purged for 5 minutes. The reaction mixture was heated at 100 °C for 3 hours. The reaction mixture was diluted with water, and a solid formed, which was filtered and dried under vacuum to give the crude solid compound. The crude compound was purified by flash chromatography eluting with 2% to 5% MeOH / ethyl acetate to give 1-(5-(4-fluorophenyl)-2-phenyloxazol-4-yl)-5-(3-hydroxypropyl)pyrimidine-2,4(1H,3H)-dione (140 mg, 328 μmol, 51.3%, 95.34% purity) as an off-white solid.

[0503] LCMS: (+esi) [M+H] = 408.1

[0504] 1 H NMR:1H NMR(400 MHz,DMSO-d6)δ=11.73(s,1H),8.13(dd,J=2.9,6.6 Hz,2H),7.75-7.68(m,2H),7.66(s,1H),7.64-7.59(m,3H),7.41(t,J=8.9 Hz,2H),4.43(t,J=5.3 Hz,1H),3.44-3.37(m,2H),2.29(br t,J=7.5 Hz,2H),1.66-1.57(m,2H).

[0505] 3-(1-(5-(4-fluorophenyl)-2-phenyloxazol-4-yl)-2,4-dioxo-1,2,3,4-tetrahydropyrimidin-5-yl)propyl methanesulfonate To a stirred solution of 1-(5-(4-fluorophenyl)-2-phenyloxazol-4-yl)-5-(3-hydroxypropyl)pyrimidine-2,4(1H,3H)-dione (130 mg, 1 equiv., 319 μmol) in pyridine (2.5 mL) at 0° C. was added MsCl (54.8 mg, 37.0 μL, 1.5 equiv., 479 μmol). The resulting reaction mixture was allowed to stir for 2 hours. The reaction mixture was then poured onto crushed ice, and the separated solid was filtered and washed with cold water and MTBE. The solid was then redissolved in 10% methanol in chloroform, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 3-(1-(5-(4-fluorophenyl)-2-phenyloxazol-4-yl)-2,4-dioxo-1,2,3,4-tetrahydropyrimidin-5-yl)propyl methanesulfonate (130 mg, 0.24 mmol, 76%, 90% purity) as an off-white solid.

[0506] LCMS: (+esi) [M+H] +: 486.1

[0507] 1H NMR(400 MHz,DMSO-d)δ=11.79-11.76(m,1H),8.13(dd,J=3.0,6.7 Hz,2H),7.76-7.71(m,3H),7.65-7.58(m,3H),7.40(t,J=8.9 Hz,2H),4.22(t,J=6.2 Hz,2H),3.15(s,3H),2.37(br t,J=7.5 Hz,2H),1.94-1.86(m,2H).

[0508] 3-(5-(4-Fluorophenyl)-2-phenyloxazol-4-yl)-3,5,6,7-tetrahydro-2H-pyrano[2,3-d]pyrimidin-2-one. Compound 185 To a stirred solution of 3-(1-(5-(4-fluorophenyl)-2-phenyloxazol-4-yl)-2,4-dioxo-1,2,3,4-tetrahydropyrimidin-5-yl)propyl methanesulfonate (130 mg, 1 equiv., 268 μmol) in THF (3.6 mL) was added a solution of DBU (61.6 mg, 405 μmol) in THF (1 mL) at room temperature, and the resulting reaction mixture was stirred at 65° C. for 30 min. The reaction mixture was cooled to room temperature, diluted with 10% methanol in dichloromethane, washed with cold water, dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to give 90 mg of crude compound, which was purified using flash chromatography eluting with methanol in ethyl acetate (3% to 5%) to give 3-(5-(4-fluorophenyl)-2-phenyloxazol-4-yl)-3,5,6,7-tetrahydro-2H-pyrano[2,3-d]pyrimidin-2-one (25 mg, 62 μmol, 23%, 96.34% purity) as an off-white solid.

[0509] LCMS: (+esi) [M+H] + = 390.0

[0510] 1 H NMR(400 MHz,DMSO-d6)δ=8.23-7.98(m,3H),7.80-7.44(m,5H),7.43-7.32(m,2H),4.46(br t,J=4.8 Hz,2H),2.60(br s,2H),1.95(br s,2H).

[0511] Synthesis of compound 92. 3-(5-(4-Fluorophenyl)-2-phenyloxazol-4-yl)-7-methyl-3,5,6,7-tetrahydro-2H-pyrano[2,3-d]pyrimidin-2-one was prepared in a similar manner to compound 75, starting from 6-methyltetrahydro-2H-pyran-2-one. LCMS: (+esi) [M+H] = 404.1 [ka]

[0512] 1-(3-fluorophenyl)-2,2-dihydroxyethan-1-one 3'-Fluoroacetophenone (2.0 g, 14.5 mmol, 1.0 equiv) was dissolved in dioxane (12.0 mL) and water (2.0 mL). Selenium dioxide (3.21 g, 29.0 mmol, 2.0 equiv) was added, and the reaction was stirred at 70 °C for 16 hours. Upon cooling to room temperature, the reaction mixture was filtered through a pad of Celite, and the filtrate was concentrated. Water (50 mL) was added to the resulting oil, and the mixture was heated to reflux for 24 hours. Upon cooling to 0 °C, a precipitate formed, which was collected by vacuum filtration. The solid was washed with cold water and dried under vacuum to give 1-(3-fluorophenyl)-2,2-dihydroxyethan-1-one (1.65 g, 9.70 mmol, 67%) as a solid. This material was used directly in the next step. [ka]

[0513] 1-[4-(difluoromethoxy)-3-fluorophenyl]ethan-1-one Under an argon inert atmosphere, 3'-fluoro-4'-hydroxyacetophenone (5.0 g, 32.4 mmol, 1.0 equiv.), sodium chlorodifluoroacetate (7.42 g, 48.7 mmol, 1.5 equiv.), and cesium carbonate (21.1 g, 64.9 mmol, 2.0 equiv.) were suspended in DMF (50.0 mL). The reaction vessel was equipped with a carbon dioxide outlet. The reaction was then stirred at 120 °C for 16 h. Upon cooling to room temperature, the resulting mixture was diluted with ethyl acetate (100 mL) and washed with brine (3 × 100 mL). The organic layers were combined, dried over sodium sulfate, and concentrated under reduced pressure. The resulting crude material was purified by flash column silica gel chromatography (DCM, 100%) to give 1-[4-(difluoromethoxy)-3-fluorophenyl]ethan-1-one (2.9 g, 14.2 mmol, 44%).

[0514] 1H NMR(300 MHz,DMSO-d6)δ ppm 7.94(dd,J=11.4,2.1 Hz,1H),7.87(ddd,J=8.5,2.1,1.1 Hz,1H),7.60-7.46(m,1H),7.41(t,J=72.7 Hz,1H),2.59(s,3H).

[0515] 1-(4-chloro-3-fluorophenyl)-2,2-dihydroxyethan-1-one 1-[4-(Difluoromethoxy)-3-fluorophenyl]ethan-1-one (2.9 g, 14.2 mmol, 1.0 equiv) was dissolved in dioxane (17.4 mL) and water (2.9 mL). Selenium dioxide (3.15 g, 28.4 mmol, 2.0 equiv) was added, and the reaction was stirred at 70 °C for 16 hours. The reaction mixture was filtered through a pad of Celite, and the filtrate was concentrated. Water (15 mL) was added to the resulting oil, and the mixture was heated to reflux for 24 hours. Upon cooling to 0 °C, a precipitate formed. This material was collected by vacuum filtration, washed with cold water, and dried under vacuum to give 1-(4-chloro-3-fluorophenyl)-2,2-dihydroxyethan-1-one (1.2 g, 5.87 mmol, 36%) as a beige solid. This material was used directly in the next reaction. [ka]

[0516] 2-[(tert-butyldimethylsilyl)oxy]propanamide To a stirred mixture of lactamide (2.0 g, 22.4 mmol, 1.0 equiv.) and imidazole (1.99 g, 29.2 mmol, 1.3 equiv.) in dry dichloromethane (30.0 mL) was added tert-butyldimethylsilyl chloride (4.40 g, 29.2 mmol, 1.3 equiv.). The resulting mixture was stirred at room temperature for 16 h. Water (25 mL) was added to the reaction mixture. The mixture was extracted with DCM (3 × 25 mL). The combined organic extracts were washed with water (2 × 20 mL) and brine (1 × 50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude material was purified by silica gel column chromatography (0–5% MeOH in DCM) to give 2-[(tert-butyldimethylsilyl)oxy]propanamide (4.3 g, 21.1 mmol, 94%).

[0517] 1 H NMR(300 MHz,Chloroform-d)δ ppm 6.60(s,1H),5.64(d,J=25.1 Hz,1H),4.21(q,J=6.8 Hz,1H),1.39(d,J=6.8 Hz,3H),0.92(s,9H),0.11(d,J=2.7 Hz,6H). [ka]

[0518] 2-Cyclopropylpyrimidine-5-carboxamide 2-Cyclopropylpyrimidine-5-carboxylic acid (2.45 g, 14.9 mmol, 1.0 equiv.) was dissolved in anhydrous THF (36.8 mL). Triethylamine (2.50 mL, 17.9 mmol, 1.2 equiv.) was added, and the resulting mixture was cooled to 0° C. Ethyl chloroformate (1.78 g, 16.4 mmol, 1.1 equiv.) was added dropwise, and the reaction mixture was stirred at 0° C. for 2 hours. 25% aqueous ammonia (2.93 mL, 74.6 mmol, 5.0 equiv.) was then added, and the mixture was warmed to room temperature and stirred for 16 hours. The reaction mixture was diluted with ethyl acetate, and the layers were separated. The organic layer was washed with saturated aqueous sodium bicarbonate and brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 2-cyclopropylpyrimidine-5-carboxamide (0.8 g, 4.81 mmol, 32%) as a pale yellow solid.

[0519] 1 H NMR(300 MHz,DMSO-d6)δ ppm 9.00(s,2H),7.92(d,J=151.5 Hz,2H),2.32-2.22(m,1H),1.19-1.00(m,4H). [ka]

[0520] 5-(trifluoromethyl)furan-2-carboxamide 5-(Trifluoromethyl)-2-furoic acid (1.0 g, 5.55 mmol, 1.0 equiv.) was dissolved in THF (15.0 mL). Triethylamine (0.937 mL, 6.66 mmol, 1.2 equiv.) was added. The mixture was cooled to 0°C, and ethyl chloroformate (0.436 mL, 6.66 mmol, 1.2 equiv.) was slowly added. At this time, 25% aqueous ammonia solution (4.75 mL, 27.8 mmol, 5.0 equiv.) was added and stirred at room temperature for 16 hours. Ethyl acetate was added to the reaction mixture, and the layers were separated. The organic phase was washed with sodium bicarbonate and brine, dried over sodium sulfate, and concentrated under reduced pressure to give 5-(trifluoromethyl)furan-2-carboxamide (0.855 g, 4.77 mmol, 77%) as a colorless solid.

[0521] 1 H NMR(300 MHz,DMSO-d6)δ ppm 8.12(s,1H),7.73(s,1H),7.36(dt,J=3.6,1.2 Hz,1H),7.29(dd,J=3.7,1.0 Hz,1H). [ka]

[0522] 4-Fluoro-3-methylbenzoic acid (2.5 g, 16.2 mmol, 1.0 equiv) was dissolved in THF (37.5 mL). Triethylamine (2.71 mL, 19.5 mmol, 1.2 equiv) was added, and the resulting mixture was cooled to 0 °C. Ethyl chloroformate (1.27 mL, 19.5 mmol, 1.2 equiv) was added dropwise, and the reaction mixture was stirred at 0 °C for 1 h. 25% aqueous ammonia solution (40 mL, 40.5 mmol, 5.0 equiv) was added, and the mixture was warmed to room temperature and stirred for 1 h. Ethyl acetate was added to the reaction mixture, and the layers were separated. The organic phase was washed with sodium bicarbonate and brine, dried over sodium sulfate, and concentrated under reduced pressure to give 4-fluoro-3-met...

Claims

1. A compound having a structure according to formula (I), or a salt or hydrate or solvate thereof: 【Chemical 1】 (In the formula, T is a substituted or unsubstituted naphthyridinone or a substituted or unsubstituted dihydronaphthyridinone; X is a substituted or unsubstituted aryl or a substituted or unsubstituted heteroaryl, X is monocyclic; Z is substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, or substituted or unsubstituted ethenyl.

2. T, 【Chemistry 2】 (In the formula, R a , R b , R c , and R d is H, halogen, substituted or unsubstituted C 1－3 Alkyl, C 2 -C 4 Alkenyl, substituted or unsubstituted C 1－3 alkoxy, and when the bond between C* and C** is a single bond, R a and R b is optionally attached to C* or C** to form a substituted or unsubstituted cyclopropyl), or a salt, hydrate or solvate thereof.

3. T, 【Chemistry 3】 ,or 【Chemistry 4】 2. The compound of claim 1, or a salt, hydrate, or solvate thereof, wherein:

4. T, 【Chemistry 5】 2. The compound of claim 1, or a salt, hydrate, or solvate thereof, wherein:

5. 5. The compound of claim 1, wherein X is substituted or unsubstituted phenyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted pyrimidinyl, or substituted or unsubstituted thienyl, or a salt, hydrate, or solvate thereof.

6. 6. The compound according to claim 1, wherein X is 4-(trifluoromethyl)phenyl, 4-fluorophenyl, 2-(trifluoromethyl)pyridin-5-yl, or 2-(1,1-difluoroethyl)pyridin-5-yl, or a salt, hydrate, or solvate thereof.

7. 7. The compound of claim 1, wherein Z is substituted or unsubstituted phenyl, substituted or unsubstituted pyridinyl, substituted or unsubstituted furan, substituted or unsubstituted thienyl, substituted or unsubstituted pyrimidinyl, or substituted or unsubstituted oxazolyl, or a salt, hydrate, or solvate thereof.

8. 8. The compound according to claim 1, wherein Z is 1-methylisoquinolin-6-yl, 2-(trifluoromethyl)pyrimidin-4-yl, 2-methylpyrimidin-4-yl, 1-methyl-6-isoquinolyl, 1-(2-hydroxyethyl)-6-isoquinolyl, 3-isoquinolyl, 6-quinolyl, 8-fluoro-3-quinolyl, 8-fluoro-7-quinolyl, 4-methyl-1,7a-diaza-2-indenyl, 1-thia-5-aza-2-indenyl, 1-(2-fluoroethyl)-1H-indazol-5-yl, 3-quinolyl, 2-cyclopropyl-2H-indazol-5-yl, 6-fluoro-1,3-benzoxazol-2-yl, 5-fluoro-2-pyridyl, 1-benzofuran-2-yl, or phenyl, or a salt, hydrate, or solvate thereof.

9. Formula (II), (III), (IV), (V), (VI), (VII), (VIII), or (IX): 【Chemistry 6】 ,or 【Chemistry 7】 9. The compound of claims 1 to 8, having a structure according to the formula:

10. 7-{5-[6-(1,1-difluoroethyl)-3-pyridyl]-2-(1-methyl-6-isoquinolyl)-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-[1-(2-hydroxyethyl)-6-isoquinolyl]-5-[6-(trifluoromethyl)-3-pyridyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-[1-(2-hydroxyethyl)-6-isoquinolyl]-5-[p-(trifluoromethyl)phenyl]-1,3-oxazole -4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-[5-(p-fluorophenyl)-2-(3-isoquinolyl)-1,3-oxazol-4-yl]-1,7-diaza-8(7H)-naphthalenone, 7-[5-(p-fluorophenyl)-2-(6-quinolyl)-1,3-oxazol-4-yl]-1,7-diaza-8(7H)-naphthalenone, 7-{2-(8-fluoro-3-quinolyl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7 -[5-(p-fluorophenyl)-2-(8-fluoro-7-quinolyl)-1,3-oxazol-4-yl]-1,7-diaza-8(7H)-naphthalenone, 7-{2-(4-methyl-1,7a-diaza-2-indenyl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-(1-thia-5-aza-2-indenyl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone non, 7-{2-[1-(2-fluoroethyl)-1H-indazol-5-yl]-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-(3-quinolyl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-(2-cyclopropyl-2H-indazol-5-yl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-(6-fluoro-1,3-benzoxazol-2-yl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{5-[p-(trifluoromethyl)phenyl]-2-[2-(trifluoromethyl)-4-pyrimidinyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, 7-{2-(5-fluoro-2-pyridyl)-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}- The compound according to claim 1, which is 7-[2-(1-benzofuran-2-yl)-5-(p-fluorophenyl)-1,3-oxazol-4-yl]-1,7-diaza-8(7H)-naphthalenone, 7-[2-(1-benzofuran-2-yl)-5-(p-fluorophenyl)-1,3-oxazol-4-yl]-1,7-diaza-8(7H)-naphthalenone, or 7-{2-phenyl-5-[p-(trifluoromethyl)phenyl]-1,3-oxazol-4-yl}-1,7-diaza-8(7H)-naphthalenone, or a salt, hydrate, or solvate thereof.

11. The compound according to claim 1, which is 7-(2-(2-methylpyrimidin-4-yl)-5-(4-(trifluoromethyl)phenyl)oxazol-4-yl)-6,7-dihydro-1,7-naphthyridin-8(5H)-one, or a salt, hydrate, or solvate thereof.

12. The compound according to claim 1, which is 7-(5-(4-(trifluoromethyl)phenyl)-2-(2-(trifluoromethyl)pyrimidin-4-yl)oxazol-4-yl)-1,7-naphthyridin-8(7H)-one, or a salt, hydrate, or solvate thereof.

13. 1. A pharmaceutical formulation comprising: a) a compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt, hydrate or solvate thereof; b) a pharmaceutically acceptable excipient; and 10. A pharmaceutical formulation comprising:

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