Isoindlinone compounds and imaging agents for imaging huntingtin protein

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 of related patent applications This application claims priority to U.S. Provisional Patent Application No. 63 / 172,617, filed 8 April 2021, which is incorporated herein by reference for all purposes.

[0002] Compounds and imaging agents, compositions thereof, and methods of using them are provided herein, which are useful for detecting, treating, or preventing diseases or conditions related to protein aggregation. [Background technology]

[0003] The emergence 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, both before symptom onset and in the clinical environment. Such measurements have broad diagnostic utility, and their use in evaluating treatment responses and supporting drug development is rapidly expanding. The introduction of high-resolution molecular imaging techniques is seen by many experts as a major breakthrough.

[0004] PET involves administering a positron-emitting radionuclide tracer to a target and subsequently detecting positron emission (annihilation) events within the body. Radionuclide tracers typically consist of target molecules that incorporate one or more types of positron-emitting radionuclides.

[0005] Molecular probes labeled with positron-emitting radionuclides and associated PET imaging assays are under development to target, detect, visualize, and quantify various extracellular and intracellular molecules and processes associated with various diseases.

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

[0007] As with any medical condition, treatment for hemodialysis (HD) should ideally be initiated at or before the onset of early disease symptoms. Therefore, early indicators of disease onset and reliable pharmacodynamic biomarkers of disease progression are highly desirable. [Overview of the project] [Problems that the invention aims to solve]

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

[0009] This 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 compounds of formula I are 11 C, 13 N, 15 O, and 18 It contains one or more positron-emitting radioactive isotopes selected from F.

[0010] In some embodiments, imaging agents are provided that include a compound of formula I, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a prodrug, a tautomer, a stereoisomer, or a mixture of stereoisomers.

[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 It contains one or more positron-emitting radionuclides selected from F.

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

[0013] In some embodiments, compounds or imaging agents are provided for use in detecting the presence or absence of proteins that are prone to aggregation in an individual, wherein use comprises administering an effective amount of the compound or imaging agent described herein to the individual and generating an image of a body part or region of the individual.

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

[0015] In some embodiments, compounds or imaging agents for use as described herein are provided, wherein the presence or absence of protein aggregates corresponds to the presence or absence of neurodegenerative disease.

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

[0017] In some embodiments, compounds or imaging agents for use as described herein are provided, wherein the effective amount of the imaging agent is about 0.1 to about 20 mCi. In some embodiments, compounds or imaging agents for use as described herein are provided, wherein the effective amount of the imaging agent is about 10 mCi.

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

[0019] In some embodiments, compounds or imaging agents for use as described herein are provided, wherein the HTT protein exists as an oligomer or aggregate, or a combination thereof. In some embodiments, compounds or imaging agents for use as described herein are provided, wherein the HTT protein is a variant.

[0020] In some embodiments, compounds or imaging agents for use as described herein are provided, wherein the body part or region is the head, spinal cord, limb, chest, or abdomen. In some embodiments, compounds or imaging agents for use as described herein are provided, wherein the body part or region is the brain. [Brief explanation of the drawing]

[0021] [Figure 1] Figure 1 shows the specific and saturated binding of the test radioligand compound over a range of concentrations determined in the cortical tissue of 12-month-old HOM zQ175, an HD mouse model. [Figure 2] Figure 2 shows the specific binding of [3H]-compounds 1-6 to various postmortem human brain tissue samples (healthy subjects [CTRL], Huntington's disease [HD], and Alzheimer's disease [AD]). [Figure 3] Figure 3 compares the specific binding of [3H]-compounds 1-6 to [3H]-comparative compound 3 and [3H]-comparative compound 4 in various postmortem human brain tissue samples (healthy subjects [CTRL], Huntington's disease [HD], and Alzheimer's disease [AD]). [Modes for carrying out the invention]

[0022] The following description provides exemplary embodiments of the Technology. However, it should be recognized that such descriptions are not intended to limit the scope of the Disclosure, but rather are provided as exemplary embodiments.

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

[0024] The compounds described herein refer to any 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 their isotopically labeled analogs, pharmaceutically acceptable salts, solvates, prodrugs, stereoisomers or mixtures of stereoisomers, or compounds described anywhere in this specification, including the examples, or the compounds in Table 1, or labeled isomers of such compounds as defined herein, or imaging agents or pharmaceutical compositions containing such compounds or labeled compounds.

[0025] A dash symbol ("-") that is not between two characters or two symbols is used to indicate the point of attachment of a substituent to the parent structure. For example, -C(O)NH2 is attached to the parent structure through a carbon atom. Dash symbols before or at the end of a chemical group are for convenience; chemical groups can be depicted with one or more dash symbols or without any dash symbols without losing their normal meaning. A wavy line drawn through a bond in a structure indicates a specified point of attachment. Unless required chemically or structurally, no directionality or stereochemistry is indicated or implied by the order in which chemical groups are written or named.

[0026] The prefix "C u~v " indicates that the following group has u to v carbon atoms, barring further substitution. For example, "C 1~6 alkyl" indicates an alkyl group having 1 to 6 carbon atoms.

[0027] References in this specification to "about" values or parameters include (and describe) embodiments that are directed to the value or parameter itself. In certain embodiments, the term "about" includes ±10% of the recited amount. In other embodiments, the term "about" includes ±5% of the recited amount. In certain other embodiments, the term "about" includes ±1% of the recited amount. Also, the term "about X" includes the recitation of "X". Also, the singular forms "a" and "the" include their plural forms unless the context clearly dictates otherwise. Thus, for example, a reference to "the compound" includes a plurality of such compounds, and a reference to "the assay" includes references to one or more assays and their equivalents known to those of skill in the art.

[0028] "Alkyl" refers to an unbranched or branched saturated hydrocarbon chain. As used herein, alkyl has 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 1~9Alkyl), 1 to 8 carbon atoms (i.e., C 1~8 Alkyl), 1 to 6 carbon atoms (i.e., C 1~6 Alkyl) or 1 to 4 carbon atoms (i.e., C 1~4 Alkyl compounds include, for example, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, and 3-methylpentyl. When an alkyl residue having a specific number of carbon atoms is named by its chemical name or specified by its molecular formula, all positional isomers having that number of carbon atoms may be included; for example, "butyl" includes n-butyl (i.e., -(CH2)3CH3), sec-butyl (i.e., -CH(CH3)CH2CH3), isobutyl (i.e., -CH2CH(CH3)2), and tert-butyl (i.e., -C(CH3)3), and "propyl" includes n-propyl (i.e., -(CH2)2CH3) and isopropyl (i.e., -CH(CH3)2).

[0029] Instead of the terms provided herein, alternative chemical names known to those skilled in the art may be used. For example, divalent groups, such as divalent "alkyl" groups and divalent "aryl" groups, may also be called "alkylene" or "arylene" groups, respectively. Furthermore, unless otherwise expressly indicated (for example by a dash), when a combination of groups is referred herein as a single part, for example, arylalkyl or aralkyl, the last group described contains the atom to which that part is bonded to the rest of the molecule.

[0030] "Alkenyl" contains at least one carbon-carbon double bond and 2 to 20 carbon atoms (i.e., C 2~20 Alkenyl), 2 to 8 carbon atoms (i.e., C 2~8 Alkenyl), 2 to 6 carbon atoms (i.e., C 2~6 Alkenyl) or 2-4 carbon atoms (i.e., C 2~4This refers to a hydrocarbon group having an alkenyl group. Examples of alkenyl groups include ethenyl, propenyl, butadienyl (including 1,2-butadienyl and 1,3-butadienyl), and isoprenyl.

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

[0032] "Alkoxy" refers to the "alkyl-O-" group. Examples of alkoxy groups include 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 "alkyl-NH-" group. Examples of alkylamino groups include methylamino, ethylamino, isopropylamino, tert-butylamino, and n-hexylamino. "Dialkylamino" refers to the "(alkyl)2N-" group. Examples of dialkylamino groups include dimethylamino, diethylamino, (isopropyl)(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)2-" group. "Alkylsulfonylalkyl" refers to the -alkyl-S(O)2-alkyl group.

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

[0036] "Amide" is -C(O)NR y R z The group referred to is the "C-amide" group, and -NR y C(O)R z The "N-amide" group refers to the group (in the formula, R y and R z These are independently hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl, each of which may be optionally substituted as defined herein, or R y and R z Together, these form a cycloalkyl or heterocycline, 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 z '' independently refers to 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 an NH2 group.

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

[0039] "Aryl" refers to an aromatic carbocyclic group having a monocyclic (e.g., monocyclic) or polycyclic (e.g., bicyclic or tricyclic) system including a condensed system. As used herein, aryl refers to a ring carbon atom (i.e., C) with 6 to 20 carbon atoms. 6~20 aryl) or 6 to 10 carbon ring atoms (i.e., C 6~10 It contains an aryl group. Examples of aryl groups include phenyl, naphthyl, fluorenyl, and anthryl. However, aryl never includes or overlaps with heteroaryls 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 "aryl-alkyl-" group.

[0041] "Carbamoyl" is -OC(O)NR y R z The group referred to is the "O-carbamoyl" group and -NR y C(O)OR z The "N-carbamoyl" group (in the formula, R) refers to the group y and R z This refers independently to both hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl, each of which may be substituted as provided herein.

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

[0043] "Cycloalkyl" refers to saturated or partially unsaturated cyclic alkyl groups having monocyclic or polycyclic structures including condensed, cross-linked, and spirocyclic systems. The term "cycloalkyl" refers to a cycloalkenyl group (i.e., a cyclic group having at least one double bond) and at least one sp 3 It comprises a carbocyclic fused ring system having ring carbon atoms (i.e., at least one non-aromatic ring). As used herein, cycloalkyl has 3 to 20 ring carbon atoms (i.e., C 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 a ring of 3-6 carbon atoms (i.e., C 3~6 It has a cycloalkyl group. 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, norborneyl, dekalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, etc. Furthermore, the term cycloalkyl is intended to encompass any non-aromatic ring system that may contain a fused aryl ring regardless of its bonding to the rest of the molecule. Also further, cycloalkyl also includes "spirocycloalkyl," such as spiro[2.5]octanyl, spiro[4.5]decanyl, or spiro[5.5]undecanyl. If, in the parent structure, there are two positions on the carbon atom relative to the substitution, the cycloalkyl as a substituent may also include spirocycloalkyl. The cycloalkyl group may be substituted at the carbon atom of the bond to the parent structure.

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

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

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

[0047] "Imino" is a -C(=NR y )R z group (where R y and R z are each independently hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl, each of which may optionally be substituted as defined herein).

[0048] "Imide" is a -C(O)NR y C(O)R z group (where R y and R z are each independently hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl, each of which may optionally be substituted as defined herein).

[0049] "Halogen" or "halo" refers to a substituted atom of Group VIIA of the periodic table, for example, fluoro, chloro, bromo or iodo.

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

[0051] A "haloalkoxy" refers to an alkoxy group as defined above, in which all hydrogen atoms and one or more hydrogen atoms (e.g., 1 to 6 or 1 to 3) including all hydrogen atoms are replaced by halogens.

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

[0053] A "heteroalkyl" refers to an alkyl group in which one or more carbon atoms (and any associated hydrogen atoms) of the alkyl chain are independently replaced by the same or different heteroatomic groups, provided that the bonding sites to the rest of the molecule are via carbon atoms. The term "heteroalkyl" includes unbranched or branched saturated chains having carbon and heteroatoms. For example, one, two, or three carbon atoms may be independently replaced by the same or different heteroatomic groups. Examples of heteroatomic groups include, but are not limited to, -NR y -, -C(O)NR y -, -NRy C(O)-, -O-, -S-, -S(O)-, and -S(O)2- (wherein R y Examples of heteroalkyl groups include hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which may be substituted as defined herein. Examples of heteroalkyl groups include, for example, ethers (e.g., -CH2OCH3, -CH(CH3)OCH3, -CH2CH2OCH3, -CH2CH2OCH2CH2OCH3, etc.), thioethers (e.g., -CH2SCH3, -CH(CH3)SCH3, -CH2CH2SCH3, -CH2CH2SCH2CH2SCH3, etc.), sulfones (e.g., -CH2S(O)2CH3, -CH(CH3)S(O)2CH3, -CH2CH2S(O)2CH3, -CH2CH2S(O)2CH3, etc.) and aminoalkyl groups (e.g., -CH2NR y CH3, -CH(CH3)NR y CH3, -CH2CH2NR y CH3, -CH2CH2NR y CH2 CH2NR y CH3, etc., here, R y Examples include hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl, each of which may be substituted as defined herein. As used herein, a heteroalkyl comprises 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] A "heteroaryl" is a ring heteroatom in which one or more ring heteroatoms are independently selected from nitrogen, oxygen, and sulfur, and one or more (e.g., 1 to 3) N-oxides (-O) - This refers to an aromatic group having a monocyclic or polycyclic fused ring that may include a portion of the ring. As used herein, heteroaryl refers to an aromatic group having 1 to 20 ring carbon atoms (i.e., C 1~20Heteroaryl), 3 to 12 ring carbon atoms (i.e., C 3~12 Heteroaryl) or 3 to 8 carbon ring atoms (i.e., C 3~8A heteroaryl comprises a heteroaryl ring and one to five ring heteroatoms, one to four ring heteroatoms, one to three ring heteroatoms, one to two ring heteroatoms, or one ring heteroatom, independently selected from nitrogen, oxygen, and sulfur. In certain cases, the heteroaryl ring comprises a 5 to 10-membered ring system, a 5 to 7-membered ring system, or a 5 to 6-membered ring system, each independently having one to four ring heteroatoms, one to three ring heteroatoms, one to two ring heteroatoms, or one ring heteroatom, 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, sinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, Examples include soquinoryl, isoxazolyl, naphthilidinyl, oxadiazolyl, oxazolyl, 1-oxidepyridinyl, 1-oxidepyrimidinyl, 1-oxidepyradinyl, 1-oxidepyradinyl, 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, though not limited to these, include benzo[d]thiazolyl, quinolinyl, isoquinolinyl, benzo[b]thiophenyl, indazolyl, benzo[d]imidazolyl, pyrazolo[1,5-a]pyridinyl, and imidazo[1,5-a]pyridinyl, where the heteroaryl can be bonded via any of the rings in the fused system. Any aromatic ring system having a single or multiple fused ring containing at least one ring heteroatom, regardless of its bond to the rest of the molecule (i.e., via any one of the fused rings), can be considered a heteroaryl. Heteroaryls do not encompass or overlap with the aryls as defined above.

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

[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 may optionally be oxidized to form 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. Heterocyclyl may be monocyclic or polycyclic, and the polycycle may be fused, bridged, or spiro. Regardless of the listed substituents, heterocyclyl may, unless otherwise expressly indicated, contain one or more (e.g., 1 to 3) oxo (=O) or N-oxide (-O - ) moieties. Heterocyclyl can be bonded through a carbon atom or a heteroatom as valence permits. Further, the term heterocyclyl encompasses any ring system containing a non-aromatic ring or ring system containing at least one heteroatom, and this ring may be fused to an aryl or heteroaryl ring regardless of the bond to the rest of the molecule. Heterocyclyl may have a charged resonance structure that is aromatic (e.g., pyridin-2(1H)-on-1-yl). As used herein, heterocyclyl has 3 to 14 ring atoms, 3 to 10 ring atoms, 3 to 6 ring atoms, or 5 to 6 ring atoms, and / or 2 to 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 3~6It may contain heterocyclyl groups, 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, benzodioxynyl, benzopyranonyl, benzofuranonyl, dioxolanyl, dihydropyranyl, hydropyranyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, furanonyl, imidazolinyl, imidazolidinyl, indolinyl, indolidinyl, isoindolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindo Examples include lyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, oxylanil, oxetanil, phenothiazinyl, phenoxadinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, tetrahydropyranyl, trithianil, 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. In the parent structure, if there are two positions on the carbon atom for a substitution, the heterocyclyl substituent may include a spiro-heterocyclyl. Examples of bridging heterocyclyl rings, but not limited to these, include 2,5-diazabicyclo[2.2.1]heptane and 2-oxa-5-azabicyclo[2.2.1]heptanyl.Examples of fused heterocyclic rings include, but are not limited to, 1,2,3,4-tetrahydroisoquinolinyl, 4,5,6,7-tetrahydrothieno[2,3-c]pyridinyl, indolinyl and isoindolinyl, and the heterocyclyl can be attached via any ring of the fused system. The "oxo-heterocyclyl" group is a heterocyclyl containing at least one oxo substituent (e.g., 1, or 1-2 oxo substituents), regardless of whether additional substituents are permitted (i.e., unsubstituted oxo-heterocyclyl contains oxo and no other substitution). In some embodiments, the oxo-heterocyclyl contains a cyclic amide moiety.

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

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

[0059] "Sulfonyl" refers to a -S(O)2R y group (wherein 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" refers to a -S(O)R y group (wherein R yis hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl, each of which may be substituted as defined herein. Examples of sulfinyl are methylsulfinyl, ethylsulfinyl, phenylsulfinyl and toluenesulfinyl.

[0061] "Sulfonamide" is -SO2NR y R z and -NR y SO2R z group (in the formula, R y and R z Each of these independently refers to hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl (each of which may be substituted as defined herein).

[0062] The terms "depending on the circumstances" or "depending on the circumstances" mean that the event or situation described thereafter may or may not occur, and that the description includes both cases in which the event or situation occurs and cases in which it does not occur. Furthermore, the term "substituted depending on the circumstances" refers to the base of either the non-substitution or the substitution.

[0063] As used herein, the term "substituted" means that one or more (e.g., 1 to 5 or 1 to 3) hydrogen atoms are substituted for a non-hydrogen group, e.g., but not limited to, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, acyl, amide, amino, amidino, aryl, arylalkyl, azide, carbamoyl, carboxyl, carboxyl ester, cyano, cycloalkyl, cycloalkylalkyl, guanidino, halo, haloalkyl, haloalkoxy, hydroxyalkyl, heteroalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, -NHNH2, =NNH2, imino, imide, hydroxy, oxo, oxime, nitro, sulfonyl, sulfinyl, alkylsulfonyl, alkylsulfinyl, thiocyanate, -S(O)OH, -S(O)2OH, sulfonamide, thiol, thioxo, N-oxide, or -Si(R y )3(in the formula, each R y This refers to a group that is independently replaced by hydrogen, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, cycloalkyl, aryl, heteroaryl, or heterocyclyl.

[0064] In certain embodiments, "substituted" means that one or more (e.g., 1 to 5 or 1 to 3) hydrogen atoms are independently deuterium, halo, cyano, hydroxyl, imino, nitro, azide, 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)Rg , -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)2R g -OS (=O) 1~2 R g -S(=O) 1~2 Ure g , -NR g S(=O) 1~2 NR g R h ,=NSO2R g 、=NOR g -S(=O) 1~2 NR g R h This refers to a group that is replaced with -SF5 or -SCF3. In certain embodiments, "substituted" also means that one or more (e.g., 1 to 5 or 1 to 3) hydrogen atoms are replaced with -C(=O)R g , -C(=O)OR g -C(=O)NR g R h ,-CH2SO2R g , or -CH2SO2NR g R h It means the base that is replaced by R. g and R h These are the same or different, independently of hydrogen, alkyl, alkenyl, alkynyl, alkoxy, thioalkyl, aryl, arylalkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, and / or heteroarylalkyl, or R g and R h These two, together with the atom to which they are bonded, form an oxo, halo, or optionally alkyl-substituted heterocyclyl ring, which may be substituted with an oxo, halo, amino, hydroxyl, or alkoxy ring.

[0065] Polymers or similar indeterminate structures achieved by defining substituents having an infinitely increasing number of further substituents (e.g., substituted aryls having a substituted alkyl that itself is substituted with a substituted aryl group (which is further substituted with a substituted heteroalkyl group, etc.)) are not intended to arise from the above definition. Unless otherwise explicitly stated, the maximum number of consecutive substitutions in the compounds described herein is three. For example, consecutive substitution of a substituted aryl group having two other substituted aryl groups is limited to ((substituted aryl)substituted aryl)substituted aryls. Similarly, the above definition is not intended to include compounds having substitution patterns that are chemically impossible or inseparable (e.g., a methyl group substituted with five fluorine atoms or a heteroaryl group having 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 as 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 described herein is intended to represent both the unlabeled form of the compound and its “isotope-enriched analogue.” The isotope-enriched form of a compound may also be referred to as “labeled.” An isotope-enriched analogue has the structure described 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, for example, respectively. 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 I are included. Isotope-enriched analogs generally include compounds having any isotope enrichment above the natural abundance of the isotope (e.g., on the surface of the earth). Various isotopically labeled compounds, such as radioisotopes, e.g., 3 H, 18 F, 11 C, and 14 those incorporating C are included in the present disclosure. 18 F, 3 H, or 11 Compounds labeled with C may be useful in metabolic studies, 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 the radiotherapy of patients.

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

[0069] Therapeutic compounds of the present disclosure that are deuterium-labeled or substituted may have improved DMPK (drug metabolism and pharmacokinetic) properties with respect to distribution, metabolism, and elimination (ADME). Substitution with heavier isotopes, such as deuterium, may result in certain therapeutic advantages, such as increased in vivo half-life, reduced dose required, and / or improved therapeutic index, due to higher metabolic stability. The isotope-labeled compounds and their prodrugs of the present disclosure can generally be prepared by substituting non-isotope labeling reagents with readily available isotope labeling reagents, by performing the procedures disclosed in the schemes described below or in the Examples and Preparations. Where a compound is described as a deuterated analog, the compound may be described with deuterium as a substituent.

[0070] The concentrations of such heavier isotopes, particularly deuterium, can be defined by the isotopic enrichment coefficient. In the compounds of this disclosure, any atom not specifically indicated as a particular isotope is meant to represent any stable isotope of that atom. Unless otherwise stated, where a position is specifically designated as "H" or "hydrogen," it is understood that this position contains hydrogen and its isotopes in their natural abundances.

[0071] In many cases, the compounds of this disclosure can form acids and / or base salts in the presence of amino and / or carboxyl groups or similar groups.

[0072] Isotope-enriched analogs, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, and mixtures of stereoisomers of the compounds described herein are also provided. "Pharmaceutically acceptable" or "physiologically acceptable" means compounds, salts, compositions, dosage forms, and other materials useful for preparing pharmaceutical compositions suitable for veterinary or human pharmaceutical use.

[0073] In this specification, the term "pharmaceutically acceptable salt" of a compound means a salt that retains the biological efficacy and properties of a given compound and is not biologically or otherwise inappropriate. "pharmaceutically acceptable salts" or "physiologically acceptable salts" of compounds described herein include, for example, acid addition salts obtained by interacting a compound having a basic functional group with an acid, and base addition salts obtained by interacting a compound having an acidic functional group with a base. When a compound is obtained as an acid addition salt, the free base can be obtained by basicizing a solution of the acidic salt. Conversely, when a compound is a free base (e.g., an amine compound), the addition salt can be produced by dissolving the free base in a suitable organic solvent and treating this 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 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, and phosphoric acid. 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-toluenesulfonic acid, and salicylic acid. Similarly, pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. Examples of salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, aluminum, ammonium, calcium, and magnesium salts.Salts derived from organic bases are not limited to these, but include primary, secondary, and tertiary amines, such as alkylamines (i.e., NH2(alkyl)), dialkylamines (i.e., HN(alkyl)2), trialkylamines (i.e., N(alkyl)3), substituted alkylamines (i.e., NH2(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., NH2(alkenyl)), dialkenylamines (i.e., HN(alkenyl)2), trialkenylamines (i.e., N(alkenyl)3), substituted alkenylamines (i.e., NH2(substituted alkenyl)), di(substituted alkenyl)amines (i.e., HN(substituted alkenyl)2), and tri(substituted alkenyl)amines (i.e., N(substituted alkyl)). Examples of suitable amines include salts of monocycloalkylamines, dicycloalkylamines or tricycloalkylamines (i.e., NH2(cycloalkyl), HN(cycloalkyl)2, N(cycloalkyl)3), monoarylamines, diarylamines or triarylamines (i.e., NH2(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, by definition only, include isopropylamine, trimethylamine, diethylamine, tri(iso-propyl)amine, tri(n-propyl)amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, and N-ethylpiperidine.

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

[0075] The compounds described herein may contain chiral centers, thus giving rise to enantiomers, diastereomers, and other stereoisomers that can be defined as (R)- or (S)-, or (D)- or (L)- with respect to an amino acid in terms of absolute stereochemistry. The compounds described herein are intended 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 they may be resolved using conventional techniques, e.g., chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor, or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high-performance liquid chromatography (HPLC). If the compounds described herein contain a double bond or other geometrically asymmetric centers, and unless otherwise specified, the compounds are intended to include both cis- and trans- or E- and Z- geometric isomers.

[0076] A "stereoisomer" refers to one of a series of compounds that have different three-dimensional structures despite being composed of the same atoms bonded together by the same bonds. Various stereoisomers and mixtures thereof are assumed to include "enantiomers," which are stereoisomer compounds that are mirror images of each other and cannot be superimposed.

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

[0078] A “prodrug” is any molecule that, when administered to a mammalian subject, releases a parent drug that is presumed to be active in vivo by the compounds described herein. A prodrug may be any form of the compounds described herein that has been modified so 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 so that the modification can be cleaved to the parent compound either by conventional procedures or in vivo. Prodrugs include the 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 a free hydroxy, amino, or sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to, esters (e.g., acetates, formates, and benzoate derivatives), amides, guanidines, and carbamates (e.g., N,N-dimethylaminocarbonyl) such as hydroxyl functional groups 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, respectively, which are incorporated herein by reference in their entirety.

[0079] In some embodiments, the term “neurodegenerative disease” refers to a disease or condition in which the function of the nervous system in question is impaired. Examples of neurodegenerative diseases are 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 organism, such as an animal or a human. In this context, the methods described herein can be used therapeutically in an organism. “Ex vivo” means outside a living organism. Examples of ex vivo cell populations include in vitro cell cultures and biological samples, including fluid or tissue samples obtained from organisms. 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 the optimal schedule and / or dosage of the compounds disclosed for a given symptom, cell type, organism, and other parameters. Information gathered from such use can be used clinically for experimental purposes or to establish 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 those skilled in the art. The selected compounds can be further characterized to investigate their safety or tolerable dose in human or non-human subjects. Such properties can be investigated using methods generally known to those skilled in the art.

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

[0082] As used herein, the terms “group,” “part,” “radical,” “substituent,” and “fragment” are synonymous and are intended to refer to a part of a molecule that can be bonded to another part of the molecule, for example, via the indicated bond or bond.

[0083] The term “active agent” is used to refer to a compound that has biological activity in the treatment, remission, or prevention of a disease or condition. In some embodiments, “active agent” is a compound with pharmaceutically useful properties or its isotope-labeled analogues, pharmaceutically acceptable salts, solvates, prodrugs, stereoisomers, or mixtures of stereoisomers. For example, an active agent may be an antineurodegenerative agent.

[0084] The term “effective dose” means the amount of a compound described herein that is sufficient to produce a desired response in an individual or patient. In relation to the use of imaging agents, the effective dose may be the amount required to produce an image that has diagnostic or therapeutic utility. The term “therapeutic effective dose” means the amount that, when administered to a human or non-human patient, is effective in conferring a therapeutic benefit such as remission of symptoms, delay of disease progression, or prevention of disease. For example, the therapeutic effective dose may be the amount sufficient to reduce the symptoms of a disease described herein. The (therapeutic) effective dose may vary depending on the subject being treated, the disease or condition, the subject’s weight and age, the severity of the disease or condition, and the method of administration, and can be readily determined by those skilled 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) located on the short (p) arm of chromosome 4 at position 16.3. More precisely, the IT gene encoding the HTT protein. 15 The genes are located on chromosome 4, specifically at base pairs 3,076,407 to 3,245,686.

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

[0087] The term “imaging agent,” as used herein, refers to a compound described herein, or a composition comprising a labeled compound, that is labeled with one or more positron-emitting isotopes or radionuclides. The positron-emitting compound only needs to be enriched with detectable isotopes to the extent that it enables detection using techniques suitable for the particular application.

[0088] The term "PET imaging" (also known as positron emission tomography imaging), as used herein, refers to the generation of images of the internal structures of a human or animal body using positron-emitting labeled compounds.

[0089] The term "positron-emitting radionuclide," as used herein, refers to a radionuclide in which protons within the nucleus emit positrons and electron neutrinos (ν). e This refers to radioactive isotopes that exhibit a specific type of radioactive decay called β+ decay, which is converted into neutrons while emitting positrons. Some examples of positron-emitting radionuclides include: 15 O, 13 N, 11 C, 18 F, 76 Br, and 124 I is included.

[0090] As used herein, the term “labeled” refers to a compound associated with one or more positron-emitting radionuclides at a greater abundance than in nature. For example, the labeled compounds described herein may contain one or more positron-emitting radionuclides in which atoms within the molecule (including atoms in the indicated substituents) exist as positron-emitting isotopes.

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

[0092] In some embodiments, the term “neurodegenerative disease” refers to a disease or condition in which the function of the nervous system in question is impaired. Examples of neurodegenerative diseases are those described herein.

[0093] "Treatment" or "to treat" means any treatment of a patient's disease condition. a) To inhibit the disease (for example, to reduce one or more symptoms resulting from the disease or condition, and / or to reduce the severity of the disease or condition); b) Delaying or cessation of the onset of clinical symptoms associated with the disease or condition (e.g., stabilizing the disease or condition, preventing or delaying the worsening or progression of the disease or condition, and / or preventing or delaying the spread of the disease or condition (e.g., metastasis)); and / or c) To alleviate the disease, that is, to bring about a regression of clinical symptoms (e.g., to bring the condition into remission, to provide partial or complete remission of the disease or condition, to enhance the action of another drug, to slow the progression of the disease, to improve the quality of life and / or to prolong survival). Includes.

[0094] "Prevention" or "prevention" means any treatment of a disease or condition that prevents the development of clinical symptoms of the disease or condition. In some embodiments, the compound may be administered to subjects (including humans) who are at risk (e.g., have genetic or epigenetic markers, have been engaged in activities related to the disease or condition, or have been exposed to environmental conditions) or who have a family history of the disease or condition.

[0095] "Subject" or "patient" refers to an animal, such as a mammal, that is or will be the subject of treatment, observation, or experimentation. The methods described herein may be useful in both human therapeutic 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 measurement for radioactivity and has a conventional meaning to those skilled in the art.

[0097] As used herein, the term "diagnostic imaging" refers to the use of electromagnetic radiation to generate images of the internal structures of a human or animal body for diagnostic purposes.

[0098] For clarity, it should be understood that some features described herein, which are described in relation to separate embodiments, may also be provided in combination in a single embodiment. Conversely, for brevity, various features described herein, which are described in relation to a single embodiment, may also be provided separately or in any suitable subcombination. All combinations of embodiments relating to the chemical group represented by the variable contained in Formula I or any other formula are incorporated herein in particular as if each and every combination were individually and explicitly enumerated, insofar as such combinations result in stable compounds (i.e., compounds that can be isolated, characterized, and tested for biological activity). Furthermore, all subcombinations of the chemical group enumerated in embodiments describing such variable, as well as all subcombinations of uses and medical applications described herein, are also incorporated herein in particular as if each and every subcombination of the chemical group and the subcombinations of uses and medical applications were individually and explicitly enumerated herein. Furthermore, some embodiments include all combinations of one or more additional agents disclosed herein, each and every combination as if they were individually and explicitly enumerated.

[0099] [Table 1] TIFF0007855011000002.tif236146TIFF0007855011000003.tif233145TIFF0007855011000004.tif231146TIFF0007855011000005.tif61146

[0100] compound This disclosure relates to compounds useful for imaging proteins prone to aggregation, such as huntingtin protein. Some embodiments use formula I:

[0101] [ka] [In the formula,

[0102] [ka] teeth,

[0103] [ka] or

[0104] [ka] and; R 1 If present, hydrogen, C 1~6 Alkyl, or C 1~6 It is a haloalkyl; R 10 If present, hydrogen, C 1~6 Alkyl, or C 1~6 It is a haloalkyl; Ring A is a 5-6 member heteroaryl; X is CR 11 or N; R 11 These are hydrogen, cyano, hydroxy, halo, and C. 1~6 Alkyl, C 1~6 Haloalkyl, or C 1~6 It is an alkoxy; Y 1 CR 12 or N; Y 2 CR 13 or N; R 12 and R 13 Each of these is hydrogen, hydroxyl, halo, and C. 1~6 Alkyl, C 1~6 Haloalkyl, or C 1~6 It is an alkoxy; R 2 is hydrogen, hydroxyl, halo, C 1~6 Alkyl, C 1~6 Haloalkyl, or C 1~6 It is an alkoxy; L is a C1-C3 alkylene that is optionally substituted with 1-6 fluorocarbons; R 3 is hydrogen, fluorocarbon, C 1~6 Alkyl, or C 1~6 It is a haloalkyl; Each R 4 These are independently cyano, hydroxy, halo, and C. 1~6 Alkyl, C 1~6 Haloalkyl, or C 1~6 It is an alkoxy; Each R 5 These are independently cyano, hydroxy, halo, and C. 1~6 Alkyl, C 1~6 Haloalkyl, or C 1~6 It is an alkoxy; R 6 These are hydrogen, cyano, hydroxy, halo, and C. 1~6 Alkyl, -SO2F, or L 1 -R 7 and; L 1 is -O-, -SO2-, or -OSO2-; R 7 is hydrogen, C 1~6 Alkyl, or C 1~6 It is a haloalkyl, R 7 C 1~6 Alkyl or C 1~6 Haloalkyls are optionally substituted with -SO2-aryl, -OSO2-aryl, 1 to 6 deuterium atoms, or combinations thereof, where -SO2-aryl or -OSO2-aryl is cyano, hydroxy, halo, C 1~6 Alkyl, C 1~6 Haloalkyl, or C 1~6 It is further sometimes substituted with alkoxy; m is 0, 1, 2, or 3; n is 0, 1, or 2. The present invention provides compounds thereof, or isotope-enriched analogs thereof, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0106] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0108] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

[0109] In some embodiments, the compound of formula I is defined as formula IV:

[0110] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

[0111] In some embodiments, the compound of formula I is defined as formula V:

[0112] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0114] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0116] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0118] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0120] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0122] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

[0123] In some embodiments, the compound of formula I is defined as formula XI:

[0124] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0126] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0128] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0130] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0132] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0134] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

[0136] [ka] The compound, or its isotope-enriched analogues, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

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

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

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

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

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

[0142] In some embodiments, m is 1.

[0143] In some embodiments, R 5 is a halo. In some embodiments, R 5 is fluoro. In some embodiments, n is 1 and R 5 It 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 is L 1 -R 7 And L 1 It is -O-.

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

[0147] In some embodiments, R 6 It 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 CH2.

[0150] In some embodiments, Y1 is N and Y 2 It 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 pyridine-2-yl.

[0152] In some embodiments,

[0153] [ka] teeth,

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

[0155] In some embodiments, the compound comprises at least one halo. In some embodiments, the compound comprises at least one fluoro. In some embodiments, the compound comprises 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 them contains a fluorine atom. In some embodiments, R 4 , R 5 , R 6 , and R 11 At least one of them contains a fluorine atom. In some embodiments, R 4 , R 5 , or R 11 One of them is fluoro. In some embodiments, one R 4 or one R 11 is fluoro. In some embodiments, R 4 or R11 One of them is fluoro.

[0156] In some embodiments, the compound of formula I is labeled with one or more radioactive isotopes.

[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 radioactive isotopes selected from F.

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

[0159] In some embodiments, imaging agents are provided that include a compound of formula I, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a prodrug, a tautomer, a stereoisomer, or a mixture of stereoisomers.

[0160] Additional compounds described herein are also provided. In some embodiments, compounds selected from Table 1, or their isotopically labeled analogs, pharmaceutically acceptable salts, solvates, prodrugs, stereoisomers, or mixtures of stereoisomers are provided.

[0161] In some embodiments, compounds selected from the compounds in Table 1 are provided, which may be labeled with one or more radioisotopes, or isotopic-enriched analogs thereof, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers.

[0162] In some embodiments, pharmaceutical compositions are provided that include compounds described herein, or isotopic-enriched analogs thereof, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers, and pharmaceutically acceptable excipients.

[0163] Nonmetallic radionuclides can be covalently bonded to the compounds described herein by reactions known from the latest technology. It is understood that if the radionuclide is a metallic positron emitter, labeling may require the use of a chelating agent. Such chelating agents are well known from the latest technology.

[0164] In some embodiments, compounds selected from the compounds described in the Examples section provided herein are provided.

[0165] Compounds selected from Table 1, or their isotope-enriched analogs, pharmaceutically acceptable salts, prodrugs, tautomers, stereoisomers, or mixtures of stereoisomers are also provided:

[0166] [Table 2] TIFF0007855011000029.tif193169TIFF0007855011000030.tif192169TIFF0007855011000031.tif192169TIFF0007855011000032.tif123168

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

[0168] A method for generating diagnostic images using positron emission tomography (PET) and a method for detecting the presence or absence of aggregating proteins are provided. PET imaging may be carried out as known to those skilled in the art, or as described below. PET imaging may involve administering a positron-emitting radionuclide tracer, e.g., a compound or imaging agent described herein, to an individual. The tracer is then allowed to bind to the target protein for a sufficient amount of time, at which point the individual is placed in a scanning apparatus equipped with a scintillation detector ring. The emitted positron travels a short (isotope-dependent) distance within the individual's tissue until it interacts with an electron. This interaction annihilates both the electron and the positron, producing a pair of photons. The photons are detected by a scintillator in the scanning apparatus. Photons that do not form a pair are ignored.

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

[0170] Therefore, 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 that are prone to aggregation, or to aggregates thereof. When the compounds or imaging agents are labeled with radioisotopes, the released particles can be detected.

[0171] In some embodiments, the compound or imaging agent is administered into the vascular system of an individual. The compound or imaging agent can cross the blood-brain barrier. Therefore, generating an image may involve generating an image of at least a portion of the individual's brain, for example, the portion to which the compound is distributed.

[0172] Also provided are methods for generating diagnostic images in a biological sample, comprising contacting the biological sample with an effective amount of a compound or imaging agent described herein, and generating an image related to the biological sample, and for detecting the presence or absence of agglutinating proteins. In some embodiments, contacting and generation may be carried out in vitro. In some embodiments, contacting is in vivo and generation is in vitro.

[0173] A method is also provided 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 exists as a monomer, oligomer, aggregate, or a combination thereof. In some embodiments, the protein prone to aggregation is huntingtin protein (HTT protein). The HTT protein may be a variant. 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 agglutinating protein, e.g., the HTT protein, is present in the brain, liver, heart, and / or muscles of an individual. In some embodiments, image generation includes positron emission tomography (PET) imaging, PET with simultaneous computed tomography imaging (PET / CT), PET with simultaneous magnetic resonance imaging (PET / MRI), single-photon emission tomography (SPECT) imaging, or a combination thereof. In some embodiments, image generation includes PET imaging. In some embodiments, the agglutinating protein, e.g., the HTT protein, is present in the basal ganglia, cortex, hippocampus, and / or brainstem of an individual's brain. In some embodiments, the agglutinating protein, e.g., the HTT protein, exists as monomers, oligomers, aggregates, or a combination thereof.

[0175] In some embodiments, individuals are found to have or be found to have Huntington's disease.

[0176] Also provided is a method for 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 is found to have or be having Alzheimer's disease (AD).

[0177] Diagnostic methods are also provided that use the compounds or imaging agents described herein to monitor disease progression in patients by quantifying changes in the levels of proteins prone to aggregation in those patients.

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

[0179] In some embodiments, the effective amount of the compound or imaging agent described herein includes about 0.1 to about 20 mCi. In some embodiments, the effective amount of the compound or imaging agent described herein includes values ​​in the range of 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 values ​​in between. In some embodiments, the effective amount of the compound or imaging agent described herein includes 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 At is one example. The radionuclides incorporated into the compound depend on the specific imaging application. In some embodiments, including PET imaging, 11 C, 18 F, 123 I, 131 I, 75 Br, 76 Br or 77 Compounds incorporating radionuclides selected from Br may be used. In specific applications, 99m The incorporation of chelated radionuclides such as Tc may also be useful. In some embodiments, 18 The longer half-life of F allows imaging to be performed for a sufficiently long time for a stronger signal to be generated, 18 F 11 C may be preferable to C. In some embodiments, the compounds or imaging agents described herein may be labeled with positron-emitting radionuclides or gamma-emitting radionuclides. Some examples of positron-emitting radionuclides include: 15 O, 13 N, 11 C, 18 F, 76 Br, and 124 It contains I, which has 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 are 3 H,11 C, or 18 They may be labeled with F. In some embodiments, the compounds or imaging agents described herein are 11 C and 18 It may be labeled with a positron emitter selected from F. 11 The method for introducing C is, 11 C) Iodomethane or [ 11 This may include, but is not limited to, alkylation with [C]methyl triflate. Carbon 11 has a half-life of approximately 20 minutes, and therefore 11 C generally needs to be produced inside a cyclotron at the site, 11 [C]Can be produced as carbon dioxide. 11 [C] Carbon dioxide is a chemical species suitable for direct labeling (generally [ 11 The radiopharmaceuticals are converted to [C]iodomethane, etc., and after the appropriate radiochemical purity and specific activity are determined, the synthesis is completed in situ and used in PET imaging studies. 18 Typical methods for introducing F include, but are not limited to, nucleophilic and electrophilic methods. Nucleophilic methods include substituting a halide, tosylate, or other leaving group with labeled cesium fluoride, potassium fluoride, tetrabutylammonium fluoride, tetramethylammonium fluoride, or potassium fluoride kryptofix-222. 18Suitable electrophiles for introducing the [F] isotope 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]octanbis(tetrafluoroborate) (Selectfluor), N-fluoropyridinium triflate, xenon fluoride, 2-pyridinesulfonyl fluoride (PyFluor), 3-pyridinesulfonyl fluoride, 4-pyridinesulfonyl fluoride, 4-chloro-2-pyridinesulfonyl fluoride, ethensulfonyl fluoride, fluorobenzoiodoxol, p-fluorophenylaminosulfur trifluoride, p-nitrophenylaminosulfur trifluoride, or pentafluorophenylaminosulfur trifluoride. General methods for introducing 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). Methods for introducing tritium can be achieved by methods known in the art, such as synthesis, recoil, or exchange reactions.

[0182] Fluorine-18 has a half-life of approximately 110 minutes, and therefore [ 18The synthesis of [Fluorine-18] radiopharmaceuticals does not necessarily need to be performed in a location with a cyclotron, nor does it need to be located near a PET imaging research center. Fluorine-18 is also expected to exhibit favorable nuclear and physical properties, including a high positron decay ratio (97%), a relatively short half-life (109.7 minutes), and a low positron energy (up to 0.635 MeV). This positron energy can correspond to a short in vivo diffusion range (<2.4 mm), which can provide an excellent resolution limit for PET images.

[0183] As is recognized, the steps of the methods described herein do not need to be performed a specific number of times or in a specific order. Further objects, advantages and novel features of this disclosure will become apparent to those skilled in the art in the description of the embodiments set forth below, but these are illustrative and not limiting.

[0184] Indications and treatment methods The compounds or imaging agents described herein may be useful for treating diseases or conditions at least partially mediated by agglutinating proteins. In some embodiments, the compounds or imaging agents described herein are useful for treating diseases or conditions at least partially mediated by HTT proteins. In some embodiments, treatment of diseases or conditions at least partially mediated by agglutinating proteins may include administration of the compounds or imaging agents described herein. The treatment may include the co-administration of the compounds or imaging agents described herein with one or more other active agents and / or therapeutic agents. Accordingly, in some embodiments, a method is provided for treating or preventing a disease or condition at least partially mediated by agglutinating proteins in a patient in need thereof, the method comprising administering to the patient a therapeutically effective amount of the compounds or imaging agents described herein.

[0185] Examples of diseases and conditions are listed below.

[0186] Huntington's disease (HD) Huntington's disease (HD) is a hereditary, 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 can spread to other subcortical brain regions. HD belongs to a group of neurodegenerative diseases in which elongated CAG repeat sequences result in long stretches of polyglutamine (polyQ) in the encoded protein. This group also includes dentatorubral-pallidoluysian atrophy (DRPLA), spinal-bulbar muscular atrophy (SBMA), and spinocerebellar ataxia (SCAs). In HD, selective neurodegeneration of gamma-aminobutyric acid-releasing spiny-projection neurons in 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, and memory and / or cognitive impairment.

[0187] The HD protein huntingtin (HTT protein) is a 348 kDa multi-domain protein containing polymorphic glutamine / proline-rich domains at its amino terminus. (Coded by IT) 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 elongation is inversely correlated with the age of disease onset, with early-onset cases characterized by more than 60 elongation repeats. Longer polyQ domains are thought to induce conformational changes in the HTT protein, which are thought to form intracellular aggregates, often appearing as nuclear inclusions. However, aggregates can also form outside the nucleus. The HTT protein is present in the nucleus, cell body, dendrites, and nerve terminals of neurons and is associated with many organelles, including the Golgi apparatus, endoplasmic reticulum, and mitochondria.

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

[0189] The basal ganglia are a group of subcortical nuclei primarily responsible for motor control, as well as other roles such as motor learning, executive function and behavior, and emotion. Damage to the basal ganglia reticular tissue is thought to lead to several motor disorders. 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 striatal complex, and the excitability of medium spiny neurons is controlled by several presynaptic and postsynaptic mechanisms and interneuronal activity, and ensured by several recurrent or internal basal ganglia circuits. The motor circuits of the basal ganglia have two input points, the striatum and the subthalamic nucleus, and one output point, the globus pallidus internal segment, which connects to the cortex via the motor thalamus.

[0190] Administration of the compounds described herein may result in a reduction, for example, of at least 10% (e.g., at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 100%) of one or more symptoms of the diseases or conditions described herein. The diseases or conditions may be diseases, conditions, or disorders of the nervous system secondary to therapies that have a primary effect on areas other than the nervous system; damage to the nervous system caused by physical, mechanical, or chemical trauma; autoimmune neurodegeneration; neurodegeneration secondary to infection; and / or ocular neurodegeneration. Symptoms of neurodegeneration include, for example, tremors, bradykinesia, ataxia, balance disorders, depression, cognitive decline, short-term memory loss, long-term memory loss, confusion, personality changes, language problems, loss of sensation, sensitivity to touch, numbness in the limbs, muscle weakness, paralysis, muscle spasms, muscle cramps, marked 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 blind spots in vision, metamorphopsia, impaired color vision, reduced 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. Examples of neurodegenerative diseases include Alexander disease, Alpers disease, Alzheimer's disease, amyotrophic lateral sclerosis, ataxia, Batten disease (also known as Spielmeiervojnt-Sjögren-Batten disease), bovine spongiform encephalopathy (BSE), Canavan disease, Cockayne syndrome, corticobasal degeneration, Creutzfeldt-Jakob disease, frontotemporal dementia, Gerstmann-Sträussler-Scheinker syndrome, Huntington's disease, dementia associated with HIV, Kennedy disease, and Krabbe disease. These include Kuru, Lewy body dementia, Machad-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 degeneration of the spinal cord secondary to pernicious anemia, schizophrenia, spinocerebellar ataxia, spinal muscular atrophy, Steele-Richardson-Olsewski 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 and bulbar muscular atrophy, spinocerebellar ataxia, spinal cord and / or brain injury, chronic pulmonary hypertension, Parkinson's disease, amyotrophic lateral sclerosis, cavernous hemangioma, cardiovascular disease, Alzheimer's disease (AD), glaucoma, multiple sclerosis (MS), corneal lesions, diabetes mellitus, chronic and / or neuropathic pain, stroke, ischemia, retinal disease, spinal muscular atrophy (SMA), erectile dysfunction, (non-hypertensive) nephropathy, hypertensive nephropathy, hypertension, optic nerve injury, hepatic fibrosis, lupus, post-transplant liver failure, encephalomyelitis, epilepsy, and neurogliablastoma.

[0193] The compounds described herein, when administered to a subject, may inhibit neuronal degeneration. In some embodiments, inhibiting neuronal degeneration may include inhibiting the degeneration of axons or neurons in neurons. Such inhibition may affect all neurons or parts thereof, for example, neuronal cell bodies, axons, and dendrites. This can be evaluated, for example, by analysis of nervous system function by methods known in the art. Administration of the compounds described herein may result in a reduction of at least 10% (e.g., at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%) of the number of degenerating neurons (or their neuronal bodies, axons, or dendrites) in a neuronal population or subject compared to the number of degenerating neurons (or their neuronal bodies, axons, or dendrites) in a neuronal population or subject that has not been administered one or more of the compounds described herein.

[0194] Neurons can transmit information from tissues and organs to the central nervous system (afferent or sensory neurons) and 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 specific examples of neuronal types that may be targeted for treatment by this 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 targeted for treatment by this disclosure include astrocytes and microglia.

[0195] Furthermore, the compounds described herein can be used to prevent or treat memory loss. Types of memory affected by loss and thus treatable by this 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 diseases, 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 and bulbar muscular atrophy, spinocerebellar ataxia, spinal cord and / or brain injury, chronic pulmonary hypertension, Parkinson's disease, amyotrophic lateral sclerosis, cavernous hemangioma, cardiovascular disease, Alzheimer's disease (AD), glaucoma, multiple sclerosis (MS), corneal lesions, diabetes mellitus, chronic and / or neuropathic pain, stroke, ischemia, retinal disease, spinal muscular atrophy (SMA), erectile dysfunction, (non-hypertensive) nephropathy, hypertensive nephropathy, hypertension, optic nerve injury, hepatic fibrosis, lupus, post-transplant hepatic failure, encephalomyelitis, epilepsy, and neurogliablastoma. 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] The use of the compounds described herein for the manufacture of pharmaceuticals for use in the diagnosis, prevention, or treatment of the diseases or conditions described herein is also provided. For example, the disease or condition may be Huntington's disease.

[0200] Imaging agents and pharmaceutical compositions Imaging agents generally include compounds described herein that are labeled with positron-emitting radionuclides. Due to the short half-life of the radionuclide, positron-emitting radionuclide-labeled imaging agents are generally administered via intravenous injection immediately after use (e.g., within one hour of synthesis). The required amount of imaging agent is usually determined by the prescribing physician. Dosages can vary, but are not limited to, the association rate of the compound, the amount emitted from the radionuclide used, the half-life of the radionuclide, the body part, area, and / or tissue to be imaged, and individual characteristics. Those skilled in the art will understand that an effective amount is generally 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 may be administered to patients in need via any preferred route. Routes of administration may include parenteral administration, such as subcutaneous, intramuscular, or intravenous administration, via a drip patch, for example. Further preferred routes of administration include, but are not limited to, oral, rectal, intranasal, topical (including oral and sublingual), infusion, vaginal, intradermal, intraperitoneal, intracranial, intrathecal, and epidural administration, or administration via oral or nasal inhalation, for example, via a spray or inhaler, or implant.

[0202] With regard to PET imaging, the administration of the compounds or imaging agents described herein to an individual may be intravenous. The pharmaceutical compositions may be in the form of sterile, injectable aqueous or oily suspensions. These suspensions may be formulated according to known techniques using the preferred dispersants or wetting agents and suspending agents listed above. Sterile injectable products may also be sterile injectable solutions or suspensions in non-toxic, parenterally acceptable vehicles, such as solutions in 1,3-butanediol. Acceptable vehicles that may be used include water, Ringer's solution, and isotonic sodium chloride solutions. In addition, sterile, non-volatile oils have been used in the prior art as solvents or suspension media. For this purpose, any non-irritating non-volatile oil, including synthetic mono or diglycerides, may be used. In addition, fatty acids such as oleic acid may be useful in the preparation of injectable products. Such solutions may be formulated with appropriate salts as 0.01% to 10% isotonic solutions, pH 5 to 7.

[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 drip infusion techniques. The compounds or imaging agents described herein may be suspended or dissolved in a vehicle, depending on the vehicle and concentration used. Advantageously, adjuvants such as local anesthetics, preservatives, and buffers may be dissolved in the vehicle. In many pharmaceutical compositions for parenteral administration, the carrier accounts for at least 90% by weight of the total composition. In some embodiments, the carrier for parenteral administration is selected from propylene glycol, ethyl oleate, pyrrolidone, ethanol, and sesame oil.

[0204] A pharmaceutical composition, such as a pharmaceutical composition for injection, may contain cyclodextrin. The cyclodextrin may be, for example, hydroxypropyl cyclodextrin or sulfobutyl ether cyclodextrin. The cyclodextrin may be, for example, α-cyclodextrin, β-cyclodextrin, or γ-cyclodextrin.

[0205] The compounds or imaging agents described herein may be administered via sustained-release formulations placed in specific tissues, including microspheres, liposomes, other particulate delivery systems, or blood. Suitable examples of sustained-release carriers include common products, such as semipermeable polymer matrices in the form of suppositories or microcapsules. Examples of the techniques and protocols described above, as well as other techniques and protocols that may be used in accordance with 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 full contents of which are incorporated herein by reference.

[0206] In some embodiments, the compounds or imaging agents described herein are administered as a pharmaceutical composition. Thus, a pharmaceutical composition comprising at least one compound or imaging agent described herein is provided, together with at least one pharmaceutically acceptable vehicle selected from carriers, adjuvants, and excipients. The compounds or imaging agents of this disclosure can be formulated into pharmaceutical compositions using techniques known to those skilled in the art.

[0207] Pharmacopoeia-acceptable vehicles must be sufficiently pure and sufficiently toxic to be suitable for administration to the animal being treated. Vehicles may be inactive, or they may have pharmaceutically beneficial properties. The amount of vehicle used with the compound or imaging agent may be sufficient to provide a practical amount of material for each dose of the compound or imaging agent.

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

[0209] An active agent may be included in the pharmaceutical composition, which does not substantially interfere with the activity of the compound or imaging agent described herein.

[0210] At least one compound or imaging agent described herein in an effective concentration is mixed with a suitable pharmaceutically acceptable vehicle. If the compound or imaging agent exhibits 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, the use of a cosolvent such as dimethyl sulfoxide (DMSO), the use of a surfactant such as TWEEN®, or dissolution in an aqueous buffer, such as sodium bicarbonate.

[0211] When the compounds or imaging agents described herein are mixed or added, the resulting mixture may be a solution, suspension, emulsion, or the like. The form of the resulting mixture depends on many factors, including the intended method of administration and the solubility of the compound or imaging agent in the selected vehicle. An effective concentration sufficient for imaging or therapy can be experimentally determined by methods known in the art.

[0212] Pharmaceutical compositions may be formulated for oral use, for example, as tablets, 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 those skilled in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents, such as sweeteners, fragrances, colorants and preservatives, to provide a simple and palatable pharmaceutical product. In some embodiments, the oral pharmaceutical composition contains 0.1 to 99% of the compounds or imaging agents described herein. In some embodiments, the oral pharmaceutical composition contains at least 5% (by weight) of the compounds or imaging agents. Some embodiments contain 25% to 50% or 5% to 75% of the compounds or imaging agents.

[0213] Pharmaceutical compositions administered orally may also include liquid solutions, emulsions, suspensions, powders, granules, elixirs, tinctures, and syrups. Suitable pharmaceutically acceptable carriers for the preparation of such compositions are well known in the art. Oral pharmaceutical compositions may contain preservatives, fragrances, sweeteners such as sucrose or saccharin, taste masking agents, and colorants.

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

[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. Furthermore, pharmaceutical compositions containing the compounds or imaging agents described herein may be presented as dry products for preparation with water or other suitable vehicles before use. Such liquid products may contain prior art 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., p-hydroxybenzoate methyl or propyl, and sorbic acid).

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

[0217] An aqueous suspension containing a compound or imaging agent is provided in a mixture with an excipient suitable for the production of an aqueous suspension. Such excipients are suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and acacia gum, and the dispersing or wetting agent may be a naturally occurring phosphatide, such as lecithin, or a condensation product of alkylene oxide with a fatty acid, such as polyoxyethylene stearate, or a condensation product of ethylene oxide with a long-chain aliphatic alcohol, such as heptadecaethyleneoxycetanol, or a condensation product of ethylene oxide with a partial ester derived from a fatty acid and hexitol, such as polyoxyethylene sorbitol substituted, or a condensation product of ethylene oxide with a partial ester derived from a fatty acid and hexitol anhydride, such as polyethylene sorbitan substituted. The aqueous suspension may also contain one or more preservatives, such as ethyl p-hydroxybenzoate or n-propyl.

[0218] Oily suspensions can be formulated by suspending a 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. The oily suspension may contain thickeners, such as beeswax, solid paraffin, or cetyl alcohol. Sweeteners and flavorings, such as those mentioned above, may be added to provide a palatable oral product. These pharmaceutical compositions may be preserved by adding antioxidants such as ascorbic acid.

[0219] The pharmaceutical composition may take 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 rubbers, such as acacia rubber or tragacanth rubber, naturally occurring phosphatides, such as soy, lecithin, and esters or partial esters derived from fatty acids and hexitol, anhydrides, such as sorbitan monooleate, and condensation products of the partial esters with ethylene oxide, such as sorbitan polyoxyethylene monooleate.

[0220] Dispersible powders and granules suitable for preparing aqueous suspensions by adding water provide the active ingredient in a mixture with a dispersant or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersants or wetting agents and suspending agents are illustrated by those already listed above.

[0221] Tablets typically contain, as inert diluents, pharmaceutically acceptable adjuvants of the prior art, 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; and lubricants, such as magnesium stearate, stearic acid, and talc. Flow enhancers, such as silicon dioxide, may be used to improve the flow properties of powder mixtures. Colorants, such as FD&C dyes, may be added for appearance. Sweeteners and flavorings, such as aspartame, saccharin, menthol, peppermint, and fruit flavors, may be useful adjuvants for chewable tablets. Capsules (including time-release and sustained-release formulations) typically contain one or more solid diluents disclosed above. The selection of carrier components is often based on secondary considerations such as taste, cost, and storage stability.

[0222] The pharmaceutical composition may be coated by conventional methods, typically by pH or time-dependent coatings, so that the compound or imaging agent is released into the gastrointestinal tract near the desired topical application or at various rates to extend the desired effect. Such dosage forms typically include, but are not limited to, one or more cellulose phthalate acetates, polyvinyl acetate phthalates, hydroxypropyl methylcellulose phthalates, ethylcellulose, Eudragit® coatings, waxes, and shellacs.

[0223] Pharmaceutical compositions for oral use may also be provided as hard gelatin capsules, where the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules, where the active ingredient is mixed with water or an oily 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 the drug. These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature, and thus melts in the rectum to release the drug. Examples of such materials include cocoa butter and polyethylene glycol.

[0225] The compounds or imaging agents described herein may be formulated for local or topical application, for example, for topical application to the skin and mucous membranes such as the eyes, in the form of gels, creams, and lotions, and for application to the eyes. 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 its isotopically labeled analogues, pharmaceutically acceptable salts, solvates, prodrugs, stereoisomers, or mixtures of stereoisomers, may be mixed with various carrier materials known in the art, such as water, alcohol, aloe vera gel, allantoin, glycerin, vitamin A and E oils, mineral oil, propylene glycol, myristyl PPG-2 propionate, and the like.

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

[0228] Typical 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, and dipropyl sebacate. Examples include 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, mineral oil, butyl myristate, isostearic acid, palmitic acid, isopropyl linoleate, lauryl lactate, myristyl lactate, decyl oleate, and myristyl myristate; as sprays, propane, butane, isobutane Examples of solvents include dimethyl ether, carbon dioxide, and nitrous oxide; examples of solvents include ethyl alcohol, methylene chloride, isopropanol, castor oil, ethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol monoethyl ether, dimethyl sulfoxide, dimethylformamide, and tetrahydrofuran; examples of humectants include glycerin, sorbitol, sodium 2-pyrrolidone-5-carboxylate, soluble collagen, dibutyl phthalate, and gelatin; and examples of powders include chalk, talc, fuller's earth, kaolin, starch, rubber, 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 transdermal patches.

[0230] The compounds or imaging agents described herein may also be administered via liposome delivery systems. Liposomes can be classified into small monolayer vesicles, large monolayer vesicles, and multilayer vesicles. Liposomes can be formed from various amphiphilic molecules, particularly phospholipids. Examples of liposome components include cholesterol, stearylamine, and / or phosphatidylcholine. Liposomes are suitable for various administration routes, including local and intra-tissue injection. Therefore, intravitreous (e.g., in the treatment of glaucoma), intraperitoneal, intravenous, intravascular, intra-articular, and intramuscular administration of liposomes is possible.

[0231] Other pharmaceutical compositions useful for achieving systemic delivery of compounds or imaging agents include sublingual, oral, and intranasal administration forms. Such pharmaceutical compositions typically contain soluble filler substances, such as sucrose, sorbitol, and mannitol, and one or more binders, such as acacia, microcrystalline cellulose, carboxymethylcellulose, and hydroxypropylmethylcellulose. The flow enhancers, lubricants, sweeteners, colorants, antioxidants, and fragrances disclosed above may also be included.

[0232] Pharmaceutical compositions for inhalation can typically be provided in the form of solutions, suspensions, or emulsions, and may be administered as a dry powder or in the form of an aerosol using a conventional spray (e.g., dichlorodifluoromethane or trichlorofluoromethane).

[0233] The pharmaceutical composition may optionally contain an activity enhancer. The activity enhancer can be selected from a wide variety of molecules that function in various ways, either enhancing or not relying on, the therapeutic effect of the compounds or imaging agents described herein. Certain classes of activity enhancers include skin penetration enhancers and absorption enhancers.

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

[0235] The dosage of the compounds or imaging agents described herein depends on various factors, among other considerations, including the specific pathological process to be treated or detected, the individual's physiology, the severity of symptoms, the route of administration, the frequency of dosing intervals, the specific compound used, the efficacy of the compound, its toxicological profile, its pharmacokinetic profile, and the presence of toxic side effects. The dosage under given circumstances is generally determined appropriately by the practitioner based on the above and other factors.

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

[0237] Kit and packaging A kit comprising the compounds described herein and appropriate packaging is also provided herein. In certain embodiments, the kit further includes instructions for use. In some embodiments, the kit includes labels and / or instructions for the use of the compounds or imaging agents described herein and the compounds in the treatment of indications including diseases or conditions described herein.

[0238] A manufactured article containing the compounds or imaging agents described herein in a suitable container is also provided herein. The container may be a vial, bottle, ampoule, pre-filled syringe, or 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. Packaged pharmaceutical compositions may include, for example, prescription information for the patient or healthcare provider, or as a label on the packaged pharmaceutical composition. Prescription information may include, for example, efficacy, dosage and administration, contraindications and adverse reaction information relating to the pharmaceutical composition.

[0240] In all the preceding descriptions, the compound or imaging agent may be administered alone, as a mixture, or in combination with other active agents.

[0241] The use of compounds or imaging agents described herein for the manufacture of pharmaceuticals for use in the diagnosis, prevention, or treatment of diseases or conditions described herein is also provided. For example, the disease or condition may be Huntington's disease.

[0242] The use of the compounds described herein for the manufacture of imaging agents for use in the diagnosis, prevention, or treatment of the diseases or conditions described herein is also provided. For example, the disease or condition may be Huntington's disease.

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

[0244] Pharmaceutical compositions are also provided that include the compounds or imaging agents 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. Similarly, also provided are pharmaceutical compositions comprising compounds or imaging agents described herein, and packaged pharmaceutical compositions comprising another composition comprising one or more additional active agents used for the treatment of Huntington's disease, for example, 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, the methods comprising administering the compounds or imaging agents described herein and one or more additional agents to a subject simultaneously or sequentially. In some embodiments, the active agent is Reminyl®, Cognex®, Aricept®, Exelon®, Akatinol®, Neotropin®, Eldepryl®, estrogen, or cryoquinol.

[0246] In some embodiments, the compounds described herein can be administered together with active agents for treating Parkinson's disease, for example, L-dopa, dopamine agonists (e.g., bromocriptine, pergolide, pramipexole, ropinirole, cabergoline, apomorphine, and rislide), 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 together with active agents for treating Alzheimer's disease, for example, acetylcholinesterase inhibitors (e.g., donepezil, galantamine, and rivastigmine), and / or NMDA receptor antagonists (e.g., memantine).

[0247] Compound synthesis The compounds described herein may be prepared using the methods disclosed herein and their usual modifications as evident from the disclosure herein and from methods well known in the art. Conventional and well known synthetic methods may be used in addition to the teachings herein. The synthesis of typical compounds described herein may be achieved as described in the following examples. Where available, reagents can be purchased commercially from, for example, 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 temperature, time, molar ratio of reactants, solvent, pressure, etc.) are given, it is understood that other process conditions may also be used unless otherwise stated. Optimal reaction conditions may vary depending on the specific reactants or solvent used, but such conditions can be determined by those skilled in the art through standard 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 undesirable reactions. Suitable protecting groups for various functional groups, as well as suitable conditions for protecting and deprotecting specific 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 the references cited herein.

[0250] Furthermore, the compounds described herein may contain one or more chiral centers. Thereafter, 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 this 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, etc. If pure or enriched compounds as enantiomers are desired, chiral chromatography and / or pure or enriched starting materials as enantiomers 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 suppliers such as Sigma Aldrich and Alfa Aesar. Others can be prepared by procedures or obvious modifications thereof described in standard references such as Fieser and Fieser's Reagents for Organic Synthesis, Vol. 1–15 (John Wiley, and Sons, 1991), Rodd's Chemistry of Carbon Compounds, Vol. 1–5 and Supplement (Elsevier Science Publishers, 1989), organic Reactions, Vol. 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 solvents that are inert under the conditions of the reaction described therein (e.g., benzene, toluene, acetonitrile, tetrahydrofuran ("THF"), dimethylformamide ("DMF"), chloroform, methylene chloride (or dichloromethane), diethyl ether, methanol, pyridine, etc.). Generally, the term inert, as used herein with respect to a solvent, refers to a material that does not undergo a reaction to form the target compound of interest by a carbon-carbon bond formation reaction. Unless otherwise specified, the solvents used in the reactions of this 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 sufficient amount to achieve the stated function, for example, to bring the solution to the desired volume (i.e., 100%).

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

[0255] The incorporation of labels into the compounds or imaging agents described herein may be carried out by reacting a suitable starting material with a reagent containing a radioisotope. The method generally follows the same principles as standard organic chemical reactions and can be carried out by any method known to those skilled in the art, including those provided herein.

[0256] Scheme 1 provides an exemplary synthetic route for the synthesis of the 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. 1R is either a protecting group (e.g., the nitrogen protecting group described in the following examples), or 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 If as defined herein, compound 1 is prepared using methods known in the art and / or converted to the compound of formula I starting from commercially available dihydroisoindlinone in one or more steps. Dihydroisoindlinone 1 is coupled with compound 2 by transition metal-based coupling (e.g., in the presence of a palladium-based reagent, e.g., Pd2(dba)3 or any other suitable reagent) in the presence of a base (e.g., potassium phosphate, cesium carbonate, or any other suitable base) 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 method described in the Examples section below. Hydroxy compound 5 is O-alkylated with compound 6 to yield the compound of formula I. LG may be any suitable leaving group, including but not limited to chloro, iodine, bromo, or triflate groups. In some embodiments, in formula I, R 1 If the atom is H, a further 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] Dihydroisoindrinone 1a can be used as a starting material (for example, from a commercial source or obtained by a process known in the art), coupled with compound 2 to produce compound 5a, and then the hydroxyl group of compound 5a may be O-alkylated with compound 6 to produce the compound of formula I.

[0264] Those skilled in the art will understand that any of compounds 1, 2, and 6 may be available from suppliers for specific embodiments. Alternatively, the synthesis of compounds 1, 2, and 6 may be as described herein or as known to those skilled in the art. [Examples]

[0265] The following embodiments are included to illustrate specific embodiments of the Disclosure. Those skilled in the art will understand that the techniques disclosed in the following embodiments represent techniques that function well in the implementation of the Disclosure and may therefore be considered to constitute a particular mode for its implementation. However, those skilled in the art will understand that many modifications can be made to the specific embodiments disclosed in light of the Disclosure to obtain similar or similar results without departing from the spirit and scope of the Disclosure.

[0266] 1. General experimental procedure Commercially available reagents and solvents (HPLC grade) were used without further purification. Spectrometers were used in a deuterated solvent with a Bruker DRX 500 MHz spectrometer, a Bruker DPX 250 MHz spectrometer, or a Bruker AVANCE 300 or Bruker AVANCE 500 spectrometer. 1¹H NMR spectra were recorded. Chemical shifts (δ) are expressed in parts per million. Flash column chromatography refers to automated purification using a Biotage Isolera system with an appropriately sized SNAP or KPNH pre-packed silica column and solvent as recorded in the experimental section, or an Isco Combiflash Rf system with an appropriately sized pre-packed silica column and solvent as recorded in the experimental section. Reverse-phase MPLC chromatography was performed using an Isco Combiflash Rf system with an appropriately sized pre-packed C18 column and solvent 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 a Biotage Isolute Flash SCX-2, loading the sample in methanol and eluting with methanol, then with 5% ammonia in methanol.

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

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

[0269] Alternatively, analytical HPLC-MS (MET-uHPLC-AB-101) was performed using a Waters Acquity UPLC system equipped with Waters PDA and ELS detectors, with 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 minutes, followed by 100% B for 0.5 minutes, 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 obtained using a Waters ZQ at a sampling rate of 2 scans per second, over the range of m / z 150–850. Data were integrated and reports were generated using OpenLynx software.

[0270] Alternatively, analytical UHPLC-MS (METCR1704) was performed using a reversed-phase system with a Waters UPLC™ BEH™ C18 column (2.1 mm × 50 mm, 1.7 μm; temperature: 40°C) at an injection volume of 1 μL, 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 minutes, followed by 100% B for 0.25 minutes. A second gradient of 100–5% B was then applied for 0.05 minutes and held for 0.1 minutes. 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 reports were generated using Waters MassLynx and OpenLynx software.

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

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

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

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

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

[0276] Alternatively, mass spectra and LCMS analyses were obtained using either 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 using a Hewlett Packard HPLC system with a Phenomenex Gemini C18 reversed-phase column (3 μm, 2.0 × 50 mm) at a column temperature of 60°C. The procedure involved a gradient of 1–100% B (A = 2 mM ammonium bicarbonate buffered in pH 10 water, B = acetonitrile) for 1.8 minutes, followed by 100% B for 0.3 minutes, with an injection volume of 3 μL and a flow rate of 1 mL / min. UV spectra were recorded at 215 nm using a Waters PDA detector. Mass spectra were obtained using a Waters ZQ at a sampling rate of two scans per second, covering the range of m / z 150–850. Data were integrated and reports were generated using OpenLynx software.

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

[0279] Alternatively, analytical UHPLC-MS (MET-uHPLC-AB-2005) was performed in reverse phase using a Waters UPLC™ BEH™ C18 column (2.1 mm × 30 mm, 1.7 μm; temperature 40°C) with an injection volume of 1 μL, flow rate = 1.0 mL / min, and a gradient of 1–100%B (A = 2 mM ammonium bicarbonate buffered in pH 10 water; B = acetonitrile) for 1.1 minutes, followed by 100%B for 0.25 minutes. A second gradient of 100–1%B was then applied for 0.05 minutes and held for 0.4 minutes. 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 reports were generated using Waters MassLynx and OpenLynx software.

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

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

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

[0283] [Table 3]

[0284] intermediate Intermediate 1: (5-(fluoromethoxy)pyridine-2-yl)methanol

[0285] [ka]

[0286] Step 1: Methylenebis(4-methylbenzenesulfonate) A mixture of p-silver 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 under reflux for 16 hours. The mixture was then cooled to ambient temperature, filtered, and the filter cake was washed with MeCN (3 × 20 mL). The filtrate was concentrated under vacuum. 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 under vacuum, and the resulting residue was recrystallized from EtOH (30 mL). The isolated product was dried under vacuum to obtain the title compound (4.09 g, 62%). 1 H NMR (300 MHz, CDCl3) 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 methylenebis(4-methylbenzenesulfonate) (4.09 g, 11.5 mmol) and MeCN (26.7 mL) was treated with 1 M TBAF (12.6 mL, 12.6 mmol) in THF, and the mixture was stirred under reflux for 2 hours. After this, the solvent was removed under vacuum, and the resulting residue was dissolved in SiO (40 mL). The solution was washed with brine (40 mL), dried over sodium sulfate, filtered, and concentrated under vacuum. The resulting residue was purified by FCC (silica, 0-50% SiO in heptane) to obtain the title compound (609 mg, 26%). 1 H NMR (300 MHz, CDCl3) 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)pyridine-2-yl)methanol A mixture of 6-(hydroxymethyl)pyridine-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 hours. After this, the mixture was cooled to ambient temperature and extracted with DCM (3 × 40 mL). The combined organic layer was dried over sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The resulting residue was purified with FCC (silica, 0-5% MeOH in DCM) to obtain the title compound (108 mg, 29%). 1 H NMR (300 MHz, DMSO-d6) 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)pyridine-3-yl)oxy)ethyl-1,1,2,2-d44-methylbenzenesulfonate

[0290] [ka]

[0291] Step 1: Ethane-1,2-diyl-d4bis(4-methylbenzenesulfonate) p-toluenesulfonyl chloride (5.77 g, 30.3 mmol) was added to a mixture of ethylene glycol-d4 (0.673 mL, 12.1 mmol) and triethylamine (8.41 mL, 60.5 mmol) in DCM (80 mL), and the mixture was stirred at room temperature for 16 hours. After this, 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 under vacuum. The resulting residue was purified with FCC (silica, 0-5% ethyl phosphate in DCM) to obtain the title compound (3.84 g, 85%). 1 H NMR (300 MHz, CDCl3) 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)pyridine-3-yl)oxy)ethyl-1,1,2,2-d44-methylbenzenesulfonate Ethane-1,2-diyl-d4-bis(4-methylbenzenesulfonate) (4.51 g, 11.8 mmol) was added to a mixture of 6-(hydroxymethyl)pyridine-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 hours. After this, the reaction mixture was cooled and filtered through diatomaceous earth. The filter cake was rinsed with ELISA (2 × 50 mL), and the filtrate was concentrated under vacuum. The resulting residue was purified by FCC (silica, 0-5% MeOH in DCM) to obtain the title compound (450 mg, 34%). 1 H NMR (300 MHz, CDCl3) 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)pyridine-3-yl)oxy)ethyl-1,1,2,2-d44-methylbenzenesulfonate Thionyl chloride (0.197 mL, 2.70 mmol) was added to a mixture of 2-((6-(hydroxymethyl)pyridine-3-yl)oxy)ethyl-1,1,2,2-d44-methylbenzenesulfonate (450 mg, 1.35 mmol) in DCM (9.4 mL) at 0°C, and the solution was stirred at 0°C for 1 hour. After this, 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 under vacuum to obtain the title compound (475 mg, 99%). 1 H NMR (300 MHz, CDCl3) 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-d4)pyridine

[0295] [ka]

[0296] Step 1: 2-Fluoroethyl-1,1,2,2-d44-methylbenzenesulfonate 1.0 M TBAF (8.97 mL, 8.97 mmol) in THF was added to ethane-1,2-diyl-d4-bis(4-methylbenzenesulfonate) (2.80 g, 7.48 mmol) in MeCN (17.4 mL), and the mixture was stirred under reflux for 2 hours. After that, the mixture was cooled, diluted with DCM (100 mL), and washed with water (40 mL). The aqueous layer was extracted with DCM (100 mL), the combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The resulting residue was purified by FCC (silica, heptane, 0-100% DCM) to obtain the title compound (601 mg, 36%). 1 H NMR (300 MHz, CDCl3) 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-d4)pyridine-2-yl)methanol A mixture of 2-fluoroethyl-1,1,2,2-d44-methylbenzenesulfonate (363 mg, 1.60 mmol), 6-(hydroxymethyl)pyridine-3-ol (200 mg, 1.60 mmol), and cesium carbonate (1.56 g, 4.80 mmol) in MeCN (20.0 mL) was stirred at 80°C for 2.5 hours. The reaction mixture was then cooled and filtered through diatomaceous earth. The filter cake was rinsed with ELISA (2 × 50 mL), and the filtrate was concentrated under vacuum. The resulting residue was purified using FCC (silica, 0-5% MeOH in DCM) to obtain the title compound (133 mg, 47%). 1H NMR (300 MHz, CDCl3) 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-d4)pyridine Thionyl chloride (0.139 mL, 1.91 mmol) was added to a mixture of (5-(2-fluoroethoxy-1,1,2,2-d4)pyridine-2-yl)methanol (167 mg, 0.953 mmol) in DCM (6.7 mL), and the mixture was stirred at 0°C for 1 hour. After that, 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 under vacuum to obtain the title compound (180 mg, 98%). 1 H NMR (300 MHz, CDCl3) 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-methoxypyridine-4-ol (50 mg, 0.32 mmol) in MeCN (2 mL), phosphorus oxychloride (0.090 mL, 0.98 mmol) was added, and the mixture was heated at 70°C for 22 hours. After this, an additional 0.090 mL, 0.98 mmol of phosphorus oxychloride was added, and heating was continued for 20 hours. Next, a third portion of phosphorus oxychloride (0.090 mL, 0.98 mmol) was added, and heating was continued for 20 hours. After this, volatile matter was removed under reduced pressure, and the resulting residue was diluted with siRNA (20 mL) and neutralized with saturated sodium bicarbonate solution. The layers were separated, and the aqueous layer was extracted with siRNA (10 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified using FCC (silica, 0-10% MeOH in DCM) to obtain the title compound (22 mg, 35%). 1 H NMR (300 MHz, CDCl3) 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-diirbis(4-methylbenzenesulfonate) A mixture of 1,1-diiodoethane (500 mg, 1.77 mmol) and p-silver toluenesulfonate (990 mg, 3.55 mmol) in MeCN (20 mL) was stirred at room temperature for 3 days. After this, volatile matter was removed under vacuum, and the resulting residue was suspended in DCM. The solid was removed by filtration, and the filtrate was concentrated under vacuum at room temperature to obtain the title compound (570 mg, 87%). 1H NMR (300 MHz, CDCl3) 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-diyrbis(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 this, the solvent was removed under reduced pressure, and the resulting residue was purified by FCC (silica, 0-10% butyl in hexane) to obtain the title compound (67 mg, 20%). 1 H NMR (300 MHz, CDCl3) 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, 5.5Hz, 3H).

[0306] Step 3: (5-(1-fluoroethoxy)pyridine-2-yl)methanol A mixture of 6-(hydroxymethyl)pyridine-3-ol (103 mg, 0.825 mmol) and potassium bicarbonate (165 mg, 1.65 mmol) in DMF (10 mL) at room temperature was mixed with 1-fluoroethyl 4-methylbenzene sulfonate (60 mg, 0.27 mmol), and the mixture was heated overnight at 100°C. After this, the solvent was removed under reduced pressure, and the resulting residue was purified by FCC (silica, 0-50% MeOH in butyl) to obtain the title compound (20 mg, 42%). 1H NMR (300 MHz, CDCl3) 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 A mixture of (5-(1-fluoroethoxy)pyridine-2-yl)methanol (16 mg, 0.093 mmol) in room temperature DCM (5 mL) was mixed with thionyl chloride (111 mg, 0.930 mmol), and the mixture was stirred at room temperature for 1 hour. After this, the solvent was removed under reduced pressure to obtain the title compound (23 mg, >99%). 1 H NMR (300 MHz, CD3OD) 8.30 (d, J = 2.5 Hz, 1H), 7.63 - 7.50 (m, 2H), 6.14 (dq, J = 61.9, 4.8 Hz, 1H), 4.67 (s, 2H), 1.65 (dd, J = 20.2, 4.8 Hz, 3H).MS (ES + ) (M+H) + 190.

[0308] Intermediate 6: 6-bromo-2-(2-fluoroethyl)pyridazine-3(2H)-one

[0309] [ka]

[0310] Potassium carbonate (79 mg, 0.57 mmol) was added to 6-bromopyridazine-3(2H)-one (50 mg, 0.29 mmol) in a reaction vial, followed by the addition of DMF (1 mL) and a solution of 1-bromo-2-fluoroethane (58 mg, 0.46 mmol) in 1,4-dioxane (2 mL). The reaction vial was sealed and heated at 130°C for 1 hour. The reaction mixture was then cooled to room temperature and concentrated under reduced pressure until dry. The resulting residue was purified by FCC (silica, hexane with 0-50% ethyl acetate) to obtain the title compound (48 mg, 76%). 1 H NMR (300 MHz, CDCl3) 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, CDCl3) -225.67. MS (ES + ) (M+H) + 221.

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

[0312] [ka]

[0313] Step 1: tert-butyl-[(5-methoxy-2-pyridyl)methoxy]-dimethyl-silane (5-methoxypyridine-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 mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with H2O (15 mL), and the organic fraction was extracted. The aqueous phase was re-extracted with DCM (10 mL), the combined organic matter was washed with brine solution (10 mL), dried over Na2SO4, filtered, and concentrated under vacuum. The crude residue was purified by FCC (silica, 0-100% ethyl phosphate in heptane) to obtain 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 hour. Next, N-fluoro-N-(phenylsulfonyl)benzenesulfonamide (647 mg, 2.05 mmol) was added, and the reaction mixture was warmed to room temperature and stirred for 1.5 hours. The reaction mixture was diluted with brine and ethyl acetate, and the organic fraction was extracted. The combined organic matter was dried over Na2SO4, filtered, and concentrated under vacuum. The crude residue was purified by FCC (silica, heptane, 0-100% ethyl acetate) to obtain 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 room temperature THF (2 mL), 1 M TBAF (0.31 mL, 0.31 mmol) in THF was added, and the reaction mixture was stirred for 1 hour. The reaction mixture was diluted with saturated ammonium chloride solution (3 mL), and the product was extracted with HCl (2 × 5 mL). The combined organic matter was dried over Na₂SO₄, filtered, and concentrated under vacuum. The crude residue was purified by column chromatography (0-100% HCl in silica and heptane) to obtain 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-methoxypyridine (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 mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated under vacuum and azeotropically reacted with DCM (×2) and Et2O to obtain 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-methoxypyridine

[0318] [ka]

[0319] Step 1: Methyl 5-(methoxymethoxy)pyridine-2-carboxylate To a room temperature RBF containing THF (60 mL) under N2, triethylamine (6.8 mL, 49.0 mmol) and methyl 5-hydroxypyridine-2-carboxylate (5.00 g, 32.6 mmol) were added sequentially (in one step), followed by the dropwise addition of chloro(methoxy)methane (3.7 mL, 49.0 mmol) over 5 minutes. The mixture was flushed with N2 to remove all fumes, and the resulting suspension was stirred overnight at room temperature. The reaction mixture was quenched by pouring it over water (50 mL). After extraction with HCl (2 × 50 mL), the combined organic matter was washed with brine (50 mL), dried, filtered, and concentrated under vacuum. The crude residue (pale yellow solid) was purified by column chromatography (Biotage Sfar Duo 50 g cartridge, 0-40% HCl in heptane, product eluted with 30% HCl) to obtain 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 Under nitrogen, a solution of methyl 5-(methoxymethoxy)pyridine-2-carboxylate (5.50 g, 27.9 mmol) in anhydrous toluene (200 mL) was cooled to -78°C, and DIBAL (1 M in heptane, 73 mL, 73 mmol) was added over 45 minutes. The reaction mixture was warmed to 0°C and stirred for 2 hours. The reaction mixture was retreated with further DIBAL (1 M in heptane, 11 mL, 11 mmol) at 0°C, and stirred for a further 1 hour. The reaction mixture was quenched by adding water (50 mL). HCl was added (200 mL). After the addition of MgSO4, the mixture was filtered, eluted with further HCl, and the filtrate was concentrated to obtain 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 mixture was stirred at room temperature for 2 hours. 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 a further 1.75 hours. The reaction mixture was diluted with water (50 mL). After separation, the aqueous phase was extracted with CH2Cl2 (2 × 50 mL), the combined organic matter was washed with brine solution (50 mL), dried, filtered, and concentrated under vacuum. The crude residue was purified by column chromatography using a Biotage Sfar Duo 2×25g cartridge and 0-40% RINKAN in heptane to obtain the title compound (3.5g, 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]-dimethylsilane (2.2 g, 7.06 mmol) in anhydrous THF (35 mL) at -78°C, N-butyllithium (2.5 M in hexane, 3.7 mL, 9.17 mmol) was added, and the reaction mixture was stirred at that temperature for 1 hour. Then, N-fluoro-N-(phenylsulfonyl)benzenesulfonamide (NFSI) (2.89 g, 9.17 mmol) was added directly as a solid over 10 seconds, and the reaction mixture was warmed to room temperature and stirred for 40 minutes. The reaction mixture was quenched by pouring it over brine solution (50 mL). After extraction with siRNA (2 × 40 mL), the combined organic extracts were dried (MgSO4), filtered, and concentrated under vacuum. The residue was purified by FCC using Biotage Isolera (Sfar Duo 25g, 2-30% ethyl acetate in heptane, loaded with DCM) to obtain the title compound (1.4g, 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) = 1.13min, m / z (ES + ) [M+H] + = 302.2, 68%.

[0323] Step 5: 6-[[tert-butyl(dimethyl)silyl]oxymethyl]-4-fluoropyridine-3-ol To a solution of tert-butyl-[[4-fluoro-5-(methoxymethoxy)-2-pyridyl]methoxy]dimethylsilane (1013 mg, 3.36 mmol) in DCM (20.26 mL), zinc dibromide (1.5 g, 6.72 mmol) and propane-1-thiol (0.61 mL, 6.72 mmol) were added. The mixture was stirred at room temperature for 3 hours. The reaction was cooled to 0°C and then quenched by adding saturated NaHCO3 aqueous solution (10 mL). The mixture was stirred at 0°C for 15 minutes. Water (25 mL) was added, and after extraction with DCM (3 × 30 mL), the combined organic extract was washed with brine (30 mL), dried, filtered, concentrated, and a dark orange oil was obtained. The residue was purified by FCC using Biotage Isolera (Sfar Duo 50g, 12-80% toluene in heptane, loaded with DCM) to obtain the title compound (388 mg, 43% yield). 1 H NMR (400 MHz, CDCl3) 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-fluoropyridine-3-ol (385 mg, 1.50 mmol) in DMF (5 mL), cesium carbonate (585 mg, 1.80 mmol) was added, followed by iodomethane (0.11 mL, 1.80 mmol). The mixture was heated to 35°C and stirred at that temperature for 3 hours. After cooling, the reaction was quenched by pouring it over saturated NaHCO3 aqueous solution (10 mL), and water (10 mL) was added. After extraction with Et2O (3 × 20 mL), the combined organic extract was washed with brine (15 mL), dried, filtered, and concentrated under vacuum. The residue was purified by FCC using Biotage Isolera (10 g, 2-30% alkyl hydroxide in heptane, loaded with DCM) to obtain the title compound (243 mg, 59% yield) as a colorless, fluid 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 6 mL of THF 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 hours. The reaction mixture was quenched by pouring it over 10 mL of saturated NaHCO3 aqueous solution. Water (10 mL) and toluene (10 mL) were added. Since separation was insufficient, brine (5 mL) was added. After separation, the aqueous phase was extracted with toluene (2 × 10 mL). The combined organic extracts were washed with brine (15 mL), dried, filtered, and concentrated under vacuum. The residue was purified by FCC using Biotage Isolera (10 g, 5-30% methanol in DCM, loaded with DCM) to obtain 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-methoxypyridine (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 mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under a flow of N2. It was triturated with Et2O(2×), the volatiles were evaporated (under a flow of N2), and then dried in a vacuum oven to obtain the title compound (33 mg, 95% yield). 1H NMR (400 MHz, CDCl3) 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 of Method 1

[0328] [ka]

[0329] Step 1: 5-Methoxy-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)isoindorin-1-one A mixture of 5-methoxy-2,3-dihydroisoindole-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 Pd2(dba)3 (153 mg, 0.167 mmol) in 1,4-dioxane (22.8 mL) was heated at 100°C for 16 hours. The reaction mixture was then cooled and water (50 mL) was added. The resulting solid was collected by filtration and dried under vacuum to obtain the title compound (691 mg, 76%). 1 H NMR (300 MHz, DMSO-d6) 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-dihydropyridazine-3-yl)isoindorin-1-one 1.0 M boron tribromide (27.6 mL, 27.6 mmol) in DCM was added to a solution of 5-methoxy-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindorin-1-one (691 mg, 2.55 mmol) in 1,2-dichloroethane (276 mL), and the mixture was stirred under reflux for 16 hours. After that, the mixture was cooled, ice was added, followed by saturated sodium bicarbonate aqueous solution (30 mL). The formed solid was collected by filtration, dried under vacuum, and then suspended in methanol (659 mL), and the mixture was stirred under reflux for 1 hour. After that, the solvent was removed under vacuum, water (100 mL) was added, and the mixture was sonicated for 10 minutes. The product was collected by filtration, dried under vacuum, and the title compound (540 mg, 82%) was obtained. 1 H NMR (300 MHz, DMSO-d6) 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-fluoropyridine-2-yl)methanol (50 mg, 0.39 mmol) in DCM (2.7 mL), and the mixture was stirred at room temperature for 20 minutes. The mixture was then poured into water (25 mL) and extracted with DCM (2 × 25 mL). The combined organic layer was dried over sodium sulfate, filtered, and concentrated under vacuum to obtain the title compound (57 mg, 99% yield), which was used in the next step without purification.

[0332] Step 4: 5-((5-fluoropyridine-2-yl)methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)isoindorin-1-one A solution of 5-hydroxy-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)isoindorin-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 hours. After this, water (20 mL) was added, and the resulting solid 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). It was then freeze-dried in 1:2 MeCN / water (10 mL) to obtain the title compound (28 mg, 39%).

[0333] Example 1-1: 5-((5-fluoropyridine-2-yl)methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)isoindorin-1-one 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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] Examples 1-2: 5-Methoxy-2-(pyridine-4-yl)-2,3-dihydro-1H-isoindole-1-one

[0336] [ka]

[0337] 1 H NMR (500 MHz, DMSO-d6) 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] Examples 1-3: 5-[(5-iodopyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0339] [ka]

[0340] 1H NMR (500 MHz, DMSO-d6) 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 points, (ES+) (M+H)+ 475, 98%.

[0341] Example 1-4 (Comparative example 2): 5-[(5-メトキシピリジン-2-イル)メトキシ]-2-(ピリジン-4-イル)-2,3-ジヒドロ-1H-イソインドール-1-オン

[0342]

change

[0343] 1 H NMR (500 MHz, DMSO-d6) 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 points, (ES) + (M+H) + 348, 99%.

[0344] Examples 1-5: 5-hydroxy-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0345] [ka]

[0346] 1 H NMR (500 MHz, DMSO-d6) 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] Examples 1-6: 5-[(5-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0348] [ka]

[0349] 1H NMR (500 MHz, DMSO-d6) 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-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0351] [ka]

[0352] 1 H NMR (500 MHz, DMSO-d6) 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). 19F NMR (282 MHz, DMSO-d6) -123.35. Tr(MET-uHPLC-001) = 4.93 min m / z (ES+) (M+H)+ 397.2, 99%.

[0353] Examples 1-8: 5-({5-[2-fluoro(1,1,2,2- 2 H4)Ethoxy]pyridine-2-yl}methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0354] [ka]

[0355] 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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-isoindole-5-yl]oxy}methyl)pyridine-3-yl]oxy}(1,1,2,2- 2 H4) Ethyl 4-methylbenzene-1-sulfonate

[0357] [ka]

[0358] 1 H NMR (500 MHz, DMSO-d6) 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] Examples 1-10: 5-{[5-(1-fluoroethoxy)pyridine-2-yl]methoxy}2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0360] [ka]

[0361] 1H NMR (500 MHz, DMSO-d6) 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-d6) -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-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0363] [ka]

[0364] 1H NMR (500 MHz, CDCl3) 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, 1H), 7.01 (d, J = 10.0 Hz, 1H), 5.21 (s, 2H), 4.85 (s, 2H), 4.03 (s, 3H), 3.76 (s, 3H). Tr(MET-uHPLC-001) = 5.45 min m / z (ES+) (M+H)+ 413.1, 98%.

[0365] Examples 1-12: 6-[(5-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0366] [ka]

[0367] 1 H NMR (500 MHz, DMSO-d6) 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-oxopyridazine-3-yl)isoindorin-1-one

[0369] [ka]

[0370] 1 H NMR (500 MHz, DMSO-d6) 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-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0372] [ka]

[0373] 1H NMR (400 MHz, DMSO-d6) 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-d6) -125.50. Tr(MET-uHPLC-AB-101) = 2.64 min m / z (ES + )(M+H) + 397.2%, 97%.

[0374] Method 2 Scheme of Method 2

[0375] [ka]

[0376] Step 1: (5-(2-fluoroethoxy)pyridine-2-yl)methanol A mixture of 6-(hydroxymethyl)pyridine-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). The resulting reaction mixture was heated in a sealed tube at 70°C for 24 hours. The reaction mixture was then cooled to room temperature, diluted with water (50 mL), and extracted with pharmaceutically acceptable ammonium compounds (3 × 50 mL). The organic layers were combined, dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the title compound (202 mg, 55%) as an orange-brown oil, which was used in the next step without purification. 1H NMR (300 MHz, DMSO-d6) 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)pyridine-2-yl]methoxy}2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one A mixture of 5-hydroxy-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindorin-1-one (80 mg, 0.31 mmol) and (5-(2-fluoroethoxy)pyridine-2-yl)methanol (106 mg, 0.622 mmol) in toluene (20 mL) was mixed with CMBP (188 mg, 0.777 mmol). The mixture was heated in a sealed tube at 120 °C for 48 hours. After this, the solvent was removed under vacuum, and the resulting residue was purified by FCC (silica, 0-10% MeOH in DCM). The product was re-purified by preparative HPLC (water-MeCN) to obtain the title compound (31 mg, 24%).

[0378] Example 2-1: 5-{[5-(2-fluoroethoxy)pyridine-2-yl]methoxy}-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one 1H NMR (500 MHz, DMSO-d6) 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-d6) -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)pyridine-2-yl]methoxy}-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0381] [ka]

[0382] 1H NMR (300 MHz, DMSO-d6) 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-d6) -151.56. Tr(MET-uHPLC-001) = 3.84 min, (ES+) (M+H)+ 397.3, 99%.

[0383] Examples 2-3: 5-[(4-chloro-5-fluoropyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0384] [ka]

[0385] 1 H NMR (500 MHz, DMSO-d6) 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). 19F NMR (282 MHz, DMSO-d6) -132.59. Tr(MET-uHPLC-001) = 5.65 min m / z (ES+) (M+H)+ 401.1, 98%.

[0386] Method 3 Scheme of Method 3

[0387] [ka]

[0388] Step 1: 5-[(5-fluoro-4-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one To a solution of 5-((4-chloro-5-fluoropyridine-2-yl)methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)isoindorin-1-one (68 mg, 0.16 mmol) in MeOH (2.0 mL) and THF (2.0 mL), a 25 wt% solution of sodium methoxide in MeOH (0.041 mL, 0.18 mmol) was added, and the reaction mixture was heated at 65°C for 16 hours. After this, 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 ethyl acetate). The collected material was purified again by FCC (silica, 0-10% MeOH in dimethyl acetate), recrystallized from MeCN, and the title compound (23 mg, 35%) was obtained.

[0389] Example 3-1: 5-[(5-fluoro-4-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2,3-dihydro-1H-isoindole-1-one 1H NMR (500 MHz, DMSO-d6) 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-d6) -152.71. Tr(MET-uHPLC-001) = 4.42 min, (ES + ) (M+H) + 397.0, 99%.

[0390] Method 4 Scheme of Method 4

[0391] [ka]

[0392] Step 1: (5-(allyloxy)pyridine-2-yl)methanol A solution of potassium carbonate (1.65 g, 11.9 mmol) in water (4 mL) was added dropwise over 15 minutes to a mixture of 6-(hydroxymethyl)pyridine-3-ol (1.00 g, 7.99 mmol) and allyl bromide (0.80 mL, 9.3 mmol) in acetone (10 mL). The reaction mixture was heated in a sealed tube at 60°C for 2 hours. After this, 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 obtain the title compound (713 mg, 54%) as a red to brown oil, which was used in the next step without purification. 1H NMR (500 MHz, DMSO-d6) 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)pyridine-2-yl)methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)isoindorin-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)isoindorin-1-one (250 mg, 0.972 mmol) and (5-(allyloxy)pyridine-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, the solvent was removed under vacuum. The resulting residue was suspended in DCM (5 mL) and heptane (5 mL) and filtered. The filtrate was washed with heptane (5 mL), and the filtrate was concentrated under vacuum. The resulting residue was purified by FCC (silica, 0-5% MeOH in DCM). The obtained product was triturated with heptane (5 mL), collected by filtration, washed with heptane (5 mL), and the title compound (45 mg, 12%) was obtained as a brown solid. 1H NMR (300 MHz, DMSO-d6) 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-hydroxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-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)-pyridine-2-yl)methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)isoindorin-1-one (74 mg, 0.18 mmol) in MeOH (6.6 mL), and the mixture was stirred at room temperature for 16 hours. After this, the solvent was removed under vacuum, and DCM (20 mL) was added. The mixture was washed with saturated sodium bicarbonate aqueous solution (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 under vacuum. The resulting residue was purified by FCC (silica, 0-5% MeOH in DCM), and the product was freeze-dried with MeCN (10 mL) and water (10 mL) to obtain the title compound (41 mg, 46%).

[0395] Example 4-1: 5-[(5-hydroxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one 1 H NMR (500 MHz, DMSO-d6) 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 min, (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-isoindole-5-yl]oxy}methyl)pyridine-3-yl]oxydansulfonic acid Chlorosulfonic acid (0.091 mL, 1.4 mmol) was added to a mixture of 5-((5-hydroxypyridine-2-yl)methoxy)-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)isoindorin-1-one (50 mg, 0.14 mmol) in pyridine (6.3 mL) at -15°C. After addition, the mixture was warmed to room temperature and stirred for 2 days. After this, the solvent was removed under vacuum and water (5.0 mL) was added. The mixture was aged for 2.5 days. After this, the formed solid was collected by filtration, washed with water (5.0 mL), and dried under vacuum. The dried solid was triturated with DCM (7.9 mL) and then purified by reverse-phase chromatography (MPLC, water-MeCN). The obtained product was freeze-dried, then triturated with water (15.0 mL) for 2 hours, collected by filtration, and dried under vacuum to obtain 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-isoindole-5-yl]oxy}methyl)pyridine-3-yl]oxydansulfonic acid 1 H NMR (300 MHz, DMSO-d6) 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, 2.1 Hz, 1H), 7.08 (d, J = 10.2 Hz, 1H), 5.29 (s, 2H), 4.87 (s, 2H), 3.64 (s, 3H). 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-dihydropyridazine-3-yl)isoindorin-1-one In a sealed tube, a mixture of 5-bromoisoindorin-1-one (700 mg, 3.30 mmol), 6-bromo-2-((2-(trimethylsilyl)ethoxy)methyl)pyridazine-3(2H)-one (1.21 g, 3.96 mmol), RuPhos (231 mg, 0.495 mmol), cesium carbonate (3.23 g, 9.90 mmol), and Pd2(dba)3 (151 mg, 0.165 mmol) in 1,4-dioxane (16.9 mL) was heated at 100°C for 16 hours under nitrogen. The reaction mixture was then combined with other batches, diluted with water (250 mL), and extracted with ethyl acetate (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% toluene in DCM) to obtain the title compound (1.438 g, >99%). 1 H NMR (300 MHz, DMSO-d6) 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-dihydropyridazine-3-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoindorin-1-one Pd(dppf)Cl2 (134 mg, 0.165 mmol) was added to a mixture of 5-bromo-2-(6-((2-(trimethylsilyl)ethoxy)methoxy)pyridazin-3-yl)isoindorin-1-one (718 mg, 1.65 mmol), bis(pinacolato)diborone (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 hours. The reaction mixture was then combined with another batch and concentrated under reduced pressure. The resulting residue was partitioned between butyl (100 mL) and water (100 mL). The layers were separated, and the aqueous layer was extracted with butyl (2 × 100 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated to obtain 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-dihydropyridazine-3-yl)isoindorin-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)isoindorin-1-one (estimated 3.29 mmol) in THF (41 mL). The reaction mixture was stirred at room temperature for 16 hours. Then, 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. The MeOH was removed under reduced pressure, and the resulting residue was suspended in water and aged at room temperature for 16 hours. The resulting solid was collected by filtration, rinsed with water, and dried. This material was purified by FCC (silica, 0-40% alkyl hydroxide in DCM, followed by 0-10% MeOH in DCM) to obtain the title compound (499 mg, 40%). 1H NMR (300 MHz, DMSO-d6) 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 Hz, 1H), 5.31 (s, 2H), 4.82 (s, 2H), 3.70 (t, J = 7.8 Hz, 2H), 0.89 (t, J = 8.1 Hz, 2H), 0.01 (s, 9H). + ) (M+H) + 374.

[0405] Step 4: 5-((5-methoxypyridin-2-yl)methoxy)-2-(6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazine-3-yl)isoindorin-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)isoindorin-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 hour. After this, 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 mixture was purified by FCC (silica, 20-80% ethyl phosphate in DCM, followed by 0-20% MeOH in DCM) to obtain the title compound (494 mg, 75%). 1H NMR (300 MHz, DMSO-d6) 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-methoxypyridine-2-yl)methoxy]-2-(6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one TFA (1.9 mL) was added to a solution of 5-((5-methoxypyridine-2-yl)methoxy)-2-(6-((2-(trimethyl-silyl)ethoxy)methoxy)pyridazin-3-yl)isoindorin-1-one (494 mg, 0.999 mmol) in DCM (5.7 mL), and the mixture was stirred at room temperature for 2 hours. After this, heptane (73 mL) was added, and volatiles were removed under reduced pressure. The resulting residue was exchanged with Depositphotos (14 mL) and heptane (73 mL), and the residue was triturated in 50:50 water / MeOH (145 mL) at room temperature for 1 hour. 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 Depositphotos, then 0-20% MeOH in DCM). The obtained product was combined with the previous batch and purified by reverse-phase chromatography (MPLC, MeCN-water, 0.1% v / v TFA). The resulting clean fraction was 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, washed with water and MeCN, and the title compound (264 mg) was obtained.

[0407] Example 6-1: 5-[(5-methoxypyridine-2-yl)methoxy]-2-(6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one 1H NMR (500 MHz, DMSO-d6) 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 min, (ES + ) (M+H) + 365.1, 100%.

[0408] Method 7 Scheme of Method 7

[0409] [ka]

[0410] Step 1: 5-[(5-methoxy-2-pyridyl)methoxy]isoindoline-1-one (5-methoxypyridine-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 stirred under N2 for 1 hour. The reaction mixture was concentrated under vacuum, and the residue was co-distilled with DCM (3 × 10 mL), concentrated under vacuum, and 2-(chloromethyl)-5-methoxypyridine was obtained and used without further purification.

[0411] 5-Hydroxyisoindorin-1-one (100 mg, 0.670 mmol), KI (111 mg, 0.670 mmol), and Cs2CO3 (0.107 mL, 1.34 mmol) were dissolved in DMF (6 mL), and the reaction mixture was stirred at room temperature for 10 minutes. 2-(chloromethyl)-5-methoxypyridine (116 mg, 0.738 mmol) was added, and the reaction mixture was stirred at room temperature overnight. The DMF was removed under vacuum, and the residue was triturated with H2O (10 mL) and EtOH (10 mL) to obtain the title compound (140 mg, 77% yield). 1 H NMR (500 MHz, DMSO-d6) 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]isoindoline-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]isoindorin-1-one (140 mg, 0.518 mmol), and the reaction mixture was heated under reflux at 70°C for 1 hour. The reaction mixture was then cooled to room temperature, and 5-(chloromethyl)-2-methoxypyridine (prepared as in 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, and it was then heated under reflux for 30 minutes. The reaction mixture was cooled to room temperature, and 5-(chloromethyl)-2-methoxypyridine (98 mg, 0.622 mmol) was added, and the reaction mixture was stirred for 3 days. The mixture was diluted with toluene (25 mL) and washed with H₂O (2 × 15 mL). The combined organic layer was dried over Na₂SO₄, filtered, and concentrated. The crude product was purified by preparative HPLC (MeCN-water, 2 mM NH₄HCO₃) to obtain the title compound (11.5 mg, 5%).

[0413] Example 7-1: 5-[(5-methoxypyridine-2-yl)methoxy]-2-[(6-methoxypyridine-3-yl)methyl]-2,3-dihydro-1H-isoindole-1-one 1 H NMR (500 MHz, DMSO-d6) 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)pyridazine-3(2H)-one To a solution of 6-bromopyridazine-3(2H)-one (150 mg, 0.857 mmol) in 1,4-dioxane (5 mL) in a reaction vial, potassium carbonate (237 mg, 1.71 mmol) was added, 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 hours. After that, the mixture was cooled to room temperature and concentrated under reduced pressure until dry. The resulting residue was adsorbed onto silica gel and purified by FCC (silica, 0-50% ethyl hexane) to obtain the title compound (90 mg, 51%). 1 H NMR (300 MHz, CDCl3) 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, CDCl3) -176.17. MS (ES+) (M + H) + 207.0.

[0417] Step 2: 2-(1-(fluoromethyl)-6-oxo-1,6-dihydropyridazine-3-yl)-5-((5-methoxypyridine-2-yl)methoxy)-isoindorin-1-one A mixture of 5-(5-methoxypyridine-2-yl)methoxy)isoindorin-1-one (35 mg, 0.13 mmol), 6-bromo-2-(fluoromethyl)pyridazine-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 minutes, and Pd2(dba)3 (6.70 mg, 0.00732 mmol) was added. The reaction vial was sealed and heated at 110°C for 4 hours. After this, the reaction mixture was concentrated until dry, and the resulting residue was purified by FCC (silica, 0-40% MeOH in ethyl acetate). The product was triturated in ethyl acetate, the solid was collected by filtration, and dried under vacuum at 50°C for 3 hours to obtain the title compound (46 mg, 90%).

[0418] Example 8-1: 2-(1-(fluoromethyl)-6-oxo-1,6-dihydropyridazine-3-yl)-5-((5-methoxypyridine-2-yl)methoxy)isoindorin-1-one 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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-methoxypyridine-2-yl)methoxy]-2,3-dihydro-1H-isoindole-1-one

[0421] [ka]

[0422] 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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]isoindorin-1-one 5-Hydroxy-2,3-dihydro-1H-isoindole-1-one (400 mg, 2.68 mmol), 2-(chloromethyl)-5-fluoropyridine (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 hours. 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 obtain the title compound (667 mg, 92% yield). 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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-bromopyridazine-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 mixture was stirred at room temperature for 20 hours. The reaction mixture was retreated with 2-(chloromethoxy)ethyl-trimethyl-silane (4.9 mL, 27.4 mmol), stirred for a further 2 hours, and concentrated until dry. DCM (75 mL) was added, and the organic phase was washed with water (2 × 50 mL) and brine (30 mL). The organic phase was dried over Na₂SO₄, filtered, and concentrated under vacuum. The crude residue was purified by column chromatography (0-100% siRNA in silica and heptane) to obtain the title compound (4.27 g, 61% yield). 1 H NMR (400 MHz, DMSO-d6) 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 min, (ES + ) (M+H) + 356.8, 358.7, 100%.

[0427] Step 3: 5-[(5-fluoro-2-pyridyl)methoxy]-2-[6-oxo-1-(2-trimethylsilylethoxymethyl)pyridazine-3-yl]isoindorin-1-one A solution of 5-[(5-fluoro-2-pyridyl)methoxy]isoindorin-1-one (200 mg, 0.774 mmol) and 6-bromo-2-(2-trimethylsilylethoxymethyl)pyridazine-3-one (260 mg, 0.852 mmol) in 1,4-dioxane (15 mL) was degassed in a pressure tube for 5 minutes, and then RuPhos (11 mg, 0.0232 mmol) and Pd2(dba)3 (21 mg, 0.023 mmol) were added. The reaction mixture was degassed for a further 5 minutes, and Cs2CO3 (0.30 g, 0.929 mmol) was added. The reaction mixture was degassed for 5 minutes and stirred at 100°C for 2 hours. The cooled reaction mixture was retreated with 5-[(5-fluoro-2-pyridyl)methoxy]isoindorin-1-one (50 mg, 0.19 mmol) and stirred overnight. The cooled reaction mixture was filtered, and the filtrate was concentrated. The residue was suspended in DCM (20 mL) and washed (10 mL). The organic phase was dried using a separator cartridge and concentrated under vacuum. The crude residue was purified by column chromatography (0-100% ethyl phosphate in silica and heptane) to obtain the title compound (281 mg, 74% yield). 1 H NMR (500 MHz, DMSO-d6) 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-pyridazine-3-yl)isoindorin-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]isoindorin-1-one (100%, 281 mg, 0.582 mmol) in DCM (9 mL), and the reaction mixture was stirred at room temperature for 2.5 hours. The reaction mixture was concentrated under vacuum, redissolved in DCM, and concentrated again (×3). The crude residue was partitioned between DCM (3 mL) and saturated NaHCO3 solution (2 mL) to separate the organic fraction, which was dried using a separator cartridge. The dried organic fraction was concentrated under vacuum to obtain the title compound (24 mg, 11% yield).

[0429] Example 9-1: 5-[(5-fluoro-2-pyridyl)methoxy]-2-(6-oxo-1H-pyridazine-3-yl)isoindorin-1-one 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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- 2H4)pyridine-2-yl)methoxy)-2-(6-oxo-1,6-dihydropyridazine-3-yl)isoindorin-1-one

[0432] [ka]

[0433] 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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-methoxypyridine-2-yl)methoxy)-2-(6-oxo-1,6-dihydropyridazine-3-yl)isoindorin-1-one

[0435] [ka]

[0436] 1H NMR (500 MHz, DMSO-d6) 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-d6) -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)isoindorin-1-one A mixture of 5-bromo-7-fluoroisoindorin-1-one (500 mg, 2.17 mmol), bis(pinacolato)diborone (828 mg, 3.26 mmol), potassium acetate (533 mg, 5.43 mmol), and Pd(dppf)Cl2 (159 mg, 0.217 mmol) in 1,4-dioxane (15.7 mL) was heated at 90°C for 2 hours. The reaction mixture was then cooled, poured into water (50 mL), and extracted with DCM (3 × 30 mL). The combined organic layer was dried over sodium sulfate, filtered, and concentrated under vacuum to obtain the title compound (870 mg, >99%). 1H NMR (300 MHz, CDCl3) 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-hydroxyisoindoline-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)isoindorin-1-one (870 mg, 3.14 mmol) in THF (53.7 mL), and the mixture was stirred at room temperature for 1 hour. After this, saturated aqueous ammonium chloride solution (200 mL) was added, and volatile matter was removed under vacuum. 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 under vacuum. The resulting residue was purified by FCC (silica, 0-10% MeOH in DCM) to obtain the title compound (111 mg, 31%). 1 H NMR (300 MHz, DMSO-d6) 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-methoxypyridine-2-yl)methoxy)isoindorin-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 hours. After this, water (100 mL) was added, and the mixture was extracted with DCM (2 × 100 mL). The combined organic layer was dried over sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The resulting residue was purified with FCC (silica, 0-5% MeOH in DCM) to obtain the title compound (200 mg, 55%). 1 H NMR (300 MHz, DMSO-d6) 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-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one A mixture of 7-fluoro-5-((5-methoxypyridine-2-yl)methoxy)isoindorin-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 Pd2(dba)3 (16 mg, 0.017 mmol) in 1,4-dioxane (11.1 mL) was heated at 100 °C for 16 hours. The reaction mixture was then cooled, combined with water (50 mL), and extracted with DCM (3 × 100 mL). The combined organic layer was dried over sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The resulting residue was purified by FCC (silica, 0-5% MeOH in DCM) to obtain the title compound (81 mg, 59%).

[0443] Example 10-1: 7-Fluoro-5-[(5-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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-methoxypyridine-2-yl)methoxy]-2-(6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0446] [ka]

[0447] 1 H NMR (500 MHz, DMSO-d6) 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). 19 F NMR (282 MHz, DMSO-d6) -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-[(pyridine-2-yl)methoxy]pyridine Pyridine-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 and ice cooling. The mixture was stirred for 10 minutes. Then, a solution of 2-chloro-5-nitropyridine (500 mg, 3.15 mmol) in tetrahydrofuran (5 mL) was slowly added. The mixture was stirred for 1 hour under ice cooling. The mixture was quenched with water (1 mL), further diluted with water (30 mL), and extracted with ethyl acetate (2 × 30 mL). The combined organic extract was dried over Na₂SO₄, filtered, and concentrated to obtain the title compound (737 mg, quantitative yield). 1 H NMR (250 MHz, DMSO-d6) 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-[(pyridine-2-yl)methoxy]pyridine-3-amine A stirred suspension of 5-nitro-2-[(pyridine-2-yl)methoxy]pyridine (729 mg, 3.15 mmol) in ethanol (15 mL) was heated to 70°C. Then, ammonium chloride (1.65 g, 31.5 mmol) in water (5 mL) was added, followed by the addition of iron powder (0.704 g, 12.6 mmol) all at once. The reaction mixture was stirred at 80°C for 1 hour. The mixture was then filtered through a glass fiber filter, and the inorganic components 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 (Na2SO4), filtered, and concentrated to obtain the title compound (0.62 g, 98% yield). 1 H NMR (500 MHz, DMSO-d6) 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, 1H), 7.02 (dd, J = 8.7, 2.9 Hz, 1H), 6.62 (d, J = 8.6 Hz, 1H), 5.23 (s, 2H), 4.79 (s, 2H). 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 under reflux for 2 hours. The reaction mixture was cooled and concentrated. The title compound (806 mg, 64%) was obtained by column chromatography of the crude residue (silica, 2-20% siRNA-heptane). 1H NMR (500 MHz, CDCl3) 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-[(pyridine-3-yl)methoxy]pyridine-3-yl}-2,3-dihydro-1H-isoindole-1-one N,N-diethylisopropylamine (0.038 mL, 0.22 mmol) was added to 6-(pyridine-3-ylmethoxy)pyridine-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 overnight in a sealed tube at 110 °C. The mixture was then diluted with water (4 mL) and filtered. The collected solid was then purified by FCC (silica, dichloromethane, 0-5% methanol) and then by preparative HPLC (water-acetonitrile) to obtain the title compound (6.1 mg, 10% yield).

[0454] Example 11-1: 5-Methoxy-2-{6-[(pyridine-3-yl)methoxy]pyridine-3-yl}-2,3-dihydro-1H-isoindole-1-one 1H NMR (500 MHz, DMSO-d6) 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), NBS (1.08 g, 6.07 mmol) and AIBN (8 mg, 0.05 mmol) were added, and the mixture was stirred overnight at 80°C. After this, volatile substances were removed under reduced pressure, and the resulting residue was purified with FCC (silica, 0-5% ethyl hexane) to obtain the title compound (720 mg, 73%). 1 H NMR (300 MHz, CDCl3) 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), diethyl phosphonate (856 mg, 6.62 mmol) was added, and the mixture was stirred at room temperature for 18 hours. After this, volatile matter was removed under reduced pressure, and the resulting residue was purified by FCC (silica, hexane with 0-5% ethyl phosphate) to obtain the title compound (346 mg, 64%). 1 H NMR (300 MHz, CDCl3) 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-dihydropyridazine-3-yl)isoindorin-1-one 6-amino-2-methylpyridazine-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 hours. The reaction mixture was then concentrated under vacuum. 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 hours. Volatile matter was then removed under vacuum, and water (10 mL) was added. The resulting solid was collected by filtration and dried under vacuum to obtain the title compound (217 mg, 62%). 1 H NMR (300 MHz, DMSO-d6) 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-dioxaborolan-2-yl)isoindorin-1-one A mixture of 5-bromo-6-fluoro-2-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)isoindorin-1-one (154 mg, 0.455 mmol), bis(pinacolato)diborone (173 mg, 0.683 mmol), potassium acetate (112 mg, 1.14 mmol), and Pd(dppf)Cl2 (33 mg, 0.046 mmol) in 1,4-dioxane (8.2 mL) was heated at 90°C for 6 hours. The reaction mixture was then cooled and concentrated under vacuum. The resulting residue was triturated in water (5 mL), the formed solid was collected by filtration, washed with water (5 mL), and dried under vacuum to obtain the title compound (172 mg, 98%). 1 H NMR (300 MHz, CDCl3) 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-dihydropyridazine-3-yl)isoindorin-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)isoindorin-1-one (172 mg, 0.447 mmol) in MeCN (12.9 mL), and the mixture was stirred at room temperature for 2 hours. Then, saturated aqueous ammonium chloride (1 mL) was added, and the mixture was concentrated under vacuum. 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 that, the formed solid was collected by filtration, dried under vacuum, and triturated in MeOH (8 mL) to obtain the title compound (95 mg, 76%). 1 H NMR (300 MHz, DMSO-d6) 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-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-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-dihydropyridazine-3-yl)isoindorin-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 hours. After this, water (20 mL) was added, and the mixture was extracted with DCM (2 × 30 mL). The combined organic layer was dried over sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The resulting residue was purified with FCC (silica, 0-5% MeOH in DCM) to obtain 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), the resulting solid was collected by filtration, washed with hexane (50 mL), and dried under vacuum to obtain the title compound (73 mg).

[0463] Example 12-1: 6-Fluoro-5-[(5-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0466] [ka]

[0467] 1 H NMR (300 MHz, DMSO-d6) 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-d6) -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-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0469] [ka]

[0470] 1H NMR (500 MHz, DMSO-d6) 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-d6) -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-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0472] [ka]

[0473] 1 H NMR (500 MHz, DMSO-d6) 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). 19F NMR (282 MHz, DMSO-d6) -125.54. Tr(MET-uHPLC-001) = 4.41 min m / z (ES+) (M+H)+ 397.0, 99%.

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

[0475] [ka]

[0476] 1 H NMR (500 MHz, DMSO-d6) 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). Tr(MET-uHPLC-001) = 4.39 min, (ES+) (M+H)+ 380.3, 99%.

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

[0478] [ka]

[0479] 1H 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-d6) -222.32. Tr(MET-uHPLC-001) = 3.23 min, (ES+) (M+H)+ 412.4, 98%.

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

[0481] [ka]

[0482] 1 H NMR (500 MHz, CDCl3) 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, CDCl3) -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-methoxypyridine-2-yl)methoxy]-7-oxo-5H,6H,7H-pyrrolo[3,4-b]pyridine-6-yl}-2-methyl-2,3-dihydropyridazine-3-one

[0484] [ka]

[0485] 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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 H4)Ethoxy]pyridine-2-yl}methoxy)-7-oxo-5H,6H,7H-pyrrolo[3,4-b]pyridine-6-yl]-2-methyl-2,3-dihydropyridazine-3-one

[0487] [ka]

[0488] 1 H NMR (500 MHz, DMSO-d6) 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, CDCl3) -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-methoxypyridine-2-yl)methoxy]-2-(1-methyl-6-oxo-1,6-dihydropyridazine-3-yl)-2,3-dihydro-1H-isoindole-1-one

[0490] [ka]

[0491] 1 H NMR (500 MHz, DMSO-d6) 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). 19F NMR (282 MHz, DMSO-d6) -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-fluoroisoindoline-1-one Ammonia gas was passed through a solution of methyl 4-bromo-2-(bromomethyl)-5-fluorobenzoate (330 mg, 1.01 mmol) in MeOH (10 mL) until saturated at room temperature, and the mixture was stirred overnight. After this, 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 obtain the title compound (217 mg, 93%). 1 H NMR (300 MHz, DMSO-d6) 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-hydroxyisoindoline-1-one A mixture of 5-bromo-6-fluoroisoindorin-1-one (210 mg, 0.913 mmol), bis(pinacorato)diborone (348 mg, 1.37 mmol), and potassium acetate (224 mg, 2.28 mmol) in 1,4-dioxane (20 mL) was mixed with Pd(dppf)Cl2 (33 mg, 0.046 mmol), and the mixture was heated at 100 °C for 18 hours. After that, 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 hours and concentrated under vacuum. The resulting residue was purified by FCC (silica, 0-20% MeOH in ethyl acetate) to obtain the title compound (107 mg, 70%). 1 H NMR (300 MHz, DMSO-d6) 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-methoxypyridine-2-yl)methoxy)isoindorin-1-one A mixture of 6-fluoro-5-hydroxyisoindorin-1-one (100 mg, 0.598 mmol) and potassium carbonate (248 mg, 1.79 mmol) in DMF (10 mL) at room temperature was mixed with 2-(chloromethyl)-5-methoxypyridine hydrochloride (174 mg, 0.897 mmol), and the mixture was heated overnight at 60°C. After this, the solvent was removed under reduced pressure, and the resulting residue was purified by FCC (silica, 0-100% MeOH in ethyl acetate) to obtain the title compound (85 mg, 49%). 1 H NMR (300 MHz, DMSO-d6) 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-dihydropyridazine-3-yl)isoindorin-1-one A mixture of 6-fluoro-5-((5-methoxypyridine-2-yl)methoxy)isoindorin-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) was mixed with Pd2(dba)3 (0.038 g, 0.042 mmol), and the mixture was heated at 100 °C overnight. Subsequently, volatile substances were removed under reduced pressure, and the resulting residue was purified by FCC (silica, 0-100% ethyl hexane) to obtain 6-fluoro-5-((5-methoxypyridin-2-yl)methoxy)-2-(6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazine-3-yl)isoindorin-1-one (0.103 g, 72%). 1 H NMR (300 MHz, DMSO-d6) 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-methoxypyridine-2-yl)methoxy)-2-(6-oxo-1,6-dihydropyridazine-3-yl)isoindorin-1-one A mixture of TFA (2.00 mL, 26.9 mmol) and 6-fluoro-5-((5-methoxypyridine-2-yl)methoxy)-2-(6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1,6-dihydropyridazin-3-yl)isoindorin-1-one (103 mg, 0.201 mmol) was stirred at room temperature for 30 minutes. After this, volatile matter was removed under vacuum, and the resulting residue was purified by preparative HPLC (MeCN-water, 0.1% v / v formic acid). The obtained product was re-purified by FCC (silica, 0-20% MeOH in ethyl acetate) to obtain the title compound (44 mg, 57%).

[0499] Example 13-1: 6-Fluoro-5-((5-methoxypyridine-2-yl)methoxy)-2-(6-oxo-1,6-dihydropyridazine-3-yl)isoindorin-1-one 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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-pyridazine-3-yl)isoindorin-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-pyridazine-3-yl)isoindorin-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), NBS (288 mg, 1.62 mmol) and AIBN (5.3 mg, 0.032 mmol) were added, and the mixture was stirred overnight at 80°C. After this, volatile matter was removed under reduced pressure, and the resulting residue was purified with FCC (silica, 0-5% ethyl hexane) to obtain the title compound (487 mg, 92%). 1 H NMR (300 MHz, CDCl3) 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-fluoroisoindoline-1-one To a solution of methyl 5-bromo-2-(bromomethyl)-4-fluorobenzoate (172 mg, 0.528 mmol) in MeOH (6 mL), 7N ammonia (0.45 mL, 3.2 mmol) in MeOH was added, followed by ammonium hydroxide (2 mL, 0.53 mmol). The mixture was stirred at room temperature for 18 hours. After this, volatile matter was 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 obtain the title compound (119 mg, 98%). 1 H NMR (300 MHz, CDCl3) 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-hydroxyisoindoline-1-one A mixture of 6-bromo-5-fluoroisoindorin-1-one (118 mg, 0.513 mmol), bis(pinacolato)diborone (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 minutes. Pd(dppf)Cl2 (38 mg, 0.051 mmol) was added, and the reaction mixture was heated in a sealed tube at 100°C for 1.5 hours. After that, the mixture was cooled to room temperature and concentrated under reduced pressure until dry. 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 minutes. After that, aqueous ammonium chloride (5 mL) was added. The volatile components were removed under reduced pressure, and the resulting residue was purified by FCC (silica, 0-20% MeOH in acetylene) to obtain the title compound (66 mg, 62%). MS (ES + ) (M+H) + 168.

[0512] Step 4: 5-Fluoro-6-((5-methoxypyridine-2-yl)methoxy)isoindorin-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 hours. After that, 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 ethyl acetate, then 0-20% MeOH in DCM) to obtain the title compound (85 mg, 92%). 1 H NMR (300 MHz, DMSO-d6) 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-bromopyrazine-2-yl)-5-fluoro-6-((5-methoxypyridine-2-yl)methoxy)isoindoline-1-one A mixture of 5-fluoro-6-((5-methoxypyridine-2-yl)methoxy)isoindorin-1-one (84 mg, 0.29 mmol), 2,5-dibromopyrazine (83 mg, 0.35 mmol), xanthophos (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 minutes. Pd2(dba)3 (8 mg, 0.009 mmol) was added, and the reaction mixture was heated in a sealed vial at 120°C for 4 hours. After this, the mixture was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by FCC (silica, 0-100% ethyl phosphate in DCM, then 0-10% MeOH in DCM) to obtain the title compound (53 mg, 41%). 1H NMR (300 MHz, DMSO-d6) 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-hydroxypyrazine-2-yl)-6-((5-methoxypyridine-2-yl)methoxy)isoindorin-1-one A mixture of 2-(5-bromopyrazine-2-yl)-5-fluoro-6-((5-methoxypyridine-2-yl)methoxy)isoindorin-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 minutes. Pd2(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 hours. After this, the mixture was cooled to room temperature, neutralized with 1N HCl, and concentrated under reduced pressure until dry. The resulting residue was purified by FCC (silica, 0-20% MeOH in ethyl acetate) to obtain the title compound (11 mg, 25%). 1 H NMR (300 MHz, DMSO-d6) 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-methoxypyridine-2-yl)methoxy)-2-(4-methyl-5-oxo-4,5-dihydropyrazine-2-yl)isoindorin-1-one and 5-Fluoro-2-(5-methoxypyrazine-2-yl)-6-((5-methoxypyridine-2-yl)methoxy)-isoindorin-1-one Methyl 4-nitrobenzene sulfonate (8 mg, 0.04 mmol) was added to a mixture of potassium carbonate (11 mg, 0.078 mmol) and 5-fluoro-2-(5-hydroxypyrazine-2-yl)-6-((5-methoxypyridine-2-yl)methoxy)isoindorin-1-one (10 mg, 0.029 mmol) in DMF (2 mL), and the mixture was stirred at room temperature for 16 hours. Subsequently, the reaction mixture was concentrated under reduced pressure until dry, and the resulting residue was absorbed onto silica gel. It was then purified by FCC (silica, 0-15% MeOH in ethyl acetate) to obtain 5-fluoro-6-((5-methoxypyridine-2-yl)methoxy)-2-(4-methyl-5-oxo-4,5-dihydropyrazine-2-yl)isoindorin-1-one (5 mg, 48%) and 5-fluoro-2-(5-methoxypyrazine-2-yl)-6-((5-methoxypyridine-2-yl)methoxy)isoindorin-1-one (5 mg, 48%).

[0516] Example 14-1: 5-Fluoro-6-((5-methoxypyridine-2-yl)methoxy)-2-(4-methyl-5-oxo-4,5-dihydropyrazine-2-yl)isoindorin-1-one 1H NMR (500 MHz, CDCl3) 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, CDCl3) -124.82. Tr(MET-uHPLC-001) = 2.90 min (ES + ) (M+H) + 397.2%, 97%.

[0517] Example 14-2: 5-Fluoro-2-(5-methoxypyrazine-2-yl)-6-((5-methoxypyridine-2-yl)-methoxy)isoindorin-1-one 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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-methoxypyridine-2-yl)methoxy)-2-(4-methyl-5-oxo-4,5-dihydropyrazine-2-yl)isoindorin-1-one

[0520] [ka]

[0521] 1 H NMR (500 MHz, DMSO-d6) 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-d6) -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 the radioligand-binding assay (RBA), the MBP-HTT(1-89)Q46-His(6×) ("exon 1-Q46") protein was generated based on a previous publication (Scherzinger et al., Cell, Vol. 90, pp. 549-558, August 8, 1997). For the experiment, 30 μM MBP-exon 1-Q46 was incubated with 150 μg / mL thrombin and 2 mM CaCl2 in assay buffer (150 mM NaCl, 50 mM Tris pH 8.0) at 37°C for 16 hours. The aggregated exon 1-Q46 was pelletized by centrifugation at 13,000 rpm for 5 minutes in a benchtop centrifuge and redissolved in the same volume of assay buffer. The test compound was prepared at 11 concentrations ranging from 63 μM to 2 nM by titration in DMSO. For RBA, exon 1-Q46 protein aggregates and test compounds were pre-incubated in assay buffer at room temperature for 20 minutes at 100 μL / well in a 96-well plate (pp, round-bottom). Then, ligand was added at 50 μL / well and incubated at 37°C for 60 minutes. The final assay concentrations were 1 μM to 30 pM of the test compound, 1 μM of exon 1-Q46 protein (equivalent monomer concentration), and 0.3 nM of ligand. 3 The sample was H3-methyl]-5-((5-methoxypyridine-2-yl)methoxy)-2-(pyrazine-2-yl)benzo[d]oxazole. The sample was transferred to a GF / B filter plate and washed twice with 200 μL PBS using a Filtermate Harvester. After drying the filter plate at 55°C for 1 hour, the back of the plate was sealed with foil, 30 μL / well of scintillation fluid (Packard MicroScint 40) was added, and the plate was incubated in the dark for 15 minutes and counted with a MicroBeta reader. For analysis, replicate data from independent assay plates were used for the vehicle's control well (0% inhibition) and 1 μM unlabeled [ 3 The results were normalized to 0% and 100% inhibition using H3-methyl]-5-((5-methoxypyridine-2-yl)methoxy)-2-(pyrazine-2-yl)benzo[d]oxazole (100% inhibition). IC 50The values ​​use normalized iterative data, and four variables (upper, lower, slope, IC) are used in the overall fit. 50 This was determined using an S-order inhibition model employing ).

[0523] The results for various example compounds are shown in the table below (+++<100nM;++100~500nM;+>500nM;ND: Undetermined):

[0524] [Table 4]

[0525] PET imaging examples The following examples illustrate a non-limiting procedure that may be used when conducting PET imaging studies on individuals in a clinical setting. The individual is either untreated or pre-treated with an unlabeled compound. The individual may fast before PET imaging and is permitted to drink water freely. 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 inside a PET camera, and a tracer dose of the imaging agent is administered via an intravenous catheter. Arterial or venous blood samples are taken at appropriate time intervals during the PET scan to analyze and quantify the proportion of unmetabolized compounds in the plasma. Images are obtained for up to 120 minutes. Within 10 minutes of radiotracer injection, and at the end of the imaging session, 1 mL blood samples are taken to determine the plasma concentration of any unlabeled imaging agent compounds (or other intervention compounds) that may have been administered prior to the PET tracer.

[0527] A tomographic image is obtained by image reconstruction. For example, a region of interest (ROI) is set on the reconstructed image to determine the distribution of the imaging agent. The region of interest in the brain image may include, for example, the striatum, cerebellum, or basal ganglia. The uptake of the imaging agent over time in these regions may be used to generate a time-radioactivity curve (TAC). The data can be expressed as radioactivity per unit time per unit volume (e.g., μCi / cc / mCi injection dose) or radioactivity per unit volume. The TAC data can be processed by various methods known in the art to obtain quantitative parameters, such as binding capacity (BP). For further description of the imaging procedure, 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] Disconnected brain fraction The compound is expressed as an unbound brain fraction (or brain free fraction, f u , brain The following was tested: It is an accepted standard in the field of CNS PET imaging that compounds with a low unbound fraction (<5%) in the brain may have a high nonspecific binding background that is undesirable for detecting target binding. As a result, f u,brainThis is an important property 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, Vol. 56 (No. 11), pp. 4568-4579; Liu et al., Imaging Mutant Huntingtin Aggregates: Development of a Potential PET Ligand. J Med Chem 2020, Vol. 63 (No. 15), pp. 8608-8633].

[0529] [Table 5] TIFF0007855011000091.tif236149TIFF0007855011000092.tif229149TIFF0007855011000093.tif225148TIFF0007855011000094.tif29148

[0530] f in the mouse brain due to equilibrium dialysis u , brain The determination was made by immersing a dialysis membrane (12-14 kDa) (HTDialysis, Connecticut) in phosphate buffer (10 mM potassium phosphate and 0.8% sodium chloride buffer, pH 7.4, 37°C) for at least 1 hour, at which point ethanol was added (final concentration of 20% v / v), and the membrane was left immersed for 30 minutes. Mouse brain tissue was diluted in phosphate buffer in a 1:4 ratio (w / v) and homogenized using Precellys 24 (Stretton Scientific, UK) to achieve a final concentration of 20% brain homogenate.

[0531] The test compound, prepared as a 0.5 mM DMSO stock, was diluted in brain homogenate to achieve a final substrate concentration of 5 μM (1% DMSO). A t=0 sample was prepared by sampling 50 μL of this homogenate into 400 μL of quench solution (acetonitrile containing 0.1% formic acid and sulfisoxazole / tolbutamide / imipramine / labetalol 200 nM). The sample was matrix-matched with 50 μL of assay buffer. HTDialysis Teflon blocks (HTDialysis, Connecticut) were assembled using pre-soaked membranes between each receiver and donor well and fixed to a stainless steel pressure plate (HTDialysis, Connecticut). 120 μL of buffer was administered to the lower compartment (acceptor side), and 120 μL of the compound in the homogenate was administered to the upper compartment (donor side). All test compounds and controls were completed in three series. The HT dialysis plates were sealed and incubated at 37°C for 6 hours while being shaken at 250 rpm.

[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 using 50 μL of alternative blank matrix. Analytical samples were diluted 1:1 with water using a Janus Robot and analyzed by LC-MS / MS.

[0533] The peak areas of the donor and acceptor sections are compared to determine the unbound percentage (%f). u The ) was determined and the average of the three consecutive measurements was reported. Sample recovery (%) was calculated using the reference T0 sample. The functionality of the assay was confirmed by comparing the control compound with historical and literature values.

[0534] If the average free brain fraction of a compound is less than 5%, the background signal increases due to undesirable indiscriminate binding. 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 adequately detect brain structures containing aggregated proteins such as mHTT. The data show that the compounds described herein have a higher free brain fraction than comparative compounds and simultaneously provide good binding to mHTT protein species. Therefore, the compounds described herein have desirable properties for in vivo use as imaging agents.

[0535] In-situ autoradiography (ARG) Using in situ autoradiography (ARG), tritium-labeled compounds 1-6 ("[ 3 The pharmacological binding properties of compound 1-6 (H) were investigated; the specificity of binding was investigated by including samples from Alzheimer's disease (AD) patients containing other pathological aggregates (e.g., A-beta-containing plaques and phosphorylated Tau-expressing tangles). Saturation binding experiments were performed in coronary brain sections; 3 The H]-compound 1-6 bonds were quantified by concentration analysis, and specific binding was determined in the cortex (CTX). For each region of interest (ROI), affinity (K D ) and the maximum number of bonding sites (B max ) was decided.

[0536] Tissue preparation and sectioning At appropriate age, HD and WT mice of different strains were sacrificed by cervical dislocation, their brains, including the cerebellum, were dissected, and washed with ice-cold PBS. The brains were then gently pressed on paper towels to remove excess moisture and transferred to 6-well plates filled with isopentane cooled to 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. This resulted in a series of 20 μm thick coronal tissue sections mounted on Superfrost slides, which were dried at room temperature for approximately 90 minutes. The slides were stored at -80°C until various experiments were performed, but not for more than 3 weeks. A similar procedure was performed using fresh-frozen human postmortem brain blocks (healthy subjects {CTRL}, Huntington's disease {HD}, and Alzheimer's disease {AD}).

[0538] Assay procedure A suitable number of slides containing each tissue section were acclimatized to room temperature for 30 minutes. The slides were then pre-incubated by immersing them in 40 mL of assay buffer at room temperature for 20 minutes. After this pre-incubation step, one slide per animal was incubated for 60 minutes at room temperature with 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 Incubate one slide by immersing it in a solution of compound 1-6 [H], and then add 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 The compounds were incubated by immersion in a 10 μM solution of unlabeled compound 1-6 with compound 1-6 [H]. A similar paradigm was applied to tritium-labeled comparative compound 3 ("[ 3 H]-Comparative compound 3) and tritium-labeled comparative compound 4 ("[ 3 The experiment was conducted using [H]-comparative compound 4). The structures of these comparative compounds and compounds 1-6 are shown below.

[0539] [Table 6]

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

[0541] Data Analysis Concentration measurement data analysis was performed using MCID Analysis 7.1 software (Interfocus Imaging Ltd.). Within each brain section, a region of interest (ROI) was defined for STR, CTX, and HPC using appropriate sample tools. The software then calculated density values ​​(molecular dynamics counts per unit area, MDC / mm²) from the gray level values ​​of all pixels within the defined ROI. 2 The following calculations were performed. For concentration measurement calibration, the optical density was correlated to a known concentration of radioactivity (fmol / mg tissue) from a calibrated tritium standard, and the mean value was calculated for each ROI of individual brains. Total binding (TB) and nonspecific binding (NSB) of the radioactive ligand were quantified, and specific binding (SB) was derived for each brain and ROI by subtracting NSB from TB (SB = TB - NSB). Subsequently, the group mean ± standard deviation (SD) was calculated for each ROI and experimental condition. The data were fitted to a one-site binding equation using a nonlinear regression method in GraphPad Prism software.

[0542] Saturated bond: HOM zQ175 HD mouse model As can be seen in Figure 1, [ 3 Compounds 1-6 of [H]- showed concentration-dependent specific binding and low nanomolar affinity to mHTT aggregates present in HOM zQ175 brain sections; in the cortex, at 1.4 nM K D The average value was obtained. B was calculated as the measured value for the concentration of the specific binding site. maxThe concentration reached 448.6 fmol / mg in the cortex. These results are thought to reflect the amount and density of mHTT aggregates expressed in this region of the brain.

[0543] Binding to WT brain sections was minimal, and detection was only possible above the lower limit of quantification (LLoQ, 0.5 fmol / mg tissue) at the two highest concentrations investigated, 3 nM and 10 nM. A binding curve could not be established. D The value is also B max The value could not be determined, and this suggests that the mHTT aggregate expressed only in brain sections of HOM zQ175 (12 months old) is [ 3 This confirmed the specific binding of compounds 1-6 [H].

[0544] Determined in the cortex, [ 3 H]-Comparative compound 4 and [ 3 Compared with H]-comparative compound 3, over a range of radioactive ligand concentrations, 3 The specific and saturated bonds of compound 1-6 are further shown in Figure 1. As shown in the table below, 3 The Bmax (448 fmol / mg) of compound 1-6 is [ 3 H]-Comparative compound 4 (149 fmol / mg) and [ 3 [H]- Compared to comparative compound 3 (169 fmol / mg), the level was significantly higher.

[0545] [Table 7]

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

[0547] Combination: Human postmortem brain section Next, to provide information on the universal convertibility of this mHTT aggregate binder, particularly its pathological specificity and species selectivity, we extended the observation of in situ binding from tissues derived from mouse HD models to postmortem human brain tissue.3 Compounds 1-6 of [H]- showed 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 cortical 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, HD vs. CTRL). 3 Compounds 1-6 of [H] showed different specific binding in the gray matter region, with very low white matter binding (≤0.5 fmol / mg tissue). These data are consistent with observations that mHTT aggregates are mainly found in cortical neurons (gray matter) and rarely expressed in white matter.

[0549] Next, [ 3 Further investigations were conducted to determine whether compound 1-6 of [H] exhibits different binding properties in different forms of the disease. To address this, a juvenile HD sample (1 case) was tested in addition to postmortem adult-onset HD brains. Juvenile HD is characterized by earlier onset and much faster progression compared to adult HD. 3 Compounds 1-6 of [H]- showed the highest binding density in juvenile HD brains (white triangles in Figures 2 and 3), suggesting that these compounds may recognize mHTT aggregates in both forms of the disease and recognize additional epitopes expressed in juvenile HD brains compared to adult-onset HD brains.

[0550] [ 3 Compound 1-6 also showed significantly higher binding in the cortex of HD patients compared with brain sections of AD patients, as seen in Figure 2 (0.1 ± 0.0 fmol / mg tissue; p < 0.01, HD vs AD comparison; no significant difference was observed when comparing CTRL vs AD). 3The low SB of compound 1-6 supports the selectivity of this radioligand for mHTT compared to 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 Although it was detectable with H]-compounds 1-6, HD binding was [ 3 H]-Comparative compound 4 or [ 3 [H]-Neither of the comparison compounds 3 is detectable at 0.3 nM, and this means that [ 3 H]-Comparative compound 4 or [ 3 [H]-Comparison compound 3 compared with radioactive ligand, 3 This suggests that compounds 1-6 of [H]- recognize different binding epitopes and / or greater affinity for mHTT aggregates expressed in the brains of HD patients.

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

[0553] moreover,[ 3 H]-compound 1-6 is, 3 H]-Comparative compound 4 or [ 3 [H] selectively bound to human HD brain sections at a radioactive ligand concentration of 0.3 nM, the same concentration at which HD binding to any of the comparative compounds 3 could not be demonstrated. This suggests novel epitope recognition and / or higher affinity binding to compounds 1-6.

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

[0555] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which this disclosure belongs.

[0556] The disclosures described herein as examples may be appropriately implemented without any one or more elements or limitations not specifically disclosed herein. Therefore, terms such as “comprising,” “including,” and “containing” should be interpreted openly and non-restrictively. Furthermore, the terms and expressions used herein are for illustrative purposes only, not limitation, and there is no intention to exclude equivalents of the illustrated and described features or parts thereof, although various modifications are permitted within the scope of this disclosure.

[0557] All publications, patent applications, patents, and other references mentioned herein are incorporated by explicit reference as well as each is incorporated by explicit reference individually. In case of any conflict, this specification, including definitions, shall prevail. The present invention encompasses the following embodiments. (Embodiment 1) Formula I: [ka] [In the formula, [ka] teeth, [ka] or

change

change

change

change

change

change

change

change

change

change

change

change

change

change

change

change

change

Claims

1. Formula I: 【Chemistry 1】 [In the formula, 【Chemistry 2】 teeth, 【Transformation 3】 or 【Chemistry 4】 and; R 1 If present, hydrogen, C 1～6 Alkyl, or C 1～6 It is a haloalkyl; R 10 If present, hydrogen, C 1～6 Alkyl, or C 1～6 It is a haloalkyl; Ring A is pyridinyl; 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 alkoxy; Y 1 CR 12 or N; Y 2 CR 13 or N; R 12 and R 13 Each of these is hydrogen, hydroxyl, halo, and C. 1～6 Alkyl, C 1～6 Haloalkyl, or C 1～6 It is an alkoxy; R 2 is hydrogen, hydroxyl, halo, C 1～6 Alkyl, C 1～6 Haloalkyl, or C 1～6 It is an alkoxy; L is C which is optionally substituted with 1 to 6 fluorocarbons. 1 ~C 3 It is alkylene; R 3 is hydrogen, fluorocarbon, C 1～6 Alkyl, or C 1～6 It is a haloalkyl; Each R 4 These are independently cyano, hydroxy, halo, and C. 1～6 Alkyl, C 1～6 Haloalkyl, or C 1～6 It is an alkoxy; Each R 5 These are independently cyano, hydroxy, halo, and C. 1～6 Alkyl, C 1～6 Haloalkyl, or C 1～6 It is an alkoxy; R 6 These are hydrogen, cyano, hydroxy, halo, and 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 It is a haloalkyl, R 7 C 1～6 Alkyl or C 1～6 Haloalkyl is -SO 2 -Aryl, -OSO 2 -Variable, optionally substituted with aryl atoms, 1 to 6 deuterium atoms, or combinations thereof, -SO 2 -aryl or -OSO 2 -aryl is cyano, hydroxy, halo, C 1～6 Alkyl, C 1～6 Haloalkyl, or C 1～6 It is further sometimes substituted with alkoxy; m is 0, 1, 2, or 3; n is 0, 1, or 2. Compounds thereof, or isotope-enriched analogs thereof, pharmaceutically acceptable salts, tautomers, stereoisomers, or mixtures of stereoisomers.

2. Formula II: 【Transformation 5】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

3. Formula III: 【Transformation 6】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

4. Formula IV: 【Transformation 7】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

5. Formula V: 【Transformation 8】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

6. Equation VI: 【Chemistry 9】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

7. Formula VII: 【Chemistry 10】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

8. Formula VIII: 【Chemistry 11】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

9. Formula IX: 【Chemistry 12】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

10. Formula X: 【Chemistry 13】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

11. Formula XI: 【Chemistry 14】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

12. Formula XII: 【Chemistry 15】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

13. Formula XIII: 【Chemistry 16】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

14. Formula XIV: 【Chemistry 17】 The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

15. Formula XV: [Chemistry 18] The compound according to claim 1, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

16. R 1 is hydrogen or C 1～6 The compound according to claim 1, wherein it is alkyl.

17. R 1 C 1～6 The compound according to claim 1, wherein it is alkyl.

18. R 1 The compound according to claim 1, wherein is methyl.

19. R 2 The compound according to claim 1, wherein the compound is hydrogen.

20. R 3 The compound according to claim 1, wherein the compound is hydrogen.

21. R 4 The compound according to claim 1, wherein the compound is a halo.

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

23. The compound according to claim 1, wherein n is 1.

24. The compound according to claim 1, wherein n is 0.

25. R 11 The compound according to claim 1, wherein the compound is hydrogen.

26. The compound according to claim 1, wherein m is 1.

27. R 5 The compound according to claim 1, wherein the compound is a halo.

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

29. The compound according to claim 1, wherein m is 0.

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

31. R 7 C 1～6 Haloalkyl or C 1～6 The compound according to claim 30, wherein it is alkyl.

32. R 7 However, C is substituted with 1 to 6 deuterium atoms. 1～6 The compound according to claim 30, which is a haloalkyl compound.

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

34. The compound according to claim 1, wherein X is N.

35. L is CH 2 The compound according to claim 1.

36. Y 1 N is Y 2 The compound according to claim 1, wherein is CH.

37. The compound according to claim 1, wherein ring A is pyridine-2-yl. 【Request Item 38】 【Chemistry 19】 but, 【Chemistry 20】 And R 10 C 1～6 The compound according to claim 1, wherein it is alkyl.

39. R 1 、R 4 、R 5 、R 6 、and R 11 The compound according to claim 1, wherein at least one of them contains a fluorine atom.

40. One R 4 or one R 11 The compound according to claim 1, wherein it is fluoro.

41. A compound selected from the following compounds, optionally labeled with a radioactive isotope, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers: Table 1-1 Table 1-2 Table 1-3 Table 1-4 Table 1-5 Table 1-6 Table 1-7 Table 1-8 Table 1-9 。

42. The compound according to claim 1, wherein the compound is labeled with a radioactive isotope.

43. The compound, 11 C, 13 N, 15 O, and 18 The compound according to claim 42, comprising a positron-emitting radioactive isotope selected from F.

44. An imaging agent comprising the compound according to claim 42 or 43, or an isotope-enriched analog thereof, a pharmaceutically acceptable salt, a tautomer, a stereoisomer, or a mixture of stereoisomers.

45. An imaging agent for detecting the presence or absence of a protein prone to aggregation in an individual by generating an image of a part or region of the body of an individual, comprising an effective amount of the compound described in claim 42.

46. The imaging agent according to claim 45, comprising generating an image of a body part or region of an individual, and detecting the presence or absence of proteins that are prone to aggregation in the image.

47. The imaging agent according to claim 46, wherein the protein that readily undergoes aggregation is huntingtin protein (HTT protein).

48. The imaging agent according to claim 47, wherein the HTT protein is found in the basal ganglia of the brain.

49. The imaging agent according to claim 46 or 47, wherein the presence or absence of protein aggregates corresponds to the presence or absence of neurodegenerative disease.

50. The imaging agent according to claim 49, wherein the neurodegenerative disease is selected from Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, Parkinson's disease, prion disease, and spinocerebellar ataxia.

51. The imaging agent according to claim 50, wherein the neurodegenerative disease is Huntington's disease (HD).

52. The imaging agent according to claim 45, wherein the effective amount of the compound contains about 0.1 to about 20 mCi.

53. The imaging agent according to claim 52, wherein the effective amount of the compound contains about 10 mCi.

54. The imaging agent according to claim 45, wherein the generation of images includes 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.

55. The imaging agent according to claim 54, wherein generating an image includes PET imaging.

56. The imaging agent according to claim 47 or 48, wherein the HTT protein exists as an oligomer, aggregate, or a combination thereof.

57. The imaging agent according to claim 47 or 48, wherein the HTT protein is a variant.

58. The imaging agent according to claim 45, wherein the body part or region is the head, spinal cord, limbs, chest, or abdomen.

59. The imaging agent according to claim 45, wherein the body part or region is the brain.

Images