Synthesis of pyruvate kinase activators

Abstract

Provided herein are methods for preparing activators of pyruvate kinase (PK), or salts or hydrates thereof.

Description

[0001] Related Applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 525,079, filed July 5, 2023, the entirety of which is incorporated by reference herein. BACKGROUND

[0003] Pyruvate kinase (PK) is a metabolic enzyme that converts phosphoenolpyruvate to pyruvate during glycolysis. There are four isoforms of PK in mammals: L and R isoforms are expressed in the liver and red blood cells, the M1 isoform is expressed in most adult tissues, and the M2 isoform is a splice variant of M1 that is expressed during embryonic development. A well-known difference between the M1 and M2 isoforms of PK is that M2 is a low-activity enzyme that is dependent on allosteric activation by the upstream glycolytic intermediate fructose-1,6-bisphosphate (FBP), whereas M1 is a constitutively active enzyme. PK activators can be used to treat a variety of different conditions, including PKD (pyruvate kinase deficiency), thalassemia (e.g., alpha and beta-thalassemia), hereditary elliptocytosis, abetalipoproteinemia or Bassen-Kornzweig syndrome, sickle cell disease, paroxysmal nocturnal hemoglobinuria, and various anemias, including congenital anemias (e.g., enzymopathies) and hemolytic anemias (e.g., hereditary and / or congenital hemolytic anemia, acquired hemolytic anemia, chronic hemolytic anemia due to phosphoglycerate kinase deficiency, anemia due to MDS (myelodysplastic syndrome), non-spherocyte hemolytic anemia, and hereditary spherocytosis). SUMMARY

[0004] Provided herein are synthetic methods for preparing a pyruvate kinase (PK) activator or a salt or hydrate thereof. Such activators can be prepared using a compound having formula (A-1) and / or formula (B-1):

[0005] and

[0006] (A-1) (B-1),

[0007] wherein X, R 1 , R 4 , and Q are as defined herein.

[0008] Also disclosed are particular PK activators and various intermediates that are prepared by and / or used in the synthetic methods described herein. BRIEF DESCRIPTION OF DRAWINGS

[0009] Figure 1The nuclear magnetic resonance (NMR) spectrum of 2-(6-((6-aminopyridin-2-yl)methyl)-4-methyl-5-oxo-5,6-dihydro-4H-thiazo[5',4':4,5]pyrrolo[2,3-d]pyridazin-2-yl)-2-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)tert-butyl acetate was depicted.

[0010] Figure 2 The nuclear magnetic resonance (NMR) spectrum of 2-chloro-6-formyl-4-methyl-4H-pyrrolo[2,3-d]thiazol-5-carboxylic acid was depicted. Detailed Implementation

[0011] In one embodiment, a preparative compound of formula (A-1) is provided:

[0012]

[0013] (A-1),

[0014] or its salt method, wherein

[0015] R 1 It is hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, C1-C6 aminoalkyl, C1-C6 cyanoalkyl, C2-C6 alkenyl, C2-C6 alkynyl or C3-C6 cycloalkyl, wherein the C3-C6 cycloalkyl is optionally substituted by 1 to 3 groups selected from the group consisting of: halogroup, hydroxyl, NH2 and CN;

[0016] X is a halogenated group;

[0017] L is a bond or a C1-C6 alkylene group; and

[0018] Q is a C3-C6 cycloalkyl, a 5-14-membered heterocyclic group, a 6-12-membered aryl group, or a 5-14-membered heteroaryl group, each of which is optionally substituted by 1 to 3 groups selected from the following: halogroup, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, hydroxyl, C1-C6 aminoalkyl, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, CN, and NO2;

[0019] This method involves making a compound of formula (A-2):

[0020]

[0021] (A-2) or its salt,

[0022] With compound of formula (A-3):

[0023]

[0024] (A-3),

[0025] The reaction forms a compound of formula (A-1) or a salt thereof.

[0026] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. The terminology used in this specification is for describing particular embodiments only and is not intended to limit this disclosure.

[0027] Unless otherwise expressly required, the use of any and all instances or exemplary language (e.g., “such as” and “for example”) provided herein is intended to better illustrate this disclosure and is not intended to limit the scope of this disclosure. Phrases such as “in one aspect”, “in one embodiment”, “in some embodiments”, “in certain embodiments”, or “in another embodiment” should not be construed as indicating that such elements occur or exist in isolation, or that such elements are not shared by other aspects or embodiments of this disclosure. Rather, it should be understood that all aspects and embodiments are freely combinable with any and all other aspects and embodiments of this disclosure as described herein. No language in this specification should be construed as indicating that any element not protected by the claims is necessary to practice this disclosure.

[0028] The term "C1-C6 alkyl" refers to a straight-chain or branched hydrocarbon group having 1 to 6 carbon atoms. Examples of C1-C6 alkyl groups include methyl (C1), ethyl (C2), propyl (C3) (e.g., n-propyl, isopropyl), butyl (C4) (e.g., n-butyl, tert-butyl, sec-butyl, isobutyl), pentyl (C5) (e.g., n-pentyl, 3-pentyl, pentyl, neopentyl, 3-methyl-2-butyryl, tert-pentyl), and hexyl (C6) (e.g., n-hexyl).

[0029] The term "C1-C6 alkylene" refers to a divalent group of a saturated, straight-chain, or branched hydrocarbon. Exemplary alkylenes include, but are not limited to, the corresponding alkylene groups of C1-C4 alkylenes, C1-C3 alkylenes, C1-C2 alkylenes, and any of the other exemplary alkyl groups described above.

[0030] When this article uses a certain range of carbon atoms, such as C 1- C6 encompasses all ranges and the individual number of carbon atoms. For example, "C 1- "C3" includes C 1- C3, C 1- C2, C 2- C3, C1, C2, and C3.

[0031] As used herein, the term "C2-C6alkenyl" refers to a straight or branched chain group having from 2 to 6 carbon atoms in the group, wherein the group includes at least one carbon-carbon double bond. Examples of C2-C6alkenyl include ethenyl (-CH=CH2; C2alkenyl), allyl (CH2-CH=CH2; C3alkenyl), propenyl (-CH=CHCH3; C3alkenyl), isopropenyl (-C(CH3)=CH2; C3alkenyl); butenyl (-CH=CHCH2CH3; C4alkenyl), sec-butenyl (-C(CH3)=CHCH3; C4alkenyl), isobutenyl (-CH=C(CH3)2; C4alkyl), and 2-butenyl (-CH2CH=CHCH3; C4alkyl).

[0032] As used herein, the term "C2-C6alkynyl" refers to a straight or branched chain group having from 2 to 6 carbon atoms in the group, and wherein the group includes at least one carbon-carbon triple bond. Examples of alkynyl include ethynyl (-CºCH; C2alkynyl), propargyl (-CH2-CºCH; C3alkynyl), propynyl (-CºCCH3; C3alkynyl), butynyl (-CºCCH2CH3; C4alkynyl), and pentynyl (-CºCCH2CH2CH3; C5alkynyl).

[0033] The term "C3-C6cycloalkyl" refers to a ring-containing non-aromatic hydrocarbon group having from 3 to 6 carbon atoms. Examples of C3-C6cycloalkyl include cyclopropyl (C3), cyclobutyl (C4), cyclopentyl (C5), and cyclohexyl (C6).

[0034] The term "halo" or "halogen," by itself or as part of another group, refers to a fluorine, chlorine, bromine, or iodine atom.

[0035] As used herein, the term "C1-C6haloalkyl" refers to an alkyl group in which one or more hydrogen atoms have been replaced by one or more halogen atoms, which can be the same or different. In some embodiments, the alkyl group is substituted with at least one halogen. In other embodiments, the alkyl group is substituted with one, two, or three F and / or Cl. Examples of haloalkyl include fluoromethyl (CH2F), 1-fluoroethyl (CH(CH3)F), 2-fluoroethyl, difluoromethyl (CHF2), trifluoromethyl (CF3), pentafluoroethyl, 1,1-difluoroethyl (C(CH3)F2), 2,2-difluoroethyl (CH2CHF2), 2,2,2-trifluoroethyl (CH2CF3), 2-fluoroprop-2-yl (C(CH3)2F), 3,3,3-trifluoropropyl, 4,4,4-trifluorobutyl, trichloromethyl, and the like.

[0036] The term "C1-C6cyanoalkyl," by itself or in combination with another term, means an alkyl group as defined herein substituted with one or more CN. In some embodiments, the alkyl group is substituted with at least one CN. In other embodiments, the alkyl group is substituted with one, two, or three CN. Examples of cyanoalkyl include CH2CN, CH2CH2CN, CH(CN)CH3, CH2CH2CH2CN, C(CH3)2CN, CH2CH(CN)CH3, CH(CN)CH2CH3, and the like.

[0037] The term "C1-C6hydroxyalkyl," by itself or in combination with another term, means an alkyl group as defined herein in which one or more of the hydrogen atoms have been replaced with one or more hydroxyl groups (i.e., -OH). In some embodiments, the hydroxyalkyl group contains one OH. In other embodiments, the hydroxyalkyl group contains two OH. In further embodiments, the hydroxyalkyl group contains three OH. Examples of hydroxyalkyl include hydroxymethyl, hydroxyethyl (e.g., 1-hydroxyethyl, 2-hydroxyethyl), 1,2-dihydroxyethyl, hydroxypropyl (e.g., 2-hydroxypropyl, 3-hydroxypropyl), hydroxybutyl (e.g., 3-hydroxybutyl, 4-hydroxybutyl), 2-hydroxy-1-methylpropyl, 1,3-dihydroxypropan-2-yl, and the like.

[0038] The term "C1-C6aminoalkyl," by itself or in combination with another term, means an alkyl group as defined herein substituted with one or more NH2. In some embodiments, the alkyl group is substituted with at least one NH2. In other embodiments, the alkyl group is substituted with one, two, or three NH2. Examples of cyanoalkyl include CH2NH2, CH2CH2NH2, CH2CH2CH2NH2, and C(CH3)2NH2.

[0039] The term "6-12 membered aryl" means a monocyclic or bicyclic aromatic hydrocarbon ring structure having 6 or 12 carbon atoms in the ring. Examples of aryl include phenyl, indenyl, naphthyl, and 1,2,3,4-tetrahydronaphthyl.

[0040] The term "5-14 membered heteroaryl" refers to a monocyclic or bicyclic aromatic ring structure having 5 to 14 ring atoms, including carbon atoms and up to four heteroatoms each independently selected from nitrogen, oxygen, or sulfur. Examples of heteroaryl groups include thienyl, benzo[b]thienyl, furanyl, benzofuranyl, pyranyl, thiophenyl, isobenzofuranyl, benzoxazolyl, chromenyl, xanthenyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyridazinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, isoquinolyl, quinolyl, quinoxalyl, phthalazinyl, naphthyridinyl, cinnolinyl, triazolyl, tetrazolyl, thiadiazolyl, oxadiazolyl, quinazolinyl, pteridinyl, pyrimidinyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, furazanyl, pyrazolo[1,5-a]pyridinyl, pyrazolo[1,5-a]pyridinyl, benzisothiazolyl, imidazo[1,5-a]pyridinyl, pyrrolo[1,2]pyridazinyl, benzo[d]thiazolyl, benzo[d]imidazolyl, benzo[d]oxazolyl, benzisoxazolyl, isothiazolyl, and tetrahydropyrazolo[1,5-a]pyridinyl.

[0041] The term "5-14 membered heterocyclyl," by itself or as part of another substituent, refers to a non-aromatic, saturated or partially unsaturated (e.g., containing one or two double bonds) cyclic group containing one, two, or three rings having five to fourteen ring members, wherein at least one carbon atom in one of the rings is replaced with a heteroatom. Each heteroatom is independently selected from an oxygen, sulfur (including sulfoxide and sulfone), and / or a nitrogen atom, which can be oxidized or quaternized. Examples of heterocyclyl groups include azetidinyl, dioxanyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, pyrrolidinyl, dihydroindolyl, oxetanyl, tetrahydrofuranyl, tetrahydrothiophenyl, azepanyl, aziridinyl, dioxolanyl, imidazolidinyl, pyrazolidinyl, thianyl, dithianyl, thiomorpholinyl, oxazepanyl, oxiranyl, tetrahydropyranyl, and the like. In some embodiments, heterocyclyl includes azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, and 6-azaspiro[2.5]octyl.

[0042] As used herein, the term "salt" or "salt form" refers to an acid addition or base addition salt of a compound described. Acid addition salts can be formed with inorganic acids and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, and the like. It will be appreciated that the term "salt" or "salt form", refers to all stoichiometrically, including, for example, bis-salts of a compound described. Such bis-salts would include, for example, bis-acidic salts (such as bis-hydrochloride or bis-hydrobromide) as well as bis-basic salts (such as bis-sodium salt).

[0043] In some instances, compounds are repeated throughout the specification but can simply be given different reference numerals in order to facilitate reading. For example, compounds 10A and 10B have the same structure but different reference numerals due to the protocol in which they appear. To the extent there is any difference, the depicted structure takes precedence over the reference numeral.

[0044] In some embodiments, R in the method of making a compound of formula (A-1) is C1-C6alkyl or C1-C6haloalkyl. 1 In some embodiments, R in the method of making a compound of formula (A-1) is C1-C6alkyl or C1-C6haloalkyl. 1 In some embodiments, R in the method of making a compound of formula (A-1) is methyl.

[0045] In some embodiments, X in the method of making a compound of formula (A-1) is Cl.

[0046] In some embodiments, L in the method of making a compound of formula (A-1) is C1-C6alkylene. In some embodiments, L in the method of making a compound of formula (A-1) is -CH2-.

[0047] In some embodiments, Q in the method of making a compound of formula (A-1) is 6-12 membered aryl or 5-14 membered heteroaryl, each of which is optionally substituted with 1 to 3 groups selected from halo, C1-C6alkyl, C1-C6haloalkyl, C1-C6hydroxyalkyl, hydroxyl, C1-C6aminoalkyl, NH2, -NH(C1-C6alkyl), N(C1-C6alkyl)2, CN, and NO2. In some embodiments, Q in the method of making a compound of formula (A-1) is 5-7 membered heteroaryl substituted with -NH2.

[0048] In some embodiments, the compound of formula (A-1) in the disclosed method is:

[0049] or a salt thereof.

[0050] (9A)

[0051] In some embodiments, the compound of Formula (A-2) in the disclosed methods is:

[0052] or a salt thereof. (4)

[0054] In some embodiments, the compound of Formula (A-2) is a salt. In some embodiments, the compound of Formula (A-2) is a bis-salt. In other embodiments, the compound of Formula (A-2) is a bis-acid salt. In other embodiments, the compound of Formula (A-2) is a bis-HCl salt.

[0055] Similarly, in some embodiments, the compound of Formula (4) is a salt. In some embodiments, the compound of Formula (4) is a bis-salt. In other embodiments, the compound of Formula (4) is a bis-acid salt. In other embodiments, the compound of Formula (4) is a bis-HCl salt.

[0056] In some embodiments, the compound of Formula (A-3) in the disclosed methods is:

[0057] or a salt thereof.

[0058] (8A)

[0059] In some embodiments, the compound of formula (A-2) or salt thereof is reacted with a compound of formula (A-3) in the presence of an acid (e.g., hydrochloric acid, acetic acid, phosphoric acid, sulfuric acid, citric acid, formic acid, methanesulfonic acid, or p-toluenesulfonic acid) and one or more solvents (e.g., a polar protic solvent such as water, methanol, ethanol, n-propanol, isopropanol, or tert-butanol, or any combination thereof). In some embodiments, the compound of formula (A-2) or salt thereof is reacted with a compound of formula (A-3) in the presence of citric acid and one or more solvents (e.g., a polar protic solvent such as water, methanol, ethanol, n-propanol, isopropanol, or tert-butanol, or any combination thereof). In some embodiments, the one or more solvents is an alcohol. In some embodiments, the compound of formula (A-2) or salt thereof is reacted with a compound of formula (A-3) in the presence of an acid (e.g., hydrochloric acid, acetic acid, phosphoric acid, sulfuric acid, citric acid, formic acid, methanesulfonic acid, or p-toluenesulfonic acid) and ethanol. In some embodiments, the compound of formula (A-2) or salt thereof is reacted with a compound of formula (A-3) in the presence of an acid (e.g., hydrochloric acid, acetic acid, phosphoric acid, sulfuric acid, citric acid, formic acid, methanesulfonic acid, or p-toluenesulfonic acid) and ethanol and at least one other solvent. In some embodiments, the compound of formula (A-2) or salt thereof is reacted with a compound of formula (A-3) in the presence of citric acid and ethanol. In other embodiments, the compound of formula (A-2) or salt thereof is reacted with a compound of formula (A-3) in the presence of citric acid and a mixture of ethanol and isopropanol (IPA). In other embodiments, the compound of formula (A-2) or salt thereof is reacted with a compound of formula (A-3) in the presence of citric acid and n-propanol. In some embodiments, the compound of formula (4) or salt thereof is reacted with a compound of formula (8A) in the presence of an acid and one or more solvents. In some embodiments, the bis-acid salt of the compound of formula (4) is reacted with a compound of formula (8A) in the presence of an acid and one or more solvents. In some embodiments, the bis-acid salt of the compound of formula (4) is reacted with a compound of formula (8A) in the presence of citric acid or hydrochloric acid and one or more alcoholic solvents. In some embodiments, the bis-HCl salt of the compound of formula (4) is reacted with a compound of formula (8A) in the presence of citric acid or hydrochloric acid and one or more of ethanol, isopropanol, and n-propanol. In other embodiments, the bis-acid salt of the compound of formula (4) is reacted with a compound of formula (8A) in the presence of one or more solvents in the absence of additional acid.

[0060] In one embodiment, provided herein is a method of preparing a compound of formula (A-3):

[0061]

[0062] (A-3),

[0063] wherein R 1is hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6aminoalkyl, C1-C6cyanoalkyl, C2-C6alkenyl, C2-C6alkynyl, or C3-C6cycloalkyl, wherein the C3-C6cycloalkyl is optionally substituted with one to three groups selected from halo, hydroxyl, NH2, and CN; and

[0064] X is halo;

[0065] The method comprises:

[0066] hydrolyzing a compound of Formula (A-6):

[0067]

[0068] (A-6)

[0069] wherein R 2 is C1-C6alkyl, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6aminoalkyl, C1-C6cyanoalkyl, C2-C6alkenyl, C2-C6alkynyl, or C3-C6cycloalkyl, wherein the C3-C6cycloalkyl is optionally substituted with one to three groups selected from halo, hydroxyl, NH2, and CN, to form a compound of Formula (A-3). In one embodiment, R 2 is C1-C6alkyl.

[0070] In some embodiments, R 1 is C1-C6alkyl or C1-C6haloalkyl. In some embodiments, R 1 is methyl.

[0071] In some embodiments, X in the method of making a compound of Formula (A-3) is Cl or Br. In one embodiment, X in the method of making a compound of Formula (A-3) is Cl.

[0072] In some embodiments, R 2 is ethyl.

[0073] In some embodiments, the compound of Formula (A-3) in the disclosed method is:

[0074]

[0075] (8A).

[0076] In some embodiments, the compound of Formula (A-6) in the disclosed method is:

[0077]

[0078] (7A).

[0079] In some embodiments, the compound of Formula (A-4) in the disclosed methods is:

[0080]

[0081] (5A).

[0082] In some embodiments, the compound of Formula (A-6) is hydrolyzed in the presence of a base (e.g., lithium hydroxide (LiOH), calcium hydroxide (Ca(OH)2), potassium hydroxide (KOH), sodium hydroxide (NaOH), or barium hydroxide (Ba(OH)2)) and one or more solvents (e.g., a mixture of water and at least one polar aprotic solvent). In some embodiments, the compound of Formula (A-6) is hydrolyzed in the presence of a base (e.g., lithium hydroxide (LiOH), calcium hydroxide (Ca(OH)2), potassium hydroxide (KOH), sodium hydroxide (NaOH), or barium hydroxide (Ba(OH)2)), water, and at least one polar aprotic solvent. In some embodiments, the compound of Formula (A-6) is hydrolyzed in the presence of a base (e.g., lithium hydroxide (LiOH), calcium hydroxide (Ca(OH)2), potassium hydroxide (KOH), sodium hydroxide (NaOH), or barium hydroxide (Ba(OH)2)), water, and at least one polar aprotic solvent selected from dimethylacetamide (DMAC), 2-methyltetrahydrofuran (2-Me THF), tetrahydrofuran (THF), acetonitrile (ACN), or dimethylsulfoxide (DMSO), or any combination thereof. In some embodiments, the compound of Formula (A-6) is hydrolyzed in the presence of sodium hydroxide (NaOH), water, and at least one polar aprotic solvent. In some embodiments, the compound of Formula (A-6) is hydrolyzed in the presence of sodium hydroxide (NaOH), water, and at least one polar aprotic solvent selected from dimethylacetamide (DMAC), 2-methyltetrahydrofuran (2-Me THF), tetrahydrofuran (THF), acetonitrile (ACN), or dimethylsulfoxide (DMSO), or any combination thereof. In some embodiments, the compound of Formula (A-6) is hydrolyzed in the presence of sodium hydroxide (NaOH), water, and tetrahydrofuran.

[0083] In some embodiments, the compound of Formula (A-6) is prepared by hydrolyzing a compound of Formula (A-5):

[0084]

[0085] (A-5),

[0086] with phosphorus oxychloride (POCI3), dimethylformamide (DMF), and one or more additional solvents (e.g., non-polar solvents such as dioxane, dichloroethane, benzene, toluene, chlorobenzene, dichlorobenzene, or xylene) to form a compound of Formula (A-6). In some embodiments, the compound of Formula (A-6) is prepared by reacting a compound of Formula (A-5) with phosphorus oxychloride (POCI3), dimethylformamide (DMF) in chlorobenzene.

[0087] In some embodiments, the compound of Formula (A-5) in the disclosed methods is:

[0088]

[0089] (6A).

[0090] In some embodiments, the compound of Formula (A-5) is prepared by reacting a compound of Formula (A-4):

[0091]

[0092] (A-4),

[0093] with a halogenating agent (e.g., elemental halogen, carbon tetrachloride, hexachloroethane, carbon tetrabromide, n-bromosuccinimide (NBS), or n-chlorosuccinimide (NCS)), a base (e.g., lithium diisopropylamide (LDA), sodium bis(trimethylsilyl)amide (NaHMDS), lithium bis(trimethylsilyl)amide (LiHMDS), potassium bis(trimethylsilyl)amide (KHMDS), and sodium hydride (NaH)), and at least one solvent (e.g., a polar aprotic solvent such as dimethylacetamide (DMCA), 2-methyltetrahydrofuran (2-Me THF), tetrahydrofuran (THF), acetone, dimethylformamide (DMF), acetonitrile (ACN), and dimethyl sulfoxide (DMSO), or any combination thereof) to form a compound of Formula (A-5).

[0094] In some embodiments, the compound of Formula (A-5) is prepared by reacting a compound of Formula (A-4) with hexachloroethane, a base (e.g., lithium diisopropylamide (LDA), sodium bis(trimethylsilyl)amide (NaHMDS), lithium bis(trimethylsilyl)amide (LiHMDS), potassium bis(trimethylsilyl)amide (KHMDS), and sodium hydride (NaH)), and at least one solvent (e.g., a polar aprotic solvent) to form the compound of Formula (A-5). In some embodiments, the compound of Formula (A-5) is prepared by reacting a compound of Formula (A-4) with hexachloroethane, lithium bis(trimethylsilyl)amide (LiHMDS), and at least one solvent (e.g., a polar aprotic solvent) to form the compound of Formula (A-5). In some embodiments, the compound of Formula (A-5) is prepared by reacting a compound of Formula (A-4) with hexachloroethane, lithium bis(trimethylsilyl)amide (LiHMDS), and at least one solvent (e.g., a polar aprotic solvent) to form the compound of Formula (A-5). In some embodiments, the compound of Formula (A-5) is prepared by reacting a compound of Formula (A-4) with hexachloroethane, lithium bis(trimethylsilyl)amide (LiHMDS) in THF to form the compound of Formula (A-5).

[0095] In some embodiments, the compound of Formula (A-4) in the disclosed methods is:

[0096]

[0097] (5A).

[0098] In one embodiment, provided herein is a method of preparing a compound of Formula (B-1):

[0099]

[0100] (B-1),

[0101] wherein

[0102] R 1 is hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6aminoalkyl, C1-C6cyanoalkyl C2-C6alkenyl, C2-C6alkynyl, or C3-C6cycloalkyl, wherein the C3-C6cycloalkyl is optionally substituted with 1 to 3 groups selected from halo, hydroxyl, NH2, and CN;

[0103] R 4 is C1-C6alkyl, C1-C6haloalkyl, 6-12 membered aryl, or 5-14 membered heteroaryl, wherein the 6-12 membered aryl or the 5-14 membered heteroaryl is optionally substituted with 1 to 3 groups selected from halo or C1-C6alkyl;

[0104] L is a bond or C1-C6alkylene; and

[0105] Q is C3-C6cycloalkyl, 5-14 membered heterocyclyl, 6-12 membered aryl, or 5-14 membered heteroaryl, each of which is optionally substituted with 1 to 3 groups selected from halo, C1-C6alkyl, C1-C6haloalkyl, C1-C6hydroxyalkyl, hydroxyl, C1-C6aminoalkyl, -NH2, -NH(C1-C6alkyl), -N(C1-C6alkyl)2, CN, and NO2;

[0106] The method comprises:

[0107] reacting a compound of Formula (A-2):

[0108]

[0109] (A-2) or a salt thereof,

[0110] with a compound of Formula (B-5):

[0111]

[0112] (B-5),

[0113] to form a compound of Formula (B-1).

[0114] In some embodiments, the compound of Formula (A-2) or a salt thereof is reacted with the compound of Formula (B-5) in the presence of an acid (e.g., hydrochloric acid, acetic acid, trifluoroacetic acid, phosphoric acid, sulfuric acid, citric acid, formic acid) and at least one solvent (e.g., a polar protic solvent such as methanol, ethanol, n-propanol, isopropanol, sec-butanol, or tert-butanol or any combination thereof). In some embodiments, the compound of Formula (A-2) or a salt thereof is reacted with the compound of Formula (B-5) in the presence of an acid (e.g., hydrochloric acid, trifluoroacetic acid, acetic acid, phosphoric acid, sulfuric acid, citric acid, formic acid, or carbonic acid) in ethanol or n-propanol or any combination thereof. In some embodiments, the compound of Formula (A-2) is a bis-salt. In some embodiments, the compound of Formula (A-2) is a bis-acid salt. In some embodiments, the compound of Formula (A-2) is a bis-HCl salt.

[0115] In some embodiments, the compound of Formula (B-5) in the disclosed methods is prepared by hydrolyzing a compound of Formula (B-4):

[0116]

[0117] (B-4),

[0118] wherein R 2C1-C6alkyl, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6aminoalkyl, C1-C6cyanoalkyl, C2- C6alkenyl, C2-C6alkynyl, or C3-C6cycloalkyl, wherein the C3-C6cycloalkyl is optionally substituted with 1 to 3 groups selected from halo, hydroxy, NH2, and CN. In another embodiment, R 2 C1-C6alkyl.

[0119] In some embodiments, the compound of formula (B-4) is hydrolyzed in the presence of a base (e.g., lithium hydroxide (LiOH), calcium hydroxide (Ca(OH)2), potassium hydroxide (KOH), sodium hydroxide (NaOH), or barium hydroxide (Ba(OH)2)) and one or more solvents (e.g., a mixture of water and at least one polar aprotic solvent). In some embodiments, the compound of formula (B-4) is hydrolyzed in the presence of a base (e.g., lithium hydroxide (LiOH), calcium hydroxide (Ca(OH)2), potassium hydroxide (KOH), sodium hydroxide (NaOH), or barium hydroxide (Ba(OH)2)), water, and at least one polar aprotic solvent. In some embodiments, the compound of formula (B-4) is hydrolyzed in the presence of a base (e.g., lithium hydroxide (LiOH), calcium hydroxide (Ca(OH)2), potassium hydroxide (KOH), sodium hydroxide (NaOH), or barium hydroxide (Ba(OH)2)), water, and at least one polar aprotic solvent selected from dimethylacetamide (DMAC), 2-methyltetrahydrofuran (2-Me THF), tetrahydrofuran (THF), dimethylformamide (DMF), acetonitrile (ACN), or dimethylsulfoxide (DMSO), or any combination thereof. In some embodiments, the compound of formula (B-4) is hydrolyzed in the presence of sodium hydroxide (NaOH), water, and at least one polar aprotic solvent. In some embodiments, the compound of formula (B-4) is hydrolyzed in the presence of sodium hydroxide (NaOH), water, and at least one polar aprotic solvent selected from dimethylacetamide (DMAC), 2-methyltetrahydrofuran (2-Me THF), tetrahydrofuran (THF), dimethylformamide (DMF), acetonitrile (ACN), or dimethylsulfoxide (DMSO), or any combination thereof. In some embodiments, the compound of formula (B-4) is hydrolyzed in the presence of sodium hydroxide (NaOH), water, and tetrahydrofuran.

[0120] In some embodiments, the compound of formula (B-4) is prepared by hydrolyzing a compound of formula (B-3):

[0121]

[0122] (B-3),

[0123] Compounds of formula (B-4) are prepared by reacting an oxidizing agent (e.g., hydrogen peroxide and sodium tungstate (Na2WO4), 39% peracetic acid, trichloroisocyanuric acid (TCCA) and (sodium hypochlorite) NaOCl, potassium persulfate (oxone), hydrogen peroxide and SeO2, or hydrogen peroxide and MoCl2O2) in at least one solvent (e.g., a nonpolar solvent, such as dichloromethane, toluene, chlorobenzene, dichlorobenzene, or xylene, or any combination thereof) to form a compound of formula (B-4). In some embodiments, compounds of formula (B-4) are prepared by reacting a compound of formula (B-3) with hydrogen peroxide and sodium tungstate (Na2WO4) in chlorobenzene to form a compound of formula (B-4). In some embodiments, the compound of formula (B-4) is prepared by reacting the compound of formula (B-3) with an oxidant selected from 39% peracetic acid, trichloroisocyanuric acid (TCCA) and (sodium hypochlorite) NaOCl, potassium persulfate, hydrogen peroxide and SeO2 or hydrogen peroxide and MoCl2O2 in at least one solvent (e.g., a polar solvent, such as ethanol, tetrahydrofuran (THF), water, acetic acid, ethyl acetate or any mixture thereof) to form the compound of formula (B-4).

[0124] In some embodiments, the method of forming the (B-4) compound further includes at least one phase transfer catalyst, such as methyltrioctyl ammonium hydrogen sulfate or tetrabutyl ammonium hydrogen sulfate. In some embodiments, the phase transfer catalyst is methyltrioctyl ammonium hydrogen sulfate.

[0125] In some embodiments, the method of forming the (B-4) compound further includes at least one acid, such as phenylphosphonic acid and acetic acid. In some embodiments, the acid is phenylphosphonic acid.

[0126] In some embodiments, the compound of formula (B-3) is produced by making the compound of formula (B-2):

[0127]

[0128] (B-2),

[0129] The compound is prepared by reacting it with phosphorus oxychloride (POCl3), dimethylformamide (DMF), and one or more solvents (such as nonpolar solvents) (e.g., toluene, chlorobenzene, dichlorobenzene, or xylene, or any combination thereof) to form a compound of formula (B-3). In some embodiments, the compound of formula (B-3) is prepared by reacting the compound of formula (B-2) with phosphorus oxychloride (POCl3) and dimethylformamide (DMF) in chlorobenzene.

[0130] In some embodiments, R in the method for preparing compounds of formula (B-1), formula (B-5), formula (B-4), formula (B-3) and / or formula (B-2) 1is C1-C6alkyl or C1-C6haloalkyl. In some embodiments, R 1 is methyl.

[0131] In some embodiments, R 2 is ethyl.

[0132] In some embodiments, L in the method of making a compound of Formula (B-1) is C1-C6alkylene. In some embodiments, L in the method of making a compound of Formula (B-1) is -CH2-.

[0133] In some embodiments, Q in the method of making a compound of Formula (B-1) is 6-12 membered aryl or 5-14 membered heteroaryl, each of which is optionally substituted with 1 to 3 groups selected from halo, C1-C6alkyl, C1-C6haloalkyl, C1-C6hydroxyalkyl, hydroxyl, C1-C6aminoalkyl, -NH2, -NH(C1-C6alkyl), N(C1-C6alkyl)2, -CN, or -NO2. In some embodiments, Q in the method of making a compound of Formula (B-1) is 5-7 membered heteroaryl substituted with -NH2.

[0134] In some embodiments, R 4 is C1-C6alkyl, C1-C6haloalkyl, 6-12 membered aryl, or 5-14 membered heteroaryl, wherein the 6-12 membered aryl or the 5-14 membered heteroaryl is optionally substituted with 1 to 3 groups selected from halo and C1-C6alkyl. In some embodiments, R 4 is 6-12 membered aryl substituted with C1-C6alkyl.

[0135] In some embodiments, the compound of Formula (B-1) in the disclosed methods is:

[0136]

[0137] (9B).

[0138] In some embodiments, the compound of Formula (B-2) in the disclosed methods is:

[0139]

[0140] (5B).

[0141] In some embodiments, the compound of Formula (B-3) in the disclosed methods is:

[0142]

[0143] (6B).

[0144] In some embodiments, the compound of Formula (B-4) in the disclosed methods is:

[0145]

[0146] (7B).

[0147] In some embodiments, the compound of Formula (B-5) in the disclosed methods is:

[0148]

[0149] (8B).

[0150] In some embodiments, the compound of Formula (A-2) in the disclosed methods is:

[0151] or a salt thereof

[0152] (4).

[0153] In some embodiments, compound 4 is a bis-salt. In some embodiments, compound 4 is a bis-acid salt. In some embodiments, compound 4 is a bis-HCl salt.

[0154] In one embodiment, a method (C) of preparing a compound of Formula (C-1)

[0155]

[0156] (C-1),

[0157] or a salt or hydrate thereof, is provided, wherein

[0158] R 1 is hydrogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6aminoalkyl, C1-C6cyanoalkyl, C2-C6alkenyl, C2-C6alkynyl, or C3-C6cycloalkyl, wherein the C3-C6cycloalkyl is optionally substituted with one to three groups selected from halo, hydroxyl, NH2, and CN;

[0159] L is a bond or C1-C6alkylene; and

[0160] Q is C3-C6cycloalkyl, 5-14 membered heterocyclyl, 6-12 membered aryl, or 5-14 membered heteroaryl, each of which is optionally substituted with 1 to 3 groups selected from halo, C1-C6alkyl, C1-C6haloalkyl, C1-C6hydroxyalkyl, hydroxyl, C1-C6aminoalkyl, -NH2, -NH(C1-C6alkyl), -N(C1-C6alkyl)2, CN, and NO2;

[0161] The method comprises the steps of:

[0162] a) deprotecting the compound of formula (C-1):

[0163]

[0164] (C-1) to form the compound of formula (C-2):

[0165] wherein

[0166] P1is a protecting group;

[0167] R 3 is C1-C6alkyl;

[0168] with a compound of formula (C-3):

[0169]

[0170] (C-3) to form the compound of formula (C-4):

[0171] wherein

[0172] X is halo or S(O)2R 4 ;

[0173] R 4 is C1-C6alkyl, C1-C6haloalkyl, 6-12 membered aryl, or 5-14 membered heteroaryl, wherein the 6-12 membered aryl or the 5-14 membered heteroaryl is optionally substituted with 1 to 3 groups selected from halo or C1-C6alkyl;

[0174] to form the compound of formula (C-4):

[0175]

[0176] (C-4); and

[0177] b) deprotecting the compound to form the compound of formula (C-1).

[0178] In some embodiments, L in the method of making the compound of formula (C-1) is C1-C6alkylene. In some embodiments, L in the method of making the compound of formula (C-1) is -CH2-.

[0179] In some embodiments, the method of preparing a compound of formula (C-1) is:

[0180] In some embodiments, the compound of formula (C-1) prepared according to the above method is:

[0181]

[0182] (13).

[0183] In some embodiments, the compound of formula (C-1) can be isolated in free base form or salt form. In some embodiments, the compound of formula (C-1) can be isolated in free base or salt form of the compound:

[0184]

[0185] (13).

[0186] In some embodiments, the method of preparing a compound of formula (C-1) is:

[0187] In some embodiments, the method of preparing a compound of formula (C-1) is: 4 and R 4 is a 6-12 membered aryl, wherein the 6-12 membered aryl is optionally substituted with 1 to 3 groups selected from halo and C1-C6 alkyl. In some embodiments, the method of preparing a compound of formula (C-1) is: 4 and R 4 is 4-methylphenyl. In some embodiments, the compound of formula (C-3) is a compound of formula (B-1). In other embodiments, the compound of formula (C-3) is a compound of formula (B-1), wherein the compound of formula (B-1) is prepared by the disclosed method.

[0188] In some embodiments, the compound of formula (C-3) in the disclosed method is:

[0189]

[0190] (9A).

[0191] In some embodiments, the compound of formula (C-3) in the disclosed method is:

[0192]

[0193] (9B).

[0194] In some embodiments, P1 in the method for preparing the (C-1) compound is tert-butoxycarbonyl (BOC), fluorenylmethoxycarbonyl (FMOC), acetate, benzyl, triphenylmethyl, tetrahydropyranyl ether (THP), neopentanoyl, or toluenesulfonate. In some embodiments, P1 is tetrahydropyranyl ether (THP).

[0195] In some embodiments, R in the method for preparing (C-1) compounds 3 It is tert-butyl.

[0196] In some embodiments, the compound of formula (C-2) is:

[0197]

[0198] (12).

[0199] In some embodiments, the compound of formula (C-4) is:

[0200]

[0201] (10B).

[0202] In some embodiments, the compound of Formula (C-2) is reacted with the compound of Formula (C-3) in the presence of a base (e.g., lithium bis(trimethylsilyl)amide (LiHMDS), lithium tert-butoxide (LiOtBu), sodium tert-butoxide (NaOtBu), potassium tert-butoxide (KOtBu), or sodium hydride (NaH)) and one or more solvents (e.g., a polar aprotic solvent such as tetrahydrofuran (THF), 2-methyltetrahydrofuran (2-MeTHF), acetone, dimethylformamide (DMF), dimethylacetamide (DMA), acetonitrile (ACN), or dimethylsulfoxide (DMSO), or any combination thereof). In some embodiments, the compound of Formula (C-2) is reacted with the compound of Formula (C-3) in the presence of lithium bis(trimethylsilyl)amide (LiHMDS) and one or more solvents (e.g., a polar aprotic solvent such as tetrahydrofuran (THF), 2-methyltetrahydrofuran (2-MeTHF), acetone, dimethylformamide (DMF), dimethylacetamide (DMA), acetonitrile (ACN), or dimethylsulfoxide (DMSO), or any combination thereof). In some embodiments, the compound of Formula (C-2) is reacted with the compound of Formula (C-3) in the presence of lithium tert-butoxide (LiOtBu) and one or more solvents (e.g., a polar aprotic solvent such as tetrahydrofuran (THF), 2-methyltetrahydrofuran (2-MeTHF), acetone, dimethylformamide (DMF), dimethylacetamide (DMA), acetonitrile (ACN), or dimethylsulfoxide (DMSO), or any combination thereof). In some embodiments, the compound of Formula (C-2) is reacted with the compound of Formula (C-3) in the presence of lithium bis(trimethylsilyl)amide (LiHMDS) and tetrahydrofuran (THF). In some embodiments, the compound of Formula (C-2) is reacted with the compound of Formula (C-3) in the presence of lithium tert-butoxide (LiOtBu) and 2-methyltetrahydrofuran (2-MeTHF) and dimethylacetamide (DMA), acetonitrile (ACN).

[0203] In some embodiments, the compound of Formula (C-4) is deprotected in the presence of an acid (e.g., hydrochloric acid, acetic acid, phosphoric acid, sulfuric acid, citric acid, trifluoroacetic acid, formic acid) and one or more solvents (e.g., a polar protic solvent such as water, methanol, ethanol, n-propanol, isopropanol, or tert-butanol, or any combination thereof) to provide a compound of Formula (C-l). In some embodiments, the compound of Formula (C-4) is deprotected in the presence of hydrochloric acid and one or more solvents (e.g., a polar protic solvent such as water, methanol, ethanol, n-propanol, isopropanol, or tert-butanol, or any combination thereof). In some embodiments, the compound of Formula (C-4) is deprotected in the presence of an acid (e.g., hydrochloric acid, acetic acid, phosphoric acid, sulfuric acid, citric acid, formic acid) and n-propanol. In some embodiments, the compound of Formula (C-4) is deprotected in the presence of hydrochloric acid and n-propanol.

[0204] In some embodiments, the compound of Formula (C-l) prepared according to the methods disclosed above is:

[0205] (13)

[0207] or a salt, tautomer, or hydrate thereof.

[0208] In one embodiment, provided herein is a compound having the structure:

[0209]

[0210] (10B)

[0211] or a salt, tautomer, or hydrate thereof.

[0212] In one embodiment, provided herein is a compound having the structure:

[0213]

[0214] (8A)

[0215] or a salt, tautomer, or hydrate thereof.

[0216] In one embodiment, provided herein is a compound having the structure:

[0217]

[0218] (9A)

[0219] or a salt, tautomer, or hydrate thereof.

[0220] In one embodiment, provided herein is a compound having the structure:

[0221]

[0222] (9B)

[0223] or a salt, tautomer, or hydrate thereof.

[0224] In one embodiment, provided herein is a compound having the following structure:

[0225]

[0226] (6B)

[0227] or a salt, tautomer, or hydrate thereof.

[0228] In one embodiment, provided herein is a compound having the following structure:

[0229]

[0230] (7B)

[0231] or a salt, tautomer, or hydrate thereof.

[0232] In one embodiment, provided herein is a compound having the following structure:

[0233]

[0234] (8B)

[0235] or a salt, tautomer, or hydrate thereof.

[0236] In one embodiment, provided herein is a bis-acid compound having the following structure:

[0237]

[0238] or a tautomer or hydrate thereof.

[0239] Examples

[0240] While several embodiments have been described, the scope of the disclosure is not limited by the specific embodiments described herein. Rather, the scope of the disclosure is defined by the appended claims, and equivalents thereto, as well as any additional claims added after the filing date of this disclosure. The content of all references (including literature references, issued patents, published patent applications, and co-pending patent applications) cited throughout this application is expressly incorporated herein by reference. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0241] Typical abbreviations used are summarized below.

[0242]

[0243]

[0244]

[0245] Route A:

[0246]

[0247] Scheme 1

[0248] Synthesis of tert-butyl (E)-2-((6-((tert-butoxycarbonyl)amino)pyridin-2- yl)methylene)hydrazine-1-carboxylate (2):

[0249]

[0250] To a room temperature mixture of (1) (1 wt) and MeOH (2 vol) was added AcOH (0.05 vol, 0.2 eq). The mixture was warmed to 50 °C then a solution of Boc-hydrazine (0.61 wt) in methanol (1 vol) was added over 2.5 h. The mixture was stirred at 50 °C until the reaction was complete. The mixture was cooled to 20 °C and water (1.5 vol) was charged over 1 h, stirred for 4 h and filtered. The wet cake was washed with 2:1 MeOH:water (1.5 vol) and dried under vacuum at 40 °C for 20 h to recover (2) as an off-white solid.

[0251] Synthesis of tert-butyl 2-((6-((tert-butoxycarbonyl)amino)pyridin-2- yl)methyl)hydrazine-1-carboxylate (3):

[0252]

[0253] A mixture of (2) (1 wt) in MeOH (8 vol) was evacuated and flushed with nitrogen three times at 25 °C. 10% Pd / C (0.04 wt) was charged and the reactor was evacuated and flushed with hydrogen three times. The mixture was heated to 50 °C and the pressure was adjusted to 15-20 psi and stirred for 6-12 h. Once complete, the reaction was cooled to 20 °C and evacuated and flushed with nitrogen three times. The mixture was rapidly filtered through celite (0.6 wt) and the solids were rinsed with MeOH (1 wt). The filtrate was added to the reactor at 25 °C. Water (7.5 vol) was added dropwise over 2 h to induce precipitation. The suspension was stirred for 3 h, filtered, washed with 4:3 MeOH:water (1 vol) and dried under vacuum at 50 °C for 24 h to give (3) as an off-white solid.

[0254] Synthesis of 6-(hydrazinylmethyl)pyridin-2-amine dihydrochloride (4):

[0255]

[0256] To a reactor containing HC1 (6 N, 5 vol) was added (3) (1.0 wt) portion-wise while maintaining the internal temperature between 10-20 °C. The mixture was heated to 50 °C for 2 h (or until complete), cooled to 20 °C, and treated with EtOH (10 vol) over 2 h to induce precipitation and held for an additional 2 h. The suspension was filtered, washed with EtOH (2 vol), and dried under vacuum at 50 °C for 24 h to give (4) as a white solid.

[0257] Synthesis of 4-methyl-4H-pyrrolo[2,3-d]thiazole-5-carboxylic acid ethyl ester (5A):

[0258]

[0259]

[0260] Scheme 6

[0261] Commercially available 2,4-thiazolidinedione was aromatized and brominated by treatment with phosphorous pentoxide and TBAB in a manner similar to that reported by Sampson (see Grubb, M.; Schmidt, M. J.; Seed, A. J.; Sampson, P. Synthesis 2012, 44, 1026-1029) to yield 2,4-dibromothiazole. Formylation at the 5 position to generate compound (6-1) was achieved by deprotonation with tetramethylpiperidin lithium (TMP) and introduction of the formyl group from ethyl formate. The formyl group was reacted with readily available N-benzoyl-glycine (hippuric acid) and acetic anhydride in an Erlenmeyer Plochl type reaction and esterified upon treatment with ethanol (see, Zhao, H. Koenig S. G. Dankwardt, J. W. Singh, S. P. OPRD 18(1): 198-204, 2013). Removal of the bromine at the 2-thiazole position was achieved by treatment with zinc metal to provide (6-2). Cyclization of (6-2) was carried out under copper catalyzed amidation and removal of the benzoyl group by treatment with ethylenediamine. The final stage of N-methylation was carried out in acetone by treatment with dimethyl sulfate to give compound (5A).

[0262] Synthesis of 2-chloro-4-methyl-4H-pyrrolo[2,3-d]thiazole-5-carboxylic acid ethyl ester (6A):

[0263]

[0264] To a reactor containing (5A) (1 wt) and THF (10 vol) at 10 °C was added hexachloroethane (1.35 wt). The mixture was cooled to -40 °C and LiHMDS (1.0 M in THF, 4.5 wt) was added dropwise over 1 h and further stirred for 2 h while maintaining the temperature between -30 °C and -50 °C. Once complete, AcOH (0.66 wt) was added at -40 °C over 30 min then the mixture was warmed to 25 °C over 1 h. Water (5 vol) was added at 25 °C over 2 h. The layers were separated. The organic layer was washed with 5% Na2SO4 (5 wt), concentrated to 2.5 vol under vacuum and DCM (6.65 wt) was added. After stirring for 20 min, the mixture was allowed to settle and the layers were separated. The organic layer was concentrated to 2.5 vol under vacuum at 30 °C. Acetone (3.85 wt) was added and the mixture was concentrated to 2.5 vol again (2x). Acetone (2.5 vol) was added and the mixture was transferred dropwise into 5 °C water (5.0 vol) over 2 h and the suspension was stirred at 5 °C for up to 10 h, filtered and washed with a 5 °C 4:3 water:acetone (4.5 vol) mixture. The wet cake was dried under vacuum at 25 °C for 24 h to give (6A) as a yellow solid.

[0265] Synthesis of 2-chloro-6-formyl-4-methyl-4H-pyrrolo[2,3-d]thiazole-5-carboxylic acid ethyl ester (7A):

[0266]

[0267] To a reactor containing (6A) (1 wt) and 1,2-dichlorobenzene (2.6 wt) at 25 °C was added POCl3(2.5 wt). The mixture was heated to 100 °C, then DMF (0.90 wt) was added over 12 h, then the mixture was stirred at 100 °C for 6 h or until complete. Once complete, the reaction was cooled to 50 °C and DMF (5.25 wt) was added over 1 h. The mixture was added to a cooled (10-20 °C) solution of EtOH (9.5 wt) and water (6.0 wt) and the temperature was kept below 30 °C during the addition. The resulting suspension was stirred between 15-30 °C for 20 h and filtered. The wet cake was washed with 1:1 v:v EtOH:H2O (2.0 vol). The wet cake was slurried with 1:1 EtOH:H2O (5 vol) at 50 °C for 2 h, cooled to 20 °C, filtered, rinsed with 4:3 H2O:EtOH (3.8 vol), and dried under vacuum at 50 °C for 24 h to give (7A) as a light brown solid.

[0268] Synthesis of 2-chloro-6-formyl-4-methyl-4H-pyrrolo[2,3-d]thiazole-5-carboxylic acid (8A):

[0269]

[0270] To a reactor containing (7A) (1 wt), THF (5 vol), and water (5 vol) at 15 °C was added NaOH (5% w / w, 5.0 vol, 1.5 eq) dropwise over 30 min while maintaining the temperature below 30 °C. After the addition, the mixture was stirred at 20-30 °C for 4 h or until complete. Once complete, HCl (1 N) was added at 15 °C until pH 1-2 and this induced precipitation. The suspension was filtered, rinsed with water (8 vol), and the wet cake was dried under vacuum at 55 °C for 24 h to give (8A) as a brown solid (see, Figure 2 ).

[0271] Synthesis of 6-((6-aminopyridin-2-yl)methyl)-2-chloro-4-methyl-4,6-dihydro-5H- thiazolo[5',4':4,5]pyrrolo[2,3-d]pyridazin-5-one (9A):

[0272]

[0273] To a reactor containing EtOH (20 vol) at 20 °C was added sodium dihydrogen citrate (1.75 wt) and anhydrous citric acid (0.39 wt) and the mixture was stirred for 30 min. Compound (8A) (1.0 wt) was added and the mixture was warmed to 75 °C for 10 h (or until completion by IPC) then cooled to 10 °C and stirred for 10 h. The suspension was filtered and the wet cake was washed with cold EtOH (2.0 vol).[Note: if needed, the following sequence can be repeated to improve purity. The wet cake was transferred to a reactor containing water (25 vol), mixed at 20 °C and treated dropwise with ammonium hydroxide (25 wt%) until the solution pH reached 8-9. The basified mixture was stirred at 20 °C for up to 8 h, filtered and the wet cake was washed with water (4 vol)]. Once the desired purity was obtained, the wet cake was dried under vacuum at 50-60 °C for 24 h to give (9A) as a light brown solid.

[0274]

[0275] Scheme 2

[0276] Synthesis of 2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-3-yl)acetic acid (11):

[0277]

[0278] A reactor containing (10) (1.0 wt), TsOH-H20 (0.12 wt), and 2-MeTHF (8.0 vol) was warmed to 60 °C at 25 °C. DHP (1.2 wt) was added dropwise at 60 °C, and the mixture was stirred at 60 °C for 4 h. After complete conversion, the mixture was cooled to 25 °C, MTBE (3.75 wt) was added, and the reaction was cooled to 10 °C. NaOH (1 N, 8.0 wt) was added dropwise at 10 °C, then the mixture was warmed to 20 °C. The layers were separated. To the aqueous layer was added DCM (5.0 vol). The biphasic mixture was cooled to 10 °C and treated with HC1 (2 N) over 1 h until the pH was 3-4. After mixing at 20 °C for 1 h, the two layers were allowed to separate, and the organic layer was retained. The aqueous layer was re-extracted twice with DCM (5.0 vol) as above. The combined organic layers were washed with water (3.0 vol), concentrated to 2.5 vol under vacuum in the reactor, EtOAc (10 vol) was added, and the mixture was concentrated to 2.5 vol. More EtOAc (5.0 vol) was added, and the mixture was concentrated to 2.5 vol again. The concentrate was warmed to 55 °C to yield a solution, cooled to 35 °C, and seeded (0.010 wt). The mixture was stirred at 35 °C for 3 h, cooled to 0 °C over 8 h, and held at that temperature for 8 h. The suspension was filtered, washed with cold EtOAc (0.5 vol), and dried under vacuum at 35 °C for 25 h to give (11) as an off-white solid.

[0279] Synthesis of tert-butyl 2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-3-yl)acetate (12):

[0280]

[0281] To a reactor containing (11) (1.0 wt) and t-BuOH (2.5 vol) at 25 °C was added DMAP (0.03 wt) and THF (2.5 vol). After 30 min, the mixture was warmed to 50 °C, then a solution of Boc20 (1.15 wt) in THF (2.0 vol) was added dropwise over at least 5 h, then further stirred for up to 2 h. Upon completion, the reaction was cooled to 25 °C, and both heptane (5.0 vol) and water (5.0 vol) were added. The mixture was stirred at 25 °C for 2 h, then the layers were separated. The organic layer was concentrated under vacuum to 2 vol. Heptane (10 vol) was added and the mixture was concentrated to 2 vol again. More heptane (10 vol) was added, and the diluted mixture was recycled through a charcoal filter for 24 h. The filtrate was concentrated to 2 vol and 2-MeTHF (6.3 vol) was added; this cycle was repeated at least two more times. This solution of (12) was used in the next step without further treatment.

[0282] 2-(6-((6-Aminopyridin-2-yl)methyl)-4-methyl-5-oxo-5,6-dihydro-4H-thiazolo[5',4':4,5]pyrrolo[2,3- d]pyridazin-2-yl)-2-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)acetic acid tert-butyl ester (10A) was synthesized as follows:

[0283]

[0284] To a reactor containing (9A) (1.0 wt, 1.0 eq) and THF (13 vol) was added (12) (1.1 eq) and more THF (2.8 vol) in 2-MeTHF. The mixture was cooled to -25 °C and LiHMDS (1.0 M in THF, 7.0 wt, 2.5 eq) was added dropwise over at least 1 h. The mixture was stirred at -25 °C for 3 h, warmed to -10 °C, and stirred at -10 °C for 3 h. Once complete, 2-MeTHF (2.0 vol) containing AcOH (0.98 wt) was added dropwise at -10 °C until pH was 7-8. The mixture was stirred at -10 °C for 2 h, warmed to 10 °C over 1 h, added water (8 vol), and further warmed to 20 °C for 1 h. The reaction was filtered through Celite (1.2 wt) and the Celite bed was washed with THF (2.0 vol). The biphasic mixture was separated and the organic layer was washed with 10% Na2SO4 (5 vol) and then concentrated in vacuo to 4 vol. Toluene (5.4 vol) was added, the mixture was concentrated to 4 vol, and the sequence was repeated at least once (KF < 0.5%). Toluene (7.0 vol) was added and the mixture was warmed to 60 °C to dissolve all solids. The reaction was cooled to 43 °C, seed crystals (0.05 wt) were added, the light suspension was stirred at 43 °C for 8 h, and then heptane (10 vol) was added dropwise over 8 h. The suspension was cooled to 33 °C for 24 h and then filtered. The wet cake was washed with heptane (2.0 vol) and then dried under vacuum at 40 °C for 24 h to give (10A) as a brown solid (see, Figure 1 ).

[0285] 2-((1H-Pyrazol-3-yl)methyl)-6-((6-aminopyridin-2-yl)methyl)-4-methyl-4,6-dihydro-5H- thiazolo[5',4':4,5]pyrrolo[2,3-d]pyridazin-5-one hydrate (14) synthesis:

[0286]

[0287] Concentrated HCl (3.0 vol, 20 eq) was added to a reactor containing (10A) (1.0 wt) and n-PrOH (10 vol) over 30 min at 25°C. The mixture was then heated to 75°C and stirred for at least 1 h (or until the reaction was complete). The reaction was then cooled to 25°C over 2 h. Isopropyl acetate (20 vol) was added, and the suspension was stirred at 25°C for 4 h, then filtered. The wet filter cake was washed with IPAc (3.0 vol), transferred to another reactor, dissolved in water (20 vol), and heated to 35°C. Ammonium hydroxide (25 wt%) was added until the solution pH was 3–4. Seed crystals (0.005 wt) were added, and after 2 h at 35°C, more ammonium hydroxide (25 wt%) was added over 2 h until the pH was 8–9. After 4 h, the suspension was filtered, and the wet filter cake was washed with water (4.0 vol) and transferred to the reactor. To the wet filter cake, n-propanol (13 vol) and water (2.5 vol) were added, and the suspension was heated to 80°C. Once complete dissolution was achieved, the solution was cooled to 65°C, and seed crystals (0.005 wt) were added. The light suspension was stirred at 65°C for 2 h, cooled to 20°C over at least 3 h, held for 2 h, filtered, and washed with n-propanol / heptane (0.1 vol:1 vol), followed by washing with heptane (2 vol). The wet filter cake was dried under vacuum at 30°C for 10 h to obtain (14).

[0288] Synthesis of 2-((1H-pyrazol-3-yl)methyl)-6-((6-aminopyridin-2-yl)methyl)-4-methyl-4,6-dihydro-5H-thiazo[5',4':4,5]pyrrolo[2,3-d]pyridazine-5-one phosphate (15):

[0289]

[0290] All solvents were degassed by bubbling nitrogen through to remove oxygen. To a reactor containing DMSO (8.0 vol) and phosphoric acid (85%, 0.31 wt) at 25 °C was added (14) (1.0 wt). The mixture was warmed to 55 °C and polished to a 50 °C reactor followed by a 1.0 vol DMSO rinse. To the 50 °C solution was added a 1 / 7.2 / 1.8 DMSO / EtOH / H20 mixture (1.3 wt) under nitrogen. Seed crystals were added (0.010 wt) and the light suspension was stirred at 50 °C for 5 h. More 1 / 7.2 / 1.8 DMSO / EtOH / H20 mixture (20 wt) was added over at least 4 h, then the suspension was cooled to 25 °C over 3 h and held at 25 °C for 7 h. The suspension was wet milled at 25 °C for at least 2 h, filtered, and rinsed with 2.5 / 4 / 1 DMSO / EtOH / H20 (2 vol) followed by 4 / 1 EtOH / H20 (2 vol) at least three times until DMSO < 4000 ppm. The wet cake was dried under vacuum at 50 °C for 20 h to give (15) as an off-white solid.

[0291] Route B:

[0292]

[0293] Scheme 3

[0294] Synthesis of tert-butyl (E)-2-((6-((tert-butoxycarbonyl)amino)pyridin-2- yl)methylene)hydrazine-1-carboxylate (2):

[0295]

[0296] To a room temperature mixture of (1) (1 wt) and MeOH (2 vol) was added AcOH (0.05 vol, 0.2 eq). The mixture was warmed to 50 °C then a solution of Boc-hydrazine (0.62 wt) in methanol (1 vol) was added over 2.5 h. The mixture was stirred at 50 °C for at least 2 h (or until IPC showed reaction completion). The mixture was charged with water (3.5 vol) over 2 h, cooled to 20 °C over 7 h, and filtered. The wet cake was washed with 2:1 MeOH:water (1.5 vol) and dried under vacuum at 60 °C for 20 h to recover (2) as an off-white solid.

[0297] Synthesis of tert-butyl 2-((6-((tert-butoxycarbonyl)amino)pyridin-2- yl)methyl)hydrazine-1-carboxylate (3):

[0298]

[0299] The mixture of (2) (1 wt) in MeOH (8 vol) was evacuated at 25°C and rinsed three times with nitrogen. 10% Pd / C (0.04 wt) was added, and the reactor was evacuated and rinsed three times with hydrogen. The mixture was heated to 50°C, and the pressure was adjusted to 15-20 psi and stirred for at least 6 h. Once complete, the reaction was cooled to 20°C, evacuated, and rinsed three times with nitrogen. The mixture was rapidly filtered through diatomaceous earth (0.6 wt), and the solid was washed with MeOH (2 vol). The filtrate was added to the reactor at 25°C. Water (7.5 vol) was added dropwise over 2 h to induce precipitation. The suspension was stirred for 12 h, filtered, washed with 4:3 MeOH:water (1 vol), and dried under vacuum at 50°C for 24 h to obtain (3) as a grayish-white solid.

[0300] Synthesis of 6-(hydrazylmethyl)pyridine-2-amine (4):

[0301]

[0302] (3) (1.0 wt) was added in batches to a reactor containing HCl (6 N, 5 vol) at 15°C while maintaining the internal temperature between 10 and 20°C. The mixture was heated to 50°C for 2 h (or until completion), cooled to 20°C, and treated with EtOH (10 vol) for 2 h to induce precipitation, and then held for another 2 h. The suspension was filtered, washed with EtOH (2 vol), and dried under vacuum at 50°C for 20 h to obtain (4) as a white solid.

[0303] Synthesis of ethyl 6-formyl-4-methyl-2-(p-tolylthio)-4H-pyrrolo[2,3-d]thiazole-5-carboxylate (6B):

[0304]

[0305] A reactor was charged with (5B) (1 wt), chlorobenzene (3.5 vol), and POCl3(1.12 vol, 4.0 eq) at 25 °C and warmed to 100 °C. DMF (0.70 vol, 3.0 eq) was added dropwise over at least 4 h and the reactor was held at 100 °C for at least an additional 4 h (or longer if necessary) to reach completion. The temperature was lowered to 20 °C and the mixture was added to a separate reactor containing 5% Na2SO4(10 vol) at 5 °C over 6 h. EtOAc (5.0 vol) was added, the biphasic mixture was stirred at 20 °C for 2 h, and the layers were separated. The organic phase was slowly neutralized to pH 7-9 by the addition of NaHCO3(about 5 vol) at 20 °C. The organic phase was separated, washed with water (5 vol), then recycled through a charcoal filter at 25 °C for 15 h. The filtrate was concentrated under vacuum to 3 V, warmed to 40 °C, and charged with heptane (5.0 vol) over 3 h, then seeded (0.005 wt). The light suspension was held at 40 °C for 3 h, then more heptane (3.0 vol) was added at 40 °C over 3 h. The suspension was slowly cooled to 0 °C over 5 h, held at 0 °C for 9 h, and filtered. The wet cake was washed, rinsed with heptane (4.5 vol), and dried under vacuum at 50 °C for at least 18 h to give (6B) as a light yellow solid.

[0306] 6-Formyl-4-methyl-2-tosyl-4H-pyrrolo[2,3-d]thiazole-5-carboxylic acid ethyl ester (7B) synthesis:

[0307]

[0308] To a reactor was added (6B) (1 wt), chlorobenzene (16 vol), sodium tungstate dihydrate (0.046 wt, 0.05 eq), phenylphosphonic acid (0.022 wt, 0.05 eq), and methyltrioctylammonium hydrogen sulfate (0.065 wt, 0.05 eq) and the mixture was stirred and cooled to 5 °C. Hydrogen peroxide (30%, 0.79 wt, 2.5 eq) was added dropwise over 2 h at 5 °C, then the reaction was warmed to 35 °C and held for at least 4 h (or until complete). The reaction was cooled to 20 °C and quenched with 10% Na2SO3 (10 vol) over 2 h at 20 °C. The biphasic mixture was stirred for 2 h at 20 °C and the layers were separated. The organic layer was stirred with water (5 vol) for 1 h, warmed to 40 °C, and heptane (16 vol) was added dropwise over 4 h. The mixture was held for 2 h, cooled to 0 °C over 4 h, held for 6 h, then filtered. The wet cake was rinsed with heptane (3 vol) at 0 °C and dried under vacuum at 45 °C for at least 24 h to give (7B) as an off-white solid.

[0309] 6-Formyl-4-methyl-2-tosyl-4H-pyrrolo[2,3-d]thiazole-5-carboxylic acid (8B) synthesis:

[0310]

[0311] To a reactor containing (7B) (1 wt) and THF (10 vol) at 10 °C was added aqueous LiOH (14%, 2.3 vol, 3.0 eq) over 2 h. The mixture was stirred at 20 °C for at least 8 h (or until reaction was complete). The reaction was cooled to 10 °C and aqueous HC1 (2 N) was added until the pH was 1-2. The reaction was warmed to 20 °C, stirred for 2 h, water (10 vol) was added, and the suspension was stirred for 7 h. The suspension was filtered, washed with water (6.0 vol), and dried under vacuum at 55 °C for 24 h to give (8B) as a yellow solid.

[0312] 6-((6-Aminopyridin-2-yl)methyl)-4-methyl-2-tosyl-4,6-dihydro-5H-thiazolo[5',4':4,5]pyrrolo[2,3-d]pyridazin-5-one (9B) synthesis:

[0313]

[0314] To a reactor containing (8B) (1.0 wt) in EtOH (12 vol) at 20 °C was added (4) (1.1 eq), EtOH (7 vol), and i-PrOH (1 vol). Sodium dihydrogen citrate (1.18 wt) and citric acid (anhydrous, 0.26 wt) were added and the mixture was warmed to 75 °C and stirred for at least 40 h (or longer until the reaction is complete). The reaction was cooled to 20 °C over 4 h and filtered. The wet cake was washed with ethanol (4.0 vol) then transferred back to the reactor and dissolved in water (25 vol) at 20 °C. Ammonium hydroxide (25%, ~1.2 vol) was added over 30 min to adjust the pH to 8-9, the suspension was stirred at 20 °C for 8 h, then filtered and washed with water (4.0 vol) and dried under vacuum at 55 °C for 24 h to give (9B) as an off-white solid.

[0315]

[0316] Scheme 4

[0317] Synthesis of tert-butyl 2-(l-(tetrahydro-2H-pyran-2-yl)-lH-pyrazol-3-yl)acetate (12):

[0318]

[0319] To a reactor containing (11) (1.0 wt) and t-BuOH (2.5 vol) at 25 °C was added DMAP (0.03 wt) and THF (2.5 vol). After 30 min, the mixture was warmed to 50 °C then a solution of Boc20 (1.15 wt) in THF (2.0 vol) was added dropwise over at least 5 h then further stirred for up to 2 h. Upon completion, the reaction was cooled to 25 °C and both heptane (5.0 vol) and water (5.0 vol) were added. The mixture was stirred at 25 °C for 2 h then the layers were separated. The organic layer was concentrated under vacuum to 2 vol. Heptane (10 vol) was added and the mixture was concentrated to 2 vol again. More heptane (10 vol) was added and the diluted mixture was circulated through a charcoal filter for 24 h. The filtrate was concentrated to 2 vol and 2-MeTHF (6.3 vol) was added; this cycle was repeated at least two more times. This solution of (12) was used in the next step without further treatment.

[0320] Alternative synthesis of tert-butyl 2-(6-((6-aminopyridin-2-yl)methyl)-4- methyl-5-oxo-5,6-dihydro-4H-thiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidin-2-yl)-2-(1- (tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)acetate (10B):

[0321]

[0322] A reactor was charged with (9B) (1.0 wt, 1.0 eq), DMAc (5.0 vol), and a solution of (12) (1.1 eq.). After mixing at 25 °C, the reaction mixture was cooled to 0 °C and lithium tert-butoxide (1 M in THF, 6.5 vol, 3.0 eq) was added dropwise over 2 h under nitrogen. The mixture was stirred at 0 °C for 4 h. If necessary, additional lithium tert-butoxide solution (0.6 wt) was added and the mixture was stirred at 0 °C for 4 h. Once complete, aqueous acetic acid (6%, ca. 5.1 vol) was added to adjust the reaction pH to 5-7. The mixture was stirred at 0 °C for 1 h, raised to 20 °C, charged with 2-MeTHF (5 vol), and the biphasic mixture was stirred for 1 h. The layers were separated. The organic layer was retained and the aqueous layer was extracted with 2-MeTHF (5.0 vol). The second organic layer was combined with the first and washed sequentially with 5% Na2SO4 (5 vol), water (5 vol), 3% NaHCO3 (5 vol, 2x), and water (5 vol). The organic layer was concentrated under vacuum to 5 vol and subjected to two cycles of dilution with toluene (10 vol) and reconcentration (to 10 vol). The solution was adjusted to 45 °C. Seed crystals (0.005 wt) were added, the light suspension was stirred for 2 h, heptane (2.0 vol) was added dropwise over 2 h, and then stirred at 45 °C for 2 h. The suspension was cooled to 25 °C over 4 h, held at 25 °C for 15 h, filtered, washed with 5:1 toluene:heptane (4 vol), then heptane (4 vol), and dried under vacuum at 55 °C for 24 h to give (10B) as a light yellow solid.

[0323] Step 10A: Recrystallization of compound (10B) from toluene / heptane solution.

[0324] Route C:

[0325]

[0326] Scheme 5

[0327] 4-methyl-2-(methylthio)-4H-pyrrolo[2,3-d]thiazole-5-carboxylic acid ethyl ester (2C) synthesis:

[0328]

[0329] To a mixture of 2-bromo-4-methyl-4H-pyrrolo[2,3-d]thiazole-5-carboxylic acid ethyl ester ((1C) 500.0 mg, 1.73 mmol) in EtOH (10.0 mL) was added NaSMe (240.0 mg, 3.5 mmol). The reaction mixture was stirred at 25 °C for 3 h, then quenched with ice water and extracted with DCM. The combined organic layers were washed with brine, dried over anhydrous Na2SO4and concentrated under reduced pressure to give (2C), which was used directly in the next step without any purification. LC-MS: m / z 257 (M+H) + .

[0330] 6-formyl-4-methyl-2-(methylthio)-4H-pyrrolo[2,3-d]thiazole-5-carboxylic acid ethyl ester (3C) synthesis:

[0331]

[0332] To a solution of 4-methyl-2-(methylthio)-4H-pyrrolo[2,3-d]thiazole-5-carboxylic acid ethyl ester ((2C) 460.0 mg, 1.8 mmol) and N-methyl-N-phenylformamide (490 mg, 3.6 mmol) in DCE (10 mL) was added POCl3(550.0 mg, 3.6 mmol). The resulting mixture was stirred at 130 °C for 3 h, then quenched with ice water and extracted with DCM. The combined organic layers were washed with brine, dried over anhydrous Na2SO4and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: PE / EtOAc = 8 / 1) to give the desired intermediate. LC-MS: m / z 285 (M+H) + To a solution of the desired intermediate (300.0 mg, 1.06 mmol) in EtOH (5.0 mL) was added N2H4-H2O (2 mL, 98% wt). The reaction mixture was stirred at room temperature for 1 h, then heated to 60 °C overnight, then cooled. The solid was collected by filtration and dried under high vacuum to give (3C). LCMS: m / z 253 (M+H) + . 1 H NMR (400 MHz, DMSO-d6) δ 12.61 (s, 1H), 8.48 (s, 1H), 4.22 (s, 3H), 2.81 (s, 3H).

[0333] 4-methyl-2-(methylsulfonyl)-4,6-dihydro-5H-thiazolo[5',4':4,5]pyrrolo[2,3- d]pyridazin-5-one (4C) synthesis:

[0334]

[0335] In a three necked flask containing 4-methyl-4,6-dihydro-5H-thiazolo[5',4':4,5]pyrrolo[2,3- d]pyridazin-5-one (3C), 30 g, 0.119 mol, 1.0 eq) in DCM (600 mL), m-CPBA (61.5 g, 3 eq) was added in three portions at 20 °C. The mixture was stirred at 30 °C overnight, LC-MS indicated 100% starting material consumption, 20% of the sulfoxide and 80% of the sulfone were formed. The mixture was cooled to room temperature, another portion of m-CPBA (1.0 eq) was added. The reaction mixture was stirred at 30 °C for 2 hours, LC-MS indicated <8% of the sulfoxide (LCMS: m / z 269 (M+H) + .) The mixture was cooled to room temperature and filtered. The filter cake was suspended in MeOH (500 mL) and stirred at room temperature for 1 hour. The solid was collected by filtration, washed with ethyl acetate, dried in vacuum to give a mixture of 5% sulfoxide and 95% sulfone. The mixture was suspended in DMSO (600 mL), heated to 120-130 °C to form a clear solution. Then cooled to room temperature, solid precipitated. The mixture was filtered and dried to give pure 4-methyl-2-(methylsulfonyl)-4,6-dihydro-5H-thiazolo[5',4':4,5]pyrrolo[2,3- d]pyridazin-5-one (4C), LCMS: m / z 285 (M+H) + 1H NMR (400 MHz, DMSO) δ 12.87 (s, 1H), 8.69 (s, 1H), 4.32 (s, 3H), 3.56 (s, 3H).

[0336] Methyl 1H-pyrazole-3-carboxylate - synthesis of (1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)methanol (6C):

[0337]

[0338] To a stirred solution of methyl 1 H-pyrazole-3-carboxylate (5C), 90 g, 0.72 mol in THF (1 L) at 0 °C under N2atmosphere was added NaH (20.7 g, 0.864 mol, 60%) dropwise. The resulting mixture was slowly warmed to room temperature and stirred for 1 h. The reaction mixture was then cooled back to 0 °C and SEMCl (151.5 mL, 0.842 mol) was added dropwise. Stirring was continued for another 2 h, then quenched with saturated NH4Cl and extracted with ethyl acetate (3x). The combined organic layers were washed with brine and dried over Na2S04. The solvent was removed under vacuum to provide the crude product which was used in the next step without purification.

[0339] To a stirred solution of methyl 1 H-pyrazole-3-carboxylate (5C), 90 g, 0.72 mol in THF (1 L) at 0 °C under N2atmosphere was added NaH (20.7 g, 0.864 mol, 60%) dropwise. The resulting mixture was slowly warmed to room temperature and stirred for 1 h. The reaction mixture was then cooled back to 0 °C and SEMCl (151.5 mL, 0.842 mol) was added dropwise. Stirring was continued for another 2 h, then quenched with saturated NH4Cl and extracted with ethyl acetate (3x). The combined organic layers were washed with brine and dried over Na2S04. The solvent was removed under vacuum to provide the crude product which was used in the next step without purification. + .

[0340] (1-((2-(trifluoromethyl)ethoxy)methyl)-1 H-pyrazol-3-yl)methanol-3-(iodomethyl)-1-((2- (trifluoromethyl)ethoxy)methyl)-1 H-pyrazole (7C) was synthesized as follows:

[0341]

[0342] To a stirred solution of (1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)methanol ((6C), 61.5 g, 0.262 mol theoretically) in THF (310 mL) was added TEA (55.42 mL, 0.393 mol) followed by MsCl (24 mL, 0.314 mol) at 0 °C under N2atmosphere. The reaction was allowed to warm to room temperature and stirred for 1 h, then NaI (196.5 g, 1.31 mol in 310 mL DMF) was introduced. The resulting mixture was stirred for another 1 h and quenched with ice water, extracted with MTBE (3x). The combined organic layers were washed with saturated Na2S203and brine, dried over Na2S04and concentrated to give (7C) which was used in the next step without purification. LC-MS: m / z 339 (M+H) + .

[0343] 3-((Benzene-sulfonyl)methyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (8C) synthesis:

[0344]

[0345] To a stirred solution of (1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)methanol ((7C) 77.5 g, 0.229 mol theoretically) in DMF (600 mL) was added sodium benzenesulfinate (53.5 g, 0.32 mol) and stirred at 0 °C for 1 h. After warming to room temperature, the reaction mixture was quenched with ice water and saturated Na2S203, extracted with ethyl acetate (3x). The combined organic layers were washed with saturated NaHC03and brine, dried over Na2S04. The solvent was removed under vacuum and the residue was purified by flash chromatography (silica gel, 20%~70% ethyl acetate / petroleum ether) to give (8C) as a light yellow oil. LCMS: [M + H] + 353.1H NMR (400 MHz, DMSO) δ 7.85–7.77 (m, 4H), 7.62 (dd, 2H), 6.19 (d, 1H), 5.35 (d, 2H), 4.70 (d, 2H), 3.44-3.38 (m, 2H), 0.88–0.77 (m, 2H), -0.01 (s, 9H).

[0346] tert-Butyl (tert-butoxycarbonyl)(3-(iodomethyl)phenyl)carbamate - tert-Butyl (tert-butoxycarbonyl)(6-((4-methyl-2-(methylsulfonyl)-5-oxo-4,5-dihydro-6H-thiazolo[5',4':4,5]pyrrolo[2,3-d]pyridazin-6-yl)methyl)pyridin-2-yl)carbamate - Synthesis of 4-methyl-2-(methylsulfonyl)-4,6-dihydro-5H-thiazolo[5',4':4,5]pyrrolo[2,3-d]pyridazin-5-one (9C):

[0347]

[0348] A mixture of 4-methyl-2-(methylsulfonyl)-4H-thiazolo[5',4':4,5]pyrrolo[2,3-d]pyridazin-5(6H)-one ((4C), 7.5 g, 26.4 mmol) and K3PO4(8.3 g, 39.3 mmol) in anhydrous MeCN (300 mL) was stirred at 70 °C under N2for 1 h. Then a solution of tert-butyl N-[(tert-butoxy)carbonyl]-N-[6-(bromomethyl)pyridin-2-yl]carbamate (11.2 g, 29.0 mmol) in MeCN (30 mL) was added. After stirring at 70 °C under N2for 2.5 h, the reaction mixture was quenched with saturated NH4Cl and extracted with EA (300 mL x 3). The combined organic layers were washed with water and brine, dried over Na2SO4, filtered and the organic phase was concentrated. The crude product was purified by flash chromatography (silica gel, 0-50% ethyl acetate / petroleum ether) to give tert-butyl N-[(tert-butoxy)carbonyl]-N-[6-({4-methylsulfonyl-7-methyl-9-oxo-3-thia-5,7,10,11-tetraazatricyclo[6.4.0.0{2,6}]dodeca-1(8),2(6),4,11-tetraen-10-yl}methyl)pyridin-2-yl]carbamate (9C). LC-MS (ESI) found: 591.1 (M+H) + .

[0349] tert-Butyl (6-((4-methyl-5-oxo-2-((phenylsulfonyl)(l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrazol-3-yl)methyl)-4,5-dihydro-6H- thiazolo[5',4':4,5]pyrrolo[2,3-d]pyridazin-6-yl)methyl)pyridin-2-yl)carbamate - tert-Butyl (6-((4-methyl-2-(methylsulfonyl)-5-oxo-4,5-dihydro-6H- thiazolo[5',4':4,5]pyrrolo[2,3-d]pyridazin-6-yl)methyl)pyridin-2-yl)carbamate - 3-((phenylsulfonyl)methyl)-l-((2-(trimethylsilyl)ethoxy)methyl)-lH-pyrazole (10C) synthesis:

[0350]

[0351] To a stirred mixture of 3-((phenylsulfonyl)methyl)-l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrazole ((9C) 11.9 g, 33.8 mmol) in anhydrous THF (200 mL) was added LiHMDS (50 mL, 1 M in THF) at -40 °C under argon. After 10 min, the mixture was warmed to 10 °C and stirred for 1 h before adding (9) (9.1 g, 15.4 mmol in 35 mL THF). The reaction was stirred at 10 °C for another 30 min. The reaction mixture was poured into aqueous NH4Cl solution, extracted with EtOAc (200 mL x 3). The combined organic layers were washed with water and brine, dried over anhydrous Na2SO4 and concentrated. The crude product was purified by flash chromatography (silica gel, 0-50% ethyl acetate / petroleum ether) to give tert-butyl (6-((4-methyl-5-oxo-2-((phenylsulfonyl)(l-((2- (trimethylsilyl)ethoxy)methyl)-lH-pyrazol-3-yl)methyl)-4H-thiazolo[5',4':4,5]pyrrolo[2,3- d]pyridazin-6(5H)-yl)methyl)pyridin-2-yl)carbamate (10C). LC-MS (ESI) found: 763.2 (M+H) + .

[0352] tert-Butyl (6-((4-methyl-5-oxo-2-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)methyl)- 4,5-dihydro-6H-thiazolo[5',4':4,5]pyrrolo[2,3-d]pyridazin-6-yl)methyl)pyridin-2-yl)carbamate - Synthesis of tert-butyl (6-((4-methyl-5-oxo-2-((phenylsulfonyl)(1-((2- (trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)methyl)-4,5-dihydro-6H-thiazolo[5',4':4,5]pyrrolo[2,3- d]pyridazin-6-yl)methyl)pyridin-2-yl)carbamate (11C):

[0353]

[0354] A solution of (10C) (6.0 g, 7.86 mmol) in EtOH / AcOH (35 mL / 50 mL) was heated to 50 °C in the presence of Zn (2.55 g, 117.9 mmol) with vigorous stirring for 40 min. Additional zinc (2.55 g, twice, the reaction was monitored by TLC / LC-MS to avoid byproducts and over-reduced products) was added every 40 min. The solution was filtered and the filter cake was washed with DCM. The filtrate was partially evaporated, neutralized with a saturated NaHC03solution, dried over MgS04, and the solvent was removed under vacuum. The crude product was purified by flash chromatography (silica gel, DCM:MeOH = 40:1) to give (11C). LC-MS (ESI) found: 623.3 (M+H) + .

[0355] tert-Butyl (6-((4-methyl-5-oxo-2-((1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-3-yl)methyl)- 4,5-dihydro-6H-thiazolo[5',4':4,5]pyrrolo[2,3-d]pyridazin-6-yl)methyl)pyridin-2-yl)carbamate - 2-((1H-pyrazol-3-yl)methyl)-6-((6-aminopyridin-2-yl)methyl)-4-methyl-4,6-dihydro-5H- thiazolo[5',4':4,5]pyrrolo[2,3-d]pyridazin-5-one (13) synthesis:

[0356]

[0357] To a mixture of (11C) (3.0 g, 4.8 mmol) in ethanol (30 mL) was added HC1 (30 mL, 4 M in dioxane). The reaction mixture was stirred at 80 °C for 40 min. The reaction mixture was cooled to room temperature, filtered and the solid was collected, suspended in water and neutralized with aqueous NaHC03solution at 10 °C. Filtration afforded the desired compound (13). LC-MS (ESI) found: 393.2 (M+H) + . 1 HNMR (400 MHz, DMSO-d6) δ 12.78 (s, 1H), 8.53 (s, 1H), 7.72(s, 1H), 7.25 (dd, 1H), 6.33–6.24 (m, 2H), 6.08 (d, 1H), 5.90 (s, 2H), 5.19(s, 2H), 4.49 (s, 2H), 4.26 (s, 3H).

Claims

1. A method of preparing a compound of Formula (A-l): or a salt thereof, wherein X is halo; L is a bond or Ci-C6alkylene; and Q is C3-C6cycloalkyl, 5-14 membered heterocyclyl, 6-12 membered aryl, or 5-14 membered heteroaryl, each of which is optionally substituted with 1 to 3 groups selected from halo, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, hydroxyl, Ci-C6aminoalkyl, -NH2, -NH(Ci-C6alkyl), -N(Ci-C6alkyl)2, CN, and NO2; the method comprising reacting a compound of Formula (A-2): (A-2) or a salt thereof, with a compound of Formula (A-3): to form the compound of Formula (A-l) or a salt thereof. (A-1)， 4. The method of any one of claims 1 to 3, wherein X is CI. R 1 is hydrogen, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, Ci-C6aminoalkyl, Ci-C6cyanoalkyl, C2-C6alkenyl, C2-C6alkynyl, or C3-C6cycloalkyl, wherein said C3-C6cycloalkyl is optionally substituted with one to three groups selected from halo, hydroxyl, NH2, and CN; 5. The method of any one of claims 1 to 4, wherein L is Ci-C6alkylene.

6. The method of any one of claims 1 to 5, wherein L is -CH2-.

7. The method of any one of claims 1 to 6, wherein Q is 6-12 membered aryl or 5-14 membered heteroaryl, each of which is optionally substituted with 1 to 3 groups selected from halo, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, hydroxyl, Ci-C6aminoalkyl, NH2, -NH(Ci-C6alkyl), N(Ci-C6alkyl)2, CN, and NO2.

8. The method of any one of claims 1 to 7, wherein Q is 5-7 membered heteroaryl substituted with -NH2.

9. The method of any one of claims 1 to 8, wherein the compound of Formula (A-l) is:

10. The method of any one of claims 1 to 9, wherein the compound of Formula (A-2) is: (A-3)， 11. The method of any one of claims 1 to 9, wherein the compound of Formula (A-2) is:

2. The method of claim 1, wherein R 1 is Ci-C6alkyl or Ci-C6haloalkyl.

3. The method of claim 1 or claim 2, wherein R 1 is methyl.

12. The method of any one of claims 1 to 11, wherein the compound of Formula (A-3) is:

13. The method of any one of claims 1 to 12, wherein the compound of Formula (A-2) or a salt thereof is reacted with the compound of Formula (A-3) in the presence of an acid and one or more solvents.

14. The method of any one of claims 1 to 13, wherein the bis-acid salt of the compound of Formula (A-3) is reacted with the compound of Formula (A-2) in one or more solvents.

15. The method of claim 11 or 12, wherein the acid is selected from the group consisting of hydrochloric acid, acetic acid, phosphoric acid, sulfuric acid, citric acid, formic acid, methanesulfonic acid, and p-toluenesulfonic acid.

16. The method of any one of claims 1 to 15, wherein at least one solvent is a polar protic solvent.

17. The method of claim 16, wherein the at least one polar protic solvent is selected from the group consisting of water, methanol, ethanol, n-propanol, isopropanol, and t-butanol, or any combination thereof. or salts thereof.

18. The method of claim 17, wherein the at least one polar protic solvent is ethanol or n-propanol. or a bisacid salt thereof. ​ or tautomers or hydrates thereof. ​ 。 ​ ​ ​ ​ ​ ​ 19. The process of claim 17, wherein the polar protic solvent is a mixture of ethanol and isopropanol.

20. A process for preparing a compound of formula (A-3), (A-3)， Where R 1 It is hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, C1-C6 aminoalkyl, C1-C6 cyanoalkyl, C2-C6 alkenyl, C2-C6 alkynyl, or C3-C6 cycloalkyl, wherein the C3-C6 cycloalkyl is optionally substituted by 1 to 3 groups selected from: halogroup, hydroxyl, NH2, and CN; and X is halo; the process comprising: hydrolyzing a compound of formula (A-6): (A-6) wherein R 2 is C1-C6alkyl to form a compound of formula (A-3).

21. The process of claim 20, wherein the compound of formula (A-6) is hydrolyzed in the presence of a base and one or more solvents.

22. The process of claim 21, wherein the base is selected from the group consisting of lithium hydroxide (LiOH), calcium hydroxide (Ca(OH)2), potassium hydroxide (KOH), sodium hydroxide (NaOH), and barium hydroxide (Ba(OH)2).

23. The process of claim 22, wherein the base is sodium hydroxide (NaOH).

24. The process of any one of claims 20 to 23, wherein the compound of formula (A-6) is hydrolyzed in the presence of water and one or more solvents.

25. The process of any one of claims 20 to 24, wherein the compound of formula (A-6) is hydrolyzed in the presence of water and at least one solvent that is a polar aprotic solvent.

26. The process of claim 25, wherein the at least one polar aprotic solvent is selected from the group consisting of dimethylacetamide (DMAC), 2-methyltetrahydrofuran (2-Me THF), tetrahydrofuran (THF), acetone, dimethylformamide (DMF), acetonitrile (ACN), and dimethylsulfoxide (DMSO), or any combination thereof.

27. The process of claim 26, wherein the at least one polar aprotic solvent is THF.

28. The process of any one of claims 20 to 27, wherein the compound of formula (A-6) is prepared by reacting a compound of formula (A-5): (A-5)， with phosphorus oxychloride (POCl3), dimethylformamide (DMF), and one or more additional solvents to form the compound of formula (A-6).

29. The process of claim 28, wherein the one or more additional solvents is a non-polar solvent.

30. The process of claim 29, wherein the non-polar solvent is selected from the group consisting of dioxane, dichloroethane, benzene, toluene, chlorobenzene, dichlorobenzene, and xylene.

31. The process of claim 30, wherein the non-polar solvent is chlorobenzene.

32. The process of any one of claims 28 to 31, wherein the compound of formula (A-5) is prepared by reacting a compound of formula (A-4): (A-4)， with a halogenating agent, a base, and at least one solvent to form the compound of formula (A-5).

33. The process of claim 32, wherein the base is selected from the group consisting of lithium diisopropylamide (LDA), sodium bis(trimethylsilyl)amide (NaHMDS), lithium bis(trimethylsilyl)amide (LiHMDS), potassium bis(trimethylsilyl)amide (KHMDS), and sodium hydride (NaH).

34. The process of claim 33, wherein the base is lithium bis(trimethylsilyl)amide (LiHMDS).

35. The method of any one of claims 31 to 34, wherein the halogenating reagent is selected from elemental halogen, carbon tetrachloride, hexachloroethane, carbon tetrabromide, n-bromosuccinimide (NBS), and n-chlorosuccinimide (NCS).

36. The method of claim 35, wherein the halogenating reagent is hexachloroethane.

37. The method of any one of claims 31 to 36, wherein at least one solvent is a polar aprotic solvent.

38. The method of claim 37, wherein the polar aprotic solvent is selected from dimethylacetamide (DMAC), 2-methyltetrahydrofuran (2-Me THF), tetrahydrofuran (THF), acetonitrile (ACN), and dimethylsulfoxide (DMSO), or any combination thereof.

39. The method of claim 38, wherein the polar aprotic solvent is THF.

40. The method of any one of claims 22 to 39, wherein R 1 is Ci-C6alkyl or Ci-C6haloalkyl.

41. The method of any one of claims 20 to 40, wherein R 1 is Ci-C6alkyl.

42. The method of any one of claims 20 to 41, wherein R 1 is methyl.

43. The method of any one of claims 20 to 42, wherein X is -Cl.

44. The method of any one of claims 20 to 43, wherein R 2 is ethyl.

45. The method of any one of claims 20 to 44, wherein the compound of Formula (A-3) is 。 46. The method of any one of claims 32 to 45, wherein the compound of Formula (A-4) is 。 47. The method of any one of claims 20 to 46, wherein the compound of Formula (A-6) is 。 48. The method of any one of claims 28 to 47, wherein the compound of Formula (A-5) is 。 49. A method of making a compound of Formula (B-1): (B-1)， wherein R 1 is hydrogen, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, Ci-C6aminoalkyl, Ci-C6cyanoalkyl, C2-C6alkenyl, C2-C6alkynyl, or C3-C6cycloalkyl, wherein said C3-C6cycloalkyl is optionally substituted with one to three groups selected from halo, hydroxyl, NH2, and CN; R 4 is C1-C6alkyl, C1-C6haloalkyl, 6-12 membered aryl, or 5-14 membered heteroaryl, wherein the 6-12 membered aryl or the 5-14 membered heteroaryl is optionally substituted with 1 to 3 groups selected from halo or C1-C6alkyl; L is a bond or C1-C6 alkylene; and Q is C3-C6 cycloalkyl, 5-14 membered heterocyclyl, 6-12 membered aryl, or 5-14 membered heteroaryl, each of which is optionally substituted with 1 to 3 groups selected from halo, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 hydroxyalkyl, hydroxyl, C1-C6 aminoalkyl, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, CN, and NO2; the method comprising: reacting a compound of Formula (A-2): (A-2) or a salt thereof, with a compound of Formula (B-5): (B-5)， to form the compound of Formula (B-1).

50. The method of claim 49, wherein the compound of Formula (A-2) or a salt thereof is reacted with the compound of Formula (B-5) in the presence of an acid and at least one solvent.

51. The method of claim 50, wherein the acid is selected from hydrochloric acid, trifluoroacetic acid, acetic acid, phosphoric acid, sulfuric acid, citric acid, formic acid, and carbonic acid.

52. The method of any one of claims 49 to 51, wherein at least one solvent is a polar protic solvent.

53. The method of claim 52, wherein the polar protic solvent is an alcohol.

54. The method of claim 53, wherein at least one polar protic solvent is selected from methanol, ethanol, n-propanol, isopropanol, sec-butanol, and tert-butanol, or any combination thereof.

55. The method of claim 54, wherein at least one polar protic solvent is ethanol, isopropanol, or n-propanol, or a mixture thereof.

56. The process of any one of claims 49 to 55, wherein the compound of formula (B-5) is prepared by hydrolysis of a compound of formula (B-4): (B-4)， wherein R 2 is C1-C6 alkyl.

57. The process of claim 56, wherein the compound of formula (B-4) is hydrolyzed in the presence of a base and one or more solvents.

58. The process of claim 57, wherein the base is selected from the group consisting of lithium hydroxide (LiOH), calcium hydroxide (Ca(OH)2), potassium hydroxide (KOH), sodium hydroxide (NaOH), and barium hydroxide (Ba(OH)2).

59. The process of claim 58, wherein the base is lithium hydroxide (LiOH) or sodium hydroxide (NaOH).

60. The process of any one of claims 56 to 59, wherein at least one solvent is water.

61. The process of any one of claims 56 to 60, wherein the compound of formula (B-4) is hydrolyzed in water and a polar aprotic solvent.

62. The process of claim 61, wherein the polar aprotic solvent is selected from the group consisting of dimethylacetamide (DMAC), 2-methyltetrahydrofuran (2-Me THF), tetrahydrofuran (THF), dimethylformamide (DMF), acetonitrile (ACN), and dimethylsulfoxide (DMSO), or any combination thereof.

63. The process of claim 62, wherein the polar aprotic solvent is THF.

64. The process of any one of claims 56 to 63, wherein the compound of formula (B-4) is prepared by reacting a compound of formula (B-3): (B-3)， with an oxidizing agent in at least one solvent to form the compound of formula (B-4).

65. The process of claim 64, wherein the oxidizing agent is hydrogen peroxide and sodium tungstate (Na2WO4).

66. The process of claims 63 to 65, wherein at least one solvent is a non-polar solvent.

67. The process of claim 66, wherein at least one non-polar solvent is selected from the group consisting of dichloromethane, toluene, chlorobenzene, dichlorobenzene, and xylene, or any combination thereof.

68. The process of claim 67, wherein the non-polar solvent is chlorobenzene.

69. The process of any one of claims 63 to 68, wherein the process further comprises at least one phase transfer catalyst.

70. The process of claim 69, wherein the phase transfer catalyst is methyltrioctylammonium hydrogen sulfate.

71. The process of any one of claims 63 to 70, wherein the process further comprises at least one acid.

72. The process of claim 71, wherein the acid is phenylphosphonic acid.

73. The process of any one of claims 63 to 72, wherein the compound of formula (B-3) is prepared by reacting a compound of formula (B-2): (B-2)， with phosphorus oxychloride (POCI3), dimethylformamide (DMF), and one or more solvents to form the compound of formula (B-3).

74. The process of claim 73, wherein one solvent is a non-polar solvent.

75. The method of claim 74, wherein the non-polar solvent is selected from toluene, chlorobenzene, dichlorobenzene, and xylene, or any combination thereof.

76. The method of claim 75, wherein the non-polar solvent is chlorobenzene.

77. The method of any one of claims 47 to 76, wherein R 1 is Ci-C6alkyl or Ci-C6haloalkyl.

78. The method of any one of claims 47 to 77, wherein R 1 is Ci-C6alkyl.

79. The method of any one of claims 47 to 78, wherein R 1 is methyl.

80. The method of any one of claims 56 to 79, wherein R 2 is ethyl.

81. The method of any one of claims 56 to 80, wherein R 4 is a 6-12 membered aryl group optionally substituted with 1 to 3 groups selected from halo and C1-C6 alkyl.

82. The method of any one of claims 56 to 81, wherein R 4 is 4-methylphenyl.

83. The method of any one of claims 49 to 82, wherein L is Ci-C6alkylene.

84. The method of any one of claims 49 to 83, wherein L is -CH2-.

85. The method of any one of claims 49 to 84, wherein Q is 6-12 membered aryl or 5-14 membered heteroaryl, each of which is optionally substituted with 1 to 3 groups selected from halo, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, hydroxyl, Ci-C6aminoalkyl, -NH2, -NH(Ci-C6alkyl), N(Ci-C6alkyl)2, -CN, and -NO2.

86. The method of any one of claims 49 to 85, wherein Q is 5-7 membered heteroaryl substituted with -NH2.

87. The method of any one of claims 49 to 86, wherein the compound of Formula (B-1) is 。 88. The method of any one of claims 73 to 87, wherein the compound of Formula (B-2) is 。 89. The method of any one of claims 64 to 88, wherein the compound of Formula (B-3) is 。 90. The method of any one of claims 56 to 89, wherein the compound of Formula (B-4) is 。 91. The method of any one of claims 49 to 90, wherein the compound of Formula (B-5) is 。 92. The method of any one of claims 49 to 91, wherein the compound of Formula (A-2) is: or salts thereof.

93. A compound having the structure: or a salt, tautomer, or hydrate thereof.

94. A compound having the structure: or a salt, tautomer, or hydrate thereof.

95. A compound having the structure: or a salt, tautomer, or hydrate thereof.

96. A compound having the structure: or a salt, tautomer, or hydrate thereof.

97. A compound having the structure: or a salt, tautomer, or hydrate thereof.

98. A compound having the structure: or a salt, tautomer, or hydrate thereof.

99. A compound having the structure: or a salt, tautomer, or hydrate thereof.

100. A compound having the structure: or a bisacid salt thereof.

101. A compound having the structure: or tautomers or hydrates thereof.

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