Isoindolinone compounds and imaging agents for imaging huntingtin protein - Patents.com

Abstract

Provided herein are certain isoindolinone compounds and imaging agents useful for detecting diseases or conditions associated with protein aggregation, compositions thereof, and methods of their use.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED PATENT APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 172,617, filed April 8, 2021, which is incorporated by reference herein for all purposes.

[0002] Provided herein are compounds and imaging agents, compositions thereof, and methods of their use that are useful for detecting, treating, or preventing diseases or conditions associated with protein aggregation. [Background technology]

[0003] The advent of molecular imaging techniques such as positron emission tomography (PET) and single photon emission computed tomography (SPECT) has enabled the measurement of molecular and cellular mechanisms throughout the body in pre-clinical and clinical settings. Such measurements have widespread diagnostic utility, and their use to assess treatment response and to aid in drug development is expanding rapidly. The introduction of high-resolution molecular imaging techniques is seen by many experts as a major breakthrough.

[0004] PET involves the administration of a positron-emitting radionuclide tracer to a subject, followed by detection of the positron-emitting (annihilation) event in the body. Radionuclide tracers typically consist of a targeting molecule with one or more types of positron-emitting radionuclides incorporated within it.

[0005] Molecular probes labeled with positron-emitting radionuclides and related PET imaging assays are being developed to target, detect, visualize, and quantify a variety of extracellular and intracellular molecules and processes associated with various diseases.

[0006] Huntington's disease (HD) is an inherited, progressive neurodegenerative disorder characterized by motor, cognitive, and psychiatric impairments and neurodegeneration, with brain atrophy beginning in the striatum and cortex and extending to other subcortical brain regions. HD is caused by an expanded CAG trinucleotide repeat in the exon-1 region of the huntingtin gene (HTT). The resulting polyglutamic acid domain expansion induces misfolding and conformational changes in mutant huntingtin (mHTT) protein, which can lead to the formation of protein aggregates. HD has a worldwide prevalence of 5-10 cases per 100,000 people, making it the most common inherited and monogenic neurodegenerative disorder.

[0007] As with other medical conditions, treatment of HD is ideally initiated at or before the earliest signs of disease, therefore early indicators of disease onset and reliable pharmacodynamic biomarkers of disease progression are highly desirable. Summary of the Invention [Problem to be solved by the invention]

[0008] In view of the central role of the accumulation of aggregated forms of proteins in the pathogenesis of neurodegenerative conditions, including HD, there is a need for molecules that bind such proteins with high sensitivity and specificity, enabling molecular imaging. [Means for solving the problem]

[0009] The present disclosure relates to compounds useful for imaging huntingtin protein. Some embodiments provide compounds of formula I as described herein, optionally labeled with one or more radioisotopes. In some embodiments, the compound of formula I is 11 C. 13 N, 15 O, and 18 It contains one or more positron-emitting radioisotopes selected from F.

[0010] In some embodiments, an imaging agent is provided comprising a compound of formula I, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0011] Also provided are imaging agents comprising the compounds described herein, wherein the compounds are labeled with one or more positron-emitting radionuclides. In some embodiments, the compounds are 11 C. 13 N, 15 O, and 18 F.

[0012] Also provided is a method of detecting the presence or absence of an aggregation-prone protein in an individual, the method comprising administering an effective amount of a compound described herein or an imaging agent comprising a compound described herein, and generating an image of a body part or region of the individual.

[0013] In some embodiments, there is provided a compound or imaging agent for use in detecting the presence or absence of an aggregation-prone protein in an individual, the use comprising administering to the individual an effective amount of a compound or imaging agent as described herein and generating an image of a body part or region of the individual.

[0014] In some embodiments, a compound or imaging agent is provided for use as described herein, wherein generating an image of a body part or region of an individual comprises detecting the presence or absence of an aggregation-prone protein in the image. In some embodiments, a compound or imaging agent is provided for use as described herein, wherein the aggregation-prone protein is huntingtin protein (HTT protein). In some embodiments, a compound or imaging agent is provided for use as described herein, wherein HTT protein is found in the basal ganglia.

[0015] In some embodiments, there is provided a compound or imaging agent for the uses described herein, wherein the presence or absence of protein aggregates corresponds to the presence or absence of a neurodegenerative disease.

[0016] In some embodiments, there is provided a compound or imaging agent for use as described herein, wherein the neurodegenerative disease is selected from Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, Parkinson's disease, prion disease, and spinocerebellar ataxia. In some embodiments, there is provided a compound or imaging agent for use as described herein, wherein the neurodegenerative disease is Huntington's disease (HD).

[0017] In some embodiments, there is provided a compound or imaging agent for a use as described herein, wherein an effective amount of the imaging agent comprises about 0.1 to about 20 mCi. In some embodiments, there is provided a compound or imaging agent for a use as described herein, wherein an effective amount of the imaging agent comprises about 10 mCi.

[0018] In some embodiments, there is provided a compound or imaging agent for use as described herein, wherein generating an image comprises positron emission tomography (PET) imaging, PET with simultaneous computed tomography imaging (PET / CT), PET with simultaneous magnetic resonance imaging (PET / MRI), single photon emission computed tomography (SPECT) imaging, or a combination thereof. In some embodiments, there is provided a compound or imaging agent for use as described herein, wherein generating an image comprises PET imaging.

[0019] In some embodiments, there is provided a compound or imaging agent for the uses described herein, wherein the HTT protein is present as an oligomer or aggregate, or a combination thereof. In some embodiments, there is provided a compound or imaging agent for the uses described herein, wherein the HTT protein is a mutant.

[0020] In some embodiments, there is provided a compound or imaging agent for a use as described herein, wherein the body part or body region is the head, spinal cord, limbs, chest, or abdomen. In some embodiments, there is provided a compound or imaging agent for a use as described herein, wherein the body part or body region is the brain. [Brief description of the drawings]

[0021] [Figure 1] FIG. 1 shows the specific and saturable binding of test radioligand compounds over a range of concentrations determined in cortical tissue of 12 month old HOM zQ175, a HD mouse model. [Diagram 2] FIG. 2 shows the specific binding of [3H]-Compounds 1-6 in various samples of postmortem human brain tissue (healthy subjects [CTRL], Huntington's disease [HD], and Alzheimer's disease [AD]). [Diagram 3] FIG. 3 compares the specific binding of [3H]-Compounds 1-6 with [3H]-Comparative Compound 3 and [3H]-Comparative Compound 4 in various samples of postmortem human brain tissue (healthy subjects [CTRL], Huntington's Disease [HD], and Alzheimer's Disease [AD]). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] The following description describes exemplary embodiments of the present technology, but it should be recognized that such description is not intended as a limitation on the scope of the present disclosure, but rather is provided as a description of exemplary embodiments.

[0023] definition As used herein, the following words, phrases and symbols are generally intended to have the meanings set forth below, except to the extent that the context in which they are used herein indicates otherwise.

[0024] A compound described herein refers to a compound of any formula described herein including those of formula I, II, III, IV, V, VI, VII, VII, IX, X, XI, XII, XIII, XIV, XV, XVI, or XVII, or an isotopically labeled analogue, pharma- ceutically acceptable salt, solvate, prodrug, stereoisomer or mixture of stereoisomers thereof, or a compound described anywhere herein including the Examples, or a compound of Table 1, or a labeled isomer of such a compound as defined herein, or an imaging agent or pharmaceutical composition comprising such a compound or labeled compound.

[0025] A dash ("-") that is not between two letters or symbols is used to indicate the point of attachment of a substituent to the parent structure. For example, -C(O)NH 2is attached to the parent structure through a carbon atom. Dashes preceding or at the end of chemical groups are for convenience; chemical groups may be illustrated with or without one or more dashes without losing their ordinary meaning. Wavy lines drawn through bonds in structures indicate specific points of attachment. Unless chemically or structurally required, no orientation or stereochemistry is indicated or implied by the order in which chemical groups are written or named.

[0026] Prefix “C” u~v " indicates that the following group has u to v carbon atoms, excluding further substitutions. For example, "C 1~6 "Alkyl" refers to an alkyl group having 1 to 6 carbon atoms.

[0027] Reference herein to "about" a value or parameter includes (and describes) embodiments that are directed to the value or parameter per se. In certain embodiments, the term "about" includes ±10% of the stated amount. In other embodiments, the term "about" includes ±5% of the stated amount. In certain other embodiments, the term "about" includes ±1% of the stated amount. Additionally, the term "about X" includes the reference to "X." Additionally, the singular forms "a" and "the" include their plurals unless the context clearly dictates otherwise. Thus, for example, reference to "the compound" includes a plurality of such compounds, and reference to "the assay" includes reference to one or more assays and equivalents thereof known to those of skill in the art.

[0028] "Alkyl" refers to an unbranched or branched saturated hydrocarbon chain. As used herein, alkyl refers to an alkyl group having 1 to 20 carbon atoms (i.e., C 1~20 alkyl), 1 to 12 carbon atoms (i.e., C 1~12 alkyl), 1 to 9 carbon atoms (i.e., C 1~9 alkyl), 1 to 8 carbon atoms (i.e., C 1~8 alkyl), 1 to 6 carbon atoms (i.e., C 1~6alkyl) or 1 to 4 carbon atoms (i.e., C 1~4 Examples of alkyl groups include, for example, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, and 3-methylpentyl. When an alkyl residue having a particular number of carbons is named by a chemical name or specified by a molecular formula, all positional isomers having that number of carbons can be included; thus, for example, "butyl" refers to n-butyl (i.e., -(CH 2 ) 3 CH 3 ), sec-butyl (i.e., -CH(CH 3 )CH 2 CH 3 ), isobutyl (i.e., -CH 2 CH(CH 3 ) 2 ) and tert-butyl (i.e., -C(CH 3 ) 3 ), and "propyl" includes n-propyl (i.e., -(CH 2 ) 2 CH 3 ) and isopropyl (i.e., -CH(CH 3 ) 2 ).

[0029] Alternative chemical names known to those skilled in the art may be used in place of the terms provided herein. For example, divalent groups, such as divalent "alkyl" groups, divalent "aryl" groups, etc., may also be referred to as "alkylene" or "arylene" groups, respectively. Also, unless expressly indicated otherwise (e.g., by a dash symbol), when a combination of groups is referred to herein as a moiety, e.g., arylalkyl or aralkyl, the last-mentioned group contains the atom that connects the moiety to the remainder of the molecule.

[0030] "Alkenyl" refers to an alkyl group containing at least one carbon-carbon double bond and having 2 to 20 carbon atoms (i.e., C 2~20alkenyl), 2 to 8 carbon atoms (i.e., C 2~8 alkenyl), 2 to 6 carbon atoms (i.e., C 2~6 alkenyl) or 2 to 4 carbon atoms (i.e., C 2~4 Alkenyl refers to a hydrocarbon group having the substituents 1,2-, 1,3-, 1,4-, 1,5-, 1,6-, 1,7-, 1,8-, 1,9-, 2,10-, 2,2-, 2,3-, 3,4-, 2,5-, 3,6-, 1,8-, 2,5-, 1,9 ...

[0031] "Alkynyl" refers to an alkynyl group containing at least one carbon-carbon triple bond and having 2 to 20 carbon atoms (i.e., C 2~20 alkynyl), 2 to 8 carbon atoms (i.e., C 2~8 alkynyl), 2 to 6 carbon atoms (i.e., C 2~6 alkynyl) or 2 to 4 carbon atoms (i.e., C 2~4 The term "alkynyl" also includes such groups having triple bonds and double bonds.

[0032] "Alkoxy" refers to the group "alkyl-O-". Example alkoxy groups include, for example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, and 1,2-dimethylbutoxy.

[0033] "Alkylamino" refers to the group "alkyl-NH-". Examples of alkylamino groups include, for example, methylamino, ethylamino, iso-propylamino, tert-butylamino, and n-hexylamino. "Dialkylamino" refers to "(alkyl) 2 "N-" group. Examples of dialkylamino groups include, for example, dimethylamino, diethylamino, (iso-propyl)(methyl)amino, (n-pentyl)(tert-butyl)amino, and di-n-hexylamino.

[0034] "Alkylthio" refers to the "alkyl-S-" group. "Alkylsulfinyl" refers to the "alkyl-S(O)-" group. "Alkylsulfonyl" refers to the "alkyl-S(O)-" group. 2 "Alkylsulfonylalkyl" refers to the -alkyl-S(O) group. 2 -refers to alkyl.

[0035] "Acyl" is -C(O)R y group (in the formula, R y refers to hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which may be optionally substituted as defined herein. Examples of acyls include, for example, formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethyl-carbonyl, and benzoyl.

[0036] "Amide" is -C(O)NR y R z -C-amide groups, which refer to the -NR y C(O)R z An "N-amide" group refers to the group, y and R z is independently hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which is optionally substituted as defined herein, or R y and R z taken together form a cycloalkyl or heterocyclyl, each of which may be optionally substituted as defined herein.

[0037] "Amino" is -NR y R z group (in the formula, R y and R zis independently hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which may be optionally substituted as defined herein. In some embodiments, "amino" refers to NH 2 Refers to the base.

[0038] "Amidino" is -C(=NR y )NR z 2 group (in the formula, R y and R z refers independently to hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which is optionally substituted as defined herein.

[0039] "Aryl" refers to an aromatic carbocyclic group having a single ring (e.g., monocyclic) or multiple rings (e.g., bicyclic or tricyclic), including fused systems. As used herein, aryl refers to a ring system having 6 to 20 ring carbon atoms (i.e., C 6~20 aryl) or 6 to 10 carbon ring atoms (i.e., C 6~10 aryl). Examples of aryl groups include, for example, phenyl, naphthyl, fluorenyl, and anthryl. However, aryl does not in any way encompass or overlap with heteroaryl, as defined below. When one or more aryl groups are fused to a heteroaryl, the resulting ring system is a heteroaryl. When one or more aryl groups are fused to a heterocyclyl, the resulting ring system is a heterocyclyl.

[0040] "Arylalkyl" or "aralkyl" refers to the group "aryl-alkyl-".

[0041] "Carbamoyl" is -OC(O)NR y R z The "O-carbamoyl" group refers to the -NR group y C(O)ORz An "N-carbamoyl" group refers to the group (wherein R y and R z is independently hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which may be optionally substituted as defined herein.

[0042] "Carboxyl ester" or "ester" refers to the -OC(O)R x and -C(O)OR x (In the formula, R x is alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which may be optionally substituted as defined herein.

[0043] "Cycloalkyl" refers to saturated or partially unsaturated cyclic alkyl groups having a single ring or multiple rings, including fused, bridged, and spiro ring systems. The term "cycloalkyl" refers to cycloalkenyl groups (i.e., cyclic groups having at least one double bond) and at least one sp 3 As used herein, cycloalkyl includes carbocyclic fused ring systems (i.e., at least one non-aromatic ring) having ring carbon atoms. 3~20 cycloalkyl), 3 to 12 ring carbon atoms (i.e., C 3~12 cycloalkyl), 3 to 10 ring carbon atoms (i.e., C 3~10 cycloalkyl), 3 to 8 ring carbon atoms (i.e., C 3~8 cycloalkyl), or 3 to 6 ring carbon atoms (i.e., C 3~6cycloalkyl). Monocyclic groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. Polycyclic groups include, for example, bicyclo[2.2.1]heptanyl, bicyclo[2.2.2]octanyl, adamantyl, norbornyl, norbornenyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Furthermore, the term cycloalkyl is intended to encompass any non-aromatic ring system, which may include fused aryl rings, regardless of the attachment to the rest of the molecule. Still further, cycloalkyl also includes "spirocycloalkyl", such as spiro[2.5]octanyl, spiro[4.5]decanyl, or spiro[5.5]undecanyl. Cycloalkyl as a substituent may include spirocycloalkyl, if there are two positions on the carbon atom for substitution in the parent structure. Cycloalkyls may be optionally substituted at the carbon atom of the bond to the parent structure.

[0044] "Cycloalkoxy" refers to the group "-O-cycloalkyl."

[0045] "Cycloalkylalkyl" refers to the group "cycloalkyl-alkyl-".

[0046] "Guanidino" is -NR y C(=NR z )NR y R z (In the formula, each R y and R z refers independently to hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which is optionally substituted as defined herein.

[0047] "Imino" is -C(=NR y )R z group (in the formula, R y and R zeach independently refers to hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which is optionally substituted as defined herein.

[0048] "Imide" is -C(O)NR y C(O)R z group (in the formula, R y and R z each independently refers to hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which is optionally substituted as defined herein.

[0049] "Halogen" or "halo" refers to a substituent atom in Group VIIA of the periodic table, e.g., fluoro, chloro, bromo, or iodo.

[0050] "Haloalkyl" refers to an unbranched or branched alkyl group as defined above in which one or more (e.g., 1-6 or 1-3) hydrogen atoms, including up to and including all hydrogen atoms, have been replaced with a halogen. For example, if a residue is substituted with more than one halogen, it can also be referred to using a prefix corresponding to the number of halogen moieties attached. Dihaloalkyl and trihaloalkyl refer to alkyl substituted with two ("di") or three ("tri") halo groups, which may, but do not necessarily, be the same halogen. Perhaloalkyl groups are haloalkyl groups in which all hydrogen substituents have been replaced with halo. Examples of haloalkyl include, for example, trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and the like.

[0051] "Haloalkoxy" refers to an alkoxy group as defined above in which one or more (e.g., 1-6 or 1-3) hydrogen atoms, up to and including all hydrogen atoms, have been replaced with a halogen.

[0052] "Hydroxyalkyl" refers to an alkyl group, as defined above, in which one or more (eg, 1 to 6 or 1 to 3) hydrogen atoms are replaced with a hydroxy group.

[0053] "Heteroalkyl" refers to an alkyl group in which one or more carbon atoms (and any associated hydrogen atoms) of the alkyl chain are each independently replaced with the same or different heteroatom group, provided that the point of attachment to the remainder of the molecule is through a carbon atom. The term "heteroalkyl" includes unbranched or branched saturated chains having carbon and heteroatoms. By way of example, one, two, or three carbon atoms may be independently replaced with the same or different heteroatom groups. Heteroatom groups include, but are not limited to, -NR y -, -C(O)NR y -, -NR y C(O)-, -O-, -S-, -S(O)-, and -S(O) 2 -(In the formula, R y is hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which may be optionally substituted as defined herein. Examples of heteroalkyl groups include, for example, ethers (e.g., -CH 2 OCH 3 , -CH(CH 3 )OCH 3 , -CH 2 CH 2 OCH 3 , -CH 2 CH 2 OCH 2 CH 2 OCH 3 etc.), thioethers (e.g., -CH 2 SCH 3 , -CH(CH3 )SCH 3 , -CH 2 CH 2 SCH 3 , -CH 2 CH 2 SCH 2 CH 2 SCH 3 etc.), sulfones (e.g., -CH 2 S(O) 2 CH 3 , -CH(CH 3 )S(O) 2 CH 3 , -CH 2 CH 2 S(O) 2 CH 3 , -CH 2 CH 2 S(O) 2 CH 2 CH 2 OCH 3 etc.) and aminoalkyl (e.g., -CH 2 NR y CH 3 , -CH(CH 3 )NR y CH 3 , -CH 2 CH 2 NR y CH 3 , -CH 2 CH 2 NR y CH 2 CH 2 NR y CH 3 etc., where R y is hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which may be optionally substituted as defined herein. As used herein, heteroalkyl contains 1 to 10 carbon atoms, 1 to 8 carbon atoms, or 1 to 4 carbon atoms and 1 to 3 heteroatoms, 1 to 2 heteroatoms, or 1 heteroatom.

[0054] "Heteroaryl" refers to an aryl group having one or more ring heteroatoms independently selected from nitrogen, oxygen, and sulfur, and one or more (e.g., 1 to 3) N-oxides (-O - As used herein, heteroaryl refers to an aromatic group having 1 to 20 ring carbon atoms (i.e., C 1~20 Heteroaryl), 3 to 12 ring carbon atoms (i.e., C 3~12 heteroaryl), or 3 to 8 carbon ring atoms (i.e., C 3~8Heteroaryl) and 1-5 ring heteroatoms, 1-4 ring heteroatoms, 1-3 ring heteroatoms, 1-2 ring heteroatoms, or 1 ring heteroatom, each independently selected from nitrogen, oxygen, and sulfur. In certain instances, heteroaryl includes a 5-10 membered ring system, a 5-7 membered ring system, or a 5-6 membered ring system having 1-4 ring heteroatoms, 1-3 ring heteroatoms, 1-2 ring heteroatoms, or 1 ring heteroatom, each independently selected from nitrogen, oxygen, and sulfur. Examples of heteroaryl groups include, for example, acridinyl, benzimidazolyl, benzothiazolyl, benzoindolyl, benzofuranyl, benzothiazolyl, benzothiadiazolyl, benzonaphthofuranyl, benzoxazolyl, benzothienyl (benzothiophenyl), benzotriazolyl, imidazo[1,2-a]pyridyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl ... Examples of such aryl groups include isoquinolyl, isoxazolyl, naphthyridinyl, oxadiazolyl, oxazolyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl, phenazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl and triazinyl. Examples of fused heteroaryl rings include, but are not limited to, benzo[d]thiazolyl, quinolinyl, isoquinolinyl, benzo[b]thiophenyl, indazolyl, benzo[d]imidazolyl, pyrazolo[1,5-a]pyridinyl and imidazo[1,5-a]pyridinyl, and the heteroaryl can be bonded through any ring of the fused system. Any aromatic ring system having a single or multiple fused rings containing at least one ring heteroatom, regardless of the bond to the rest of the molecule (i.e., through any one of the fused rings), is considered heteroaryl. Heteroaryl does not encompass or overlap with aryl as defined above.

[0055] "Heteroarylalkyl" refers to the group "heteroaryl-alkyl-".

[0056] "Heterocyclyl" refers to a saturated or partially unsaturated cyclic alkyl group having one or more ring heteroatoms independently selected from nitrogen, oxygen, and sulfur, where the nitrogen or sulfur atom is optionally oxidized to an N-oxide, sulfinyl (-S(O)-), or sulfoxide (-S(O)-). 2 -). The term "heterocyclyl" includes heterocycloalkenyl groups (i.e., heterocyclyl groups having at least one double bond), bridged heterocyclyl groups, fused heterocyclyl groups, spiro-heterocyclyl groups, and oxo-heterocyclyl groups. Heterocyclyls may be monocyclic or polycyclic, which polycyclic rings may be fused, bridged, or spiro. Regardless of the enumerated substituents, a heterocyclyl may include one or more (e.g., 1-3) oxo (=O) or N-oxide (-O) groups, unless otherwise specified. - ) moiety. Heterocyclyl can be attached via a carbon atom or a heteroatom, where valence allows. Additionally, the term heterocyclyl encompasses any ring system, including non-aromatic rings or ring systems, containing at least one heteroatom, which ring may be fused to an aryl or heteroaryl ring regardless of attachment to the remainder of the molecule. Heterocyclyl may have charged resonance structures that are aromatic (e.g., pyridin-2(1H)-one-1-yl). As used herein, heterocyclyl refers to a ring having 3-14 ring atoms, 3-10 ring atoms, 3-6 ring atoms, or 5-6 ring atoms, and / or 2-12 ring carbon atoms (i.e., C 2~12 heterocyclyl), 2 to 10 ring carbon atoms (i.e., C 2~10 heterocyclyl), 2 to 8 ring carbon atoms (i.e., C 2~8 heterocyclyl), 3 to 12 ring carbon atoms (i.e., C 3~12 heterocyclyl), 3 to 8 ring carbon atoms (i.e., C 3~8 heterocyclyl), or 3 to 6 ring carbon atoms (i.e., C 3~6Heterocyclyl) and may have 1 to 5 ring heteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom. Examples of heterocyclyl groups include, for example, azetidinyl, azepinyl, benzodioxolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzopyranyl, benzodioxinyl, benzopyranonyl, benzofuranonyl, dioxolanyl, dihydropyranyl, hydropyranyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, furanonyl, imidazolinyl, imidazolidinyl, indolinyl, indolizinyl, isoindolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, and the like. Heterocyclyl includes aryl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, oxiranyl, oxetanyl, phenothiazinyl, phenoxazinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, tetrahydropyranyl, trithianyl, tetrahydroquinolinyl, thiophenyl (i.e., thienyl), tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. The term "heterocyclyl" also includes "spiro-heterocyclyl." Examples of spiro-heterocyclyl rings include, for example, bicyclic and tricyclic ring systems, such as 2-oxa-7-azaspiro[3.5]nonanyl, 2-oxa-6-azaspiro[3.4]octanyl, and 6-oxa-1-azaspiro[3.3]heptanyl. Heterocyclyl as a substituent may include spiro-heterocyclyl when there are two positions on the carbon atom for substitution in the parent structure. Examples of bridged heterocyclyl rings include, but are not limited to, 2,5-diazabicyclo[2.2.1]heptane, 2-oxa-5-azabicyclo[2.2.1]heptanyl.Examples of fused heterocyclyl rings include, but are not limited to, 1,2,3,4-tetrahydroisoquinolinyl, 4,5,6,7-tetrahydrothieno[2,3-c]pyridinyl, indolinyl, and isoindolinyl, where the heterocyclyl can be attached via either ring of the fused system. An "oxo-heterocyclyl" group is a heterocyclyl that includes at least one oxo substituent (e.g., 1, or 1-2 oxo substituents), regardless of whether additional substituents are allowed (i.e., an unsubstituted oxo-heterocyclyl includes oxo and no other substitutions). In some embodiments, the oxo-heterocyclyl includes a cyclic amide moiety.

[0057] "Heterocyclylalkyl" refers to the group "heterocyclyl-alkyl-".

[0058] "Oxime" is -CR y (=NOH) group (wherein R y refers to hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which may be optionally substituted as defined herein.

[0059] "Sulfonyl" is -S(O) 2 R y group (in the formula, R y is hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which may be optionally substituted as defined herein. Examples of sulfonyl are methylsulfonyl, ethylsulfonyl, phenylsulfonyl, and toluenesulfonyl.

[0060] "Sulfinyl" is -S(O)R y group (in the formula, R yis hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which may be optionally substituted as defined herein. Examples of sulfinyl are methylsulfinyl, ethylsulfinyl, phenylsulfinyl, and toluenesulfinyl.

[0061] "Sulfonamide" is -SO 2 NR y R z and -NR y SO 2 R z group (in the formula, R y and R z each independently refers to hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which is optionally substituted as defined herein.

[0062] The terms "optionally" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and that the description includes instances in which said event or circumstance occurs and instances in which it does not occur. Also, the term "optionally substituted" refers to a group that is unsubstituted or substituted.

[0063] As used herein, the term "substituted" refers to any one or more (e.g., 1-5 or 1-3) hydrogen atoms being replaced with a non-hydrogen group, such as, but not limited to, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, acyl, amido, amino, amidino, aryl, arylalkyl, azido, carbamoyl, carboxyl, carboxyl ester, cyano, cycloalkyl, cycloalkylalkyl, guanidino, halo, haloalkyl, haloalkoxy, hydroxyalkyl, heteroalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, -NHNH 2 , =NNH 2, imino, imido, hydroxy, oxo, oxime, nitro, sulfonyl, sulfinyl, alkylsulfonyl, alkylsulfinyl, thiocyanate, -S(O)OH, -S(O) 2 OH, sulfonamide, thiol, thioxo, N-oxide or -Si(R y ) 3 (In the formula, each R y refers to a group that is replaced by (which is independently hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, aryl, heteroaryl, or heterocyclyl).

[0064] In certain embodiments, "substituted" means that one or more (e.g., 1-5 or 1-3) hydrogen atoms are independently replaced with deuterium, halo, cyano, hydroxyl, imino, nitro, azido, oxo, thioxo, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, thioalkyl, haloalkoxy, cycloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, -NR g R h , -NR g C(=O)R h , -NR g C(=O)NR g R h , -NR g C(=O)OR h , -NR g S(=O) 1~2 R h , -C(=O)R g , -C(=O)OR g , -OC(=O)OR g , -OC(=O)R g , -C(=O)NR g R h , -OC(=O)NR g R h , -OR g , -SR g , -S(=O)R g , -S(=O) 2 R g , -OS(=O) 1~2 Rg , -S(=O) 1~2 OR g , -NR g S(=O) 1~2 NR g R h , =NSO 2 R g , =NOR g , -S(=O) 1~2 NR g R h ,-SCIENCE FICTION 5 , or -SCF 3 In certain embodiments, "substituted" also refers to a group in which one or more (e.g., 1-5 or 1-3) hydrogen atoms have been replaced with -C(=O)R. g , -C(=O)OR g , -C(=O)NR g R h , -CH 2 SO 2 R g , or -CH 2 SO 2 NR g R h In the above, R g and R h are the same or different and are independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, thioalkyl, aryl, arylalkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, and / or heteroarylalkyl; or R g and R h two of these, together with the atom to which they are attached, form a heterocyclyl ring optionally substituted with oxo, halo, or alkyl optionally substituted with oxo, halo, amino, hydroxyl, or alkoxy.

[0065] Polymers or similar indefinite structures arrived at by defining a substituent with infinitely more additional substituents (e.g., a substituted aryl with a substituted alkyl that is itself substituted with a substituted aryl group (which is further substituted with a substituted heteroalkyl group, etc.) are not intended to result from the above definition. Unless otherwise specified, the maximum number of consecutive substitutions in the compounds described herein is three. For example, consecutive substitution of a substituted aryl group with two other substituted aryl groups is limited to ((substituted aryl) substituted aryl) substituted aryl. Similarly, the above definition is not intended to encompass compounds with chemically infeasible or inseparable substitution patterns (e.g., a methyl substituted with five fluorines or a heteroaryl group with three consecutive oxygen ring atoms). Such unacceptable substitution patterns are well known to those skilled in the art. When used to modify a chemical group, the term "substituted" may describe other chemical groups defined herein.

[0066] In certain embodiments, as used herein, the phrase "one or more" refers to 1 to 5. In certain embodiments, as used herein, the phrase "one or more" refers to 1 to 3.

[0067] Any compound or structure provided herein is intended to represent unlabeled forms of the compound as well as "isotopically enriched analogs." Isotopically enriched forms of the compound may also be referred to as "labeled." Isotopically enriched analogs have the structure depicted herein except that one or more atoms are enriched with an isotope having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into the compounds described herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 89, 90, 91, 92, 93, 94, 95, 9 2 H, 3 H, 11 C. 13 C. 14 C. 13 N, 15 N, 15 O. 17 O. 18 O.31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I, and 125 Isotopically enriched analogs generally include compounds having any isotopic enrichment above the natural abundance of the isotope (e.g., at the Earth's surface). Various isotopically labeled compounds, such as radioisotopes, e.g., 3 H, 18 F, 11 C, and 14 C and the like are included in the present disclosure. 18 F, 3 H, or 11 Compounds labeled with C may be useful in metabolic studies, reaction kinetic studies, detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT) including drug or substrate tissue distribution assays, or in radiation treatment of patients.

[0068] The term "isotopically enriched analog" includes "deuterated analogs" of compounds described herein in which one or more hydrogens, e.g., hydrogens on carbon atoms, are replaced by deuterium. Such compounds can exhibit increased resistance to metabolism and are therefore useful for increasing the half-life of any compound when administered to a mammal, particularly a human. See, for example, Foster, "Deuterium Isotope Effects in Studies of Drug Metabolism," Trends Pharmacol. Sci. 5(12):524-527 (1984). Such compounds are synthesized by means well known in the art, e.g., by utilizing starting materials in which one or more hydrogens have been replaced by deuterium.

[0069] The therapeutic compounds of the present disclosure that are deuterium labeled or substituted can have improved DMPK (drug metabolism and pharmacokinetic) properties in terms of distribution, metabolism and excretion (ADME). Substitution with heavier isotopes, e.g., deuterium, can result in certain therapeutic advantages resulting from higher metabolic stability, e.g., increased in vivo half-life, reduced dose requirements and / or improved therapeutic index. The isotopically labeled compounds of the present disclosure and their prodrugs can generally be prepared by carrying out the procedures disclosed in the schemes described below or in the examples and preparations, by substituting readily available isotopically labeled reagents for non-isotopically labeled reagents. Where a compound is described as a deuterated analog, the compound can be depicted with deuterium as a substituent.

[0070] The concentration of such heavier isotopes, particularly deuterium, can be defined by the isotopic enrichment factor. In the compounds of the present disclosure, any atom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom. Unless otherwise stated, when a position is specifically designated as "H" or "hydrogen", it is understood that this position has hydrogen and its isotopes at their natural abundance.

[0071] In many cases, the compounds of the present disclosure are capable of forming acid and / or base salts by virtue of the presence of amino and / or carboxyl groups or groups similar thereto.

[0072] Also provided are isotopically enriched analogs, pharma- ceutically acceptable salts, prodrugs, tautomers, stereoisomers, and mixtures of stereoisomers of the compounds described herein. "Pharmaceutically acceptable" or "physiologically acceptable" refers to compounds, salts, compositions, dosage forms, and other materials useful in the preparation of pharmaceutical compositions suitable for veterinary or human pharmaceutical use.

[0073] The term "pharmaceutically acceptable salt" of a compound described herein refers to a salt that retains the biological effectiveness and properties of a given compound and is not biologically or otherwise unsuitable. A "pharmaceutically acceptable salt" or "physiologically acceptable salt" of a compound described herein includes, for example, an acid addition salt obtained by interacting a compound having a basic functional group with an acid, and a base addition salt obtained by interacting a compound having an acidic functional group with a base. If a compound is obtained as an acid addition salt, the free base can be obtained by basifying a solution of the acid salt. Conversely, if the compound is a free base (e.g., an amine compound), an addition salt can be generated by dissolving the free base in a suitable organic solvent and treating the solution with an acid. Those skilled in the art will recognize various synthetic methods that may be used to prepare non-toxic, pharmaceutically acceptable addition salts. Pharmaceutically acceptable acid addition salts of the compounds described herein can be prepared from inorganic and organic acids. Suitable inorganic acids include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Suitable organic acids include, for example, acetic acid, propionic acid, gluconic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid, salicylic acid, and the like. Similarly, pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. Salts derived from inorganic bases include, by way of example only, sodium, potassium, lithium, aluminum, ammonium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, primary, secondary, and tertiary amines, such as alkyl amines (i.e., NH 2 (alkyl)), dialkylamines (i.e., HN(alkyl) 2 ), trialkylamines (i.e., N(alkyl) 3 ), substituted alkylamines (i.e., NH 2 (substituted alkyl)), di(substituted alkyl)amines (i.e., HN(substituted alkyl) 2), tri(substituted alkyl)amines (i.e., N(substituted alkyl) 3 ), alkenylamines (i.e., NH 2 (alkenyl)), dialkenylamines (i.e., HN(alkenyl) 2 ), trialkenylamines (i.e., N(alkenyl) 3 ), substituted alkenylamines (i.e., NH 2 (substituted alkenyl)), di(substituted alkenyl)amines (i.e., HN(substituted alkenyl) 2 ), tri(substituted alkenyl)amines (i.e., N(substituted alkenyl) 3 , monocycloalkylamines, dicycloalkylamines or tricycloalkylamines (i.e., NH 2 (cycloalkyl), HN(cycloalkyl) 2 , N(cycloalkyl) 3 ), monoarylamines, diarylamines or triarylamines (i.e., NH 2 (aryl), HN(aryl) 2 , N(aryl) 3 ), cyclic amines (e.g., piperidine, piperazine, 1,4-diazabicyclo[2.2.2]octane), aromatic amines (e.g., pyridine, quinoline), or mixed amines. Specific examples of suitable amines include, by way of example only, isopropylamine, trimethylamine, diethylamine, tri(iso-propyl)amine, tri(n-propyl)amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, N-ethylpiperidine, and the like.

[0074] Some compounds described herein may exist as tautomers. For example, if a compound is depicted as containing an amide, the compound may exist as an imidic acid tautomer, and if a compound is depicted as containing a ketone, the compound may also exist as an enol tautomer. Regardless of which tautomer is depicted and regardless of the nature of the equilibrium between tautomers, the compound is understood by those skilled in the art to include both tautomers. Thus, for example, compounds containing amides are understood to include these imidic acid tautomers, and compounds containing imidic acids are understood to include these amide tautomers.

[0075] The compounds described herein may contain asymmetric centers and thus may give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined in terms of absolute stereochemistry as (R)- or (S)-, or (D)- or (L)- with respect to amino acids. The compounds described herein are meant to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents or resolved using conventional techniques, such as chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from suitable optically pure precursors, or resolution of the racemates (or racemates of salts or derivatives) using, for example, chiral high performance liquid chromatography (HPLC). Where the compounds described herein contain double bonds or other centers of geometric asymmetry, and unless otherwise specified, the compounds are intended to include both cis- and trans- or E- and Z-geometric isomers.

[0076] "Stereoisomer" refers to one of a set of compounds made up of the same atoms joined by the same bonds but with different three-dimensional structures. The various stereoisomers and mixtures thereof are intended to include "enantiomers," which refer to stereoisomeric compounds that are non-superimposable mirror images of one another.

[0077] A "diastereomer" is one of a series of stereoisomers that have at least two asymmetric atoms which are not mirror images of one another.

[0078] A "prodrug" is any molecule that releases a parent drug that is presumed to be active with the compounds described herein in vivo when such prodrug is administered to a mammalian subject. A prodrug may be a form of a compound described herein that has been modified such that the modification can be cleaved in vivo to release the parent compound. Prodrugs can be prepared by modifying functional groups present in the compounds described herein such that the modification can be cleaved to the parent compound either in routine manipulation or in vivo. Prodrugs include compounds described herein in which a hydroxy, amino, carboxyl, or sulfhydryl group in the compounds described herein is bonded to any group that can be cleaved in vivo to regenerate the free hydroxy, amino, or sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to, esters (e.g., acetate, formate, and benzoate derivatives), amides, guanidines, carbamates (e.g., N,N-dimethylaminocarbonyl) of hydroxy functional groups, etc., in the compounds described herein. The preparation, selection and use of prodrugs are discussed in T. Higuchi and V. Stella, "Pro-drugs as Novel Delivery Systems," Vol. 14 of the ACS Symposium Series; "Design of Prodrugs," edited by H. Bundgaard, Elsevier, 1985; and Bioreversible Carriers in Drug Design, edited by Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, each of which is incorporated herein by reference in its entirety.

[0079] In some embodiments, the term "neurodegenerative disease" refers to a disease or condition in which the function of a subject's nervous system is impaired. Examples of neurodegenerative diseases include those described herein.

[0080] The methods described herein can be applied to cell populations in vivo or ex vivo. "In vivo" means within a living individual, such as within an animal or human. In this context, the methods described herein can be used therapeutically in an individual. "Ex vivo" means outside a living individual. Examples of ex vivo cell populations include in vitro cell cultures and biological samples, including fluid or tissue samples obtained from an individual. Such samples can be obtained by methods well known in the art. Exemplary biological fluid samples include blood, cerebrospinal fluid, urine, and saliva. In this context, the compounds and compositions described herein can be used for a variety of purposes, including therapeutic and experimental purposes. For example, the compounds and compositions described herein can be used ex vivo to determine optimal schedules and / or dosing of administration of the disclosed compounds for a given indication, cell type, individual, and other parameters. Information gleaned from such uses can be used in the clinic for experimental purposes or to set up protocols for in vivo treatment. Other ex vivo uses to which the compounds and compositions described herein can be adapted are described below or will be apparent to one of skill in the art. Selected compounds can be further characterized to determine safety or tolerable doses in human or non-human subjects. Such properties can be determined using methods commonly known to those of skill in the art.

[0081] The above listed terms also include in vitro and ex vivo methods.

[0082] As used herein, the terms "group," "moiety," "radical," "substituent," and "fragment" are synonymous and are intended to indicate a portion of a molecule that can be attached to another portion of the molecule, for example, through a designated point of attachment or bond.

[0083] The term "active agent" is used to refer to a compound that has biological activity in the treatment, amelioration, or prevention of a disease or condition. In some embodiments, an "active agent" is a compound having pharmaceutical utility or an isotopically labeled analog, pharma- ceutically acceptable salt, solvate, prodrug, stereoisomer, or mixture of stereoisomers thereof. For example, an active agent may be an anti-neurodegenerative therapeutic agent.

[0084] The term "effective amount" refers to an amount of a compound, e.g., as described herein, sufficient to produce a desired response in an individual or patient. In the context of the use of an imaging agent, an effective amount may be the amount required to produce an image having diagnostic or therapeutic utility. The term "therapeutically effective amount" refers to an amount effective to confer a therapeutic benefit, such as amelioration of symptoms, delay of disease progression, or prevention of disease, when administered to a human or non-human patient, e.g., a therapeutically effective amount may be an amount sufficient to reduce the symptoms of a disease as described herein. A (therapeutically) effective amount may vary depending on the subject and disease or condition being treated, the weight and age of the subject, the severity of the disease or condition, and the mode of administration, and can be readily determined by one of skill in the art.

[0085] The term "huntingtin protein" or "HTT protein" as used herein refers to the protein encoded by the human huntingtin gene (HTT gene), which is located on the short (p) arm of chromosome 4 at position 16.3. More precisely, the IT that encodes the HTT protein 15 The gene is located on chromosome 4 from base pair 3,076,407 to base pair 3,245,686.

[0086] The term "protein aggregates" as used herein refers to aggregates of proteins, which may be, for example, insoluble fibrillar amyloids comprising misfolded HTT protein molecules ("HTT protein aggregates") or misfolded β-amyloid protein molecules ("β-amyloid aggregates"). An "aggregation-prone protein" is a protein that is capable of forming such aggregates in its wild-type or mutant form.

[0087] The term "imaging agent" as used herein refers to a compound described herein that is labeled with one or more positron-emitting isotopes or radionuclides, or a composition that includes the labeled compound. The positron-emitter labeled compound need only be enriched with a detectable isotope to a degree that allows detection using a technique appropriate for the particular application.

[0088] The term "PET imaging" (which may also be referred to as positron emission tomography imaging), as used herein, refers to the production of images of internal structures of the human or animal body using positron emitter-labeled compounds.

[0089] The term “positron-emitting radionuclide,” as used herein, refers to a type of radionuclide in which protons inside the nucleus emit positrons and electron neutrinos (ν e β+ decay refers to a radioisotope that exhibits a particular type of radioactive decay called β+ decay, in which the isotope is converted into a neutron while emitting a positron. Some examples of positron-emitting radionuclides include: 15 O. 13 N, 11 C. 18 F, 76 Br, and 124 Contains I.

[0090] The term "labeled," as used herein, refers to a compound that is associated with one or more positron-emitting radionuclides that are greater than their natural abundance. For example, the labeled compounds described herein may contain one or more positron-emitting radionuclides in which atoms within the molecule (including those within the indicated substituents) are present as positron-emitting isotopes.

[0091] The term "tomography" as used herein refers to a segmental imaging technique. The images can be viewed individually as a series of two-dimensional slices, or together as a computer-generated three-dimensional display.

[0092] In some embodiments, the term "neurodegenerative disease" refers to a disease or condition in which the function of a subject's nervous system is impaired. Examples of neurodegenerative diseases include those described herein.

[0093] "Treatment" or "treating" means any treatment of a disease condition in a patient; a) inhibiting the disease (e.g., reducing one or more symptoms resulting from the disease or condition and / or lessening the severity of the disease or condition); b) delaying or halting the onset of clinical symptoms associated with the disease or condition (e.g., stabilizing the disease or condition, preventing or slowing the worsening or progression of the disease or condition, and / or preventing or slowing the spread (e.g., metastasis) of the disease or condition); and / or c) palliating the disease, i.e., causing regression of clinical symptoms (e.g., causing remission of the disease state, providing partial or total relief of a disease or condition, enhancing the action of another drug, slowing the progression of a disease, improving quality of life and / or prolonging survival). Includes.

[0094] "Prevention" or "preventing" refers to any treatment of a disease or condition that keeps the clinical symptoms of the disease or condition from developing. The compounds, in some embodiments, can be administered to subjects (including humans) who are at risk (e.g., have genetic or epigenetic markers, have engaged in activities or have been exposed to environmental conditions associated with the disease or condition) or have a family history of the disease or condition.

[0095] "Subject" or "patient" refers to an animal, e.g., a mammal, who is or will be the object of treatment, observation or experiment. The methods described herein can be useful in both human therapy and veterinary applications. In some embodiments, the subject or patient is a mammal. In some embodiments, the subject or patient is a human.

[0096] The term "curie" (Ci) is a unit of measure of radioactivity and has its conventional meaning to those of ordinary skill in the art.

[0097] The term "diagnostic imaging," as used herein, refers to the production of pictures of internal structures of the human or animal body using electromagnetic radiation for diagnostic purposes.

[0098] It is to be understood that some features described herein that are described in the context of separate embodiments for clarity can also be provided in the form of a combination in a single embodiment. Conversely, various features described herein that are described in the context of a single embodiment for brevity can also be provided separately or in any suitable subcombination. All combinations of the embodiments with respect to the chemical groups represented by the variables contained in Formula I or any other formula, so long as such combinations result in stable compounds (i.e., compounds that can be isolated, characterized, and tested for biological activity), are specifically included herein as if each and every combination were individually and explicitly recited. Furthermore, all subcombinations of the chemical groups recited in the embodiments describing such variables, as well as all subcombinations of the uses and medical applications described herein, are also specifically included herein as if each and every subcombination of the chemical groups and the subcombinations of the uses and medical applications were individually and explicitly recited herein. Furthermore, some embodiments include any combination of one or more additional agents disclosed herein, as if each and every combination were individually and explicitly recited herein.

[0099] [Table 1] TIFF2024513913000002.tif236146TIFF2024513913000003.tif233145TIFF2024513913000004.tif231146TIFF2024513913000005.tif61146

[0100] compound The present disclosure relates to compounds useful for imaging aggregation-prone proteins, such as huntingtin proteins. Some embodiments have the formula I:

[0101] [ka] [In the formula,

[0102] [ka] teeth,

[0103] [ka] or

[0104] [ka] and; R 1 is hydrogen, if present, C 1~6 Alkyl or C 1~6 is haloalkyl; R 10 is hydrogen, if present, C 1~6 Alkyl or C 1~6 is haloalkyl; Ring A is a 5-6 membered heteroaryl; X is CR 11 or N; R 11 is hydrogen, cyano, hydroxy, halo, C 1~6 Alkyl, C 1~6 Haloalkyl, or C 1~6 is alkoxy; Y 1 CR 12 or N; Y 2 CR 13 or N; R 12 and R 13 Each of these is hydrogen, hydroxy, halo, C 1~6 Alkyl, C 1~6 Haloalkyl, or C 1~6 is alkoxy; R 2 is hydrogen, hydroxy, halo, C 1~6 Alkyl, C 1~6 Haloalkyl, or C 1~6 is alkoxy; L is C optionally substituted with 1 to 6 fluoro. 1 ~C 3 is alkylene; R 3 is hydrogen, fluoro, C 1~6 Alkyl or C 1~6 is haloalkyl; Each R 4 are independently cyano, hydroxy, halo, C 1~6 Alkyl, C 1~6 Haloalkyl or C 1~6 is alkoxy; Each R 5 are independently cyano, hydroxy, halo, C 1~6 Alkyl, C 1~6 Haloalkyl or C 1~6 is alkoxy; R 6 is hydrogen, cyano, hydroxy, halo, C 1~6 Alkyl, -SO 2 F or L 1 -R 7 and; L 1 -O-, -SO 2 - or -OSO 2 - and; R 7 is hydrogen, C 1~6 Alkyl or C 1~6 haloalkyl, R 7 C 1~6 Alkyl or C 1~6 Haloalkyl is -SO 2 -aryl, -OSO 2 -aryl, optionally substituted with 1 to 6 deuterium atoms, or combinations thereof; 2 -aryl or -OSO 2 -Aryl is cyano, hydroxy, halo, C 1~6 Alkyl, C 1~6 Haloalkyl, or C 1~6 is further optionally substituted with alkoxy; m is 0, 1, 2, or 3; n is 0, 1, or 2. or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0105] In some embodiments, the compound of formula I has formula II:

[0106] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0107] In some embodiments, the compound of formula I has formula III:

[0108] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0109] In some embodiments, the compound of formula I has formula IV:

[0110] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0111] In some embodiments, the compound of formula I has formula V:

[0112] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0113] In some embodiments, the compound of formula I has formula VI:

[0114] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0115] In some embodiments, the compound of formula I has formula VII:

[0116] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0117] In some embodiments, the compound of formula I has formula VIII:

[0118] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0119] In some embodiments, the compound of formula I has formula IX:

[0120] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0121] In some embodiments, the compound of formula I has formula X:

[0122] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0123] In some embodiments, the compound of formula I has formula XI:

[0124] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0125] In some embodiments, the compound of formula I has formula XII:

[0126] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0127] In some embodiments, the compound of formula I has formula XIII:

[0128] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0129] In some embodiments, the compound of formula I has formula XIV:

[0130] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0131] In some embodiments, the compound of formula I has formula XV:

[0132] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0133] In some embodiments, the compound of formula I has formula XVI:

[0134] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0135] In some embodiments, the compound of formula I has formula XVII:

[0136] [ka] or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0137] In some embodiments, R 1 is hydrogen or C 1~6 In some embodiments, R 1 is C 1~6 In some embodiments, R 1 is C 1~6 In some embodiments, R 1 is methyl.

[0138] In some embodiments, R 2 is hydrogen. In some embodiments, R 3 is hydrogen.

[0139] In some embodiments, R 4 is halo. In some embodiments, R 4 is fluoro.

[0140] In some embodiments, n is 1. In some embodiments, n is 0.

[0141] In some embodiments, R 11 is hydrogen.

[0142] In some embodiments, m is 1.

[0143] In some embodiments, R 5 is halo. In some embodiments, R 5 In some embodiments, n is 1 and R 5 is fluoro.

[0144] In some embodiments, m is 0.

[0145] In some embodiments, R 6 Is F or L 1 -R 7 In some embodiments, R 6 L 1 -R 7 And L 1 is -O-.

[0146] In some embodiments, R 7 is C 1~6 Haloalkyl or C 1~6 In some embodiments, R 7 is C substituted with 1 to 6 deuterium atoms 1~6 It is haloalkyl.

[0147] In some embodiments, R 6 is methoxy.

[0148] In some embodiments, X is N. In some embodiments, X is CR 11 In some embodiments, X is CH.

[0149] In some embodiments, L is CH 2 It is.

[0150] In some embodiments, Y 1 is N and Y 2 is CH.

[0151] In some embodiments, ring A is a heteroaryl containing one or two ring nitrogen atoms. In some embodiments, ring A is a heteroaryl containing one ring nitrogen atom. In some embodiments, ring A is pyridinyl. In some embodiments, ring A is pyridin-2-yl.

[0152] In some embodiments,

[0153] [ka] teeth,

[0154] [ka] and R 10 is C 1~6 It is an alkyl.

[0155] In some embodiments, the compound contains at least one halo. In some embodiments, the compound contains at least one fluoro. In some embodiments, the compound contains one fluoro. In some embodiments, the compound is substituted with at least one fluoro. In some embodiments, R 1 , R 4 , R 5 , R 6 , and R 11 At least one of R 4 , R 5 , R 6 , and R 11 At least one of R 4 , R5 , or R 11 In some embodiments, one of R is fluoro. 4 Or one R 11 is fluoro. In some embodiments, R 4 or R 11 One of them is fluoro.

[0156] In some embodiments, the compounds of formula I are labeled with one or more radioisotopes.

[0157] In some embodiments, the compound of formula I is 11 C. 13 N, 15 O, and 18 It contains one or more positron-emitting radioisotopes selected from F.

[0158] In some embodiments, the compound of formula I, or any compound described herein, is 3 H, 11 C. 13 N, 15 O, or 18 In some embodiments, the compound of formula I or any compound described herein is labeled with F. 3 H, 11 C, or 18 It is labeled with F.

[0159] In some embodiments, an imaging agent is provided comprising a compound of formula I, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0160] Also provided are additional compounds described herein. In some embodiments, a compound selected from Table 1, or an isotopically labeled analog, pharma- ceutically acceptable salt, solvate, prodrug, stereoisomer, or mixture of stereoisomers thereof is provided.

[0161] In some embodiments, a compound selected from the compounds of Table 1 is provided, optionally labeled with one or more radioactive isotopes, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

[0162] In some embodiments, a pharmaceutical composition is provided that includes a compound described herein, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof, and a pharma- ceutically acceptable excipient.

[0163] Non-metallic radionuclides can be covalently bound to the compounds described herein by reactions known from the state of the art. It is understood that if the radionuclides are metallic positron emitters, labeling may require the use of chelating agents. Such chelating agents are well known from the state of the art.

[0164] In some embodiments, a compound is provided that is selected from the compounds described in the Examples section provided herein.

[0165] Also provided is a compound selected from Table 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof:

[0166] [Table 2] TIFF2024513913000029.tif193169TIFF2024513913000030.tif192169TIFF2024513913000031.tif192169TIFF2024513913000032.tif123168

[0167] Diagnostic Methods and Uses In some embodiments, a method of detecting the presence or absence of an aggregation-prone protein in an individual is provided, comprising administering to the individual an effective amount of a compound or imaging agent described herein, and generating an image of a body part or region of the individual. Generating an image of a body part or region of the individual may include generating an image and detecting the presence or absence of the aggregation-prone protein in the image. Thus, the compounds disclosed herein are useful for detecting diseases or conditions mediated at least in part by proteins prone to protein aggregation. In some embodiments, the presence or absence of protein aggregates corresponds to the presence or absence of a neurodegenerative disease. In some embodiments, the neurodegenerative disease is selected from Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, Parkinson's disease, prion disease, and spinocerebellar ataxia.

[0168] Methods are provided for producing diagnostic images using positron emission tomography (PET) and for detecting the presence or absence of proteins prone to aggregation. PET imaging may be performed as known to those skilled in the art or as follows. PET imaging may involve administration of a positron-emitting radionuclide tracer, such as a compound or imaging agent described herein, to an individual. The tracer is then allowed to bind to the protein of interest for a sufficient period of time, at which point the individual is placed in a scanning device equipped with a scintillation detector ring. The emitted positron travels a short (isotope-dependent) distance within the tissues of the individual until it interacts with an electron. This interaction annihilates both the electron and the positron, generating a pair of photons. The photons are detected by a scintillator in the scanning device. Photons that do not result in a pair are ignored.

[0169] Methods of producing diagnostic images, including PET with simultaneous computed tomography imaging (PET / CT), PET with simultaneous magnetic resonance imaging (PET / MRI), or single photon emission computed tomography (SPECT) imaging, and methods of detecting the presence or absence of aggregation-prone proteins are also provided. Generally, computed tomography uses x-rays or gamma rays to detect brain structures, while magnetic resonance imaging uses magnetic fields and radio waves.

[0170] Thus, the compounds or imaging agents described herein can be administered by methods known in the art, including those described herein. The compounds or imaging agents can enter the circulation and bind to proteins prone to aggregation, or aggregates thereof. If the compounds or imaging agents are labeled with a radioisotope, the emitted particles can be detected.

[0171] In some embodiments, the compound or imaging agent is administered to the vascular system of the individual. The compound or imaging agent is capable of crossing the blood-brain barrier. Thus, generating an image can include generating an image of at least a portion of the brain of the individual, for example, a portion into which the compound is distributed.

[0172] Also provided are methods of generating diagnostic images in biological samples, and methods of detecting the presence or absence of aggregation-prone proteins, comprising contacting the biological sample with an effective amount of a compound or imaging agent described herein and generating an image associated with the biological sample. In some embodiments, the contacting and generating may be performed in vitro. In some embodiments, the contacting is in vivo and the generating is in vitro.

[0173] Also provided is a method for detecting the presence or absence of a pathological process associated with a protein prone to protein aggregation, such as huntingtin protein (HTT protein), in an individual, comprising administering an effective amount of a compound or imaging agent described herein; generating an image and detecting the presence or absence of huntingtin protein (HTT protein) in the image; and detecting the presence or absence of a pathological process, such as a neurodegenerative disease. In some embodiments, the HTT protein is present as a monomer, oligomer, or aggregate, or a combination thereof. In some embodiments, the protein prone to aggregation is huntingtin protein (HTT protein). The HTT protein may be a mutant. In some embodiments, the HTT protein is found in the brain, for example, in the basal ganglia.

[0174] In some embodiments, the body part or region is selected from the head, spinal cord, limbs, chest, and / or abdomen. In some embodiments, the body part or region is the brain. In some embodiments, the HTT protein is found in the basal ganglia. In some embodiments, the aggregation-prone protein, e.g., HTT protein, is present in the brain, liver, heart, and / or muscle of the individual. In some embodiments, generating an image comprises positron emission tomography (PET) imaging, PET with simultaneous computed tomography imaging (PET / CT), PET with simultaneous magnetic resonance imaging (PET / MRI), single photon emission computed tomography (SPECT) imaging, or a combination thereof. In some embodiments, generating an image comprises PET imaging. In some embodiments, the aggregation-prone protein, e.g., HTT protein, is present in the basal ganglia, cortex, hippocampus, and / or brainstem of the brain of the individual. In some embodiments, the aggregation-prone protein, e.g., HTT protein, is present as a monomer, oligomer, or aggregate, or a combination thereof.

[0175] In some embodiments, the individual has or is found to have Huntington's disease.

[0176] Also provided are methods of detecting the presence or absence of a pathological process associated with β-amyloid protein in an individual, comprising administering an effective amount of a compound or imaging agent described herein; generating an image of a body part or region of the individual; and detecting the presence or absence of the pathological process. In some embodiments, the individual has or is found to have Alzheimer's disease (AD).

[0177] Also provided are diagnostic methods using the compounds or imaging agents described herein to monitor disease progression in a patient by quantifying changes in the levels of aggregation-prone proteins in the patient.

[0178] In some embodiments, compounds are provided that have a binding rate for protein aggregates, such as HTT protein aggregates or β-amyloid protein aggregates, suitable for functioning as imaging agents. Thus, the compounds described herein may be characterized by one or more of: 1) a high affinity for such protein aggregates; 2) a low affinity for adjacent structures; and / or 3) a slow dissociation rate from such protein aggregates. The dissociation rate can be calculated according to the following formula, where A and B refer to the protein aggregate and the imaging agent, and k assn is the association rate constant), the dissociation rate constant k diss It can be expressed as: d[AB] / dt = k assn [A][B] - k diss [AB]

[0179] In some embodiments, an effective amount of a compound or imaging agent described herein comprises about 0.1 to about 20 mCi. In some embodiments, an effective amount of a compound or imaging agent described herein comprises about 0.1, about 0.3, about 0.5, about 0.7, about 1, about 3, about 5, about 7, about 10, about 15, or about 20 mCi, or any range therebetween. In some embodiments, an effective amount of a compound or imaging agent described herein comprises about 10 mCi.

[0180] Suitable radionuclides that can be incorporated into the compounds described herein include, but are not limited to, 3 H (also written as T), 11 C. 18 F, 35 S, 123 I, 125 I, 75 Br, 76 Br, 77 Br, 82 Br, 131 I, 15 O. 13 N, and 211 The radionuclide incorporated into the compound depends on the particular imaging application. In some embodiments, including PET imaging, 11 C. 18 F, 123 I, 131 I, 75 Br, 76 Br or 77 Compounds incorporating a radionuclide selected from: 99m Incorporation of chelated radionuclides, such as Tc, may also be useful. 18 Due to the longer half-life of F, imaging can be performed for a long enough time for a stronger signal to develop. 18 F 11 In some embodiments, the compounds or imaging agents described herein may be labeled with a positron-emitting radionuclide or a gamma-emitting radionuclide. Some examples of positron-emitting radionuclides include: 15 O. 13 N, 11 C. 18 F, 76 Br, and 124 I, which have half-lives of approximately 2, 10, 20, 110 minutes, 16 hours, and 4.2 days, respectively.

[0181] In some embodiments, the compounds or imaging agents described herein include 3 H,11 C, or 18 In some embodiments, the compounds or imaging agents described herein may be labeled with F. 11 C and 18 The compound may be labeled with a positron emitter selected from F. 11 The method for introducing C is 11 C] iodomethane or [ 11 Carbon-11 has a half-life of approximately 20 minutes, and thus 11 C typically needs to be produced in an on-site cyclotron, 11 C] can be produced as carbon dioxide. 11 C] Carbon dioxide is a suitable species for direct labeling (generally [ 11 C]iodomethane, and the synthesis of the radiopharmaceutical is completed on-site for use in PET imaging studies after appropriate radiochemical purity and specific activity have been determined. 18 Exemplary methods for introducing F include, but are not limited to, nucleophilic and electrophilic methods. Nucleophilic methods include replacing halides, tosylates, or other leaving groups with labeled cesium fluoride, potassium fluoride, tetrabutylammonium fluoride, tetramethylammonium fluoride, or potassium fluoride kryptofix-222. 18Electrophiles that may be suitable for introducing [F] isotopes include labeled diethylaminosulfur trifluoride (DAST), bis(2-methoxyethyl)aminosulfur trifluoride (Deoxofluor), N-fluorobenzenesulfonimide (NFSI), N-fluoropyridinium salts, 1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate) (Selectfluor), N-fluoropyridinium triflate, xenon fluoride, 2-pyridinesulfonyl fluoride (PyFluor), 3-pyridinesulfonyl fluoride, 4-pyridinesulfonyl fluoride, 4-chloro-2-pyridinesulfonyl fluoride, ethenesulfonyl fluoride, fluoro-benzoiodoxol, p-fluorophenylaminosulfur trifluoride, p-nitrophenylaminosulfur trifluoride, or pentafluorophenylaminosulfur trifluoride. General methods for the introduction of positron emitters are described in the literature (see, for example, Miller et al., Angewandte Chemie International Edition, Vol. 47 (2008), pp. 8998-9033; Jacobson, O. et al., Bioconjugate Chem., Vol. 26 (2015), pp. 1-18; Deng, X. et al., Angewandte Chemie International Edition, Vol. 58 (No. 9), (2019), pp. 2580-2605). The method of introducing tritium can be achieved according to methods known in the art, such as synthetic methods, recoil, or exchange reactions.

[0182] Fluorine-18 has a half-life of approximately 110 minutes, so [ 18Synthesis of the [F] radiopharmaceutical does not necessarily have to be performed at the site of a cyclotron or in close proximity to a PET imaging research center. Fluorine-18 is also believed to exhibit favorable nuclear and physical properties, including a high positron decay ratio (97%), a relatively short half-life (109.7 minutes), and low positron energy (up to 0.635 MeV). This positron energy can accommodate a short diffusion range in vivo (<2.4 mm), which may provide an excellent resolution limit for PET imaging.

[0183] As will be appreciated, the steps of the methods described herein do not have to be performed a particular number of times or in a particular order. Additional objects, advantages and novel features of the present disclosure will become apparent to those skilled in the art upon examination of the following examples, which are intended to be illustrative and not limiting.

[0184] Indications and Treatment Methods The compounds or imaging agents described herein may be useful for treating a disease or condition mediated at least in part by an aggregation-prone protein. In some embodiments, the compounds or imaging agents described herein are useful for treating a disease or condition mediated at least in part by an HTT protein. In some embodiments, the treatment of a disease or condition mediated at least in part by an aggregation-prone protein may include administration of a compound or imaging agent described herein. The treatment may include co-administration of a compound or imaging agent described herein with one or more other active agents and / or therapies. Thus, in some embodiments, a method is provided for treating or preventing a disease or condition mediated at least in part by an aggregation-prone protein in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound or imaging agent described herein.

[0185] Exemplary diseases and conditions are provided below.

[0186] Huntington's Disease (HD) Huntington's disease (HD) is an inherited progressive neurodegenerative disorder characterized by motor, cognitive, and psychiatric impairments as well as neurodegeneration and brain atrophy. Atrophy begins in the striatum and cortex and may extend to other subcortical brain regions. HD belongs to a group of neurodegenerative disorders in which expanded CAG repeats result in long stretches of polyglutamine (polyQ) in the encoded protein. The group also includes dentatorubral-pallidoluysian atrophy (DRPLA), spinal-bulbar muscular atrophy (SBMA), and spinocerebellar ataxias (SCAs). In HD, selective neurodegeneration of gamma-aminobutyric acid releasing spiny-projection neurons of the striatum has been observed, but neuronal loss in many other brain regions has also been reported. Symptoms of HD include loss of motor control, psychiatric symptoms, memory and / or cognitive impairment.

[0187] The HD protein huntingtin (HTT protein) is a 348 kDa multidomain protein that contains a polymorphic glutamine / proline-rich domain at its amino terminus. 15 The number of CAG repeats in the gene varies from 6 to 35 in healthy individuals, with 36 or more repeats defining the HD allele. The length of the CAG expansion is inversely correlated with the age of disease onset, with early-onset cases characterized by more than 60 repeats of the expansion. It is believed that longer polyQ domains cause conformational changes in the HTT protein that lead to the formation of intracellular aggregates that often appear as nuclear inclusions; however, aggregates can also form outside of the nucleus. The HTT protein is present in the nucleus, cell body, dendrites, and nerve terminals of neurons, and is also associated with many organelles, including the Golgi apparatus, endoplasmic reticulum, and mitochondria.

[0188] The parts of the brain most affected by HD, and therefore most likely to contain HTT protein abnormalities, are a group of nerve cells at the base of the brain collectively known as the basal ganglia. The basal ganglia organize muscle-propelled body movements, or "motor activity." The main components of the basal ganglia are the caudate nucleus and putamen (together known as the striatum) and the globus pallidus (external and internal regions). The substantia nigra and subthalamic nucleus are often included as part of the basal ganglia as well.

[0189] The basal ganglia are a group of subcortical nuclei that are primarily responsible for motor control, as well as other roles such as motor learning, executive function and behavior, and emotion. It is believed that disruption of the basal ganglia reticular system leads to several motor disorders. The normal function of the basal ganglia requires fine-tuning of neuronal excitability within each nucleus to determine the degree of motor facilitation or inhibition at any given time. This is mediated by the complex organization of the striatum, where the excitability of medium spiny neurons is controlled by several pre- and post-synaptic mechanisms as well as interneuron activity, and is ensured by several recurrent or internal basal ganglia circuits. The motor circuit of the basal ganglia has two input points, the striatum and the subthalamic nucleus, and one output point, the internal pallidal segment, which connects to the cortex via the motor thalamus.

[0190] Administration of a compound described herein may result in a reduction, e.g., at least a 10% reduction (e.g., at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 100%), in one or more symptoms of a disease or condition described herein. The disease or condition may be a nervous system disorder secondary to a disease, condition, or therapy that has a primary effect outside the nervous system; damage to the nervous system caused by physical, mechanical, or chemical trauma; autoimmune neurodegeneration; neurodegeneration secondary to an infection; and / or ocular neurodegeneration. Symptoms of neurodegeneration include, for example, tremors, bradykinesia, ataxia, loss of balance, depression, cognitive decline, short-term memory loss, long-term memory loss, confusion, personality changes, speech problems, loss of sensation, sensitivity to touch, numbness in the limbs, muscle weakness, muscle paralysis, muscle spasms, muscle cramps, significant changes in eating habits, excessive anxiety or worry, insomnia, delusions, hallucinations, fatigue, back pain, chest pain, indigestion, headache, rapid heart rate, dizziness, blurred vision, shadows or missing areas in vision, metamorphopsia, impairment in color vision, decreased recovery of visual function after exposure to bright light, and loss of visual contrast sensitivity.

[0191] Neurodegenerative diseases are diseases or conditions that impair the function of the nervous system of a subject. Examples of neurodegenerative diseases include, for example, Alexander's disease, Alper's disease, Alzheimer's disease, amyotrophic lateral sclerosis, ataxia telangiectasia, Batten disease (also known as Spielmeyer-Vogt-Sjogren-Batten disease), bovine spongiform encephalopathy (BSE), Canavan disease, Cockayne syndrome, corticobasal degeneration, Creutzfeldt-Jakob disease, frontotemporal dementia, Gerstmann-Straussler-Scheinker syndrome, Huntington's disease, HIV-associated dementia, Kennedy's disease, and Krabbe's disease. , kuru, dementia with Lewy bodies, Machado-Joseph disease (spinocerebellar ataxia type 3), multiple sclerosis, multiple system atrophy, narcolepsy, neuroborreliosis, Parkinson's disease, Pelizaeus-Merzbacher disease, Pick's disease, primary lateral sclerosis, prion disease, Refsum disease, Sandhoff disease, Schilder's disease, subacute combined spinal cord degeneration secondary to pernicious anemia, schizophrenia, spinocerebellar ataxia, spinal muscular atrophy, Steele-Richardson-Olszewski disease, insulin resistance, or tabes dorsalis.

[0192] In some embodiments, the disease or condition is selected from Huntington's disease (HD), dentatorubral-pallidoluysian atrophy, spinal-bulbar muscular atrophy, spinocerebellar ataxia, spinal cord and / or brain injury, chronic pulmonary hypertension, Parkinson's disease, amyotrophic lateral sclerosis, cerebral cavernous hemangioma, cardiovascular disease, Alzheimer's disease (AD), glaucoma, multiple sclerosis (MS), corneal lesions, diabetes, chronic and / or neuropathic pain, stroke, ischemia, retinal disease, spinal muscular atrophy (SMA), erectile dysfunction, (non-hypertensive) nephropathy, hypertensive nephropathy, high blood pressure (hypertension), optic nerve damage, liver fibrosis, lupus, post-transplant liver failure, encephalomyelitis, epilepsy, and glioblastoma.

[0193] The compounds described herein may inhibit neuronal degeneration when administered to a subject. In some embodiments, inhibiting neuronal degeneration may include inhibiting the degeneration of axons or neurons in a neuron. Such inhibition may involve whole neurons or portions thereof, such as neuronal cell bodies, axons, and dendrites. This may be assessed, for example, by analyzing nervous system function by methods known in the art. Administration of a compound described herein may result in at least a 10% decrease (e.g., at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%) in the number of neurons (or their neuronal bodies, axons, or dendrites) that degenerate in a neuronal population or subject compared to the number of neurons (or their neuronal bodies, axons, or dendrites) that degenerate in a neuronal population or subject to which one or more compounds described herein are not administered.

[0194] Neurons can convey information from tissues and organs to the central nervous system (afferent or sensory neurons) and transmit signals from the central nervous system to effector cells (efferent or motor neurons). Other neurons, designated interneurons, connect neurons within the central nervous system (brain and spinal column). Certain illustrative examples of neuronal types that may be subject to treatment according to the present disclosure include cerebellar granule neurons, dorsal root ganglion neurons, PNS neurons (e.g., sensory neurons), and cortical neurons. Other examples of cell types that may be subject to treatment according to the present disclosure include astrocytes and microglia.

[0195] Additionally, the compounds described herein can be used to prevent or treat memory loss. Types of memory that are affected by loss and thus can be treated by the present disclosure include episodic memory, semantic memory, short-term memory, and long-term memory.

[0196] In some embodiments, the disease or condition is a neurodegenerative disease selected from Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, Parkinson's disease, prion disease, and spinocerebellar ataxia. In some embodiments, the neurodegenerative disease is classified as a triplet repeat disease. In some embodiments, the triplet repeat disease is classified as belonging to category I, category II, or category III.

[0197] In some embodiments, the pathological process is associated with or caused by a disease or condition selected from Huntington's disease (HD), dentatorubral-pallidoluysian atrophy, spinal-bulbar muscular atrophy, spinocerebellar ataxia, spinal cord and / or brain injury, chronic pulmonary hypertension, Parkinson's disease, amyotrophic lateral sclerosis, cerebral cavernous hemangioma, cardiovascular disease, Alzheimer's disease (AD), glaucoma, multiple sclerosis (MS), corneal lesions, diabetes, chronic and / or neuropathic pain, stroke, ischemia, retinal disease, spinal muscular atrophy (SMA), erectile dysfunction, (non-hypertensive) nephropathy, hypertensive nephropathy, high blood pressure (hypertension), optic nerve damage, liver fibrosis, lupus, post-transplant liver failure, encephalomyelitis, epilepsy, and glioblastoma. In some embodiments, the pathological process is a neurodegenerative disease selected from Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, Parkinson's disease, prion disease, and spinocerebellar ataxia. In some embodiments, the neurodegenerative disease is classified as a triplet repeat disease. In some embodiments, the triplet repeat disease is classified as belonging to category I, category II, or category III.

[0198] In some embodiments, the neurodegenerative disease is Huntington's disease.

[0199] Also provided is the use of a compound described herein for the manufacture of a medicament for use in the diagnosis, prevention, or treatment of a disease or condition described herein. For example, the disease or condition can be Huntington's disease.

[0200] Imaging agents and pharmaceutical compositions Imaging agents generally include compounds described herein labeled with a positron-emitting radionuclide. Imaging agents labeled with a positron-emitting radionuclide are generally administered via intravenous injection immediately (e.g., within 1 hour of synthesis) due to the short half-life of the radionuclide. The amount of imaging agent required is typically determined by the prescribing physician. Dosage may vary depending on a variety of factors, including, but not limited to, the association rate of the compound, the amount of emission from the radionuclide used, the half-life of the radionuclide, the part of the body, region of the body, and / or tissue to be imaged, and individual characteristics. One of skill in the art will appreciate that an effective amount will generally be an amount of labeled compound sufficient to produce an emission in the range of about 0.1 to about 20 mCi, or about 1 to about 5 mCi. The mass of the labeled compound in an effective amount of imaging agent may be about 0.1 to about 500 mg.

[0201] In general, the compounds or imaging agents described herein can be administered to a patient in need thereof via any suitable route. Routes of administration can include parenteral administration, such as subcutaneous, intramuscular, intravenous, for example, via a drip patch. Further suitable routes of administration include, but are not limited to, oral, rectal, intranasal, topical (including buccal and sublingual), injection, vaginal, intradermal, intraperitoneal, intracranial, intrathecal, and epidural administration, for example, via a spray or inhaler, or via implant, or via oral or nasal inhalation.

[0202] For PET imaging, administration of the compounds or imaging agents described herein to an individual may be intravenous. The pharmaceutical compositions may take the form of a sterile injectable aqueous or oleaginous suspension. The suspension may be formulated according to known techniques using those suitable dispersing or wetting agents and suspending agents listed above. The sterile injectable product may also be a sterile injectable solution or suspension in a non-toxic, parenterally acceptable vehicle, for example a solution in 1,3-butanediol. Among the acceptable vehicles that may be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as solvents or suspending media. For this purpose, any non-irritating fixed oil may be employed, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid may be useful in the preparation of injectables. Such solutions may be formulated as 0.01% to 10% isotonic solutions, pH 5 to 7, with appropriate salts.

[0203] The compounds or imaging agents described herein may be administered parenterally in a sterile medium. Parenteral administration includes subcutaneous injection, intravenous, intramuscular, intrathecal injection or infusion techniques. The compounds or imaging agents described herein may be suspended or dissolved in the vehicle, depending on the vehicle and concentration used. Advantageously, adjuvants such as local anesthetics, preservatives and buffers can be dissolved in the vehicle. In most pharmaceutical compositions for parenteral administration, the carrier accounts for at least 90% by weight of the total amount of the composition. In some embodiments, the carrier for parenteral administration is selected from propylene glycol, ethyl oleate, pyrrolidone, ethanol and sesame oil.

[0204] The pharmaceutical composition, e.g., an injectable pharmaceutical composition, can include a cyclodextrin. The cyclodextrin can be, for example, hydroxypropyl cyclodextrin or sulfobutylether cyclodextrin. The cyclodextrin can be, for example, α-cyclodextrin, β-cyclodextrin, or γ-cyclodextrin.

[0205] The compounds or imaging agents described herein may be administered via microspheres, liposomes, other microparticulate delivery systems, or sustained release formulations that are placed in specific tissues, including blood.Suitable examples of sustained release carriers include semipermeable polymer matrices in the form of co-medical products, such as suppositories or microcapsules.Examples of the techniques and protocols mentioned above and other techniques and protocols that can be used according to the present invention can be found in Remington's Pharmaceutical Sciences, 18th Edition, Gennaro, AR, Lippincott Williams & Wilkins; 20th Edition (December 15, 2000) ISBN 0-912734-04-3 and Pharmaceutical Dosage Forms and Drug Delivery Systems, Ansel, NC et al., 7th Edition, ISBN 0-683305-72-7, the entire contents of which are incorporated herein by reference.

[0206] In some embodiments, the compound or imaging agent described herein is administered as a pharmaceutical composition.Therefore, a pharmaceutical composition is provided that comprises at least one compound or imaging agent described herein together with at least one pharma- ceutically acceptable vehicle selected from carrier, adjuvant, and excipient.The compound or imaging agent of the present disclosure can be formulated into a pharmaceutical composition using techniques known to those skilled in the art.

[0207] Pharmaceutically acceptable vehicles must be of sufficiently high purity and sufficiently low toxicity to make them suitable for administration to the animal being treated. The vehicle can be inert or it can have a pharmaceutical benefit. The amount of vehicle used with the compound or imaging agent can be sufficient to provide a practical amount of material for administration per dose of the compound or imaging agent.

[0208] Exemplary pharma- ceutically acceptable carriers or components thereof are sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and methylcellulose; powdered tragacanth; malt; gelatin; talc; solid lubricants, such as stearic acid and magnesium stearate; calcium sulfate; synthetic oils; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil, and corn oil; polyols, such as propylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol; alginic acid; phosphate buffer solutions; emulsifiers, such as TWEEN®; wetting agents, such as sodium lauryl sulfate; colorants; flavorings; tableting agents; stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline; and phosphate buffer solutions.

[0209] Optional active agents may be included in the pharmaceutical compositions, which do not substantially interfere with the activity of the compounds or imaging agents described herein.

[0210] At least one compound or imaging agent described herein in effective concentration is mixed with a suitable pharma- ceutically acceptable vehicle.If the compound or imaging agent shows insufficient solubility, methods for solubilizing the compound may be used.Such methods are known to those skilled in the art and include, but are not limited to, using cosolvents such as dimethylsulfoxide (DMSO), using surfactants such as TWEEN®, or dissolving in aqueous buffer, for example sodium bicarbonate.

[0211] Upon mixing or addition of the compounds or imaging agents described herein, the resulting mixture may be a solution, suspension, emulsion, etc. The form of the resulting mixture will depend on many factors, including the intended method of administration and the solubility of the compound or imaging agent in the selected vehicle. Effective concentrations sufficient for imaging or treatment may be determined empirically by methods known in the art.

[0212] The pharmaceutical compositions may be formulated for oral use, such as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Pharmaceutical compositions intended for oral use may be prepared according to any method known to one skilled in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents, such as sweeteners, flavoring agents, coloring agents, and preservatives, to provide a medicamentically simple and palatable product. In some embodiments, the oral pharmaceutical composition contains 0.1-99% of the compound or imaging agent described herein. In some embodiments, the oral pharmaceutical composition contains at least 5% (by weight) of the compound or imaging agent. Some embodiments contain 25%-50% or 5%-75% of the compound or imaging agent.

[0213] Orally administered pharmaceutical compositions also include liquid solutions, emulsions, suspensions, powders, granules, elixirs, tinctures, syrups, etc. Suitable pharma- ceutically acceptable carriers for preparing such compositions are well known in the art. Oral pharmaceutical compositions may also contain preservatives, flavoring agents, sweeteners such as sucrose or saccharin, taste-masking agents, and coloring agents.

[0214] Typical components of carriers for syrups, elixirs, emulsions and suspensions include ethanol, glycerol, propylene glycol, polyethylene glycol, liquid sucrose, sorbitol and water. Syrups and elixirs may be formulated with sweeteners, such as glycerol, propylene glycol, sorbitol or sucrose. Such pharmaceutical compositions may also contain a demulcent.

[0215] The compounds or imaging agents described herein may be incorporated into oral liquid products such as aqueous or oily suspensions, solutions, emulsions, syrups, or elixirs. Additionally, pharmaceutical compositions containing the compounds or imaging agents described herein may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid products may contain conventional additives such as suspending agents (e.g., sorbitol syrup, methylcellulose, glucose / sugar, syrup, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel, and hydrogenated edible fats), emulsifiers (e.g., lecithin, sorbitan monooleate, or acacia), non-aqueous vehicles, which may include edible oils (e.g., almond oil, fractionated coconut oil, silyl esters, propylene glycol, and ethyl alcohol), and preservatives (e.g., methyl or propyl p-hydroxybenzoate, and sorbic acid).

[0216] For suspensions, typical suspending agents include methylcellulose, sodium carboxymethylcellulose, Avicel® RC-591, tragacanth, and sodium alginate, typical wetting agents include lecithin and polysorbate 80, and typical preservatives include methylparaben and sodium benzoate.

[0217] Aqueous suspensions are provided that contain compound or imaging agent in admixture with excipients suitable for producing aqueous suspensions.Such excipients are suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing or wetting agents can be naturally occurring phosphatides, such as lecithin, or condensation products of alkylene oxides with fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, such as heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitol replacements, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyethylene sorbitan replacements.Aqueous suspensions can also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate.

[0218] Oily suspensions can be formulated by suspending the compound or imaging agent in a vegetable oil, such as peanut oil, olive oil, sesame oil or coconut oil, or in a mineral oil, such as liquid paraffin. Oily suspensions can contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents, such as those set forth above, and flavoring agents can be added to provide a palatable oral product. These pharmaceutical compositions can be preserved by the addition of an antioxidant, such as ascorbic acid.

[0219] The pharmaceutical composition may be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, such as olive oil or peanut oil, or a mineral oil, such as liquid paraffin, or a mixture thereof. Suitable emulsifiers may be naturally occurring gums, such as gum acacia or gum tragacanth, naturally occurring phosphatides, such as soybean, lecithin, and esters or partial esters derived from fatty acids and hexitols, anhydrides, such as sorbitan monooleate, and the condensation products of said partial esters with ethylene oxide, such as sorbitan monooleate polyoxyethylene.

[0220] Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above.

[0221] Tablets typically contain, as inert diluents, conventional pharma- ceutically acceptable adjuvants, such as calcium carbonate, sodium carbonate, mannitol, lactose and cellulose; binders, such as starch, gelatin and sucrose; disintegrants, such as starch, alginic acid and croscarmellose; lubricants, such as magnesium stearate, stearic acid and talc. Glidants, such as silicon dioxide, can be used to improve the flow properties of the powder mixture. Coloring agents, such as FD&C dyes, can be added for appearance. Sweeteners and flavoring agents, such as aspartame, saccharin, menthol, peppermint, and fruit flavors, can be useful adjuvants for chewable tablets. Capsules (including time-release and sustained-release formulations) typically contain one or more solid diluents as disclosed above. The selection of carrier components is often driven by secondary considerations, such as taste, cost, and storage stability.

[0222] The pharmaceutical compositions may be coated by conventional methods, typically with pH or time dependent coatings, so that the compound or imaging agent is released in the gastrointestinal tract near the desired local application or at various times to extend the desired effect. Such dosage forms typically include, but are not limited to, one or more of cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate, ethylcellulose, Eudragit® coatings, waxes and shellac.

[0223] Pharmaceutical compositions for oral use may also be presented as hard gelatin capsules, in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules, in which the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin, or olive oil.

[0224] The compounds or imaging agents described herein may be administered in the form of suppositories for rectal administration of drugs.These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at normal temperature but liquid at rectal temperature, and therefore melts in the rectum to release the drug.Such materials include cocoa butter and polyethylene glycol.

[0225] The compounds or imaging agents described herein may be formulated for local or topical application, e.g., for topical application to the skin and mucous membranes, such as in the eye, in the form of gels, creams, and lotions, as well as for application to the eye. Topical pharmaceutical compositions may be in any form, including, for example, solutions, creams, ointments, gels, lotions, emulsions, cleansers, moisturizers, sprays, skin patches, and the like.

[0226] Topical pharmaceutical compositions comprising at least one compound described herein, or an isotopically labeled analog, pharma- ceutically acceptable salt, solvate, prodrug, stereoisomer, or mixture of stereoisomers thereof, may be mixed with a variety of carrier materials known in the art, such as water, alcohol, aloe vera gel, allantoin, glycerin, vitamin A and E oil, mineral oil, propylene glycol, PPG-2 myristyl propionate, and the like.

[0227] Other materials suitable for use in topical carriers include, for example, emollients, solvents, humectants, thickeners, and powders. Examples of each of these types of materials that can be used alone or in mixtures with one or more other materials are as follows:

[0228] Representative softening agents include stearyl alcohol, glyceryl monoricinoleate, glyceryl monostearate, propane-1,2-diol, butane-1,3-diol, mink oil, cetyl alcohol, isopropyl isostearate, stearic acid, isobutyl palmitate, isocetyl stearate, oleyl alcohol, isopropyl laurate, hexyl laurate, decyl oleate, octadecane-2-ol, isocetyl alcohol, cetyl palmitate, dimethylpolysiloxane, diisopropyl sebacic acid, and ethylhexyl stearate. -n-butyl, isopropyl myristate, isopropyl palmitate, isopropyl stearate, butyl stearate, polyethylene glycol, triethylene glycol, lanolin, sesame oil, coconut oil, peanut oil, castor oil, acetylated lanolin alcohol, petroleum oil, mineral oil, butyl myristate, isostearic acid, palmitic acid, isopropyl linoleate, lauryl lactate, myristyl lactate, decyl oleate, and myristyl myristate; propellants include propane, butane, isobutane solvents include ethyl alcohol, methylene chloride, isopropanol, castor oil, ethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol monoethyl ether, dimethyl sulfoxide, dimethylformamide, tetrahydrofuran; humectants include glycerin, sorbitol, sodium 2-pyrrolidone-5-carboxylate, soluble collagen, dibutyl phthalate, and gelatin; and powders include chalk, talc, fuller's earth, kaolin, starch, gum, colloidal silicon dioxide, sodium polyacrylate, tetraalkylammonium smectite, trialkylarylammonium smectite, chemically modified magnesium aluminum silicate, organically modified montmorillonite clay, hydrated aluminum silicate, fumed silica, carboxyvinyl polymer, sodium carboxymethylcellulose, and ethylene glycol monostearate.

[0229] The compounds or imaging agents described herein may also be formulated for transdermal administration as a transdermal patch.

[0230] The compounds or imaging agents described herein may also be administered in liposome delivery systems. Liposomes can be classified into small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of amphiphilic molecules, especially phospholipids. Liposome constituents include cholesterol, stearylamine, and / or phosphatidylcholine. Liposomes are suitable for a variety of routes of administration, including topical and injection into various tissues. Thus, intravitreal (e.g., in the treatment of glaucoma), intraperitoneal, intravenous, intravascular, intraarticular, and intramuscular administration of liposomes is contemplated.

[0231] Other pharmaceutical compositions useful for achieving systemic delivery of compound or imaging agent include sublingual, buccal and intranasal dosage forms.Such pharmaceutical compositions typically include soluble filler materials such as sucrose, sorbitol and mannitol, and binders such as one or more of acacia, microcrystalline cellulose, carboxymethylcellulose and hydroxypropylmethylcellulose.The glidants, lubricants, sweeteners, colorants, antioxidants and flavoring agents disclosed above may also be included.

[0232] Pharmaceutical compositions for inhalation can typically be provided in the form of a solution, suspension or emulsion, which can be administered as a dry powder or in the form of an aerosol using conventional propellants such as dichlorodifluoromethane or trichlorofluoromethane.

[0233] The pharmaceutical compositions may optionally include an activity enhancer. Activity enhancers may be selected from a wide variety of molecules that function in a variety of ways that enhance or are independent of the therapeutic effect of the compounds or imaging agents described herein. Particular classes of activity enhancers include skin penetration enhancers and absorption enhancers.

[0234] The pharmaceutical composition may contain additional active agents, which may be selected from a wide variety of molecules that may function in various ways to enhance the therapeutic effect of the compounds or imaging agents described herein. These optional other active agents, when present, are typically utilized in the pharmaceutical composition at levels ranging from 0.01% to 15%. In some embodiments, they comprise 0.1% to 10% by weight of the composition. In other embodiments, they comprise 0.5% to 5% by weight of the composition.

[0235] The dosage of the compounds or imaging agents described herein will depend on a variety of factors, including, among other considerations, the particular pathological process to be treated or detected, the physiology of the individual, the severity of the symptoms, the route of administration, the frequency of dosing intervals, the particular compound utilized, the efficacy of the compound, the toxicological profile, the pharmacokinetic profile, and the presence of toxic side effects. The dosage for a given situation will generally be determined appropriately by the practitioner based on these and other factors.

[0236] The compounds or imaging agents described herein are typically administered in a manner determined by a practitioner, such as a physician, at dosage levels. For example, the compounds or imaging agents may be administered in single or multiple doses, generally at dosage levels of 0.001-100 mg / kg, such as 0.01-100 mg / kg, such as 0.1-70 mg / kg, such as 0.5-10 mg / kg. The doses may be, for example, for once-daily or twice-daily administration. A unit dosage form may generally contain 0.01-1000 mg, such as 0.1-50 mg, of the compounds or imaging agents described herein. For intravenous administration, the compounds or imaging agents may be administered in single or multiple doses, generally at dosage levels of 0.001-50 mg / kg, such as 0.001-10 mg / kg, such as 0.01-1 mg / kg. A unit dosage form may contain, for example, 0.1 to 10 mg of compound or imaging agent.

[0237] Kits and Packaging Also provided herein is a kit comprising the compound described herein and suitable packaging. In certain embodiments, the kit further comprises instructions for use. In some embodiments, the kit comprises a compound or imaging agent described herein and a label and / or instructions for use of the compound in treating an indication, including a disease or condition described herein.

[0238] Also provided herein is an article of manufacture comprising a compound or imaging agent described herein in a suitable container, which may be a vial, bottle, ampoule, pre-filled syringe, and intravenous bag.

[0239] Packaged pharmaceutical compositions are also provided. Such packaged compositions include a pharmaceutical composition comprising a compound or imaging agent described herein, and instructions for using the composition to treat a subject (typically a human patient). In some embodiments, the instructions are for using the pharmaceutical composition to detect a disease or condition described herein. The packaged pharmaceutical composition may include prescribing information, for example, to a patient or health care provider, or as a label on the packaged pharmaceutical composition. Prescribing information may include, for example, efficacy, dosage and administration, contraindications, and adverse reaction information regarding the pharmaceutical composition.

[0240] In all of the foregoing, the compounds or imaging agents may be administered alone, as mixtures, or in combination with other active agents.

[0241] Also provided is the use of a compound or imaging agent described herein for the manufacture of a medicament for use in the diagnosis, prevention, or treatment of a disease or condition described herein. For example, the disease or condition may be Huntington's disease.

[0242] Also provided is the use of a compound described herein for the manufacture of an imaging agent for use in the diagnosis, prevention, or treatment of a disease or condition described herein. For example, the disease or condition may be Huntington's disease.

[0243] Combination treatment The methods described herein include methods for detecting, treating or preventing a disease or condition described herein, comprising administering to a subject a compound or imaging agent described herein and one or more additional active agents simultaneously or sequentially. For example, the disease or condition may be Huntington's disease. In methods using simultaneous administration, the agents may be in a combined composition or may be administered separately. When used in combination with one or more additional active agents, the compound or imaging agent described herein may be administered before, simultaneously with, or after administration of the additional active agents. Administration may be by the same route or by different routes.

[0244] Also provided are pharmaceutical compositions comprising a compound or imaging agent described herein and one or more additional active agents used in the treatment of Huntington's disease, such as, but not limited to, carbamazepine, clonazepam, diazepam, fluoxetine, escitalopram, valproate, lamotrigine, amitriptyline, imipramine, desipramine, nortriptyline, paroxetine, fluoxetine, sertraline, tetrabenazine, haloperidol, chlorpromazine, thioridazine, sulpiride, quetiapine, clozapine, and risperidone. Also provided are packaged pharmaceutical compositions comprising a pharmaceutical composition comprising a compound or imaging agent described herein and another composition comprising one or more additional active agents used in the treatment of Huntington's disease, such as, but not limited to, carbamazepine, clonazepam, diazepam, fluoxetine, escitalopram, valproate, lamotrigine, amitriptyline, imipramine, desipramine, nortriptyline, paroxetine, fluoxetine, sertraline, tetrabenazine, haloperidol, chlorpromazine, thioridazine, sulpiride, quetiapine, clozapine, and risperidone. In some embodiments, the active agent is carbamazepine, clonazepam, diazepam, fluoxetine, escitalopram, valproate, lamotrigine, amitriptyline, imipramine, desipramine, nortriptyline, paroxetine, fluoxetine, sertraline, tetrabenazine, haloperidol, chlorpromazine, thioridazine, sulpiride, quetiapine, clozapine, or risperidone.

[0245] Also provided are methods for treating or preventing Alzheimer's disease, including treating memory and / or cognitive impairment associated with Alzheimer's disease, comprising administering to a subject, simultaneously or sequentially, a compound or imaging agent described herein and one or more additional agents. In some embodiments, the active agent is Reminyl®, Cognex®, Aricept®, Exelon®, Akatinol®, Neotropin™, Eldepryl®, estrogen, or clioquinol.

[0246] In some embodiments, the compounds described herein can be administered with an active agent for treating Parkinson's disease, for example, L-dopa, dopamine agonists (e.g., bromocriptine, pergolide, pramipexole, ropinirole, cabergoline, apomorphine, and lisuride), dopa decarboxylase inhibitors (e.g., levodopa, benserazide, and carbidopa), and / or MAO-B inhibitors (e.g., selegiline and rasagiline). In some embodiments, the compounds described herein can be administered with an active agent for treating Alzheimer's disease, for example, acetylcholinesterase inhibitors (e.g., donepezil, galantamine, and rivastigmine) and / or NMDA receptor antagonists (e.g., memantine).

[0247] Synthesis of compounds The compounds described herein can be prepared using the methods disclosed herein and routine variations thereof that are evident from the disclosures herein and methods well known in the art. Conventional and well known synthetic methods can be used in addition to the teachings herein. The synthesis of typical compounds described herein can be achieved as described in the following examples. Where available, reagents can be purchased commercially, for example from Sigma Aldrich or other chemical suppliers.

[0248] The compounds described herein can be prepared, for example, from readily available starting materials using the following general methods and procedures. Where typical or preferred process conditions (i.e., reaction temperatures, times, molar ratios of reactants, solvents, pressures, etc.) are given, it is understood that other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the specific reactants or solvents used, but such conditions can be determined by one of ordinary skill in the art by routine optimization procedures.

[0249] In addition, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. Suitable protecting groups for various functional groups and suitable conditions for protecting and deprotecting certain functional groups are well known in the art. For example, numerous protecting groups are described in Wuts, PGM, Greene, TW, & Greene, TW (2006), Greene's protective groups in organic synthesis. Hoboken, NJ, Wiley-Interscience and references cited therein.

[0250] In addition, the compounds described herein may contain one or more asymmetric ("chiral") centers. Thus, if desired, such compounds may be prepared or isolated as pure stereoisomers, i.e., as individual enantiomers or diastereomers, or as stereoisomer-enriched mixtures. All such stereoisomers (and enriched mixtures) are included within the scope of the present disclosure, unless otherwise indicated. Pure stereoisomers (or enriched mixtures) may be prepared, for example, using optically active starting materials or stereoselective reagents well known in the art. Alternatively, racemic mixtures of such compounds may be separated, for example, using chiral column chromatography, supercritical fluid chromatography, chiral resolving agents, and the like. When enantiomerically pure or enriched compounds are desired, chiral chromatography and / or enantiomerically pure or enriched starting materials may be utilized as conventionally used in the art or as described in the Examples.

[0251] The starting materials for the following reactions are generally known compounds or can be prepared by known procedures or obvious modifications thereof. For example, many starting materials are available from commercial sources, such as Sigma Aldrich, Alfa Aesar, etc. Others can be prepared by procedures or obvious modifications thereof described in standard references such as Fieser and Fieser's Reagents for Organic Synthesis, volumes 1-15 (John Wiley, and Sons, 1991), Rodd's Chemistry of Carbon Compounds, volumes 1-5 and supplements (Elsevier Science Publishers, 1989) organic Reactions, volumes 1-40 (John Wiley, and Sons, 1991), March's Advanced Organic Chemistry, (John Wiley, and Sons, 5th edition, 2001) and Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989).

[0252] The terms "solvent", "inert organic solvent" and "inert solvent" refer to a solvent that is inert under the conditions of the reaction being described therewith (e.g., benzene, toluene, acetonitrile, tetrahydrofuran ("THF"), dimethylformamide ("DMF"), chloroform, methylene chloride (or dichloromethane), diethyl ether, methanol, pyridine, and the like). In general, the term inert, as used herein with respect to a solvent, refers to a material that does not undergo reaction to form the target compound of interest via a carbon-carbon bond forming reaction. Unless otherwise specified, the solvents used in the reactions of the present disclosure are inert organic solvents and the reactions are carried out under an inert gas, preferably nitrogen or argon.

[0253] The term "qs" means adding a quantity sufficient to achieve a stated function, e.g., to bring a solution to the desired volume (i.e., 100%).

[0254] It is also understood that the addition of any of the substituents in each of the schemes below may result in the production of multiple isomeric products (including but not limited to enantiomers or one or more diastereomers), any or all of which can be isolated and purified using conventional techniques.

[0255] Incorporation of labels into the compounds or imaging agents described herein may be accomplished by reacting a suitable starting material with a reagent containing a radioisotope. The methods generally follow the same principles as standard organic chemistry reactions and can be accomplished by any method known to those of skill in the art, including those provided in this disclosure.

[0256] Scheme 1 provides an exemplary synthetic route for the synthesis of compounds provided herein (e.g., compounds of formula I). ​​Compounds of formula I, or other formulas or compounds disclosed herein, are typically prepared by first preparing, for example, compound 1 or compound 1a, and then attaching the desired substituents using suitable conditions (e.g., nucleophilic addition or cross-coupling).

[0257] In some embodiments, the synthesis of the compounds described herein proceeds according to Scheme 1.

[0258] [ka]

[0259] In Scheme 1, R 2 , R 3 , R 4 , R 5 , R 6 , R 10 , Y 1 , Y 2 , L, X, m, n, and ring A are as defined herein. R is R as defined herein. 1or R is a protecting group (e.g., a nitrogen protecting group as described in the Examples below). LG is a leaving group (e.g., a halogen, triflate, mesylate, tosylate, or any other suitable leaving group).

[0260] In scheme 1, X is CR 11 and R 11 is as defined herein, compound 1 is converted to a compound of formula I in one or more steps starting from a dihydroisoindolinone, which is prepared using methods known in the art and / or is commercially available. Dihydroisoindolinone 1 can be ... 2 (dba) 3 or in the presence of any other suitable reagent) to yield compound 3. It will be understood that the leaving group in compounds 1, 2, or 3 (illustrated as bromo in each example) may be any other suitable leaving group (e.g., triflate). Compound 3 is converted to boronate 4, which is then converted to hydroxy compound 5 using the methods described in the Examples section below. Hydroxy compound 5 is O-alkylated with compound 6 to yield a compound of formula I. LG may be any suitable leaving group, including but not limited to a chloro, iodo, bromo, or triflate group. In some embodiments, R 1 When is H, an additional deprotection step may be required to remove the protecting group on the nitrogen atom.

[0261] In some embodiments, the synthesis of the compounds described herein proceeds according to Scheme 2.

[0262] [ka]

[0263] Dihydroisoindolinone 1a can be used as a starting material (e.g., obtained from a commercial source or by processes known in the art) and coupled with compound 2 to give compound 5a, and the hydroxy group of compound 5a can then be O-alkylated with compound 6 to give a compound of formula I.

[0264] One of ordinary skill in the art will appreciate that any of compounds 1, 2, and 6 may be available from commercial sources for certain embodiments. Alternatively, the synthesis of compounds 1, 2, and 6 may be as described herein or known to one of ordinary skill in the art. EXAMPLES

[0265] The following examples are included to illustrate specific embodiments of the present disclosure. It should be understood by those skilled in the art that the techniques disclosed in the following examples represent techniques that work well in implementing the present disclosure, and therefore can be considered to constitute specific modes for its implementation. However, those skilled in the art should understand in light of the present disclosure that many changes can be made to the specific embodiments disclosed and still obtain the same or similar results without departing from the spirit and scope of the present disclosure.

[0266] 1. General Experimental Procedure Commercially available reagents and solvents (HPLC grade) were used without further purification. Chromatography was performed in deuterated solvents on a Bruker DRX 500 MHz spectrometer, or a Bruker DPX 250 MHz spectrometer, or a Bruker AVANCE 300 or Bruker AVANCE 500 spectrometer. 1H NMR spectra were recorded. Chemical shifts (δ) are expressed in parts per million. Flash column chromatography refers to automated purification on a Biotage Isolera system using appropriately sized SNAP or KPNH pre-packed silica columns and solvents as recorded in the experimental section, or an Isco Combiflash Rf system using appropriately sized pre-packed silica columns and solvents as recorded in the experimental section. Reverse-phase MPLC chromatography was performed on an Isco Combiflash Rf system using appropriately sized pre-packed C18 columns and solvents as recorded in the experimental section. Thin-layer chromatography (TLC) analysis was performed on Kieselgel 60 F254 (Merck) plates and visualized using UV light. SCX chromatography was performed using an Isolute Flash SCX-2 from Biotage, loading samples in methanol and eluting with methanol and then 5% ammonia in methanol.

[0267] 2.Analysis method Acid phase HPLC method Analytical HPLC-MS (METCR1410) was performed on a Shimadzu LCMS-2010EV system using a Kinetix Core-Shell C18 reversed-phase column (5 μm, 2.1 × 50 mm) at a column temperature of 40 °C, a gradient of 5 to 100% B (A = water / 0.1% formic acid, B = acetonitrile / 0.1% formic acid) for 1.2 min, then 100% B for 0.1 min, injection volume 3 μL, flow rate = 1.2 mL / min. All other aspects of the method remained unchanged.

[0268] Alternatively, analytical HPLC-MS was performed on a Shimadzu LCMS-2010EV system using an Atlantis dC18 reversed-phase column (3 μm, 2.1 × 50 mm) with a gradient of 5-100% B (A = water / 0.1% formic acid, B = acetonitrile / 0.1% formic acid) for 3 min, injection volume of 3 μL, and flow rate = 1.0 mL / min (METCR1278). UV spectra were recorded using an SPD-M20A photodiode array detector at 215 nm. Mass spectra were acquired over the range m / z 150-850 using an LCMS2010EV with a sampling rate of 2 scans per second. Data were integrated and reported using Shimadzu LCMS-Solutions and PsiPort software.

[0269] Alternatively, analytical HPLC-MS (MET-uHPLC-AB-101) was performed on a Waters Acquity UPLC system equipped with a Waters PDA and ELS detector using a Phenomenex Kinetex-XB C-18 column (1.7 μm, 2.1 mm × 100 mm) at a column temperature of 40 °C with a gradient of 5-100% B (A = water / 0.1% formic acid; B = acetonitrile / 0.1% formic acid) for 5.3 min, followed by 100% B for 0.5 min, at a flow rate of 0.6 mL / min. UV spectra were recorded at 215 nm using a Waters Acquity PDA detector. Mass spectra were acquired over the range m / z 150-850 using a Waters ZQ with a sampling rate of 2 scans per second. Data were integrated and reported using OpenLynx software.

[0270] Alternatively, analytical UHPLC-MS was performed on a reversed-phase system using a Waters UPLC™ BEH™ C18 column (2.1 mm x 50 mm, 1.7 μm; temperature: 40 °C) with an injection volume of 1 μL, a flow rate of 0.9 mL / min, and a gradient of 5-100% B (A = 0.1% formic acid in water; B = 0.1% formic acid in acetonitrile) for 1.1 min, then 100% B for 0.25 min (METCR1704). A second gradient of 100-5% B was then applied for 0.05 min and held for 0.1 min. UV spectra were recorded at 215 nm, spectral range: 200-400 nm. Mass spectra were obtained using a Waters SQD or QDA detector; ionization mode: electrospray positive or negative. Data were integrated and reported using Waters MassLynx and OpenLynx software.

[0271] Alternatively, UHPLC (MET-uHPLC-001) was performed on a Waters Acquity H-Class system using an Acquity UPLC BEH C18 column (1.7 μm, 2.1 × 75 mm) with a gradient of 5 to 100% B (A = water / 0.1% trifluoroacetic acid, B = acetonitrile / 0.1% trifluoroacetic acid) for 6.0 min, followed by 100% B for 2.0 min, flow rate = 0.5 mL / min at ambient column temperature (approximately 22 °C). UV spectra were recorded at 254 and 215 nm.

[0272] Alternatively, UHPLC (MET-uHPLC-002) was performed on a Waters Acquity H-Class system using an Acquity UPLC BEH C18 column (1.7 μm, 2.1 × 75 mm) with a gradient of 5 to 100% B (A = water / 0.1% trifluoroacetic acid, B = acetonitrile / 0.1% trifluoroacetic acid) for 6.0 min, followed by 100% B for 2.0 min, flow rate = 0.4 mL / min at ambient column temperature (approximately 22 °C). UV spectra were recorded at 254 and 215 nm.

[0273] Alternatively, analytical HPLC (MET-uHPLC-003) was performed on a Varian Pro Star 210 system using an XBridge C18 column (3.5 μm, 4.6 × 150 mm) with a gradient of 5 to 100% B (A = water / 0.1% trifluoroacetic acid, B = acetonitrile / 0.1% trifluoroacetic acid) for 20.0 min at ambient column temperature (approximately 22 °C), followed by 100% B for 5.0 min at a flow rate of 1.0 mL / min. UV spectra were recorded using a Varian Pro Star 330 (PDA) detector at 254 and 215 nm.

[0274] Alternatively, analytical HPLC (MET-uHPLC-004) was performed on a Varian Pro Star 210 system using a Luna C18(2) column (5 μm, 4.6 × 250 mm) with a gradient of 5 to 100% B (A = water / 0.1% trifluoroacetic acid, B = acetonitrile / 0.1% trifluoroacetic acid) for 20.0 min at ambient column temperature (approximately 22 °C), followed by 100% B for 5.0 min at a flow rate of 1.5 mL / min. UV spectra were recorded using a Varian Pro Star 330 (PDA) detector at 254 nm.

[0275] Alternatively, analytical HPLC (MET-uHPLC-005) was performed on a Varian Pro Star 210 system using a Luna C18(2) column (5 μm, 4.6 × 150 mm) with a gradient of 5 to 90% B (A = water / 0.1% trifluoroacetic acid, B = acetonitrile / 0.1% trifluoroacetic acid) for 15.0 min at ambient column temperature (approximately 22 °C), followed by 90% B for 5.0 min at a flow rate of 1.15 mL / min. UV spectra were recorded using a Varian Pro Star 330 (PDA) detector at 254 and 215 nm.

[0276] Alternatively, mass spectra and LCMS analyses were obtained using a Waters Acquity SQD (ESI, UP-LCMS) system or an Agilent G6100A SQ LCMS system.

[0277] Basic Phase HPLC Method Analytical HPLC-MS (METCR0990) was performed on a Hewlett Packard HPLC system using a Phenomenex Gemini C18 reversed-phase column (3 μm, 2.0 × 50 mm) at a column temperature of 60 °C, a gradient of 1-100% B (A = 2 mM ammonium bicarbonate in water buffered to pH 10, B = acetonitrile) for 1.8 min, then 100% B for 0.3 min, injection volume of 3 μL, and flow rate = 1 mL / min. UV spectra were recorded using a Waters PDA detector at 215 nm. Mass spectra were acquired using a Waters ZQ over the range m / z 150-850 at a sampling rate of 2 scans per second. Data were integrated and reported using OpenLynx software.

[0278] Analytical HPLC-MS (METCR1600) was performed on a Hewlett Packard HPLC system using a Phenomenex Gemini C18 reversed-phase column (3 μm, 2.0 × 100 mm) with a gradient of 5-100% B (A = 2 mM ammonium bicarbonate in water buffered to pH 10, B = acetonitrile) for 5.5 min, followed by 100% B for 0.4 min, injection volume 3 μL, flow rate = 0.5 mL / min. UV spectra were recorded using a Waters PDA detector at 215 nm. Mass spectra were acquired using a Waters ZQ over the range m / z 150-850 with a sampling rate of 2 scans per second. Data were integrated and reported using OpenLynx software.

[0279] Alternatively, analytical UHPLC-MS was performed in reverse phase using a Waters UPLC™ BEH™ C18 column (2.1 mm x 30 mm, 1.7 μm; temperature 40° C.) with an injection volume of 1 μL, flow rate = 1.0 mL / min, gradient 1-100% B (A = 2 mM ammonium bicarbonate in water buffered to pH 10; B = acetonitrile) for 1.1 min, then 100% B for 0.25 min (MET-uHPLC-AB-2005). A second gradient 100-1% B was then applied for 0.05 min and held for 0.4 min. UV spectra were recorded at 215 nm, spectral range: 200-400 nm. Mass spectra were obtained using a Waters Quattro Premier XE mass detector or a Waters SQD2; ionization mode: electrospray positive or negative. Data were integrated and reported using Waters MassLynx and OpenLynx software.

[0280] Alternatively, UHPLC (MET-uHPLC-006) was performed on a Waters Acquity H-Class system using an Acquity UPLC BEH C18 column (1.7 μm, 2.1 × 75 mm) with a gradient of 5 to 100% B (10 mM ammonium formate in water buffered to pH 10 with ammonium hydroxide, B = 95:5 acetonitrile / water) for 6.0 min, then 100% B for 2.0 min, flow rate = 0.4 mL / min at ambient column temperature (approximately 22 °C). UV spectra were recorded at 254 and 215 nm.

[0281] All example compounds exhibit LC purity >95% unless otherwise stated.

[0282] Preparative HPLC method Preparative HPLC separations were performed on a Varian Prep HPLC system using a Varian SD-1 preparative LC pump and a ProStar 325 UV / Vis detector. An XBridge Prep C18 OBD column (5 μm, 19×250 mm) was used and eluted according to the solvent gradient method 2.

[0283] [Table 3]

[0284] Intermediates Intermediate 1: (5-(fluoromethoxy)pyridin-2-yl)methanol

[0285] [ka]

[0286] Step 1: Methylene bis(4-methylbenzenesulfonate) A mixture of silver p-toluenesulfonate (11.5 g, 41.1 mmol) and MeCN (43.4 mL) was treated with diiodomethane (5.00 g, 18.7 mmol) and the mixture was stirred at reflux for 16 h. After this time, the mixture was cooled to ambient temperature, filtered, and the filter cake was washed with MeCN (3×20 mL). The filtrate was concentrated in vacuo. DCM (40 mL) was added to the residue, the suspension was filtered, and the filter cake was washed with DCM (3×20 mL). The filtrate was concentrated in vacuo and the resulting residue was recrystallized from EtOH (30 mL). The isolated product was dried in vacuo to give the title compound (4.09 g, 62%). 1 H NMR (300 MHz, CDCl 3 ) 7.59 (d, J = 8.4 Hz, 4H), 7.25 (d, J = 8.7 Hz, 4H), 5.81 (s, 2H), 2.45 (s, 6H).

[0287] Step 2: Fluoromethyl 4-methylbenzenesulfonate A mixture of methylene bis(4-methylbenzenesulfonate) (4.09 g, 11.5 mmol) and MeCN (26.7 mL) was treated with 1M TBAF in THF (12.6 mL, 12.6 mmol) and the mixture was stirred at reflux for 2 h. After this time, the solvent was removed in vacuo and the resulting residue was dissolved in EtOAc (40 mL). The solution was washed with brine (40 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-50% EtOAc in heptane) to give the title compound (609 mg, 26%). 1 H NMR (300 MHz, CDCl 3 ) 7.84 (d, J = 8.4 Hz, 2H), 7.36 (d, J = 8.1 Hz, 2H), 5.74 (d, J = 51.0 Hz, 2H), 2.64 (s, 3H).

[0288] Step 3: (5-(fluoromethoxy)pyridin-2-yl)methanol A mixture of 6-(hydroxymethyl)pyridin-3-ol (300 mg, 2.40 mmol), fluoromethyl 4-methylbenzenesulfonate (588 mg, 2.88 mmol), and acetone (9.0 mL) was treated with potassium carbonate (994 mg, 7.19 mmol) and the mixture was heated at 70° C. for 16 h. After this time, the mixture was cooled to ambient temperature and extracted with DCM (3×40 mL). The combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-5% MeOH in DCM) to give the title compound (108 mg, 29%). 1 H NMR (300 MHz, DMSO-d 6 ) 8.32 (d, J = 3.0 Hz, 1H), 7.58 (dd, J = 8.7, 2.7 Hz, 1H), 7.46 (d, J = 8.7 Hz, 1H), 5.90 (d, J = 54.0 Hz, 2H), 5.41 (t, J = 5.7 Hz, 1H), 4.52 (d, J = 5.7 Hz, 2H).

[0289] Intermediate 2: 2-((6-(chloromethyl)pyridin-3-yl)oxy)ethyl-1,1,2,2-d 4 4-Methylbenzenesulfonate

[0290] [ka]

[0291] Step 1: Ethan-1,2-diyl-d 4 Bis(4-methylbenzenesulfonate) p-Toluenesulfonyl chloride (5.77 g, 30.3 mmol) was dissolved in ethylene glycol-d 4 (0.673 mL, 12.1 mmol) and triethylamine (8.41 mL, 60.5 mmol) and the mixture was stirred at room temperature for 16 h. After this time, DCM (40 mL) was added and the mixture was washed with water (100 mL). The aqueous layer was extracted with DCM (100 mL) and the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-5% EtOAc in DCM) to give the title compound (3.84 g, 85%). 1 H NMR (300 MHz, CDCl 3 ) 7.74 (d, J = 8.4 Hz, 4H), 7.34 (d, J = 7.8 Hz, 4H), 2.46 (s, 6H). MS (ES + ) (M+H) + 375.

[0292] Step 2: 2-((6-(hydroxymethyl)pyridin-3-yl)oxy)ethyl-1,1,2,2-d 4 4-Methylbenzenesulfonate Ethane-1,2-diyl-d 4Bis(4-methylbenzenesulfonate) (4.51 g, 11.8 mmol) was added to a mixture of 6-(hydroxymethyl)pyridin-3-ol (492 mg, 3.93 mmol) and cesium carbonate (3.84 g, 11.8 mmol) in MeCN (49.3 mL) and the mixture was stirred at 80° C. for 2.5 h. After this time, the reaction mixture was cooled and filtered through diatomaceous earth. The filter cake was rinsed with EtOAc (2×50 mL) and the filtrate was concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-5% MeOH in DCM) to give the title compound (450 mg, 34%). 1 H NMR (300 MHz, CDCl 3 ) 8.13 (dd, J = 2.4, 0.9 Hz, 1H), 7.83 - 7.80 (m, 2H), 7.35 (d, J = 7.8 Hz, 2H), 7.18 - 7.11 (m, 2H), 4.70 (s, 2H), 3.34 (br s, 1H), 2.46 (s, 3H).

[0293] Step 3: 2-((6-(chloromethyl)pyridin-3-yl)oxy)ethyl-1,1,2,2-d 4 4-Methylbenzenesulfonate Thionyl chloride (0.197 mL, 2.70 mmol) was dissolved in 2-((6-(hydroxymethyl)pyridin-3-yl)oxy)ethyl-1,1,2,2-d 4 A mixture of 4-methylbenzenesulfonate (450 mg, 1.35 mmol) was added and the solution was stirred at 0° C. for 1 h. After this time, water (25 mL) was added, the layers were separated and the aqueous layer was extracted with DCM (2×25 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to give the title compound (475 mg, 99%). 1 H NMR (300 MHz, CDCl 3 ) 8.13 (d, J = 2.7 Hz, 1H), 7.83 - 7.80 (m, 2H), 7.37 - 7.34 (m, 3H), 7.12 (dd, J = 8.4, 3.0 Hz, 1H), 4.63 (s, 2H), 2.46 (s, 3H).

[0294] Intermediate 3: 2-(chloromethyl)-5-(2-fluoroethoxy-1,1,2,2-d 4 ) Pyridine

[0295] [ka]

[0296] Step 1: 2-Fluoroethyl-1,1,2,2-d 4 4-Methylbenzenesulfonate 1.0 M TBAF in THF (8.97 mL, 8.97 mmol) was dissolved in ethane-1,2-diyl-d 4 Bis(4-methyl-benzenesulfonate) (2.80 g, 7.48 mmol) was added and the mixture was stirred at reflux for 2 h. After this time the mixture was cooled, diluted with DCM (100 mL) and washed with water (40 mL). The aqueous layer was extracted with DCM (100 mL) and the combined organic layers were dried over sodium sulfate, filtered and the filtrate was concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-100% DCM in heptane) to give the title compound (601 mg, 36%). 1 H NMR (300 MHz, CDCl 3 ) 7.36 (d, J = 8.1 Hz, 2H), 7.82 (d, J = 8.4 Hz, 2H), 2.46 (s, 3H).

[0297] Step 2: (5-(2-fluoroethoxy-1,1,2,2-d 4 )Pyridin-2-yl)methanol 2-Fluoroethyl-1,1,2,2-d in MeCN (20.0 mL) 4A mixture of 4-methylbenzenesulfonate (363 mg, 1.60 mmol), 6-(hydroxymethyl)pyridin-3-ol (200 mg, 1.60 mmol) and cesium carbonate (1.56 g, 4.80 mmol) was stirred at 80° C. for 2.5 h. After this time, the reaction mixture was cooled and filtered through diatomaceous earth. The filter cake was rinsed with EtOAc (2×50 mL) and the filtrate was concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-5% MeOH in DCM) to give the title compound (133 mg, 47%). 1 H NMR (300 MHz, CDCl 3 ) 8.29 (d, J = 2.7 Hz, 1H), 7.29 - 7.19 (m, 2H), 4.72 (s, 2H), 3.39 (br s, 1H).

[0298] Step 3: 2-(chloromethyl)-5-(2-fluoroethoxy-1,1,2,2-d 4 ) Pyridine Thionyl chloride (0.139 mL, 1.91 mmol) was dissolved in DCM (6.7 mL) to give 5-(2-fluoroethoxy-1,1,2,2-dichloromethane). 4 (pyridin-2-yl)methanol (167 mg, 0.953 mmol) and the mixture was stirred at 0° C. for 1 h. After this time, the mixture was poured into water (25 mL), the layers were separated, and the aqueous layer was extracted with DCM (2×25 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to give the title compound (180 mg, 98%). 1 H NMR (300 MHz, CDCl 3 ) 8.30 (d, J = 2.7 Hz, 1H), 7.48 (d, J = 8.7 Hz, 1H), 7.36 (dd, J = 8.7, 3.0 Hz, 1H), 4.72 (s, 2H).

[0299] Intermediate 4: 4-Chloro-2-(chloromethyl)-5-methoxypyridine

[0300] [ka]

[0301] To a solution of 2-(hydroxymethyl)-5-methoxypyridin-4-ol (50 mg, 0.32 mmol) in MeCN (2 mL) was added phosphorus oxychloride (0.090 mL, 0.98 mmol) and the mixture was heated at 70° C. for 22 h. After this time, additional phosphorus oxychloride (0.090 mL, 0.98 mmol) was added and heating was continued for 20 h. A third portion of phosphorus oxychloride (0.090 mL, 0.98 mmol) was then added and heating was continued for 20 h. After this time, the volatiles were removed under reduced pressure and the resulting residue was diluted with EtOAc (20 mL) and neutralized with saturated sodium bicarbonate solution. The layers were separated and the aqueous layer was extracted with EtOAc (10 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by FCC (silica, 0-10% MeOH in DCM) to give the title compound (22 mg, 35%). 1 H NMR (300 MHz, CDCl 3 ) 8.22 (s, 1H), 7.49 (s, 1H), 4.61 (s, 2H), 4.01 (s, 3H). MS (ES + ) (M+H) + 192.

[0302] Intermediate 5: 2-(chloromethyl)-5-(1-fluoroethoxy)pyridine hydrochloride

[0303] [ka]

[0304] Step 1: Ethane-1,1-diylbis(4-methylbenzenesulfonate) A mixture of 1,1-diiodoethane (500 mg, 1.77 mmol) and silver p-toluenesulfonate (990 mg, 3.55 mmol) in MeCN (20 mL) was stirred at room temperature for 3 days. After this time, the volatiles were removed in vacuo and the resulting residue was suspended in DCM. The solids were removed by filtration and the filtrate was concentrated in vacuo at room temperature to give the title compound (570 mg, 87%).1 H NMR (300 MHz, CDCl 3 ) 7.70 (d, J = 8.3 Hz, 4H), 7.31 (d, J = 8.1 Hz, 4H), 6.39 (q, J = 5.3 Hz, 1H), 2.45 (s, 6H), 1.54 (d, J = 5.3 Hz, 3H).

[0305] Step 2: 1-Fluoroethyl 4-methylbenzenesulfonate A mixture of ethane-1,1-diylbis(4-methylbenzenesulfonate) (550 mg, 1.48 mmol) and TBAF (1 M in THF, 1.63 mL, 1.63 mmol) in THF (30 mL) was stirred at room temperature for 5 days, after which the solvent was removed under reduced pressure and the resulting residue was purified by FCC (silica, 0-10% EtOAc in hexanes) to give the title compound (67 mg, 20%). 1 H NMR (300 MHz, CDCl 3 ) 7.82 (d, J = 8.3 Hz, 2H), 7.35 (d, J = 8.4 Hz, 2H), 6.23 (dq, J = 57.0, 5.0 Hz, 1H), 2.45 (s, 3H), 1.56 (dd, J = 20.6, 5.5 Hz, 3H).

[0306] Step 3: (5-(1-fluoroethoxy)pyridin-2-yl)methanol To a mixture of 6-(hydroxymethyl)pyridin-3-ol (103 mg, 0.825 mmol) and potassium bicarbonate (165 mg, 1.65 mmol) in DMF (10 mL) at room temperature, 1-fluoroethyl 4-methylbenzene-sulfonate (60 mg, 0.27 mmol) was added and the mixture was heated overnight at 100° C. After this time, the solvent was removed under reduced pressure and the resulting residue was purified by FCC (silica, 0-50% MeOH in EtOAc) to give the title compound (20 mg, 42%). 1 H NMR (300 MHz, CDCl 3) 8.38 (d, J = 2.5 Hz, 1H), 7.42 (dd, J = 8.3, 2.2 Hz, 1H), 7.24 (d, J = 8.6 Hz, 1H), 5.94 (dq, J = 62.3, 4.8 Hz, 1H), 4.73 (s, 2H), 1.69 (dd, J = 20.0, 4.9 Hz, 3H). MS (ES + ) (M+H) + 172.

[0307] Step 4: 2-(Chloromethyl)-5-(1-fluoroethoxy)pyridine hydrochloride To a mixture of (5-(1-fluoroethoxy)pyridin-2-yl)methanol (16 mg, 0.093 mmol) in DCM (5 mL) at room temperature was added thionyl chloride (111 mg, 0.930 mmol) and the mixture was stirred at room temperature for 1 h, after which the solvent was removed under reduced pressure to give the title compound (23 mg, >99%). 1 H NMR (300 MHz, CD 3 MS (ES) + ) (M+H) + 190.

[0308] Intermediate 6: 6-Bromo-2-(2-fluoroethyl)pyridazin-3(2H)-one

[0309] [ka]

[0310] To 6-bromopyridazin-3(2H)-one (50 mg, 0.29 mmol) in a reaction vial was added potassium carbonate (79 mg, 0.57 mmol), followed by a solution of 1-bromo-2-fluoroethane (58 mg, 0.46 mmol) in DMF (1 mL) and 1,4-dioxane (2 mL). The reaction vial was sealed and heated at 130° C. for 1 h. After this time, the reaction mixture was cooled to room temperature and concentrated to dryness under reduced pressure. The resulting residue was purified by FCC (silica, 0-50% EtOAc in hexanes) to give the title compound (48 mg, 76%). 1 H NMR (300 MHz, CDCl 3 ) 7.29 (d, J = 9.9 Hz, 1H), 6.84 (d, J = 9.6 Hz, 1H), 4.79 (dt, J = 47.1, 5.0 Hz, 2H), 4.44 (dt, J = 24.6, 5.0 Hz, 2H). 19 F NMR (282 MHz, CDCl 3 ) -225.67. MS (ES + ) (M+H) + 221.

[0311] Intermediate 7: 6-(chloromethyl)-2-fluoro-3-methoxy-pyridine

[0312] [ka]

[0313] Step 1: tert-Butyl-[(5-methoxy-2-pyridyl)methoxy]-dimethyl-silane (5-Methoxypyridin-2-yl)methanol (750 mg, 5.39 mmol) and imidazole (404 mg, 5.93 mmol) were dissolved in DCM (20 mL) and tert-butyl(chloro)dimethylsilane (2.03 g, 13.5 mmol) was added. The reaction was stirred at room temperature for 4 h. The reaction mixture was stirred at H 2The organic fraction was extracted with 20 mL of 1H2O (15 mL). The aqueous phase was re-extracted with DCM (10 mL) and the combined organics were washed with brine solution (10 mL) and diluted with Na 2 SO 4 It was dried at 40° C., filtered and concentrated in vacuo. The crude residue was purified by FCC (silica, 0-100% EtOAc in heptane) to give the title compound (1.26 g, 83% yield). 1 H NMR (400 MHz, DMSO) δ 8.19 (d, J = 2.5 Hz, 1H), 7.41 (dd, J = 8.6, 2.9 Hz, 1H), 7.38 - 7.31 (m, 1H), 4.68 (s, 2H), 3.81 (s, 3H), 0.90 (s, 9H), 0.07 (s, 6H). Tr(METCR1704) = 1.00 min, m / z (ES)+ [M+H]+ = 254.2, 100%.

[0314] Step 2: tert-Butyl-[(6-fluoro-5-methoxy-2-pyridyl)methoxy]-dimethyl-silane To a solution of tert-butyl-[(5-methoxy-2-pyridyl)methoxy]-dimethyl-silane (400 mg, 1.58 mmol) in anhydrous THF (10 mL) at -78 °C, 2.5 M butyllithium (0.82 mL, 2.05 mmol) was added and the reaction mixture was stirred for 1 h. N-fluoro-N-(phenylsulfonyl)benzenesulfonamide (647 mg, 2.05 mmol) was then added and the reaction mixture was allowed to warm to room temperature and stirred for 1.5 h. The reaction mixture was diluted with brine solution and EtOAc and the organic fraction was extracted. The combined organics were washed with Na 2 SO 4 It was dried at 40° C., filtered and concentrated in vacuo. The crude residue was purified by FCC (silica, 0-100% EtOAc in heptane) to give the title compound (84 mg, 18% yield). 1H NMR (400 MHz, DMSO) δ 7.66 (dd, J = 10.6, 8.1 Hz, 1H), 7.29 (d, J = 8.1 Hz, 1H), 4.60 (s, 2H), 3.86 (s, 3H), 0.90 (s, 9H), 0.08 (s, 6H). Tr(METCR1704) = 1.18 min, m / z (ES)+ [M+H]+ = 272.1, 90%.

[0315] Step 3: (6-Fluoro-5-methoxy-2-pyridyl)methanol To a solution of tert-butyl-[(6-fluoro-5-methoxy-2-pyridyl)methoxy]-dimethyl-silane (84 mg, 0.31 mmol) in THF (2 mL) at room temperature was added 1M TBAF in THF (0.31 mL, 0.31 mmol) and the reaction mixture was stirred for 1 h. The reaction mixture was diluted with saturated ammonium chloride solution (3 mL) and the product was extracted with EtOAc (2×5 mL). The combined organics were washed with Na 2 SO 4 It was dried at 40° C., filtered and concentrated in vacuo. The crude residue was purified by column chromatography (silica, 0-100% EtOAc in heptane) to give the title compound (40 mg, 77% yield). Tr(METCR1704) = 0.45 min, m / z (ES)+ [M+H]+ = 158.0, 94%.

[0316] Step 4: 6-(chloromethyl)-2-fluoro-3-methoxy-pyridine (6-Fluoro-5-methoxy-2-pyridyl)methanol (40 mg, 0.255 mmol) was dissolved in DCM (2 mL) and thionyl chloride (0.19 mL, 2.55 mmol) was added. The reaction was stirred at room temperature for 3 h. The reaction mixture was concentrated in vacuo and diluted with DCM (×2) and Et 2 Azeotroping with O gave the title compound (50 mg, 98% yield). Tr(METCR1704) = 0.74 min, m / z (ES)+ [M+H]+ = 176.0, 177.9, 88%.

[0317] Intermediate 8: 2-(chloromethyl)-4-fluoro-5-methoxy-pyridine

[0318] [ka]

[0319] Step 1: Methyl 5-(methoxymethoxy)pyridine-2-carboxylate N containing THF (60 mL) 2 To the RBF at room temperature was successively added triethylamine (6.8 mL, 49.0 mmol), methyl 5-hydroxypyridine-2-carboxylate (5.00 g, 32.6 mmol) (in one portion), followed by the dropwise addition of chloro(methoxy)methane (3.7 mL, 49.0 mmol) over 5 min. The mixture was purged with N 2 All fumes were expelled by flushing with ethyl acetate and the resulting suspension was stirred at room temperature overnight. The reaction was quenched by pouring onto water (50 mL). After extraction with EtOAc (2×50 mL), the combined organics were washed with brine (50 mL) and dried (MgSO 4 ), filtered and concentrated in vacuo. The crude residue (light yellow solid) was purified by column chromatography (Biotage Sfar Duo 50 g cartridge, 0-40% EtOAc in heptane, product eluted with 30% EtOAc) to give the title compound (5.50 g, 83% yield). 1 H NMR (500 MHz, DMSO) δ 8.43 (d, J = 2.8 Hz, 1H), 8.04 (d, J = 8.7 Hz, 1H), 7.59 (dd, J = 8.7, 2.9 Hz, 1H), 5.36 (s, 2H), 3.85 (s, 3H), 3.41 (s, 3H). Tr(METCR1704) = 0.55 min, m / z (ES + ) [M+H] + = 198.0, 97%.

[0320] Step 2: [5-(methoxymethoxy)-2-pyridyl]methanol A solution of methyl 5-(methoxymethoxy)pyridine-2-carboxylate (5.50 g, 27.9 mmol) in anhydrous toluene (200 mL) under nitrogen was cooled to -78°C and DIBAL (1M in heptane, 73 mL, 73 mmol) was added over 45 min. The reaction was warmed to 0°C and stirred for 2 h. The reaction was re-treated with additional DIBAL (1M in heptane, 11 mL, 11 mmol) at 0°C and stirring was continued for a further 1 h. The reaction was quenched by the addition of water (50 mL). EtOAc was added (200 mL). MgSO 4 After the addition of, the mixture was filtered, eluted with additional EtOAc, and the filtrate was concentrated to give the title compound (2.50 g, 48% yield). 1 H NMR (500 MHz, DMSO) δ 8.23 ​​(d, J = 2.7 Hz, 1H), 7.46 (dd, J = 8.6, 2.8 Hz, 1H), 7.39 (d, J = 8.6 Hz, 1H), 5.31 (t, J = 5.8 Hz, 1H), 5.23 (s, 2H), 4.49 (d, J = 5.8 Hz, 2H), 3.38 (s, 3H). Tr(MET-uHPLC-AB-2005) = 0.38 min m / z (ES + )(M+H) + 470, 91%.

[0321] Step 3: tert-Butyl-[[5-(methoxymethoxy)-2-pyridyl]methoxy]-dimethyl-silane [5-(methoxymethoxy)-2-pyridyl]methanol (2.50 g, 14.8 mmol) and 1H-imidazole (1.1 g, 16.3 mmol) were dissolved in DCM (100 mL) and tert-butyl(chloro)dimethylsilane (2.90 g, 19.2 mmol) was added. The reaction was stirred at room temperature for 2 h. Further 1H-imidazole (250 mg, 3.7 mmol) and tert-butyl(chloro)dimethylsilane (500 mg, 3.3 mmol) were added and stirring was continued at room temperature for another 1.75 h. The reaction mixture was diluted with water (50 mL). After separation, the aqueous phase was diluted with CH 2 Cl 2(2×50 mL) and the combined organics were washed with brine solution (50 mL) and dried (MgSO 4 ), filtered and concentrated in vacuo. The crude residue was purified by column chromatography using a Biotage Sfar Duo 2×25 g cartridge, 0-40% EtOAc in heptane to give the title compound (3.5 g, 84%). 1 H NMR (500 MHz, DMSO) δ 8.25 (d, J = 2.7 Hz, 1H), 7.49 (dd, J = 8.6, 2.9 Hz, 1H), 7.36 (d, 1H), 5.23 (s, 2H), 4.68 (s, 2H), 3.38 (s, 3H), 0.90 (s, 9H), 0.08 (s, 6H). Tr(METCR1704) = 1.03min, m / z (ES + ) [M+H] + = 284.2, 100%.

[0322] Step 4: tert-Butyl-[[4-fluoro-5-(methoxymethoxy)-2-pyridyl]methoxy]-dimethyl-silane To a solution of tert-butyl-[[5-(methoxymethoxy)-2-pyridyl]methoxy]-dimethyl-silane (2.2 g, 7.06 mmol) in anhydrous THF (35 mL) at -78 °C, N-butyllithium (2.5 M in hexanes, 3.7 mL, 9.17 mmol) was added and the reaction mixture was stirred at that temperature for 1 h. N-fluoro-N-(phenylsulfonyl)benzenesulfonamide (NFSI) (2.89 g, 9.17 mmol) was then added directly as a solid over 10 s and the reaction mixture was allowed to warm to room temperature and stirred for 40 min. The reaction was quenched by pouring onto a brine solution (50 mL). After extraction with EtOAc (2 x 40 mL), the combined organic extracts were dried (MgSO 4 ), filtered and concentrated in vacuo. The residue was purified by FCC using a Biotage Isolera (Sfar Duo 25 g, 2-30% EtOAc in heptane, loaded with DCM) to give the title compound (1.4 g, 55% yield). 1 H NMR (400 MHz, CDCl3 ) δ 8.39 (d, J = 10.0 Hz, 1H), 7.28 - 7.25 (m, 1H), 5.21 (s, 2H), 4.75 (d, J = 0.7 Hz, 2H), 3.54 (s, 3H), 0.95 (s, 9H), 0.12 (s, 6H). Tr(METCR1704) = 1.13 min, m / z (ES + ) [M+H] + = 302.2, 68%.

[0323] Step 5: 6-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-fluoro-pyridin-3-ol To a solution of tert-butyl-[[4-fluoro-5-(methoxymethoxy)-2-pyridyl]methoxy]-dimethyl-silane (1013 mg, 3.36 mmol) in DCM (20.26 mL) was added zinc dibromide (1.5 g, 6.72 mmol) and propane-1-thiol (0.61 mL, 6.72 mmol). The mixture was stirred at room temperature for 3 h. The reaction was cooled to 0 °C and then washed with saturated NaHCO 3 It was quenched by the addition of aqueous solution (10 mL). It was stirred at 0° C. for 15 min. Water (25 mL) was added and extracted with DCM (3×30 mL), then the combined organic extracts were washed with brine (30 mL) and dried (MgSO 4 ), filtered and concentrated to give a dark orange oil. The residue was purified by FCC using a Biotage Isolera (Sfar Duo 50 g, 12-80% EtOAc in heptane, loaded with DCM) to give the title compound (388 mg, 43% yield). 1 H NMR (400 MHz, CDCl 3 ) δ 8.24 (d, J = 10.3 Hz, 1H), 7.27 (d, J = 11.3 Hz, 1H), 4.75 (s, 2H), 0.95 (s, 9H), 0.11 (s, 6H). Tr(METCR1704) = 0.91 min, m / z (ES + ) [M+H] + = 258.2, 94%.

[0324] Step 6: tert-Butyl-[(4-fluoro-5-methoxy-2-pyridyl)methoxy]-dimethyl-silane To a solution of 6-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-fluoro-pyridin-3-ol (385 mg, 1.50 mmol) in DMF (5 mL) was added cesium carbonate (585 mg, 1.80 mmol) followed by iodomethane (0.11 mL, 1.80 mmol). The mixture was heated to 35° C. and stirred at that temperature for 3 h. After cooling, the reaction was diluted with saturated NaHCO 3 The reaction was quenched by pouring onto aqueous solution (10 mL) and water (10 mL) was added. 2 After extraction with O (3 × 20 mL), the combined organic extracts were washed with brine (15 mL) and dried (MgSO 4 ), filtered and concentrated in vacuo. The residue was purified by FCC using a Biotage Isolera (10 g, 2-30% EtOAc in heptane, loaded with DCM) to give the title compound (243 mg, 59% yield) as a colorless, mobile oil. 1 H NMR (400 MHz, CDCl3) δ 8.21 (d, J = 10.1 Hz, 1H), 7.26 - 7.23 (m, 1H), 4.74 (s, 2H), 3.96 (s, 3H), 0.95 (s, 9H), 0.12 (s, 6H). Tr(METCR1704) = 1.11 min, m / z (ES + ) [M+H] + = 272.2, 98%.

[0325] Step 7: (4-Fluoro-5-methoxy-2-pyridyl)methanol To a solution of tert-butyl-[(4-fluoro-5-methoxy-2-pyridyl)methoxy]-dimethyl-silane (98%, 243 mg, 0.877 mmol) in THF (6 mL) at 5 °C, TBAF (1 M in THF, 1.1 mL, 1.05 mmol) was added and the solution was stirred at 5-10 °C for 1.5 h. The reaction was cooled to 0 °C with saturated NaHCO 3The mixture was quenched by pouring onto aqueous solution (10 mL). Water (10 mL) and EtOAc (10 mL) were added. Due to incomplete separation, brine (5 mL) was added. After separation, the aqueous phase was extracted with EtOAc (2×10 mL). The combined organic extracts were washed with brine (15 mL) and dried (MgSO 4 ), filtered and concentrated in vacuo. The residue was purified by FCC using a Biotage Isolera (10 g, 5-30% methanol in DCM, loaded with DCM) to give the title compound (125 mg, 86% yield). 1 H NMR (400 MHz, CDCl3) δ 8.26 (d, J = 9.9 Hz, 1H), 7.03 (d, J = 11.0 Hz, 1H), 4.69 (d, J = 3.8 Hz, 2H), 3.98 (s, 3H), 3.34 (s, 1H). Tr(MET-uHPLC-AB-2005) = 0.38 min m / z (ES + ) (M+H) + 158.1, 95%.

[0326] Step 8: 2-(Chloromethyl)-4-fluoro-5-methoxy-pyridine (4-Fluoro-5-methoxy-2-pyridyl)methanol (30 mg, 0.191 mmol) was dissolved in DCM (1.5 mL) and thionyl chloride (0.07 mL, 0.955 mmol) was added. The reaction was stirred at room temperature for 2 h. The reaction mixture was cooled to 5° C. for 2 h. 2 The mixture was concentrated under reflux. 2 Triturate with O (2x) and evaporate volatiles (N 2 under reflux) followed by drying in a vacuum oven to give the title compound (33 mg, 95% yield). 1 H NMR (400 MHz, CDCl 3 ) δ 8.42 (s, 1H), 7.67 (d, J = 8.5 Hz, 1H), 5.08 (s, 2H), 4.12 (s, 3H). Tr(METCR1704) = 0.66 min, m / z (ES + ) (M+H) + = 176.0, 178.0, 96%.

[0327] method Method 1 Scheme for method 1

[0328] [ka]

[0329] Step 1: 5-Methoxy-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one 5-Methoxy-2,3-dihydroisoindol-1-one (546 mg, 3.35 mmol), 6-bromo-2-methyl-3(2H)-pyridazinone (759 mg, 4.02 mmol), RuPhos (234 mg, 0.502 mmol), cesium carbonate (3.27 g, 10.0 mmol), and Pd in ​​1,4-dioxane (22.8 mL). 2 (dba) 3 (153 mg, 0.167 mmol) was heated at 100° C. for 16 h. After this time, the reaction mixture was cooled and water (50 mL) was added. The solid that formed was collected by filtration and dried in vacuum to give the title compound (691 mg, 76%). 1 H NMR (300 MHz, DMSO-d 6 ) 8.58 (d, J = 10.2 Hz, 1H), 7.72 (d, J = 8.4 Hz, 1H), 7.25 (d, J = 1.8 Hz, 1H), 7.11-7.07 (m, 2H), 4.88 (s, 2H), 3.87 (s, 3H), 3.64 (s, 3H).

[0330] Step 2: 5-Hydroxy-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one 1.0 M Boron tribromide in DCM (27.6 mL, 27.6 mmol) was added to a solution of 5-methoxy-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one (691 mg, 2.55 mmol) in 1,2-dichloroethane (276 mL) and the mixture was stirred at reflux for 16 h. After this time, the mixture was cooled and ice was added, followed by saturated aqueous sodium bicarbonate (30 mL). The solid that formed was collected by filtration, dried in vacuum, then suspended in methanol (659 mL) and the mixture was stirred at reflux for 1 h. After this time, the solvent was removed in vacuum, water (100 mL) was added and the mixture was sonicated for 10 min. The product was collected by filtration and dried in vacuum to give the title compound (540 mg, 82%). 1 H NMR (300 MHz, DMSO-d 6 ) 10.47 (br s, 1H), 8.58 (d, J = 9.9 Hz, 1H), 7.62 (d, J = 8.4 Hz, 1H), 7.07 (d, J = 9.9 Hz, 1H), 7.00 (s, 1H), 6.91 (d, J = 8.4 Hz, 1H), 4.83 (s, 2H), 3.63 (s, 3H).

[0331] Step 3: 2-(Chloromethyl)-5-fluoropyridine Thionyl chloride (0.057 mL, 0.79 mmol) was added to a mixture of (5-fluoropyridin-2-yl)methanol (50 mg, 0.39 mmol) in DCM (2.7 mL) and the mixture was stirred at room temperature for 20 min. After this time, the mixture was poured into water (25 mL) and extracted with DCM (2×25 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to give the title compound (57 mg, 99% yield), which was used in the next step without purification.

[0332] Step 4: 5-((5-fluoropyridin-2-yl)methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one A solution of 5-hydroxy-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one (50 mg, 0.19 mmol), 2-(chloromethyl)-5-fluoropyridine (50 mg, 0.34 mmol), and potassium carbonate (81 mg, 0.58 mmol) in DMSO (2 mL) was heated at 70° C. for 20 h. After this time, water (20 mL) was added and the solid that formed was collected by filtration and triturated in MeOH (10 mL). This material was purified by FCC (silica, 0-5% MeOH in DCM) and the collected product was triturated in MeCN (10 mL) and lyophilized from 1:2 MeCN / water (10 mL) to give the title compound (28 mg, 39%).

[0333] Example 1-1: 5-((5-fluoropyridin-2-yl)methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one 1 H NMR (500 MHz, DMSO-d 6 ) 8.61 (d, J = 3.0 Hz, 1H), 8.57 (d, J = 10.0 Hz, 1H), 7.81 (td, J = 8.5, 3.0 Hz, 1H), 7.73 (d, J = 9.0 Hz, 1H), 7.66 (dd, J = 9.0, 4.5 Hz, 1H), 7.35 (d, J = 2.0 Hz, 1H), 7.19 (dd, J = 8.5, 2.0 Hz, 1H), 7.08 (d, J = 10.0 Hz, 1H), 5.30 (s, 2H), 4.88 (s, 2H), 3.64 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -128.32. Tr(MET-uHPLC-001) = 4.90 min, (ES + ) (M+H) + 367, 99%.

[0334] The following additional compounds were prepared by Method 1:

[0335] Example 1-2: 5-Methoxy-2-(pyridin-4-yl)-2,3-dihydro-1H-isoindol-1-one

[0336] [ka]

[0337] 1 H NMR (500 MHz, DMSO-d 6 ) 8.58 - 8.47 (m, 2H), 7.91 - 7.82 (m, 2H), 7.74 (d, J = 8.5 Hz, 1H), 7.23 (d, J = 1.9 Hz, 1H), 7.11 (dd, J = 8.5, 2.3 Hz, 1H), 4.98 (s, 2H), 3.89 (s, 3H). Tr (MET-uHPLC-AB-101) = 1.27 min, (ES+) (M+H)+ 241, 99%.

[0338] Example 1-3: 5-[(5-iodopyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0339] [ka]

[0340] 1 H NMR (500 MHz, DMSO-d 6) 8.83 (d, J = 2.1 Hz, 1H), 8.57 (d, J = 10.0 Hz, 1H), 8.24 (dd, J = 8.2, 2.2 Hz, 1H), 7.73 (d, J = 8.5 Hz, 1H), 7.41 (d, J = 8.2 Hz, 1H), 7.33 (d, J = 2.0 Hz, 1H), 7.18 (dd, J = 8.5, 2.2 Hz, 1H), 7.08 (d, J = 10.0 Hz, 1H), 5.26 (s, 2H), 4.87 (s, 2H), 3.63 (s, 3H). Tr(MET-uHPLC-AB-101) = 3.12 min, (ES+) (M+H)+ 475, 98%.

[0341] Examples 1-4 (Comparative Example 2): 5-[(5-methoxypyridin-2-yl)methoxy]-2-(pyridin-4-yl)-2,3-dihydro-1H-isoindol-1-one

[0342] [ka]

[0343] 1 H NMR (500 MHz, DMSO-d 6 ) 8.56 - 8.48 (m, 2H), 8.31 (d, J = 2.9 Hz, 1H), 7.90 - 7.85 (m, 2H), 7.74 (d, J = 8.5 Hz, 1H), 7.52 (d, J = 8.6 Hz, 1H), 7.44 (dd, J = 8.6, 2.9 Hz, 1H), 7.31 (d, J = 1.6 Hz, 1H), 7.19 (dd, J = 8.5, 2.1 Hz, 1H), 5.24 (s, 2H), 4.97 (s, 2H), 3.84 (s, 3H). Tr(MET-uHPLC-AB-101) = 1.59 min, (ES + ) (M+H) + 348, 99%.

[0344] Example 1-5: 5-Hydroxy-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0345] [ka]

[0346] 1 H NMR (500 MHz, DMSO-d 6 ) 10.46 (s, 1H), 8.58 (d, J = 10.0 Hz, 1H), 7.62 (d, J = 8.3 Hz, 1H), 7.07 (d, J = 10.0 Hz, 1H), 7.01 (s, 1H), 6.91 (dd, J = 8.3, 1.6 Hz, 1H), 4.83 (s, 2H), 3.63 (s, 3H). Tr(MET-uHPLC-AB-101) = 1.66 min m / z (ES+)(M+H)+ 258.1, 96%.

[0347] Example 1-6: 5-[(5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0348] [ka]

[0349] 1 H NMR (500 MHz, DMSO-d 6) 8.57 (d, J = 10.0 Hz, 1H), 8.30 (d, J = 2.7 Hz, 1H), 7.72 (d, J = 8.4 Hz, 1H), 7.51 (d, J = 8.6 Hz, 1H), 7.44 (dd, J = 8.6, 2.9 Hz, 1H), 7.34 (d, J = 2.0 Hz, 1H), 7.17 (dd, J = 8.4, 2.2 Hz, 1H), 7.08 (d, J = 9.9 Hz, 1H), 5.22 (s, 2H), 4.87 (s, 2H), 3.84 (s, 3H), 3.63 (s, 3H).Tr(MET-HPLC-004) = 11.42 min m / z (ES+) (M+H)+ 379.0, 99%.

[0350] Example 1-7: 5-[(3-fluoro-5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0351] [ka]

[0352] 1 H NMR (500 MHz, DMSO-d 6 ) 8.57 (d, J = 10.0 Hz, 1H), 8.22 (d, J = 2.5 Hz, 1H), 7.72 (d, J = 8.5 Hz, 1H), 7.53 (dd, J = 11.5, 2.5 Hz, 1H), 7.36 (d, J = 2.0 Hz, 1H), 7.18 (dd, J = 8.5, 2.5 Hz, 1H), 7.08 (d, J = 10.0 Hz, 1H), 5.26 (d, J = 1.5 Hz, 2H), 4.88 (s, 2H), 3.88 (s, 3H), 3.64 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6) -123.35. Tr(MET-uHPLC-001) = 4.93 min m / z (ES+) (M+H)+ 397.2, 99%.

[0353] Example 1-8: 5-({5-[2-fluoro(1,1,2,2- 2 H 4 )Ethoxy]pyridin-2-yl}methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0354] [ka]

[0355] 1 H NMR (500 MHz, DMSO-d 6 ) 8.57 (d, J = 10.0 Hz, 1H), 8.34 (dd, J = 3.0, 0.5 Hz, 1H), 7.72 (d, J = 8.5 Hz, 1H), 7.52 (d, J = 8.5 Hz, 1H), 7.48 (dd, J = 8.5, 3.0 Hz, 1H), 7.34 (d, J = 2.0 Hz, 1H), 7.18 (dd, J = 8.5, 2.0 Hz, 1H), 7.08 (d, J = 10.0 Hz, 1H), 5.23 (s, 2H), 4.88 (s, 2H), 3.64 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -224.24. Tr(MET-uHPLC-001) = 3.85 min m / z (ES+) (M+H)+ 415.2, 98%.

[0356] Example 1-9: 2-{[6-({[2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-yl]oxy}methyl)pyridin-3-yl]oxy}(1,1,2,2- 2 H 4) Ethyl 4-methylbenzene-1-sulfonate

[0357] [ka]

[0358] 1 H NMR (500 MHz, DMSO-d 6 ) 8.57 (d, J = 10.0 Hz, 1H), 8.19 (d, J = 3.0 Hz, 1H), 7.79 (d, J = 8.5 Hz, 2H), 7.72 (d, J = 8.5 Hz, 1H), 7.49 - 7.45 (m, 3H), 7.37 - 7.33 (m, 2H), 7.17 (dd, J = 8.5, 2.0 Hz, 1H), 7.07 (d, J = 10.0 Hz, 1H), 5.21 (s, 2H), 4.88 (s, 2H), 3.64 (s, 3H), 2.41 (s, 3H). Tr(MET-uHPLC-001) = 5.02 min m / z (ES+) (M+H)+ 567.2, 99%.

[0359] Example 1-10: 5-{[5-(1-fluoroethoxy)pyridin-2-yl]methoxy}-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0360] [ka]

[0361] 1 H NMR (500 MHz, DMSO-d 6) 8.57 (d, J = 10.0 Hz, 1H), 8.41 (d, J = 2.7 Hz, 1H), 7.73 (d, J = 8.4 Hz, 1H), 7.62 (dd, J = 8.3, 2.5 Hz, 1H), 7.58 (d, J = 8.5 Hz, 1H), 7.34 (d, J = 1.9 Hz, 1H), 7.18 (dd, J = 8.5, 2.2 Hz, 1H), 7.08 (d, J = 10.0 Hz, 1H), 6.31 (dq, J = 62.2, 4.8 Hz, 1H), 5.26 (s, 2H), 4.88 (s, 2H), 3.63 (s, 3H), 1.62 (dd, J = 20.5, 4.8 Hz, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -117.10. Tr(MET-uHPLC-001) = 3.34 min m / z (ES+) (M+H)+ 411.1, 99%.

[0362] Example 1-11: 5-[(4-chloro-5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0363] [ka]

[0364] 1 H NMR (500 MHz, CDCl 3) 8.74 (d, J = 10.5 Hz, 1H), 8.27 (s, 1H), 7.82 (d, J = 8.5 Hz, 1H), 7.55 (s, 1H), 7.12 (dd, J = 8.5, 2.5 Hz, 1H), 7.07 (d, J = 1.5 Hz, Tr(MET-uHPLC-001) = 5.45 min m / z (ES+) (M+H)+ 413.1, 98%.

[0365] Example 1-12: 6-[(5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0366] [ka]

[0367] 1 H NMR (500 MHz, DMSO-d 6 ) 8.56 (d, J = 9.9 Hz, 1H), 8.30 (d, J = 2.5 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.49 (d, J = 8.6 Hz, 1H), 7.43 (dd, J = 8.5, 2.7 Hz, 1H), 7.36 (d, J = 7.3 Hz, 2H), 7.09 (d, J = 10.0 Hz, 1H), 5.21 (s, 2H), 4.86 (s, 2H), 3.83 (s, 3H), 3.64 (s, 3H). Tr(MET-uHPLC-001) = 4.00 min m / z (ES+) (M+H)+ 379.2, 98%.

[0368] Example 1-13: 5-[(6-fluoro-5-methoxy-2-pyridyl)methoxy]-2-(1-methyl-6-oxo-pyridazin-3-yl)isoindolin-1-one

[0369] [ka]

[0370] 1 H NMR (500 MHz, DMSO-d 6 ) 8.57 (d, J = 10.0 Hz, 1H), 7.75 - 7.66 (m, 2H), 7.48 (d, J = 8.1 Hz, 1H), 7.33 (d, J = 1.9 Hz, 1H), 7.18 (dd, J = 8.5, 2.3 Hz, 1H), 7.08 (d, J = 10.0 Hz, 1H), 5.15 (s, 2H), 4.88 (s, 2H), 3.89 (s, 3H), 3.64 (s, 3H). 19 F NMR (471 MHz, DMSO-d6) -85.49 (d, J = 10.5 Hz). Tr(MET-uHPLC-AB-101) = 2.78 min m / z (ES+)(M+H)+ 397.2, 96%.

[0371] Example 1-14: 5-[(4-fluoro-5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0372] [ka]

[0373] 1 H NMR (400 MHz, DMSO-d 6) 8.57 (d, J = 9.3 Hz, 1H), 8.50 (d, J = 9.8 Hz, 1H), 7.71 (d, J =7.6 Hz, 1H), 7.52 (d, J = 10.2 Hz, 1H), 7.34 (s, 1H), 7.19 (d, J = 6.2 Hz, 1H), 7.07 (d, J = 10.1 Hz, 1H), 5.22 (s, 2H), 4.87 (s, 2H), 3.97 (s, 3H), 3.64 (s, 3H). 19 F NMR (376 MHz, DMSO-d 6 ) -125.50. Tr(MET-uHPLC-AB-101) = 2.64 min m / z (ES + )(M+H) + 397.2, 97%.

[0374] Method 2 Scheme for method 2

[0375] [ka]

[0376] Step 1: (5-(2-fluoroethoxy)pyridin-2-yl)methanol A mixture of 6-(hydroxymethyl)pyridin-3-ol (270 mg, 2.16 mmol) and potassium carbonate (447 mg, 3.23 mmol) in anhydrous MeCN (6 mL) was treated with 1-bromo-2-fluoroethane (0.32 mL, 4.3 mmol) and the resulting reaction mixture was heated in a sealed tube at 70° C. for 24 h. After this time, the reaction mixture was cooled to room temperature, diluted with water (50 mL) and extracted with EtOAc (3×50 mL). The organic layers were combined, dried over sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to afford the title compound (202 mg, 55%) as an orange-brown oil which was used in the next step without purification. 1 H NMR (300 MHz, DMSO-d 6) 8.22 (dd, J = 2.7, 0.6 Hz, 1H), 7.45 - 7.37 (m, 2H), 5.32 (t, J = 5.7 Hz, 1H), 4.85 - 4.82 (m, 1H), 4.69 - 4.66 (m, 1H), 4.49 (d, J = 5.7 Hz, 2H), 4.37 - 4.34 (m, 1H), 4.27 - 4.24 (m, 1H). MS (ES + ) (M+H) + 172.

[0377] Step 2: 5-{[5-(2-fluoroethoxy)pyridin-2-yl]methoxy}-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one To a mixture of 5-hydroxy-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one (80 mg, 0.31 mmol) and (5-(2-fluoroethoxy)pyridin-2-yl)methanol (106 mg, 0.622 mmol) in toluene (20 mL) was added CMBP (188 mg, 0.777 mmol). The mixture was heated at 120° C. in a sealed tube for 48 h. After this time, the solvent was removed under vacuum and the resulting residue was purified by FCC (silica, 0-10% MeOH in DCM). The product was repurified by preparative HPLC (water-MeCN) to give the title compound (31 mg, 24%).

[0378] Example 2-1: 5-{[5-(2-fluoroethoxy)pyridin-2-yl]methoxy}-2-(1-methyl-6-oxo-1,6-dihydro-pyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one 1 H NMR (500 MHz, DMSO-d 6) 8.58 (d, J = 10.0 Hz, 1H), 8.34 (d, J = 2.8 Hz, 1H), 7.72 (d, J = 8.4 Hz, 1H), 7.52 (d, J = 8.5 Hz, 1H), 7.49 (dd, J = 8.6, 2.8 Hz, 1H), 7.34 (d, J = 1.8 Hz, 1H), 7.18 (dd, J = 8.4, 2.1 Hz, 1H), 7.08 (d, J = 10.0 Hz, 1H), 5.23 (s, 2H), 4.87 (s, 2H), 4.82 - 4.71 (m, 2H), 4.38 - 4.31 (m, 2H), 3.63 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -222.34. Tr(MET-HPLC-005) = 8.78 min, (ES + ) (M+H) + 411.1, 99%.

[0379] The following additional compounds were prepared by Method 2:

[0380] Example 2-2: 5-{[5-(fluoromethoxy)pyridin-2-yl]methoxy}-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0381] [ka]

[0382] 1 H NMR (300 MHz, DMSO-d 6) 8.59 (d, J = 10.0 Hz, 1H), 8.45 (d, J = 2.7 Hz, 1H), 7.73 (d, J = 8.5 Hz, 1H), 7.69 - 7.57 (m, 2H), 7.35 (d, J = 1.9 Hz, 1H), 7.18 (dd, J = 8.5, 2.3 Hz, 1H), 7.09 (d, J = 10.0 Hz, 1H), 5.95 (d, J = 53.7 Hz, 2H), 5.27 (s, 2H), 4.88 (s, 2H), 3.64 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -151.56. Tr(MET-uHPLC-001) = 3.84 min, (ES+) (M+H)+ 397.3, 99%.

[0383] Example 2-3: 5-[(4-chloro-5-fluoropyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0384] [ka]

[0385] 1 H NMR (500 MHz, DMSO-d 6 ) 8.75 (d, J = 1.0 Hz, 1H), 8.57 (d, J = 9.5 Hz, 1H), 7.90 (d, J = 6.0 Hz, 1H), 7.74 (d, J = 8.5 Hz, 1H), 7.36 (d, J = 2.5 Hz, 1H), 7.22 (dd, J = 8.5, 2.5 Hz, 1H), 7.07 (d, J = 10.0 Hz, 1H), 5.29 (s, 2H), 4.88 (s, 2H), 3.64 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6) -132.59. Tr(MET-uHPLC-001) = 5.65 min m / z (ES+) (M+H)+ 401.1, 98%.

[0386] Method 3 Scheme for method 3

[0387] [ka]

[0388] Step 1: 5-[(5-fluoro-4-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one To a solution of 5-((4-chloro-5-fluoropyridin-2-yl)methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one (68 mg, 0.16 mmol) in MeOH (2.0 mL) and THF (2.0 mL) was added a 25 wt% solution of sodium methoxide in MeOH (0.041 mL, 0.18 mmol) and the reaction mixture was heated at 65° C. for 16 h. After this time, the solvent was removed under reduced pressure and the resulting residue was triturated in water and purified twice by FCC (silica, 0-15% MeOH in EtOAc). The collected material was purified again by FCC (silica, 0-10% MeOH in DCM) and recrystallized from MeCN to give the title compound (23 mg, 35%).

[0389] Example 3-1: 5-[(5-fluoro-4-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one 1 H NMR (500 MHz, DMSO-d 6) 8.57 (d, J = 10.0 Hz, 1H), 8.44 (d, J = 3.0 Hz, 1H), 7.73 (d, J = 8.5 Hz, 1H), 7.44 (d, J = 7.0 Hz, 1H), 7.36 (d, J = 2.0 Hz, 1H), 7.21 (dd, J = 8.5, 2.0 Hz, 1H), 7.07 (d, J = 10.0 Hz, 1H), 5.22 (s, 2H), 4.89 (s, 2H), 3.96 (s, 3H), 3.64 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -152.71. Tr(MET-uHPLC-001) = 4.42 min, (ES + ) (M+H) + 397.0, 99%.

[0390] Method 4 Method 4 scheme

[0391] [ka]

[0392] Step 1: (5-(allyloxy)pyridin-2-yl)methanol A solution of potassium carbonate (1.65 g, 11.9 mmol) in water (4 mL) was added dropwise over 15 min to a mixture of 6-(hydroxymethyl)pyridin-3-ol (1.00 g, 7.99 mmol) and allyl bromide (0.80 mL, 9.3 mmol) in acetone (10 mL) and the reaction mixture was heated at 60° C. for 2 h in a sealed tube. After this time, the mixture was cooled to room temperature and extracted with MTBE (3×100 mL). The organic layers were combined, dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound (713 mg, 54%) as a red-brown oil, which was used in the next step without purification. 1 H NMR (500 MHz, DMSO-d 6) 8.19 (dd, J = 2.5, 0.5 Hz, 1H), 7.49 - 7.35 (m, 2H), 6.07 - 5.99 (m, 1H), 5.40 (dq, J = 17.5, 1.5 Hz, 1H), 5.29 - 5.25 (m, 2H), 4.63 (dt, J = 5.0, 1.5 Hz, 2H), 4.48 (d, J = 5.5 Hz, 2H).

[0393] Step 2: 5-((5-(allyloxy)pyridin-2-yl)methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one CMBP (586 mg, 2.43 mmol) was added to a solution of 5-hydroxy-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one (250 mg, 0.972 mmol) and (5-(allyloxy)pyridin-2-yl)methanol (321 mg, 1.94 mmol) in toluene (36.0 mL) and the solution was heated at 120° C. for 3 days. After this time, the solvent was removed in vacuo. The resulting residue was suspended in DCM (5 mL) and heptane (5 mL) and filtered. The filter cake was washed with heptane (5 mL) and the filtrate was concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-5% MeOH in DCM). The resulting product was triturated with heptane (5 mL), collected by filtration and washed with heptane (5 mL) to give the title compound (45 mg, 12%) as a brown solid. 1 H NMR (300 MHz, DMSO-d 6) 8.58 (d, J = 9.9 Hz, 1H), 8.32 (d, J = 2.4 Hz, 1H), 7.72 (d, J = 8.4 Hz, 1H), 7.51 (d, J = 8.7 Hz, 1H), 7.45 (dd, J = 8.4, 2.7 Hz, 1H), 7.34 (d, J = 1.8 Hz, 1H), 7.18 (dd, J = 8.4, 2.1 Hz, 1H), 7.09 (d, J = 10.2 Hz, 1H), 6.11 - 5.99 (m, 1H), 5.42 (dq, J = 17.1, 1.5, Hz, 1H), 5.29 (dd, J = 10.5, 1.5 Hz, 1H), 5.22 (s, 2H), 4.88 (s, 2H), 4.67 (dt, J = 5.1, 1.2 Hz, 2H), 3.64 (s, 3H).

[0394] Step 3: 5-[(5-hydroxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one 1,3-Dimethylbarbituric acid (57 mg, 0.37 mmol) and tetrakis(triphenylphosphine)palladium(0) (10.5 mg, 0.00915 mmol) were added to a solution of 5-((5-(allyloxy)-pyridin-2-yl)methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one (74 mg, 0.18 mmol) in MeOH (6.6 mL) and the mixture was stirred at room temperature for 16 h. After this time the solvent was removed in vacuo and DCM (20 mL) was added. The mixture was washed with saturated aqueous sodium bicarbonate (20 mL), the layers were separated and the aqueous layer was extracted with DCM (2×30 mL). The combined organic layers were dried over sodium sulfate, filtered and the filtrate was concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-5% MeOH in DCM) and the product was lyophilized from MeCN (10 mL) and water (10 mL) to give the title compound (41 mg, 46%).

[0395] Example 4-1: 5-[(5-hydroxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one 1 H NMR (500 MHz, DMSO-d 6 ) 10.01 (br s, 1H), 8.57 (d, J = 10.0 Hz, 1H), 8.14 (d, J = 3.0 Hz, 1H), 7.71 (d, J = 8.5 Hz, 1H), 7.39 (d, J = 8.5 Hz, 1H), 7.33 (d, J = 2.0 Hz, 1H), 7.20 (dd, J = 8.5, 3.0 Hz, 1H), 7.16 (dd, J = 8.0, 2.0 Hz, 1H), 7.08 (d, J = 10.0 Hz, 1H), 5.16 (s, 2H), 4.87 (s, 2H), 3.64 (s, 3H). Tr(MET-HPLC-003) = 10.48 minutes, (ES + ) (M+H) + 365.2, 99%.

[0396] Method 5 Scheme of method 5

[0397] [ka]

[0398] Step 1: [6-({[2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-yl]oxy}methyl)pyridin-3-yl]oxydansulfonic acid Chlorosulfonic acid (0.091 mL, 1.4 mmol) was added to a mixture of 5-((5-hydroxypyridin-2-yl)methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one (50 mg, 0.14 mmol) in pyridine (6.3 mL) at -15°C. After the addition was complete, the mixture was allowed to warm to room temperature and stirred for 2 days. After this time, the solvent was removed in vacuo and water (5.0 mL) was added. The mixture was aged for 2.5 days. After this time, the solid that had formed was collected by filtration, washed with water (5.0 mL) and dried in vacuum. The dried solid was triturated with DCM (7.9 mL) and then purified by reverse phase chromatography (MPLC, water-MeCN). The resulting product was lyophilized and then triturated with water (15.0 mL) for 2 h, collected by filtration and dried under vacuum to give the title compound (27 mg, 44%).

[0399] Example 5-1: [6-({[2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-yl]oxy}methyl)pyridin-3-yl]oxydansulfonic acid 1 H NMR (300 MHz, DMSO-d 6 ) 8.58 (d, J = 9.9 Hz, 1H), 8.44 (d, J = 2.4 Hz, 1H), 7.79 - 7.72 (m, 2H), 7.59 (d, J = 8.7 Hz, 1H), 7.36 (s, 1H), 7.20 (dd, J = 8.7, Tr(MET-uHPLC-001) = 2.90 min, (ES - ) (MH) - 443.3, 98%.

[0400] Method 6 Scheme of method 6

[0401] [ka]

[0402] Step 1: 5-Bromo-2-(6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-3-yl)isoindolin-1-one In a sealed tube, 5-bromoisoindolin-1-one (700 mg, 3.30 mmol), 6-bromo-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazin-3(2H)-one (1.21 g, 3.96 mmol), RuPhos (231 mg, 0.495 mmol), cesium carbonate (3.23 g, 9.90 mmol), and Pd were added in 1,4-dioxane (16.9 mL). 2 (dba) 3 A mixture of (151 mg, 0.165 mmol) was heated at 100° C. under nitrogen for 16 h. After this time, the reaction mixture was combined with the other batches, diluted with water (250 mL), and extracted with EtOAc (3×250 mL). The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by FCC (silica, 0-40% EtOAc in DCM) to give the title compound (1.438 g, >99%). 1 H NMR (300 MHz, DMSO-d 6 ) 8.62 (d, J = 9.9 Hz, 1H), 8.01 (s, 1H), 7.78 - 7.77 (m, 2H), 7.17 (d, J = 9.9 Hz, 1H), 5.35 (s, 2H), 4.95 (s, 2H), 3.72 (t, J = 8.1 Hz, 2H), 0.91 (t, J = 8.1 Hz, 2H), 0.02 (s, 9H); MS (ES + ) (M+H) + 437.

[0403] Step 2: 2-(6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-1-one Pd(dppf)Cl 2 (134 mg, 0.165 mmol) was added to a mixture of 5-bromo-2-(6-((2-(trimethylsilyl)ethoxy)methoxy)pyridazin-3-yl)isoindolin-1-one (718 mg, 1.65 mmol), bis(pinacolato)diboron (627 mg, 2.47 mmol), potassium acetate (404 mg, 4.11 mmol), and 1,4-dioxane (14 mL) in a microwave vial. The suspension was sparged with argon, the vial was sealed, and heated at 90° C. for 2 h. After this time, the reaction mixture was combined with another batch and concentrated under reduced pressure. The resulting residue was partitioned between EtOAc (100 mL) and water (100 mL). The layers were separated and the aqueous layer was extracted with EtOAc (2×100 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated to give the title compound (2.947 g), which was used in the next step without purification.

[0404] Step 3: 5-Hydroxy-2-(6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-3-yl)isoindolin-1-one A suspension of sodium perborate tetrahydrate (1.27 g, 8.23 ​​mmol) in water (21 mL) was added to a suspension of crude 2-(6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-1-one (estimated 3.29 mmol) in THF (41 mL). The reaction mixture was stirred at room temperature for 16 h. After this time, saturated aqueous ammonium chloride solution (75 mL) was added and the mixture was concentrated under reduced pressure. The resulting residue was diluted with MeOH and the pH was adjusted to 3 with 2N HCl. MeOH was removed under reduced pressure and the resulting residue was suspended in water and aged at room temperature for 16 h. The resulting solid was collected by filtration, rinsed with water and dried. This material was purified by FCC (silica, 0-40% EtOAc in DCM, then 0-10% MeOH in DCM) to give the title compound (499 mg, 40%). 1 H NMR (300 MHz, DMSO-d 6 ) 10.49 (s, 1H), 8.61 (d, J = 10.2 Hz, 1H), 7.63 (d, J = 8.4 Hz, 1H), 7.11 (d, J = 10.2 Hz, 1H), 7.01 (s, 1H), 6.91 (dd, J = 8.4, 2.1 MS (ES) + ) (M+H) + 374.

[0405] Step 4: 5-((5-methoxypyridin-2-yl)methoxy)-2-(6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-3-yl)isoindolin-1-one Potassium carbonate (554 mg, 4.01 mmol) was added to a mixture of 5-hydroxy-2-(6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-3-yl)isoindolin-1-one (499 mg, 1.34 mmol) and 2-(chloromethyl)-5-methoxypyridine (253 mg, 1.60 mmol) in DMF (17 mL) and the mixture was heated at 70° C. for 1 h. After this time, the solvent was removed under reduced pressure and the resulting residue was dissolved in 80:20 DCM / MeOH and filtered. The filtrate was concentrated under reduced pressure and the crude product mixture was purified by FCC (silica, 20-80% EtOAc in DCM, then 0-20% MeOH in DCM) to give the title compound (494 mg, 75%). 1 H NMR (300 MHz, DMSO-d 6 ) 8.63 (d, J = 10.2 Hz, 1H), 8.33 (d, J = 2.7 Hz, 1H), 7.75 (d, J = 8.4 Hz, 1H), 7.54 (d, J = 8.7 Hz, 1H), 7.46 (dd, J = 8.4, 2.7 Hz, 1H), 7.37 (s, 1H), 7.20 (dd, J = 8.4, 1.8 Hz, 1H), 7.14 (d, J = 9.9 Hz, 1H), 5.34 (s, 2H), 5.24 (s, 2H), 4.89 (s, 2H), 3.86 (s, 3H), 3.72 (t, J = 7.8 Hz, 2H), 0.91 (t, J = 8.1 Hz, 2H), 0.02 (s, 9H). MS (ES + ) (M+H) + 495.

[0406] Step 5: 5-[(5-methoxypyridin-2-yl)methoxy]-2-(6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one TFA (1.9 mL) was added to a solution of 5-((5-methoxypyridin-2-yl)methoxy)-2-(6-((2-(trimethyl-silyl)ethoxy)methoxy)pyridazin-3-yl)isoindolin-1-one (494 mg, 0.999 mmol) in DCM (5.7 mL) and the mixture was stirred at room temperature for 2 h. After this time, heptane (73 mL) was added and the volatiles were removed under reduced pressure. The resulting residue was solvent swapped with EtOAc (14 mL) and heptane (73 mL) and the residue was triturated in 50:50 water / MeOH (145 mL) for 1 h at room temperature. The mixture was aged for 3 days and the solid was collected by filtration and washed with water (28 mL), MeOH (14 mL), and heptane (27 mL). The resulting solid was purified by FCC (silica, 0-20% MeOH in EtOAc, then 0-20% MeOH in DCM). The resulting product was combined with the previous batch and purified by reverse phase chromatography (MPLC, MeCN-water, 0.1% v / v TFA). The resulting clean fractions were poured into saturated sodium bicarbonate. The pH of the solution was adjusted to 6 with 2N HCl and the resulting precipitate was collected by filtration and washed with water and MeCN to give the title compound (264 mg).

[0407] Example 6-1: 5-[(5-methoxypyridin-2-yl)methoxy]-2-(6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one 1 H NMR (500 MHz, DMSO-d 6) 12.77 (s, 1H), 8.56 (d, J = 10.5 Hz, 1H), 8.30 (d, J = 3.0 Hz, 1H), 7.71 (d, J = 8.5 Hz, 1H), 7.51 (d, J = 8.5 Hz, 1H), 7.44 (dd, J = 9.0, 3.0 Hz, 1H), 7.33 (d, J = 2.0 Hz, 1H), 7.17 (dd, J = 8.5, 2.0 Hz, 1H), 7.01 (d, J = 10.5 Hz, 1H), 5.22 (s, 2H), 4.86 (s, 2H), 3.84 (s, 3H). Tr(MET-uHPLC-001) = 3.36 minutes, (ES + ) (M+H) + 365.1, 100%.

[0408] Method 7 Scheme of method 7

[0409] [ka]

[0410] Step 1: 5-[(5-methoxy-2-pyridyl)methoxy]isoindolin-1-one (5-Methoxypyridin-2-yl)methanol (100 mg, 0.719 mmol) was dissolved in DCM (2 mL). Thionyl chloride (0.104 mL, 1.44 mmol) was added and the reaction mixture was diluted with N 2 The reaction mixture was concentrated in vacuo and the residue was co-distilled with DCM (3×10 mL) and concentrated in vacuo to give 2-(chloromethyl)-5-methoxy-pyridine, which was used without further purification.

[0411] 5-Hydroxyisoindolin-1-one (100 mg, 0.670 mmol), KI (111 mg, 0.670 mmol), and Cs 2 CO 3(0.107 mL, 1.34 mmol) was dissolved in DMF (6 mL) and the reaction mixture was stirred at room temperature for 10 min. 2-(Chloromethyl)-5-methoxy-pyridine (116 mg, 0.738 mmol) was added and the reaction mixture was stirred at room temperature overnight. DMF was removed in vacuo and the residue was purified by H 2 Trituration with 2H2O (10 mL) and EtOH (10 mL) afforded the title compound (140 mg, 77% yield). 1 H NMR (500 MHz, DMSO-d 6 ) 8.29 (d, J = 2.8 Hz, 2H), 7.56 (d, J = 8.4 Hz, 1H), 7.49 (d, J = 8.6 Hz, 1H), 7.43 (dd, J = 8.6, 2.9 Hz, 1H), 7.20 (d, J = 1.9 Hz, 1H), 7.09 (dd, J = 8.4, 2.2 Hz, 1H), 5.18 (s, 2H), 4.30 (s, 2H), 3.84 (s, 3H). Tr(METCR1600) = 3.07 min, (ES + ) (M+H) + 271.1, 100%.

[0412] Step 2: 5-[(5-methoxy-2-pyridyl)methoxy]-2-[(6-methoxy-3-pyridyl)methyl]isoindolin-1-one NaH (60% in oil, 41 mg, 1.04 mmol) and THF (6 mL) were added to a solution of 5-[(5-methoxy-2-pyridyl)methoxy]isoindolin-1-one (140 mg, 0.518 mmol) and the reaction mixture was heated to 70° C. under reflux for 1 h. The reaction mixture was then cooled to room temperature and 5-(chloromethyl)-2-methoxy-pyridine (prepared as for step 1) (98 mg, 0.622 mmol) in 2:1 THF:DMF (6 mL) was added. The reaction mixture was stirred overnight at room temperature. Additional NaH (60% in oil, 41 mg, 1.036 mmol) was added to the reaction mixture, which was then heated under reflux for 30 min. The reaction mixture was cooled to room temperature and 5-(chloromethyl)-2-methoxy-pyridine (98 mg, 0.622 mmol) was added and the reaction mixture was stirred for 3 days. The mixture was diluted with EtOAc (25 mL) and H 2 The combined organic layers were washed with NaO (2×15 mL). 2 SO 4 The crude product was purified by preparative HPLC (MeCN-water, 2 mM NH 4 HCO 3 ) to give the title compound (11.5 mg, 5%).

[0413] Example 7-1: 5-[(5-methoxypyridin-2-yl)methoxy]-2-[(6-methoxypyridin-3-yl)methyl]-2,3-dihydro-1H-isoindol-1-one 1 H NMR (500 MHz, DMSO-d 6) 8.28 (d, J = 2.9 Hz, 1H), 8.12 (d, J = 2.3 Hz, 1H), 7.63 - 7.58 (m, 2H), 7.47 (d, J = 8.6 Hz, 1H), 7.42 (dd, J = 8.6, 2.9 Hz, 1H), 7.19 (d, J = 2.0 Hz, 1H), 7.11 (dd, J = 8.4, 2.2 Hz, 1H), 6.79 (d, J = 8.5 Hz, 1H), 5.16 (s, 2H), 4.63 (s, 2H), 4.30 (s, 2H), 3.83 (s, 3H), 3.83 (s, 3H). Tr(MET-uHPLC-AB-101) = 2.49 min, (ES + ) (M+H) + 392, 96%.

[0414] Method 8 Scheme of method 8

[0415] [ka]

[0416] Step 1: 6-Bromo-2-(fluoromethyl)pyridazin-3(2H)-one To a solution of 6-bromopyridazin-3(2H)-one (150 mg, 0.857 mmol) in 1,4-dioxane (5 mL) in a reaction vial was added potassium carbonate (237 mg, 1.71 mmol) followed by DMF (3 mL). The reaction vial was sealed and a cold solution of bromofluoromethane (2.0 M in MeCN, 0.69 mL, 1.4 mmol) was added. The mixture was heated at 150° C. for 1.5 h. After this time, the mixture was cooled to room temperature and concentrated under reduced pressure to dryness. The resulting residue was adsorbed onto silica gel and purified by FCC (silica, 0-50% EtOAc in hexanes) to give the title compound (90 mg, 51%). 1 H NMR (300 MHz, CDCl 3) 7.30 (d, J = 9.6 Hz, 1H), 6.87 (d, J = 9.6 Hz, 1H), 5.98 (d, J = 50.7 Hz, 2H). 19 F NMR (282 MHz, CDCl 3 ) -176.17. MS (ES+) (M+H) + 207.0.

[0417] Step 2: 2-(1-(fluoromethyl)-6-oxo-1,6-dihydropyridazin-3-yl)-5-((5-methoxypyridin-2-yl)methoxy)-isoindolin-1-one A mixture of 5-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one (35 mg, 0.13 mmol), 6-bromo-2-(fluoromethyl)pyridazin-3(2H)-one (32 mg, 0.16 mmol), RuPhos (9 mg, 0.02 mmol), and cesium carbonate (127 mg, 0.390 mmol) in 1,4-dioxane (2 mL) was purged with nitrogen for 2 min and purified by Pd 2 (dba) 3 (6.70 mg, 0.00732 mmol) was added. The reaction vial was sealed and heated at 110° C. for 4 h. After this time, the reaction mixture was concentrated to dryness and the resulting residue was purified by FCC (silica, 0-40% MeOH in EtOAc). The product was triturated in ethyl acetate and the solid was collected by filtration and dried under vacuum at 50° C. for 3 h to give the title compound (46 mg, 90%).

[0418] Example 8-1: 2-(1-(fluoromethyl)-6-oxo-1,6-dihydropyridazin-3-yl)-5-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one 1 H NMR (500 MHz, DMSO-d 6) 8.68 (d, J = 10.0 Hz, 1H), 8.31 (d, J = 2.5 Hz, 1H), 7.74 (d, J = 8.5 Hz, 1H), 7.52 (d, J = 8.5 Hz, 1H), 7.44 (dd, J = 9.0, 3.0 Hz, 1H), 7.35 (d, J = 2.0 Hz, 1H), 7.19-7.17 (m, 2H), 5.98 (d, J = 51.5 Hz, 2H), 5.22 (s, 2H), 4.88 (s, 2H), 3.84 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -174.59. Tr(MET-uHPLC-001) = 2.81 min, (ES + ) (M+H) + 397.0, 100%.

[0419] The following additional compounds were prepared by Method 8:

[0420] Example 8-2: 2-[1-(2-fluoroethyl)-6-oxo-1,6-dihydropyridazin-3-yl]-5-[(5-methoxypyridin-2-yl)methoxy]-2,3-dihydro-1H-isoindol-1-one

[0421] [ka]

[0422] 1 H NMR (500 MHz, DMSO-d 6) 8.59 (d, J = 10.0 Hz, 1H), 8.31 (d, J = 3.0 Hz, 1H), 7.73 (d, J = 8.5 Hz, 1H), 7.52 (d, J = 8.5 Hz, 1H), 7.44 (dd, J = 8.5, 3.0 Hz, 1H), 7.34 (d, J = 2.0 Hz, 1H), 7.18 (dd, J = 8.5, 2.0 Hz, 1H), 7.11 (d, J = 10.0 Hz, 1H), 5.22 (s, 2H), 4.89 (s, 2H), 4.82 (dt, J = 47.0, 5.0 Hz, 2H), 4.35 (dt, J = 26.5, 5.0 Hz, 2H), 3.84 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -224.07. Tr(MET-uHPLC-001) = 2.75 min m / z (ES+) (M+H)+ 411.1, 100%.

[0423] Method 9 Scheme of method 9

[0424] [ka]

[0425] Step 1: 5-[(5-fluoro-2-pyridyl)methoxy]isoindolin-1-one 5-Hydroxy-2,3-dihydro-1H-isoindol-1-one (400 mg, 2.68 mmol), 2-(chloromethyl)-5-fluoro-pyridine (468 mg, 3.22 mmol), and potassium carbonate (1.11 g, 8.05 mmol) were combined in DMF (50 mL) and stirred at 70° C. for 3 h. The reaction mixture was concentrated and partitioned between DCM (30 mL) and water (10 mL). The resulting precipitate was collected and dried by vacuum filtration to give the title compound (667 mg, 92% yield). 1 H NMR (500 MHz, DMSO-d 6) 8.60 (d, J = 2.9 Hz, 1H), 8.34 (s, 1H), 7.80 (td, J = 8.7, 2.9 Hz, 1H), 7.64 (dd, J = 8.7, 4.5 Hz, 1H), 7.59 (d, J = 8.4 Hz, 1H), 7.22 (d, J = 1.9 Hz, 1H), 7.12 (dd, J = 8.4, 2.3 Hz, 1H), 5.26 (s, 2H), 4.31 (s, 2H). 19 F NMR (471 MHz, DMSO-d 6 ) -128.46 (dd, J = 8.8, 4.5 Hz). Tr(METCR1410) = 0.94 min, (ES + ) (M+H) + 259.0, 96%.

[0426] Step 2: 2-[(6-bromopyridazin-3-yl)oxymethoxy]ethyl-trimethyl-silane 2-(Chloromethoxy)ethyl-trimethyl-silane (8.0 mL, 45.7 mmol) was added to a solution of 3-bromo-1H-pyridazin-6-one (4.00 g, 22.9 mmol) and potassium carbonate (9.48 g, 68.6 mmol) in DMF (80 mL) at room temperature. The reaction was stirred at room temperature for 20 h. The reaction mixture was re-treated with 2-(chloromethoxy)ethyl-trimethyl-silane (4.9 mL, 27.4 mmol), stirred for an additional 2 h, and concentrated to dryness. DCM (75 mL) was added and the organic phase was washed with water (2×50 mL) and brine (30 mL). The organic phase was diluted with Na 2 SO 4 It was dried at 40° C., filtered and concentrated in vacuo. The crude residue was purified by column chromatography (silica, 0-100% EtOAc in heptane) to give the title compound (4.27 g, 61% yield). 1 H NMR (400 MHz, DMSO-d 6) 7.62 (d, J = 9.7 Hz, 1H), 6.97 (d, J = 9.7 Hz, 1H), 5.28 (s, 2H), 3.68 - 3.58 (m, 2H), 0.91 - 0.82 (m, 2H), -0.04 (s, 9H). Tr(METCR1410) = 1.31 minutes, (ES + ) (M+H) + 356.8, 358.7, 100%.

[0427] Step 3: 5-[(5-fluoro-2-pyridyl)methoxy]-2-[6-oxo-1-(2-trimethylsilylethoxymethyl)pyridazin-3-yl]isoindolin-1-one A solution of 5-[(5-fluoro-2-pyridyl)methoxy]isoindolin-1-one (200 mg, 0.774 mmol) and 6-bromo-2-(2-trimethylsilylethoxymethyl)pyridazin-3-one (260 mg, 0.852 mmol) in 1,4-dioxane (15 mL) was degassed in a pressure tube for 5 min and then added RuPhos (11 mg, 0.0232 mmol) and Pd 2 (dba) 3 (21 mg, 0.023 mmol) was added. The reaction was degassed for an additional 5 min and Cs 2 CO 3 (0.30 g, 0.929 mmol) was added. The reaction was degassed for 5 min and stirred at 100 °C for 2 h. The cooled reaction mixture was re-treated with 5-[(5-fluoro-2-pyridyl)methoxy]isoindolin-1-one (50 mg, 0.19 mmol) and stirred overnight. The cooled reaction mixture was filtered and the filtrate concentrated. The residue was suspended in DCM (20 mL) and washed (10 mL). The organic phase was dried using a separator cartridge and concentrated in vacuo. The crude residue was purified by column chromatography (silica, 0-100% EtOAc in heptane) to give the title compound (281 mg, 74% yield). 1 H NMR (500 MHz, DMSO-d 6) 8.65 - 8.57 (m, 2H), 7.80 (td, J = 8.7, 2.9 Hz, 1H), 7.74 (d, J = 8.5 Hz, 1H), 7.66 (dd, J = 8.7, 4.5 Hz, 1H), 7.35 (d, J = 1.9 Hz, 1H), 7.20 (dd, J = 8.5, 2.3 Hz, 1H), 7.12 (d, J = 10.1 Hz, 1H), 5.38 - 5.24 (m, 4H), 4.87 (s, 2H), 3.74 - 3.66 (m, 2H), 0.92 - 0.87 (m, 2H), -0.03 (s, 9H). Tr(METCR1410) = 1.38 minutes, (ES + ) (M+H) + 483.0, 100%.

[0428] Step 4: 5-[(5-fluoro-2-pyridyl)methoxy]-2-(6-oxo-1H-pyridazin-3-yl)isoindolin-1-one TFA (1.1 mL, 14.6 mmol) was added to a solution of 5-[(5-fluoro-2-pyridyl)methoxy]-2-[6-oxo-1-(2-trimethylsilylethoxymethyl)pyridazin-3-yl]isoindolin-1-one (100%, 281 mg, 0.582 mmol) in DCM (9 mL) and the reaction was stirred at room temperature for 2.5 h. The reaction mixture was concentrated in vacuo, redissolved in DCM, and concentrated again (×3). The crude residue was dissolved in DCM (3 mL) and saturated NaHCO 3 The organic fraction was separated and dried using a separatory cartridge. The dried organic fraction was concentrated in vacuo to give the title compound (24 mg, 11% yield).

[0429] Example 9-1: 5-[(5-fluoro-2-pyridyl)methoxy]-2-(6-oxo-1H-pyridazin-3-yl)isoindolin-1-one 1 H NMR (500 MHz, DMSO-d 6) 12.78 (s, 1H), 8.61 (d, J = 2.9 Hz, 1H), 8.60 - 8.53 (m, 1H), 7.81 (td, J = 8.8, 2.9 Hz, 1H), 7.73 (d, J = 8.5 Hz, 1H), 7.67 (dt, J = 8.8, 4.6 Hz, 1H), 7.36 (d, J = 1.9 Hz, 1H), 7.24 - 7.16 (m, 1H), 7.05 (m, 1H), 5.31 (s, 2H), 4.87 (m, 2H). 19 F NMR (471 MHz, DMSO-d 6 ) -128.31 (ddd, J = 13.3, 8.8, 4.6 Hz). Tr(MET-uHPLC-AB-101) = 2.26 min m / z (ES + )(M+H) + 353.1, 99%.

[0430] The following additional compounds were prepared by Method 9:

[0431] Example 9-2: 5-((5-(2-fluoroethoxy-1,1,2,2- 2 H 4 )pyridin-2-yl)methoxy)-2-(6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one

[0432] [ka]

[0433] 1 H NMR (500 MHz, DMSO-d 6) 12.77 (s, 1H), 8.56 (d, J = 10.5 Hz, 1H), 8.34 (d, J = 2.5 Hz, 1H), 7.72 (d, J = 8.5 Hz, 1H), 7.52 (d, J = 8.5 Hz, 1H), 7.48 (dd, J = 8.5, 2.5 Hz, 1H), 7.34 (d, J = 2.0 Hz, 1H), 7.17 (dd, J = 8.5, 2.0 Hz, 1H), 7.02 (d, J = 10.5 Hz, 1H), 5.23 (s, 2H), 4.86 (s, 2H). 19 F NMR (282 MHz, DMSO-d 6 ) -224.22. Tr(MET-uHPLC-006) = 3.83 min m / z (ES+) (M+H)+ 401.1, 99%.

[0434] Example 9-3: 5-((5-fluoro-4-methoxypyridin-2-yl)methoxy)-2-(6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one

[0435] [ka]

[0436] 1 H NMR (500 MHz, DMSO-d 6 ) 12.77 (s, 1H), 8.57 (d, J = 10.2 Hz, 1H), 8.43 (d, J = 3.2 Hz, 1H), 7.73 (d, J = 8.4 Hz, 1H), 7.44 (d, J = 7.0 Hz, 1H), 7.36 (d, J = 1.6 Hz, 1H), 7.21 (dd, J = 8.4, 2.1 Hz, 1H), 7.02 (d, J = 10.1 Hz, 1H), 5.22 (s, 2H), 4.87 (s, 2H), 3.95 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6) -152.73. Tr(MET-uHPLC-006) = 3.88 min m / z (ES+) (M+H)+ 383.1, 99%.

[0437] Method 10 Scheme of method 10

[0438] [ka]

[0439] Step 1: 7-Fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-1-one 5-Bromo-7-fluoroisoindolin-1-one (500 mg, 2.17 mmol), bis(pinacolato)diboron (828 mg, 3.26 mmol), potassium acetate (533 mg, 5.43 mmol), and Pd(dppf)Cl in 1,4-dioxane (15.7 mL). 2 (159 mg, 0.217 mmol) was heated at 90° C. for 2 h. After this time, the reaction mixture was cooled, poured into water (50 mL) and extracted with DCM (3×30 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to give the title compound (870 mg, >99%). 1 H NMR (300 MHz, CDCl 3 ) 7.67 (s, 1H), 7.52 (d, J = 9.6 Hz, 1H), 6.53 (br s, 1H), 4.45 (s, 2H), 1.24 (s,12H). MS (ES + ) (M+H) + 278.

[0440] Step 2: 7-Fluoro-5-hydroxyisoindolin-1-one A solution of sodium perborate tetrahydrate (580 mg, 3.77 mmol) in water (53.6 mL) was added to a solution of 7-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-1-one (870 mg, 3.14 mmol) in THF (53.7 mL) and the mixture was stirred at room temperature for 1 h. After this time, saturated aqueous ammonium chloride (200 mL) was added and the volatiles were removed in vacuo. The remaining aqueous mixture was extracted with 3:1 chloroform / IPA (3×100 mL) and the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-10% MeOH in DCM) to give the title compound (111 mg, 31%). 1 H NMR (300 MHz, DMSO-d 6 ) 10.56 (br s, 1H), 8.22 (br s, 1H), 6.73 (d, J = 1.5 Hz, 1H), 6.55 (dd, J = 11.7, 1.8 Hz, 1H), 4.27 (s, 2H). MS (ES + ) (M+H) + 168.

[0441] Step 3: 7-Fluoro-5-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one Potassium carbonate (521 mg, 3.77 mmol) was added to a solution of 7-fluoro-5-hydroxyisoindolin-1-one (210 mg, 1.26 mmol) and 2-(chloromethyl)-5-methoxypyridine (198 mg, 1.26 mmol) in DMF (9.7 mL) and the mixture was stirred at room temperature for 16 h. After this time, water (100 mL) was added and the mixture was extracted with DCM (2×100 mL). The combined organic layers were dried over sodium sulfate, filtered and the filtrate was concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-5% MeOH in DCM) to give the title compound (200 mg, 55%). 1 H NMR (300 MHz, DMSO-d 6) 8.37 (br s, 1H), 8.30 (d, J = 2.4 Hz, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.44 (dd, J = 8.7, 3.0 Hz, 1H), 7.05 (d, J = 1.8 Hz, 1H), 6.94 (dd, J = 11.4, 1.8 Hz, 1H), 5.18 (s, 2H), 4.32 (s, 2H), 3.84 (s, 3H).

[0442] Step 4: 7-Fluoro-5-[(5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one 7-Fluoro-5-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one (100 mg, 0.347 mmol), 6-bromo-2-methyl-3(2H)-pyridazinone (79 mg, 0.42 mmol), RuPhos (24 mg, 0.052 mmol), cesium carbonate (339 mg, 1.04 mmol), and Pd in ​​1,4-dioxane (11.1 mL). 2 (dba) 3 A mixture of (16 mg, 0.017 mmol) was heated at 100° C. for 16 h. After this time, the reaction mixture was cooled, combined with water (50 mL) and extracted with DCM (3×100 mL). The combined organic layers were dried over sodium sulfate, filtered and the filtrate was concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-5% MeOH in DCM) to give the title compound (81 mg, 59%).

[0443] Example 10-1: 7-Fluoro-5-[(5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one 1 H NMR (500 MHz, DMSO-d 6) 8.50 (d, J = 10.0 Hz, 1H), 8.31 (d, J = 2.5 Hz, 1H), 7.53 (d, J = 8.5 Hz, 1H), 7.45 (dd, J = 8.5, 2.5 Hz, 1H), 7.19 (s, 1H), 7.09 - 7.05 (m, 2H), 5.22 (s, 2H), 4.88 (s, 2H), 3.85 (s, 3H), 3.63 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -116.43. Tr(MET-uHPLC-001) = 3.65 min, (ES + ) (M+H) + 397.1, 100%.

[0444] The following additional compounds were prepared by Method 10:

[0445] Example 10-2: 7-Fluoro-5-[(5-methoxypyridin-2-yl)methoxy]-2-(6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0446] [ka]

[0447] 1 H NMR (500 MHz, DMSO-d 6 ) 12.80 (s, 1H), 8.49 (d, J = 10.0 Hz, 1H), 8.31 (d, J = 2.5 Hz, 1H), 7.52 (d, J = 8.5 Hz, 1H), 7.45 (dd, J = 9.0, 3.0 Hz, 1H), 7.18 (d, J = 2.0 Hz, 1H), 7.05 (dd, J = 11.5, 2.0 Hz, 1H), 7.02 (dd, J = 10.0, 1.5 Hz, 1H), 5.22 (s, 2H), 4.86 (s, 2H), 3.84 (s, 3H). 19F NMR (282 MHz, DMSO-d 6 ) -116.59. Tr(MET-uHPLC-001) = 2.50 min m / z (ES+) (M+H)+ 383.1, 100%.

[0448] Method 11 Scheme of method 11

[0449] [ka]

[0450] Step 1: 5-Nitro-2-[(pyridin-2-yl)methoxy]pyridine Pyridin-3-ylmethanol (0.31 mL, 3.15 mmol) was added dropwise to a suspension of sodium hydride (132 mg, 3.31 mmol) in tetrahydrofuran (10 mL) under nitrogen with ice cooling. The mixture was stirred for 10 min. Then a solution of 2-chloro-5-nitropyridine (500 mg, 3.15 mmol) in tetrahydrofuran (5 mL) was added slowly. The mixture was stirred with ice cooling for 1 h. The mixture was quenched using water (1 mL), diluted with more water (30 mL) and extracted with ethyl acetate (2×30 mL). The combined organic extracts were washed with Na 2 SO 4 Dry at 40° C., filter and concentrate to give the title compound (737 mg, quantitative yield). 1 H NMR (250 MHz, DMSO-d 6 ) 9.18 - 9.07 (m, 1H), 8.71 (d, J = 1.7 Hz, 1H), 8.56 (dd, J = 4.8, 1.6 Hz, 1H), 8.52 (dd, J = 9.1, 2.9 Hz, 1H), 7.91 (dt, J = 7.8, 2.0 Hz, 1H), 7.48 - 7.39 (m, 1H), 7.12 (dd, J = 9.1, 0.5 Hz, 1H), 5.53 (s, 2H). Tr(METCR1278) = 1.16 min, (ES + ) (M+H) + 232, 100%.

[0451] Step 2: 6-[(pyridin-2-yl)methoxy]pyridin-3-amine A stirred suspension of 5-nitro-2-[(pyridin-2-yl)methoxy]pyridine (729 mg, 3.15 mmol) in ethanol (15 mL) was heated to 70° C. Ammonium chloride (1.65 g, 31.5 mmol) in water (5 mL) was then added, followed by iron powder (0.704 g, 12.6 mmol) in one portion. The reaction was stirred at 80° C. for 1 h. The mixture was then filtered through a glass fiber filter and the inorganics were washed with ethyl acetate (20 mL) and water (20 mL). The filtrate was then partitioned between ethyl acetate (80 mL) and water (80 mL). The aqueous extract was then further extracted with ethyl acetate (80 mL). The combined organic extracts were dried (Na 2 SO 4 ), filtered and concentrated to give the title compound (0.62 g, 98% yield). 1 H NMR (500 MHz, DMSO-d 6 ) 8.62 (d, J = 1.7 Hz, 1H), 8.50 (dd, J = 4.8, 1.5 Hz, 1H), 7.81 (d, J = 7.8 Hz, 1H), 7.50 (d, J = 2.6 Hz, 1H), 7.38 (dd, J = 7.8, 4.8 Hz, Tr(METCR1278) = solvent front, (ES + ) (M+H) + 202.

[0452] Step 3: Ethyl 2-(bromomethyl)-4-methoxybenzoate Ethyl 4-methoxy-2-methylbenzoate (900 mg, 4.63 mmol) and NBS (907 mg, 5.10 mmol) were dissolved in DCE (35 mL). AIBN (76 mg, 0.46 mmol) was added to the reaction mixture and the resulting solution was heated to reflux for 2 h. The reaction mixture was cooled and concentrated. Column chromatography of the crude residue (silica, 2-20% EtOAc-heptane) afforded the title compound (806 mg, 64%). 1 H NMR (500 MHz, CDCl 3 ) 7.99 (d, J = 8.75 Hz, 1H), 6.96 (d, J = 2.63 Hz, 1H), 6.86 (dd, J = 2.64, 8.76 Hz, 1H), 4.96 (s, 2H), 4.37 (q, J = 7.13 Hz, 2H), 3.86 (s, 3H), 1.41 (t, J = 7.14 Hz, 3H). Tr(METCR1278) = 2.17 min, (ES + ) (M+H) + 273, 275, 99%.

[0453] Step 4: 5-Methoxy-2-{6-[(pyridin-3-yl)methoxy]pyridin-3-yl}-2,3-dihydro-1H-isoindol-1-one N,N-Diethylisopropylamine (0.038 mL, 0.22 mmol) was added to 6-(pyridin-3-ylmethoxy)pyridin-3-amine (37 mg, 0.18 mmol) and ethyl 2-(bromomethyl)-4-methoxybenzoate (50 mg, 0.18 mmol) in ethanol (2 mL) under nitrogen. The mixture was heated at 110 °C in a sealed tube overnight. The mixture was then diluted with water (4 mL) and filtered. The collected solid was then purified by FCC (silica, 0-5% methanol in dichloromethane) and then by preparative HPLC (water-acetonitrile) to give the title compound (6.1 mg, 10% yield).

[0454] Example 11-1: 5-Methoxy-2-{6-[(pyridin-3-yl)methoxy]pyridin-3-yl}-2,3-dihydro-1H-isoindol-1-one 1 H NMR (500 MHz, DMSO-d 6 ) 8.69 (d, J = 1.7 Hz, 1H), 8.61 - 8.48 (m, 2H), 8.30 (dd, J = 9.0, 2.8 Hz, 1H), 7.88 (dt, J = 7.8, 2.0 Hz, 1H), 7.69 (d, J = 8.4 Hz, 1H), 7.42 (dd, J = 7.8, 4.8 Hz, 1H), 7.21 (d, J = 1.9 Hz, 1H), 7.09 (dd, J = 8.4, 2.3 Hz, 1H), 7.00 (d, J = 9.0 Hz, 1H), 5.40 (s, 2H), 4.96 (s, 2H), 3.87 (s, 3H). Tr(MET-uHPLC-AB-101) = 1.87 min, (ES + ) (M+H) + 348, 97%

[0455] Method 12 Scheme of method 12

[0456] [ka]

[0457] Step 1: Methyl 4-bromo-2-(dibromomethyl)-5-fluorobenzoate To a solution of methyl 4-bromo-5-fluoro-2-methylbenzoate (600 mg, 2.43 mmol) in carbon tetrachloride (40 mL) was added NBS (1.08 g, 6.07 mmol) and AIBN (8 mg, 0.05 mmol) and the mixture was stirred overnight at 80° C. After this time, the volatiles were removed under reduced pressure and the resulting residue was purified by FCC (silica, 0-5% EtOAc in hexanes) to give the title compound (720 mg, 73%). 1 H NMR (300 MHz, CDCl 3) 8.37 (d, J = 6.5 Hz, 1H), 7.97 (s, 1H), 7.65 (d, J = 8.8 Hz, 1H), 3.96 (s, 3H).

[0458] Step 2: Methyl 4-bromo-2-(bromomethyl)-5-fluorobenzoate To a solution of methyl 4-bromo-2-(dibromomethyl)-5-fluorobenzoate (670 mg, 1.65 mmol) and DIPEA (91 mg, 6.6 mmol) in THF (70 mL) was added diethyl phosphonate (856 mg, 6.62 mmol) and the mixture was stirred at room temperature for 18 h. After this time, the volatiles were removed under reduced pressure and the resulting residue was purified by FCC (silica, 0-5% EtOAc in hexanes) to give the title compound (346 mg, 64%). 1 H NMR (300 MHz, CDCl 3 ) 7.77-7.66 (m, 2H), 4.89 (s, 2H), 3.95 (s, 3H).

[0459] Step 3: 5-Bromo-6-fluoro-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one 6-Amino-2-methylpyridazin-3(2H)-one (286 mg, 2.29 mmol) and DIPEA (0.906 mL, 5.20 mmol) were added to a solution of methyl 4-bromo-2-(bromomethyl)-5-fluorobenzoate (339 mg, 1.04 mmol) in DMF (14.6 mL) and the mixture was stirred at 80° C. for 16 h. After this time, the reaction mixture was concentrated in vacuo. The resulting residue was suspended in THF (7.3 mL) and EtOH (7.3 mL) and lithium hydroxide (65.0 mg, 2.70 mmol) was added. The mixture was stirred at room temperature for 2 h. After this time, the volatiles were removed in vacuo and water (10 mL) was added. The solid that formed was collected by filtration and dried in vacuo to give the title compound (217 mg, 62%). 1 H NMR (300 MHz, DMSO-d 6) 8.54 (d, J = 10.2 Hz, 1H), 8.12 (d, J = 6.0 Hz, 1H), 7.80 (d, J = 7.8 Hz, 1H), 7.12 (d, J = 10.2 Hz, 1H), 4.92 (s, 2H), 3.65 (s, 3H). MS (ES + ) (M+H) + 339.

[0460] Step 4: 6-Fluoro-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxa-borolan-2-yl)isoindolin-1-one 5-Bromo-6-fluoro-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one (154 mg, 0.455 mmol), bis(pinacolato)diboron (173 mg, 0.683 mmol), potassium acetate (112 mg, 1.14 mmol), and Pd(dppf)Cl in 1,4-dioxane (8.2 mL). 2 (33 mg, 0.046 mmol) was heated at 90° C. for 6 h. After this time, the reaction mixture was cooled and concentrated in vacuo. The resulting residue was triturated in water (5 mL) and the solid that formed was collected by filtration, washed with water (5 mL) and dried in vacuo to give the title compound (172 mg, 98%). 1 H NMR (300 MHz, CDCl 3 ) 8.72 (d, J = 9.9 Hz, 1H), 7.89 (d, J = 4.5 Hz, 1H), 7.53 (d, J = 8.1 Hz, 1H), 7.04 (d, J = 9.9 Hz, 1H), 4.87 (s, 2H), 3.78 (s, 3H), 1.40 (s, 12H). MS (ES + ) (M+H) + 386.

[0461] Step 5: 6-Fluoro-5-hydroxy-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one A solution of sodium perborate tetrahydrate (172 mg, 1.12 mmol) in water (2.6 mL) was added to a solution of 6-fluoro-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindolin-1-one (172 mg, 0.447 mmol) in MeCN (12.9 mL) and the mixture was stirred at room temperature for 2 hours. After this time, saturated aqueous ammonium chloride solution (1 mL) was added and the mixture was concentrated in vacuo. Water (10 mL) was added, the pH was adjusted to 5 with 2N hydrochloric acid and the mixture was allowed to stand for 16 hours. After this time the solid that formed was collected by filtration, dried in vacuo and triturated in MeOH (8 mL) to give the title compound (95 mg, 76%). 1 H NMR (300 MHz, DMSO-d 6 ) 11.02 (br s, 1H), 8.55 (d, J = 10.2 Hz, 1H), 7.54 (d, J = 9.9 Hz, 1H), 7.22 (d, J = 7.5 Hz, 1H), 7.08 (d, J = 9.9 Hz, 1H), 4.82 (s, 2H), 3.63 (s, 3H). MS (ES + ) (M+H) + 276.

[0462] Step 6: 6-Fluoro-5-[(5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one Potassium carbonate (143 mg, 1.04 mmol) was added to a solution of 6-fluoro-5-hydroxy-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one (95 mg, 0.35 mmol) and 2-(chloromethyl)-5-methoxypyridine (65 mg, 0.41 mmol) in DMF (6.3 mL) and the mixture was stirred at 30° C. for 16 h. After this time, water (20 mL) was added and the mixture was extracted with DCM (2×30 mL). The combined organic layers were dried over sodium sulfate, filtered and the filtrate was concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-5% MeOH in DCM) to give the title compound (83 mg, 61%) as an off-white solid. The product was combined with another lot (19 mg) and dissolved in DCM (10 mL). The solution was added dropwise to hexane (75 mL) and the solid that formed was collected by filtration, washed with hexane (50 mL) and dried in vacuo to give the title compound (73 mg).

[0463] Example 12-1: 6-Fluoro-5-[(5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one 1 H NMR (500 MHz, DMSO-d 6 ) 8.69 (d, J = 10.0 Hz, 1H), 8.31 (d, J = 3.0 Hz, 1H), 7.57 (d, J = 9.5 Hz, 1H), 7.49 (d, J = 8.5 Hz, 1H), 7.27 - 7.24 (m, 1H), 7.17 (d, J = 7.0 Hz, 1H), 7.01 (d, J = 10.0 Hz, 1H), 5.31 (s, 2H), 4.81 (s, 2H), 3.88 (s, 3H), 3.75 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -131.86. Tr(MET-uHPLC-001) = 4.53 min, (ES + ) (M+H) + 397.1, 100%.

[0464] The following additional compounds were prepared by Method 12:

[0465] Example 12-2: 4-Fluoro-5-[(5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0466] [ka]

[0467] 1 H NMR (300 MHz, DMSO-d 6 ) 8.55 (d, J = 10.0 Hz, 1H), 8.31 (d, J = 2.5 Hz, 1H), 7.62 (d, J = 8.3 Hz, 1H), 7.58-7.42 (m, 3H), 7.10 (d, J = 10.0 Hz, 1H), 5.31 (s, 2H), 4.98 (s, 2H), 3.84 (s, 3H), 3.65 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -141.12. Tr(MET-uHPLC-006) = 4.28 min m / z (ES+) (M+H)+ 397.0, 99%.

[0468] Example 12-3: 4-Fluoro-6-[(5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0469] [ka]

[0470] 1 H NMR (500 MHz, DMSO-d 6) 8.52 (d, J = 10.0 Hz, 1H), 8.30 (d, J = 3.0 Hz, 1H), 7.50 (d, J = 8.5 Hz, 1H), 7.43 (dd, J = 8.5, 3.0 Hz, 1H), 7.30 (dd, J = 10.5, 2.0 Hz, 1H), 7.27 (d, J = 1.5 Hz, 1H), 7.10 (d, J = 10.0 Hz, 1H), 5.23 (s, 2H), 4.92 (s, 2H), 3.83 (s, 3H), 3.65 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -117.70. Tr(MET-uHPLC-001) = 4.33 min m / z (ES+) (M+H)+ 397.2, 99%.

[0471] Example 12-4: 5-Fluoro-6-[(5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0472] [ka]

[0473] 1 H NMR (500 MHz, DMSO-d 6 ) 8.54 (d, J = 9.9 Hz, 1H), 8.30 (d, J = 2.6 Hz, 1H), 7.61 (d, J = 9.0 Hz, 1H), 7.59 (d, J = 6.3 Hz, 1H), 7.51 (d, J = 8.6 Hz, 1H), 7.45 (dd, J = 8.6, 2.9 Hz, 1H), 7.09 (d, J = 9.9 Hz, 1H), 5.30 (s, 2H), 4.85 (s, 2H), 3.83 (s, 3H), 3.63 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6) -125.54. Tr(MET-uHPLC-001) = 4.41 min m / z (ES+) (M+H)+ 397.0, 99%.

[0474] Example 12-5: 6-{3-[(5-methoxypyridin-2-yl)methoxy]-7-oxo-5H,6H,7H-pyrrolo[3,4-b]pyridin-6-yl}-2-methyl-2,3-dihydropyridazin-3-one

[0475] [ka]

[0476] 1 H NMR (500 MHz, DMSO-d 6 ) 8.58 (d, J = 10.0 Hz, 1H), 8.54 (d, J = 3.0 Hz, 1H), 8.32 (d, J = 3.0 Hz, 1H), 7.82 (d, J = 2.5 Hz, 1H), 7.56 (d, J = 8.5 Hz, 1H), 7.46 (dd, J = 8.6, 2.7 Hz, 1H), 7.10 (d, J = 10.0 Hz, 1H), 5.29 (s, 2H), 4.89 (s, 2H), 3.84 (s, 3H), 3.65 (s, 3H), (ES+) (M+H)+ 380.3, 99%.

[0477] Example 12-6: 6-(3-{[5-(2-fluoroethoxy)pyridin-2-yl]methoxy}-7-oxo-5H,6H,7H-pyrrolo[3,4-b]pyridin-6-yl)-2-methyl-2,3-dihydropyridazin-3-one

[0478] [ka]

[0479] 1 H NMR (500 MHz, DMSO-d6 ) 8.58 (d, J = 10.0 Hz, 1H), 8.54 (d, J = 2.6 Hz, 1H), 8.35 (d, J = 2.9 Hz, 1H), 7.82 (d, J = 2.6 Hz, 1H), 7.57 (d, J = 8.5 Hz, 1H), 7.50 (dd, J = 8.5, 2.9 Hz, 1H), 7.09 (d, J = 10.0 Hz, 1H), 5.31 (s, 2H), 4.89 (s, 2H), 4.77 (dt, J = 47.7, 3.9 Hz, 2H), 4.36 (dt, J = 31.0, 4.0 Hz, 2H), 3.65 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -222.32. Tr(MET-uHPLC-001) = 3.23 min, (ES+) (M+H)+ 412.4, 98%.

[0480] Example 12-7: 6-{3-[(5-fluoropyridin-2-yl)methoxy]-7-oxo-5H,6H,7H-pyrrolo[3,4-b]pyridin-6-yl}-2-methyl-2,3-dihydropyridazin-3-one

[0481] [ka]

[0482] 1 H NMR (500 MHz, CDCl 3 ) 8.78 (d, J = 10.0 Hz, 1H), 8.60 (d, J = 2.5 Hz, 1H), 8.49 (d, J = 2.5 Hz, 1H), 7.55 (dd, J = 9.0, 4.5 Hz, 1H), 7.49 (td, J = 8.0, 3.0 Hz, 1H), 7.41 (d, J = 2.5 Hz, 1H), 7.04 (d, J = 10.0 Hz, 1H), 5.33 (s, 2H), 4.88 (s, 2H), 3.76 (s, 3H). 19F NMR (282 MHz, CDCl 3 ) -126.65. Tr(MET-uHPLC-001) = 3.76 min m / z (ES+) (M+H)+ 368.1, 100%.

[0483] Example 12-8: 6-{3-[(3-fluoro-5-methoxypyridin-2-yl)methoxy]-7-oxo-5H,6H,7H-pyrrolo[3,4-b]pyridin-6-yl}-2-methyl-2,3-dihydropyridazin-3-one

[0484] [ka]

[0485] 1 H NMR (500 MHz, DMSO-d 6 ) 8.58 (d, J = 10.0 Hz, 1H), 8.53 (d, J = 2.5 Hz, 1H), 8.23 ​​(d, J = 2.0 Hz, 1H), 7.86 (d, J = 2.5 Hz, 1H), 7.55 (dd, J = 11.5, 2.5 Hz, 1H), 7.10 (d, J = 10.0 Hz, 1H), 5.36 (d, J = 2.0 Hz, 2H), 4.90 (s, 2H), 3.89 (s, 3H), 3.66 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -123.19. Tr(MET-uHPLC-001) = 3.96 min m / z (ES+) (M+H)+ 398.4, 99%.

[0486] Example 12-9: 6-[3-({5-[2-fluoro(1,1,2,2- 2 H 4 )Ethoxy]pyridin-2-yl}methoxy)-7-oxo-5H,6H,7H-pyrrolo[3,4-b]pyridin-6-yl]-2-methyl-2,3-dihydropyridazin-3-one

[0487] [ka]

[0488] 1 H NMR (500 MHz, DMSO-d 6 ) 8.58 (d, J = 10.0 Hz, 1H), 8.54 (d, J = 2.5 Hz, 1H), 8.35 (d, J = 3.0 Hz, 1H), 7.82 (d, J = 3.0 Hz, 1H), 7.57 (d, J = 8.5 Hz, 1H), 7.50 (dd, J = 8.5, 3.0 Hz, 1H), 7.10 (d, J = 10.0 Hz, 1H), 5.30 (s, 2H), 4.89 (s, 2H), 3.65 (s, 3H). 19 F NMR (282 MHz, CDCl 3 ) -224.24. Tr(MET-uHPLC-002) = 2.67 min m / z (ES+) (M+H)+ 416.2, 100%.

[0489] Example 12-10: 7-Fluoro-6-[(5-methoxypyridin-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindol-1-one

[0490] [ka]

[0491] 1 H NMR (500 MHz, DMSO-d 6) 8.49 (d, J = 10.0 Hz, 1H), 8.29 (d, J = 2.9 Hz, 1H), 7.61 (t, J = 8.0 Hz, 1H), 7.51 (d, J = 8.6 Hz, 1H), 7.44 (dd, J = 8.6, 2.9 Hz, 1H), 7.39 (d, J = 8.3 Hz, 1H), 7.09 (d, J = 10.0 Hz, 1H), 5.25 (s, 2H), 4.85 (s, 2H), 3.83 (s, 3H), 3.63 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -141.33. Tr(MET-uHPLC-002) = 2.92 min m / z (ES+) (M+H)+ 397.0, 99%.

[0492] Method 13 Scheme of method 13

[0493] [ka]

[0494] Step 1: 5-Bromo-6-fluoroisoindolin-1-one Ammonia gas was bubbled through a solution of methyl 4-bromo-2-(bromomethyl)-5-fluorobenzoate (330 mg, 1.01 mmol) in MeOH (10 mL) at room temperature until saturated, and the mixture was stirred overnight. After this time, the volatiles were removed under reduced pressure, and the resulting residue was suspended in 1:1 MeOH / water and filtered. The resulting solid was washed with water and dried under vacuum to give the title compound (217 mg, 93%). 1 H NMR (300 MHz, DMSO-d 6 ) 8.81 (s, 1H), 7.99 (d, J = 5.9 Hz, 1H), 7.61 (d, J = 7.6 Hz, 1H), 4.36 (s, 2H).

[0495] Step 2: 6-Fluoro-5-hydroxyisoindolin-1-one To a mixture of 5-bromo-6-fluoroisoindolin-1-one (210 mg, 0.913 mmol), bis(pinacolato)diboron (348 mg, 1.37 mmol) and potassium acetate (224 mg, 2.28 mmol) in 1,4-dioxane (20 mL), Pd(dppf)Cl 2 (33 mg, 0.046 mmol) was added and the mixture was heated at 100° C. for 18 h. After this time, the mixture was cooled to room temperature and a solution of sodium perborate tetrahydrate (351 mg, 2.28 mmol) in water (6 mL) was added. The mixture was stirred for 2 h and concentrated in vacuo. The resulting residue was purified by FCC (silica, 0-20% MeOH in EtOAc) to give the title compound (107 mg, 70%). 1 H NMR (300 MHz, DMSO-d 6 ) 10.58 (s, 1H), 8.36 (s, 1H), 7.36 (d, J = 9.9 Hz, 1H), 7.09 (d, J = 7.5 Hz, 1H), 4.24 (s, 2H).

[0496] Step 3: 6-Fluoro-5-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one To a mixture of 6-fluoro-5-hydroxyisoindolin-1-one (100 mg, 0.598 mmol) and potassium carbonate (248 mg, 1.79 mmol) in DMF (10 mL) at room temperature, 2-(chloromethyl)-5-methoxypyridine hydrochloride (174 mg, 0.897 mmol) was added and the mixture was heated at 60° C. overnight. After this time, the solvent was removed under reduced pressure and the resulting residue was purified by FCC (silica, 0-100% MeOH in EtOAc) to give the title compound (85 mg, 49%). 1 H NMR (300 MHz, DMSO-d 6 ) 8.50 (s, 1H), 8.31 (d, J = 2.2 Hz, 1H), 7.53-7.40 (m, 4H), 5.24 (s, 2H), 4.30 (s, 2H), 3.84 (s, 3H). MS (ES + ) (M+H) + 289.

[0497] Step 4: 6-Fluoro-5-((5-methoxypyridin-2-yl)methoxy)-2-(6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-3-yl)isoindolin-1-one To a mixture of 6-fluoro-5-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one (80 mg, 0.28 mmol), 3-bromo-6-((2-(trimethylsilyl)ethoxy)methoxy)pyridazine (0.127 g, 0.416 mmol), RuPhos (0.013 g, 0.028 mmol), and cesium carbonate (0.271 g, 0.833 mmol) in 1,4-dioxane (10 mL), Pd 2 (dba) 3 (0.038 g, 0.042 mmol) was added and the mixture was heated at 100° C. overnight. After this time, the volatiles were removed under reduced pressure and the resulting residue was purified by FCC (silica, 0-100% EtOAc in hexanes) to give 6-fluoro-5-((5-methoxypyridin-2-yl)methoxy)-2-(6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-3-yl)isoindolin-1-one (0.103 g, 72%). 1 H NMR (300 MHz, DMSO-d 6 ) 8.58 (d, J = 10.0 Hz, 1H), 8.32 (d, J = 2.4 Hz, 1H), 7.64 (d, J = 17.5 Hz, 1H), 7.64 (s, 1H), 7.55 (d, J = 8.5 Hz, 1H), 7.47 (dd, J = 8.5, 2.9 Hz, 1H), 7.13 (d, J = 10.0 Hz, 1H), 5.31 (s, 2H), 5.29 (s, 2H), 4.85 (s, 2H), 3.84 (s, 3H), 3.69 (t, J = 15.9 Hz, 2H), 0.88 (t, J = 7.8 Hz, 2H), -0.024 (s, 9H). MS (ES + ) (M+H) + 513.

[0498] Step 5: 6-Fluoro-5-((5-methoxypyridin-2-yl)methoxy)-2-(6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one A mixture of TFA (2.00 mL, 26.9 mmol) and 6-fluoro-5-((5-methoxypyridin-2-yl)methoxy)-2-(6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-3-yl)isoindolin-1-one (103 mg, 0.201 mmol) was stirred at room temperature for 30 min. After this time, the volatiles were removed in vacuo and the resulting residue was purified by preparative HPLC (MeCN-water, 0.1% v / v formic acid). The resulting product was repurified by FCC (silica, 0-20% MeOH in EtOAc) to give the title compound (44 mg, 57%).

[0499] Example 13-1: 6-Fluoro-5-((5-methoxypyridin-2-yl)methoxy)-2-(6-oxo-1,6-dihydropyridazin-3-yl)isoindolin-1-one 1 H NMR (500 MHz, DMSO-d 6 ) 12.78 (br s, 1H), 8.53 (d, J = 10.2 Hz, 1H), 8.31 (d, J = 2.8 Hz, 1H), 7.65-7.60 (m, 2H), 7.54 (d, J = 8.5 Hz, 1H), 7.46 (dd, J = 8.6, 2.9 Hz, 1H), 7.02 (d, J = 10.1 Hz, 1H), 5.29 (s, 2H), 4.85 (s, 2H), 3.84 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -133.41. Tr(MET-uHPLC-002) = 2.94 min, (ES + ) (M+H) + 383.1, 98%.

[0500] The following additional compounds were prepared by Method 13:

[0501] Example 13-2: 5-Fluoro-6-[(5-methoxy-2-pyridyl)methoxy]-2-(6-oxo-1H-pyridazin-3-yl)isoindolin-1-one

[0502] [ka]

[0503] 1 H NMR (500 MHz, DMSO-d6) 12.87 (s, 1H), 8.51 (d, J = 10.2 Hz, 1H), 8.30 (d, J = 2.8 Hz, 1H), 7.51 (d, J = 8.6 Hz, 1H), 7.43 (dd, J = 8.6, 2.9 Hz, 1H), 7.30 (dd, J = 10.6, 1.9 Hz, 1H), 7.27 (d, J = 2.0 Hz, 1H), 7.05 (d, J = 10.2 Hz, 1H), 5.24 (s, 2H), 4.91 (s, 2H), 3.84 (s, 3H).19F NMR (471 MHz, DMSO-d6) -117.76 (d, J = 10.5 Hz). Tr(MET-uHPLC-AB-101) = 2.21 min m / z (ES+)(M+H)+ 383.2, 98%.

[0504] Example 13-3: 4-Fluoro-6-[(5-methoxy-2-pyridyl)methoxy]-2-(6-oxo-1H-pyridazin-3-yl)isoindolin-1-one

[0505] [ka]

[0506] 1H NMR (400 MHz, DMSO-d6) 12.87 (s, 1H), 8.51 (d, J = 10.2 Hz, 1H), 8.30 (d, J = 2.8 Hz, 1H), 7.51 (d, J = 8.6 Hz, 1H), 7.43 (dd, J = 8.6, 2.9 19F NMR (376 MHz, DMSO-d6) -117.76. Tr(MET-uHPLC-AB-101) = 2.26 minutes m / z (ES+)(M+H)+ 383.2, 100%.

[0507] Method 14 Scheme of method 14

[0508] [ka]

[0509] Step 1: Methyl 5-bromo-2-(bromomethyl)-4-fluorobenzoate To a solution of methyl 5-bromo-4-fluoro-2-methylbenzoate (400 mg, 1.62 mmol) in carbon tetrachloride (15 mL) was added NBS (288 mg, 1.62 mmol) and AIBN (5.3 mg, 0.032 mmol) and the mixture was stirred overnight at 80° C. After this time, the volatiles were removed under reduced pressure and the resulting residue was purified by FCC (silica, 0-5% EtOAc in hexanes) to give the title compound (487 mg, 92%). 1 H NMR (300 MHz, CDCl 3 ) 8.23 ​​(d, J = 7.0 Hz, 1H), 7.26 (d, J = 8.7 Hz, 1H), 4.89 (s, 2H), 3.94 (s, 3H).

[0510] Step 2: 6-Bromo-5-fluoroisoindolin-1-one To a solution of methyl 5-bromo-2-(bromomethyl)-4-fluorobenzoate (172 mg, 0.528 mmol) in MeOH (6 mL) was added 7N ammonia in MeOH (0.45 mL, 3.2 mmol), followed by ammonium hydroxide (2 mL, 0.53 mmol) and the mixture was stirred at room temperature for 18 h. After this time, the volatiles were removed under reduced pressure. The resulting residue was suspended in water (15 mL) and the mixture was neutralized with 1N HCl. The resulting solid was collected by filtration and dried under high vacuum to give the title compound (119 mg, 98%). 1 H NMR (300 MHz, CDCl 3 ) 8.08 (d, J = 6.3 Hz, 1H), 7.25 (d, J = 8.4 Hz, 1H), 6.51 (br s, 1H), 4.42 (s, 2H).

[0511] Step 3: 5-Fluoro-6-hydroxyisoindolin-1-one A mixture of 6-bromo-5-fluoroisoindolin-1-one (118 mg, 0.513 mmol), bis(pinacolato)diboron (195 mg, 0.768 mmol), and potassium acetate (126 mg, 1.28 mmol) in 1,4-dioxane (7 mL) was purged with nitrogen for 2 min. Pd(dppf)Cl 2 (38 mg, 0.051 mmol) was added and the reaction mixture was heated in a sealed tube at 100° C. for 1.5 h. After this time, the mixture was cooled to room temperature and concentrated to dryness under reduced pressure. The resulting residue was suspended in THF (5.0 mL) and water (5.0 mL), sodium perborate tetrahydrate (197 mg, 1.28 mmol) was added and the mixture was stirred at room temperature for 30 min. After this time, aqueous ammonium chloride (5 mL) was added. Volatiles were removed under reduced pressure and the resulting residue was purified by FCC (silica, 0-20% MeOH in EtOAc) to give the title compound (66 mg, 62%). MS (ES + ) (M+H) + 168.

[0512] Step 4: 5-Fluoro-6-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one Potassium carbonate (133 mg, 0.962 mmol) was added to a solution of 5-fluoro-6-hydroxyisoindolin-1-one (66 mg, 0.32 mmol) and 2-(chloromethyl)-5-methoxypyridine (71 mg, 0.45 mmol) in DMF (5 mL) and the mixture was stirred at 70° C. for 2 h. After this time, the mixture was cooled to room temperature and the solvent was removed under reduced pressure. The resulting residue was purified by FCC (silica, 0-20% MeOH in EtOAc, then 0-20% MeOH in DCM) to give the title compound (85 mg, 92%). 1 H NMR (300 MHz, DMSO-d 6 ) 8.56 (s, 1H), 8.30 (d, J = 2.1 Hz, 1H), 7.50-7.42 (m, 4H), 5.25 (s, 2H), 4.28 (s, 2H), 3.83 (s, 3H). MS (ES + ) (M+H) + 289.

[0513] Step 5: 2-(5-bromopyrazin-2-yl)-5-fluoro-6-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one A mixture of 5-fluoro-6-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one (84 mg, 0.29 mmol), 2,5-dibromopyrazine (83 mg, 0.35 mmol), Xantphos (15 mg, 0.026 mmol), and cesium carbonate (285 mg, 0.875 mmol) in 1,4-dioxane (10 mL) was purged with nitrogen for 2 min. 2 (dba) 3 (8 mg, 0.009 mmol) was added and the reaction mixture was heated in a sealed vial at 120° C. for 4 h. After this time, the mixture was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by FCC (silica, 0-100% EtOAc in DCM, then 0-10% MeOH in DCM) to give the title compound (53 mg, 41%). 1 H NMR (300 MHz, DMSO-d 6) 9.52 (s, 1H), 8.73 (s, 1H), 8.32 (d, J = 2.7 Hz, 1H), 7.67 (d, J = 4.2 Hz, 1H), 7.64 (s, 1H), 7.53 (d, J = 9.0 Hz, 1H), 7.45 (dd, J = 8.7, 3.0 Hz, 1H), 5.32 (s, 2H), 4.96 (s, 2H), 3.84 (s, 3H). MS (ES + ) (M+H) + 445.

[0514] Step 6: 5-Fluoro-2-(5-hydroxypyrazin-2-yl)-6-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one A mixture of 2-(5-bromopyrazin-2-yl)-5-fluoro-6-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one (52 mg, 0.12 mmol), tBuXPhos (5.9 mg, 0.014 mmol), and freshly ground KOH (13 mg, 0.23 mmol) in 1,4-dioxane (1.5 mL) and water (1.5 mL) was purged with nitrogen for 2 min. 2 (dba) 3 (6.4 mg, 0.0070 mmol) was added and the reaction mixture was heated in a sealed vial at 100° C. for 2 h. After this time, the mixture was cooled to room temperature, neutralized with 1N HCl, and concentrated to dryness under reduced pressure. The resulting residue was purified by FCC (silica, 0-20% MeOH in EtOAc) to give the title compound (11 mg, 25%). 1 H NMR (300 MHz, DMSO-d 6 ) 8.81 (br s, 1H), 8.31 (d, J = 2.7 Hz, 1H), 7.98 (s, 1H), 7.63-7.56 (m, 2H), 7.52 (d, J = 8.4 Hz, 1H), 7.44 (dd, J = 8.5, 2.7 Hz, 1H), 5.30 (s, 2H), 4.91 (s, 2H), 3.84 (s, 3H). MS (ES + ) (M+H) + 383.

[0515] Step 7: 5-Fluoro-6-((5-methoxypyridin-2-yl)methoxy)-2-(4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)isoindolin-1-one and 5-Fluoro-2-(5-methoxypyrazin-2-yl)-6-((5-methoxypyridin-2-yl)methoxy)-isoindolin-1-one Methyl 4-nitrobenzenesulfonate (8 mg, 0.04 mmol) was added to a mixture of potassium carbonate (11 mg, 0.078 mmol) and 5-fluoro-2-(5-hydroxypyrazin-2-yl)-6-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one (10 mg, 0.029 mmol) in DMF (2 mL) and the mixture was stirred at room temperature for 16 h. After this time, the reaction mixture was concentrated to dryness under reduced pressure and the resulting residue was absorbed onto silica gel and purified by FCC (silica, 0-15% MeOH in EtOAc) to give 5-fluoro-6-((5-methoxypyridin-2-yl)methoxy)-2-(4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)isoindolin-1-one (5 mg, 48%) and 5-fluoro-2-(5-methoxypyrazin-2-yl)-6-((5-methoxypyridin-2-yl)methoxy)isoindolin-1-one (5 mg, 48%).

[0516] Example 14-1: 5-Fluoro-6-((5-methoxypyridin-2-yl)methoxy)-2-(4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)isoindolin-1-one 1 H NMR (500 MHz, CDCl 3) 8.61 (d, J = 1.0 Hz, 1H), 8.32 (d, J = 2.5 Hz, 1H), 8.04 (d, J = 1.5 Hz, 1H), 7.53 (d, J = 7.5 Hz, 1H), 7.48 (d, J = 8.5 Hz, 1H), 7.26 (m, 1H), 7.24 (dd, J = 8.5, 3.0 Hz, 1H), 5.28 (s, 2H), 4.88 (s, 2H), 3.87 (s, 3H), 3.62 (s, 3H). 19 F NMR (282 MHz, CDCl 3 ) -124.82.Tr(MET-uHPLC-001) = 2.90min (ES + ) (M+H) + 397.2, 97%.

[0517] Example 14-2: 5-Fluoro-2-(5-methoxypyrazin-2-yl)-6-((5-methoxypyridin-2-yl)-methoxy)isoindolin-1-one 1 H NMR (500 MHz, DMSO-d 6 ) 9.23 (d, J = 1.0 Hz, 1H), 8.31 (d, J = 3.0 Hz, 1H), 8.22 (d, J = 1.5 Hz, 1H), 7.63-7.59 (m, 2H), 7.52 (d, J = 9.0 Hz, 1H), 7.45 (dd, J = 8.5, 3.0 Hz, 1H), 5.31 (s, 2H), 4.95 (s, 2H), 3.93 (s, 3H), 3.84 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -126.16. Tr(MET-uHPLC-001) = 3.56 min, (ES + ) (M+H) + 397.0, 98%.

[0518] The following additional compounds were prepared by Method 14:

[0519] Example 14-3: 6-Fluoro-5-((5-methoxypyridin-2-yl)methoxy)-2-(4-methyl-5-oxo-4,5-dihydropyrazin-2-yl)isoindolin-1-one

[0520] [ka]

[0521] 1 H NMR (500 MHz, DMSO-d 6 ) 8.53 (d, J = 1.1 Hz, 1H), 8.31 (d, J = 2.8 Hz, 1H), 8.00 (d, J = 1.1 Hz, 1H), 7.63 (d, J = 7.3 Hz, 1H), 7.61 (d, J = 9.8 Hz, 1H), 7.54 (d, J = 8.5 Hz, 1H), 7.46 (dd, J = 8.5, 2.9 Hz, 1H), 5.28 (s, 2H), 4.90 (s, 2H), 3.84 (s, 3H), 3.54 (s, 3H). 19 F NMR (282 MHz, DMSO-d 6 ) -133.49. Tr(MET-uHPLC-006) = 4.02 min m / z (ES+) (M+H)+ 397.0, 97%.

[0522] Biological assays Exon 1-Q46 radioligand binding assay For radioligand binding assays (RBA), MBP-HTT(1-89)Q46-His(6x) ("exon1-Q46") protein was produced based on a previous publication (Scherzinger et al., Cell, vol. 90, pp. 549-558, Aug. 8, 1997). For the experiments, 30 μM MBP-exon1-Q46 was diluted with 150 μg / mL thrombin and 2 mM CaCl in assay buffer (150 mM NaCl, 50 mM Tris pH 8.0). 2The aggregates were incubated with exon 1-Q46 for 16 hours at 37°C. The aggregated exon 1-Q46 was pelleted by centrifugation at 13,000 rpm for 5 minutes in a tabletop centrifuge and redissolved in an equal volume of assay buffer. Test compounds were prepared at 11 concentrations ranging from 63 μM to 2 nM by titration in DMSO. For the RBA, exon 1-Q46 protein aggregates and test compounds were pre-incubated at 100 μL / well in 96-well plates (pp, round bottom) in assay buffer for 20 minutes at room temperature. Ligand was then added at 50 μL / well and incubated at 37°C for 60 minutes. The final assay concentrations were 1 μM to 30 pM test compound, 1 μM exon 1-Q46 protein (equivalent monomer concentration) and 0.3 nM ligand [ 3 H 3 The antibody was 5'-methyl-5-((5-methoxypyridin-2-yl)methoxy)-2-(pyrazin-2-yl)benzo[d]oxazole. Samples were transferred onto GF / B filter plates and washed twice with 200 μL PBS using a Filtermate harvester. Filter plates were dried at 55° C. for 1 hour before sealing the plates back with foil and adding 30 μL / well of scintillation fluid (Packard MicroScint 40), incubating for 15 minutes in the dark, and counting on a MicroBeta reader. For analysis, replicate data from independent assay plates were compared against vehicle control wells (0% inhibition) and 1 μM unlabeled [ 3 H 3 IC was normalized to 0% inhibition and 100% inhibition using [(5-methoxypyridin-2-yl)methoxy]-2-(pyrazin-2-yl)benzo[d]oxazole (100% inhibition). 50 Values ​​were calculated using normalized replicate data and four variables in the global fit (upper, lower, slope, IC 50 ) was determined using the S-order inhibition model.

[0523] The results for various example compounds were as provided in the table below (+++<100nM; ++100-500nM; +>500nM; ND: Undetermined):

[0524] [Table 4]

[0525] PET Imaging Examples The following example illustrates an exemplary, non-limiting procedure that may be utilized when performing a PET imaging study on an individual in a clinical setting. The individual is unmedicated or pre-medicated with an unlabeled compound. The individual may fast prior to PET imaging and is allowed to have water ad libitum. A 20G 2-inch intravenous catheter is inserted into the contralateral ulnar vein for administration of the imaging agent.

[0526] A human subject is placed in the PET camera and administered a tracer dose of the imaging agent via an intravenous catheter. Arterial or venous blood samples are obtained at appropriate time intervals during the PET scan to analyze and quantify the proportion of unmetabolized compound in plasma. Images are acquired for up to 120 minutes. Within 10 minutes of injection of the radiotracer, and at the end of the imaging session, 1 mL blood samples are obtained to determine the plasma concentration of any unlabeled imaging agent compound (or other intervention compounds) that may have been administered prior to the PET tracer.

[0527] Tomographic images are obtained by image reconstruction. Regions of interest (ROIs) are set on the reconstructed images, for example, to determine the distribution of imaging agents. Regions of interest in brain images may include, for example, the striatum, cerebellum, or basal ganglia. The imaging agent uptake over time in these regions may be used to generate time activity curves (TACs). Data can be expressed as activity per unit time per unit volume (e.g., μCi / cc / mCi injected dose) or activity per unit volume. TAC data can be processed by various methods known in the art to obtain quantitative parameters, such as binding potential (BP). For further description of imaging procedures, see, for example, Waxman AD et al., Society of Nuclear Medicine Procedure Guideline for FDG PET Brain Imaging, ver.1.0, (February 8, 2009).

[0528] Unconnected brain fraction The compounds were divided into unbound brain fraction (or brain free fraction, f u , brain ) were tested. It is an accepted standard in the CNS PET imaging field that compounds with low unbound fractions in the brain (<5%) are likely to have high nonspecific binding background that is undesirable for detection of target binding. As a result, u,brainare important properties in the design of PET ligands for brain imaging [see Zhang et al., Design and selection parameters to accelerate the discovery of novel central nervous system positron emission tomography (PET) ligands and their application in the development of a novel phosphodiesterase 2A PET ligand. J Med Chem 2013;56(11):4568-79; Liu et al., Imaging Mutant Huntingtin Aggregates: Development of a Potential PET Ligand. J Med Chem 2020;63(15):8608-8633].

[0529] [Table 5] TIFF2024513913000091.tif236149TIFF2024513913000092.tif229149TIFF2024513913000093.tif225148TIFF2024513913000094.tif29148

[0530] f in mouse brain by equilibrium dialysis u , brain Dialysis membranes (12-14 kDa) (HTDialysis, Connecticut) were soaked in phosphate buffer (10 mM potassium phosphate and 0.8% sodium chloride buffer, pH 7.4, 37 °C) for a minimum of 1 h, at which point ethanol was added (final concentration of 20% v / v) and the membranes continued to soak for 30 min. Mouse brain tissue was diluted in phosphate buffer at a 1:4 ratio (w / v) and homogenized using Precellys 24 (Stretton Scientific, UK) to achieve a final concentration of 20% brain homogenate.

[0531] Test compounds prepared as 0.5 mM DMSO stocks were diluted into brain homogenate to achieve a final substrate concentration of 5 μM (1% DMSO) and t=0 samples were prepared by sampling 50 μL into 400 μL of quench solution (acetonitrile containing 0.1% formic acid and sulfisoxazole / tolbutamide / imipramine / labetalol 200 nM). Samples were matrix matched by adding 50 μL of assay buffer. HTDialysis Teflon blocks (HTDialysis, Connecticut) were assembled with presoaked membranes between each receiver and donor well and secured to a stainless steel pressure plate (HTDialysis, Connecticut). The lower compartment (acceptor side) was dosed with 120 μL of buffer and the upper compartment (donor side) was dosed with 120 μL of compound in homogenate. All test compounds and controls were completed in triplicate. The HT dialysis plate was sealed and incubated at 37° C. with shaking at 250 rpm for 6 hours.

[0532] After incubation, 50 μL of sample was removed from all donor and acceptor wells and transferred to a quench plate containing 400 μL of quench solution. All samples were matrix matched with 50 μL of alternative blank matrix. Analytical samples were diluted 1:1 with water using the Janus Robot and analyzed by LC-MS / MS.

[0533] The peak areas of the donor and acceptor compartments were compared to determine the percent unbound (%f u ) was determined and reported as the average of triplicate measurements. Sample recovery (%) was calculated using a reference TO sample. Control compounds were compared to historical and literature values ​​to ensure the functionality of the assay.

[0534] If the average free brain fraction of a compound is less than 5%, undesired indiscriminate binding will increase background signal.Therefore, those skilled in the art will understand that an average free brain fraction of more than 5% is desirable for imaging PET tracers to properly detect brain structures containing aggregated proteins such as mHTT.The data shows that the compounds described herein have higher free brain fractions than comparative compounds, while also providing good binding to mHTT protein species.Therefore, the compounds described herein have favorable properties for in vivo use as imaging agents.

[0535] In Situ Autoradiography (ARG) In situ autoradiography (ARG) was used to detect the expression of tritium-labeled compounds 1-6 ([ 3 The pharmacological binding properties of [H]-compounds 1-6" were investigated; the specificity of the binding was investigated by including samples from Alzheimer's disease (AD) patients containing other pathological aggregates (e.g., Abeta-containing plaques and phosphorylated Tau-expressing tangles). Saturation binding experiments were performed in coronal brain slices; 3 [H]-Compound 1-6 binding was quantified by densitometric analysis and specific binding was determined in the cortex (CTX). For each region of interest (ROI), affinity (K D ) and the maximum number of binding sites (B max ) was decided.

[0536] Tissue preparation and sectioning At the appropriate age, HD and WT mice of different strains were sacrificed by cervical dislocation, and the brains including the cerebellum were dissected and washed with ice-cold PBS. The brains were then patted dry on paper towels and transferred to 6-well plates filled with isopentane cooled to a temperature between -30°C and -40°C. The frozen brains were stored at -80°C until sectioning.

[0537] Unfixed and unembedded frozen brains were sectioned using a cryostat at -18°C. Serial coronal tissue sections of 20 μm thickness were then mounted on superfrost slides and dried at room temperature for approximately 90 minutes. Slides were stored at -80°C until the various experiments were performed, but not for more than 3 weeks. Similar procedures were performed with fresh frozen human postmortem brain blocks (healthy subjects {CTRL}, Huntington's disease {HD}, and Alzheimer's disease {AD}).

[0538] Assay procedure An appropriate number of slides containing each tissue section were allowed to acclimate to room temperature for 30 minutes. Slides were then pre-incubated by immersing them in 40 mL assay buffer at room temperature for 20 minutes. After this pre-incubation step, one slide per animal was immersed in 30 mL of 10 nM, 3 nM, 1 nM, 0.3 nM, 0.1 nM, 0.03 nM, 0.01 nM, or 0.003 nM [ 3 H]-compounds 1-6 solution, and one slide was incubated by immersion in 30 mL of 10 nM, 3 nM, 1 nM, 0.3 nM, 0.1 nM, 0.03 nM, 0.01 nM, or 0.003 nM [ 3 H]-Compound 1-6 + 10 μM unlabeled Compound 1-6 solution. A similar paradigm was performed with tritium-labeled comparative compound 3 ("[ 3 H]-Comparative Compound 3") and tritium-labeled Comparative Compound 4 ("[ 3 H]-Comparative Compound 4). The structures of these comparative compounds and Compounds 1-6 are shown below.

[0539] [Table 6]

[0540] The slides were then washed three times for 10 min with 200 mL of ice-cold wash buffer at 4°C and immersed in ice-cold distilled water for 3 s to remove buffer salts. The slides were dried for 3 h at 30°C and exposed to a Fuji BAS-TR 2015 tritium phosphor screen for 96 h together with calibrated tritium standards ART 0123C and ART 0123B. The radiant energy deposited on the screen was scanned using a phosphorimager Typhoon FLA 7000.

[0541] Data analysis Densitometric data analysis was performed using MCID Analysis 7.1 software (Interfocus Imaging Ltd.). Within each brain section, regions of interest (ROIs) were defined for STR, CTX, and HPC using the appropriate sample tool, and the software calculated density measurements (molecular dynamics counts per unit area, MDC / mm2) from the gray level values ​​of all pixels within the defined ROI. 2 ) was calculated. Densitometric calibration related the optical density to known concentrations of radioactivity (fmol / mg tissue) from calibrated tritium standards, and mean values ​​were calculated for each ROI of individual brains. Total binding (TB) and nonspecific binding (NSB) of the radioligand were quantified, and specific binding (SB) was derived by subtracting NSB from TB (SB=TB-NSB) for each brain and ROI, respectively. Group means ± standard deviations (SD) were then calculated for each ROI and experimental condition. Data were fitted to a one-site binding equation using nonlinear regression methods in GraphPad Prism software.

[0542] Saturable binding: HOM zQ175 HD mouse model As can be seen in Figure 1, 3 H]-Compounds 1-6 showed concentration-dependent specific binding and low nanomolar affinity to mHTT aggregates present in HOM zQ175 brain slices; in the cortex, the K D The average value was obtained. B was calculated as a measure for the concentration of specific binding sites. maxreached 448.6 fmol / mg tissue in the cortex. These results may reflect the amount and density of mHTT aggregates expressed in this region of the brain.

[0543] Binding to WT brain slices was negligible and only detectable above the lower limit of quantification (LLoQ, 0.5 fmol / mg tissue) at the two highest concentrations investigated, 3 nM and 10 nM. Binding curves could not be established and the K D The value is also B max The value of [ 3 3H]-confirmed the specific binding of compounds 1-6.

[0544] Determined in the cortex, 3 H]-Comparative Compound 4 and [ 3 H]-compared to comparative compound 3, over a range of radioligand concentrations, 3 The specific and saturation binding of [H]-compounds 1-6 is further shown in Figure 1. As shown in the table below, 3 H]-Compound 1-6 had a Bmax of 448 fmol / mg. 3 H]-Comparative compound 4 (149 fmol / mg) and [ 3 3H]-significantly higher compared to comparative compound 3 (169 fmol / mg).

[0545] [Table 7]

[0546] This data would suggest that compounds 1-6 bind to more and / or different mHTT epitopes than comparative compound 4 and comparative compound 3.

[0547] Binding: Human postmortem brain sections The in situ binding observations from tissue from a mouse HD model were then extended to post-mortem human brain tissue to provide information on the general translatability of this mHTT aggregate-binding agent, and in particular on its pathology specificity and species selectivity.3 H]-Compounds 1–6 demonstrated mHTT-specific binding in postmortem human HD brain.

[0548] As can be seen in Figure 2, 3 [H]-Compounds 1-6 showed lower but significantly higher binding to frontal cortex sections from HD donors compared to CTRL tissue (0.8±0.2 fmol / mg tissue in HD vs. 0.0±0.0 in CTRL; p<0.01 comparing HD vs. CTRL). 3 [H]-compounds 1-6 showed differential specific binding in gray matter regions with very low white matter binding (≦0.5 fmol / mg tissue). These data are consistent with the observation that mHTT aggregates are found primarily in cortical neurons (gray matter) and are rarely expressed in white matter.

[0549] Next, 3 Further investigations were carried out to determine whether [H]-compounds 1-6 would exhibit different binding properties in different forms of the disease. To address this, juvenile HD samples (one case) were tested in addition to postmortem adult-onset HD brains. Juvenile HD is characterized by an earlier onset and a much faster progression compared to adult HD. 3 [H]-Compounds 1-6 showed the highest binding density to early-onset HD brains (open triangles in Figures 2 and 3), indicating that the compounds recognize mHTT aggregates in both forms of the disease and may recognize additional epitopes expressed in early-onset HD brains versus adult-onset HD brains.

[0550] [ 3 [H]-Compounds 1-6 also showed significantly higher binding in the cortex of HD patients compared to brain sections of AD patients, as seen in Figure 2 (0.1 ± 0.0 fmol / mg tissue; p < 0.01, comparing HD vs. AD; significance was not evident when comparing CTRL vs. AD). 3The low SB of [H]-compounds 1-6 confirms the selectivity of this radioligand for mHTT versus Aβ and PHF-tau, both of which are expressed in these AD samples (by IHC; data not shown).

[0551] As can be seen in Figure 3, significant specific HD binding was observed at 0.3 nM [ 3 H]- was detectable in compounds 1-6, whereas H-D bonds were 3 H]-Comparative Compound 4 or [ 3 3H]- was not detectable at 0.3 nM for either of the comparative compounds 3, indicating that 3 H]-Comparative Compound 4 or [ 3 H]-Comparative Compound 3 radioligand, 3 These results suggest recognition of distinct binding epitopes and / or greater affinity by [H]-compounds 1-6 for mHTT aggregates expressed in the brains of HD patients.

[0552] conclusion [ 3 [H]-Compounds 1-6 show concentration-dependent specific binding to the cortex of HOM zQ175 HD mice. Binding to WT brain slices is negligible and only detectable above the lower limit of quantification (LLoQ, 0.5 fmol / mg tissue) at the two highest concentrations, 3 nM and 10 nM. 3 H]-Compounds 1-6 bind to mHTT aggregates in HD mouse brain with low nanomolar affinity, with a K of 1.4 nM, consistent with in vitro binding data generated using recombinant exon 1-Q46 protein. D Indicates the value.

[0553] moreover,[ 3 H]-Compounds 1-6 are 3 H]-Comparative Compound 4 or [ 3 3H]-selectively bound to human HD brain slices at 0.3 nM, a concentration of radioligand that failed to reveal HD binding for any of the comparative compounds 3, suggesting novel epitope recognition and / or higher affinity binding for compounds 1-6.

[0554] lastly,[ 3 H]-Compounds 1-6 demonstrated selectivity for binding to brain slices from HD patients, but little or no binding to either healthy control or Alzheimer's disease brain slices.

[0555] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0556] The disclosure illustratively described herein can be suitably implemented without any element or elements, or limitation or limitations not specifically disclosed herein. Thus, for example, terms such as "comprising", "including", "containing" and the like should be interpreted openly and without limitation. Furthermore, the terms and expressions used herein are used as terms of description and not of limitation, and in the use of such terms and expressions, there is no intention to exclude the equivalents of the illustrated and described features or portions thereof, but it is recognized that various modifications are possible within the scope of the present disclosure.

[0557] All publications, patent applications, patents, and other references mentioned herein are expressly incorporated by reference in their entirety, as if each was individually incorporated by reference. In the case of conflict, the present specification, including definitions, will control.

Claims

1. Formula I: 【Chemistry 1】 [In the formula, 【Chemistry 2】 teeth, 【Chemistry 3】 or 【Chemistry 4】 and; R 1 is hydrogen, if present, C 1～6 Alkyl or C 1～6 is haloalkyl; R 10 is hydrogen, if present, C 1～6 Alkyl or C 1～6 is haloalkyl; Ring A is a 5-6 membered heteroaryl; X is CR 11 or N; R 11 is hydrogen, cyano, hydroxy, halo, C 1～6 Alkyl, C 1～6 Haloalkyl or C 1～6 is alkoxy; Y 1 CR 12 or N; Y 2 CR 13 or N; R 12 and R 13 Each of these is hydrogen, hydroxy, halo, C 1～6 Alkyl, C 1～6 Haloalkyl or C 1～6 is alkoxy; R 2 is hydrogen, hydroxy, halo, C 1～6 Alkyl, C 1～6 Haloalkyl or C 1～6 is alkoxy; L is C optionally substituted with 1 to 6 fluoro 1 ~C 3 is alkylene; R 3 is hydrogen, fluoro, C 1～6 Alkyl or C 1～6 is haloalkyl; Each R 4 are independently cyano, hydroxy, halo, C 1～6 Alkyl, C 1～6 Haloalkyl or C 1～6 is alkoxy; Each R 5 are independently cyano, hydroxy, halo, C 1～6 Alkyl, C 1～6 Haloalkyl or C 1～6 is alkoxy; R 6 is hydrogen, cyano, hydroxy, halo, C 1～6 Alkyl, -SO 2 F or L 1 -R 7 and; L 1 -O-, -SO 2 - or -OSO 2 - and; R 7 is hydrogen, C 1～6 Alkyl or C 1～6 haloalkyl, R 7 C 1～6 Alkyl or C 1～6 Haloalkyl is -SO 2 -aryl, -OSO 2 -aryl, optionally substituted with 1 to 6 deuterium atoms, or combinations thereof; 2 -aryl or -OSO 2 -Aryl is cyano, hydroxy, halo, C 1～6 Alkyl, C 1～6 Haloalkyl or C 1～6 is further optionally substituted with alkoxy; m is 0, 1, 2, or 3; n is 0, 1, or 2. or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

2. Formula II: 【Chemistry 5】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

3. Formula III: 【Chemistry 6】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

4. Formula IV: 【Chemistry 7】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

5. Formula V: 【Chemistry 8】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

6. Formula VI: 【Chemistry 9】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

7. Formula VII: 【Chemistry 10】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

8. Formula VIII: 【Chemistry 11】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

9. Formula IX: 【Chemistry 12】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

10. Formula X: 【Chemistry 13】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

11. Formula XI: 【Chemistry 14】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

12. Formula XII: 【Chemistry 15】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

13. Formula XIII: 【Chemistry 16】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

14. Formula XIV: 【Chemistry 17】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

15. Formula XV: 【Chemistry 18】 2. The compound of claim 1, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

16. R 1 is hydrogen or C 1～6 The compound of claim 1 , wherein the aryl group is an alkyl group.

17. R 1 C 1～6 The compound of claim 1 , wherein the aryl group is an alkyl group.

18. R 1 The compound of claim 1 , wherein is methyl.

19. R 2 The compound of claim 1 , wherein is hydrogen.

20. R 3 The compound of claim 1 , wherein is hydrogen.

21. R 4 The compound of claim 1 , wherein is halo.

22. R 4 The compound of claim 1 , wherein is fluoro.

23. The compound of claim 1, wherein n is 1.

24. The compound of claim 1, wherein n is 0.

25. R 11 The compound of claim 1 , wherein is hydrogen.

26. The compound of claim 1, wherein m is 1.

27. R 5 The compound of claim 1 , wherein is halo.

28. R 5 The compound of claim 1 , wherein is fluoro.

29. The compound of claim 1, wherein m is 0.

30. R 6 L 1 -R 7 And L 1 The compound of claim 1 , wherein is -O-.

31. R 7 C 1～6 Haloalkyl or C 1～6 31. The compound of claim 30, which is alkyl.

32. R 7 is replaced by 1 to 6 deuterium atoms 1～6 31. The compound of claim 30 which is a haloalkyl.

33. R 6 The compound of claim 1 , wherein is methoxy.

34. 2. The compound of claim 1, wherein X is N.

35. L is CH 2 2. The compound of claim 1 ,

36. Y 1 is N and Y 2 The compound of claim 1 , wherein is CH.

37. The compound of claim 1 , wherein Ring A is pyridinyl.

38. The compound of claim 1 , wherein Ring A is pyridin-2-yl.

39. 【Chemical 19】 but, 【Chemistry 20】 and R 10 C 1～6 The compound of claim 1 , wherein the aryl group is an alkyl group.

40. R 1 , R 4 , R 5 , R 6 , and R 11 The compound of claim 1 , wherein at least one of the following groups contains a fluorine atom:

41. One R 4 or one R 11 The compound of claim 1 , wherein is fluoro.

42. A compound selected from the compounds of Table 1, optionally labeled with a radioactive isotope, or an isotopically enriched analog, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

43. The compound of claim 1 , wherein the compound is labeled with a radioactive isotope.

44. The compound is 11 C. 13 N, 15 O, and 18 44. The compound of claim 43, containing a positron emitting radioisotope selected from F.

45. 45. An imaging agent comprising the compound of claim 43 or 44, or an isotopically enriched analogue, pharma- ceutically acceptable salt, prodrug, tautomer, stereoisomer, or mixture of stereoisomers thereof.

46. 44. A method of detecting the presence or absence of an aggregation-prone protein in an individual comprising administering to the individual an effective amount of a compound of claim 43 and generating an image of a body part or region of the individual.

47. 47. The method of claim 46, wherein generating an image of a body part or region of an individual comprises generating an image and detecting the presence or absence of aggregation-prone proteins in the image.

48. 48. The method of claim 47, wherein the protein prone to aggregation is huntingtin protein (HTT protein).

49. 49. The method of claim 48, wherein the HTT protein is found in the basal ganglia.

50. 48. The method of claim 47, wherein the presence or absence of protein aggregates corresponds to the presence or absence of a neurodegenerative disease.

51. 51. The method of claim 50, wherein the neurodegenerative disease is selected from Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, Parkinson's disease, prion diseases, and spinocerebellar ataxias.

52. 52. The method of claim 51, wherein the neurodegenerative disease is Huntington's disease (HD).

53. 47. The method of claim 46, wherein the effective amount of the compound or imaging agent comprises from about 0.1 to about 20 mCi.

54. 54. The method of claim 53, wherein the effective amount of the compound or imaging agent comprises about 10 mCi.

55. 47. The method of claim 46, wherein generating the image comprises positron emission tomography (PET) imaging, PET with simultaneous computed tomography imaging (PET / CT), PET with simultaneous magnetic resonance imaging (PET / MRI), single photon emission computed tomography (SPECT) imaging, or a combination thereof.

56. 56. The method of claim 55, wherein generating an image comprises PET imaging.

57. 50. The method of claim 48 or 49, wherein the HTT protein is present as an oligomer or aggregate, or a combination thereof.

58. 50. The method of claim 48 or 49, wherein the HTT protein is mutant.

59. 47. The method of claim 46, wherein the body part or region is the head, spine, limb, thorax, or abdomen.

60. 47. The method of claim 46, wherein the body part or body region is the brain.

Images