WO2026030116A1 - Modulators of mitochondrial dna replication - Google Patents

Abstract

The present invention provides novel DNA polymerase γ (POL γ) modulators and pharmaceutical compositions thereof.

Description

MODULATORS OF MITOCHONDRIAL DNA REPLICATION

CROSS-REFERENCE TO RELATED APPLICATION

[0001] The present application claims priority to US provisional application number 63/676,628 filed on July 29, 2024, the entirety of which is incorporated herein.

FIELD OF THE INVENTION

[0002] The present disclosure relates to novel DNA polymerase y (POL y) modulators, their pharmaceutically acceptable salts, and pharmaceutical compositions thereof.

BACKGROUND OF THE INVENTION

[0003] Human mitochondria contain multiple copies of a circular, double-stranded (ds) DNA genome (mtDNA), and a dedicated DNA replication machinery is required for maintaining the mitochondrial genome. DNA polymerase y (POLy) is the replicative polymerase essential for maintaining the mtDNA. POLy is a heterotrimer in human cells with one catalytic subunit (POLyA) and two accessory subunits (POLyB). POLyA belongs to the family A DNA polymerases. POLyA is 1239 amino acids in length and consists of three distinct regions: an N- terminal exonuclease domain connected by a linker domain to the C-terminal polymerase domain. The accessory POLyB is 485 amino acids in length, and the crystal structures of both mouse and human POLyB have revealed the protein as a dimer with high similarities to aminoacyl tRNA synthetases. POLyB acts as a processivity factor, which increases the affinity of the polymerase for DNA and promotes tighter nucleotide binding, thereby increasing the polymerase rate. The accessory subunit B of DNA polymerase y is required for mitochondrial replisome function.

[0004] A primary biological role of POLy is to replicate the mitochondrial genome. However, POLy cannot replicate the double stranded (ds) mtDNA alone. It acts together with a DNA helicase (TWINKLE) and the mitochondrial single-stranded DNA-binding protein (mtSSB). TWINKLE is 684 amino acids in length and forms a hexamer in solution.

[0005] TWINKLE travels in front of POLy during mtDNA replication, unwinding the dsDNA and creating a single-stranded (ss) DNA template that POLy can utilize. The mtSSB is 148 amino acids in length and is active as a tetramer. It binds to ssDNA, protects this DNA against nucleases, and prevents secondary structure formation so POLy can use ssDNA as a template to synthesize dsDNA. The mtSSB enhances mtDNA synthesis by increasing the processivity of POLy and also stimulates TWINKLE’S helicase activity. POLy cannot initiate DNA synthesis de novo, as it needs a short RNA primer to initiate DNA synthesis. In mitochondria, the mitochondrial RNA polymerase (POLRMT) has a dual function; it acts as an RNA polymerase involved in mtDNA transcription but it also synthesizes the primers needed to initiate mtDNA replication from mitochondrial origins of replication.

[0006] The mitochondrial genome encodes subunits of the oxidative phosphorylation (OXPHOS) system. The OXPHOS system is composed of four respiratory chain complexes, which are responsible for electron transport and generation of the proton gradient across the mitochondrial inner membrane. ATP synthase uses this proton gradient to produce ATP. The biogenesis of the OXPHOS system is under dual genetic control and requires the concerted expression of nuclear DNA and mtDNA encoded genes. Mitochondria contain multiple copies of ds mtDNA, which encodes 2 ribosomal RNAs (mt-rRNAs), 22 transfer RNAs (mt-tRNAs), and 11 messenger RNAs (mt-mRNAs) producing 13 protein subunits of OXPHOS complexes I, III, IV, and ATP synthase (sometime referred to as complex V). The biogenesis of the OXPHOS system is critically dependent on the mtDNA-encoded subunits as they typically have key catalytic roles or are core subunits for OXPHOS assembly. Similar to the nuclear genome, expression of mammalian mtDNA requires several essential steps, including genome maintenance, replication, transcription, RNA maturation, and translation. All proteins involved in these processes are encoded in the nuclear genome, translated in the cytosol, and imported into the mitochondrial network. It is estimated that approximately one quarter of the ~1200 nucleus- encoded mitochondrial proteins are devoted to the control of mtDNA gene expression in mammals. POLy is required for mtDNA synthesis and is thus essential for biogenesis of the OXPHOS system, resulting in ATP production. ATP production is in turn vital for energy homeostasis in the cell.

[0007] Mutations affecting POLy are among the most frequent causes of mitochondrial disease. More than 300 disease-causing variants have been identified in POLy, causing a broad clinical spectrum of neurodegenerative and mitochondrial diseases such as Alpers syndrome, stroke-like episodes, and chronic progressive external ophthalmoplegia. Pathogenic variants in the gene encoding POLyA, namely POLG, are now known to cause a spectrum of overlapping phenotypes. These POLy mutations are linked to the accumulation of damaged mtDNA, including multiple deletions, but can also lead to loss of mtDNA (depletion). Many diseasecausing variants of POLy are associated with decreased replication processivity of the mtDNA replication machinery, leading to replication stalling. [0008] In view of the numerous and varied roles of POLy, the need exists for potent and specific modulators of POLy.

SUMMARY OF INVENTION

[0009] Other aspects and iterations of the present disclosure are detailed below.

[0010] In some aspects, the present disclosure is directed to a compound, or a pharmaceutically acceptable salt thereof, according to Formula (I): wherein:

A is substituted diazole, oxazole, substituted oxazole, thiazole, substituted thiazole, thiadiazole, substituted thiadiazole, oxadiazole, substituted oxadiazole, triazole, substituted triazole, pyridine, or substituted pyridine; wherein when the substituted diazole is a pyrazole the substitution is not an unsubstituted phenyl.

[0011] In some aspects, A is substituted diazole and with any one of Formula (l-a) or Formula (l-b):

wherein:

R1-R3 are each independently hydrogen or C1-C6 alkyl; and

B is substituted C1-C6 alkyl, C3-C6 cycloalkyl, substituted C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, substituted C3-C6 heterocycloalkyl, C3-C6 heterocyclyl, substituted C3-C6 heterocyclyl, substituted aryl, amide, or substituted amide.

[0012] In some aspects, B is substituted C1-C6 alkyl.

[0013] In some aspects, the substituted C1-C6 alkyl is substituted with one or more of benzyl, fluorine, or methyl. In some aspects, the substituted C1-C6 alkyl is methylbenzene, ethylbenzene, trifluoroethyl, difluoroethyl, or N,N-dimethylethanamine.

[0014] In some aspects, wherein B is substituted aryl and the compound is substituted pyrazole with Formula (l-a-1 ): wherein:

R1-R2 are each independently hydrogen or methyl;

R3 is hydrogen;

R4 is hydrogen or halogen;

R5 and Re are each independently hydrogen, halogen, C1-C6 alkyl, C1-C6 substituted alkyl, carboxylic acid, substituted carboxylic acid, sulfonyl, substituted sulfonyl, alkoxy, amide, substituted amide, amine, substituted amine, C3-C6 heterocycloalkyl, substituted C3-C6 heterocycloalkyl, C3-C6 cycloalkyl, or substituted C3-C6 cycloalkyl;

R₇ and Rs is are each independently hydrogen or alkoxy; and R₄-RS can not be hydrogen at the same time.

[0015] In some aspects, R1 and R₂ are hydrogen, and with Formula (l-a-2): wherein:

R₄ is hydrogen or fluorine;

R₅ is hydrogen, fluorine, bromine, chlorine, methyl, trifluoromethyl, carboxylic acid, methyl substituted carboxylic acid, methoxy, substituted ethyl, amide, methyl amide, dimethyl amide, methyl sulfonyl, N-methyl sulfonyl amide, or substituted C1-C6 alkyl;

Re is hydrogen, fluorine, methoxy, methoxy methyl, methoxy ethyl, substituted ethyl, amide, methyl amide, amide, methyl amide, dimethyl amide, substituted alkyl, substituted sulfonyl, cyclobutyl, substituted cyclobutyl, oxetane, substituted oxetane;

R₇ and R_s are each independently hydrogen or methoxy; and

R₄-RS can not be hydrogen at the same time.

[0016] In some aspects, R₆ is substituted oxetane. In some aspects, the substituted oxetane is oxetan-3-ol.

[0017] In some aspects, R6 is substituted cyclobutane. In some aspects, the substituted cyclobutane is 3,3-difluorocyclobutan-1-ol.

[0018] In some aspects, the substituted sulfonyl is with Formula (l-a-3):

wherein R₉ is methyl, ethyl, trifluoromethyl, cyclopropyl, or methyl amide.

[0019] In some aspects, wherein the substituted alkyl is methyl or ethyl substituted with one or more of methyl, amine, methyl amine, dimethyl amide, hydroxyl, or fluorine.

[0020] In some aspects, B is C3-C6 cycloalkyl or substituted C3-C6 cycloalkyl, with Formula (I- a-4): wherein R' and R" are each independently hydrogen or fluorine.

[0021] In some aspects, wherein B is C3-C6 heterocyclyl or substituted C3-C6 heterocyclyl, and with any one of Formula (l-a-5), Formula (l-a-6), Formula (l-a-7), Formula (l-a-8), Formula (l-a- 9), Formula (l-a-10), or Formula (l-a-11):

wherein:

R4 and R7-Ra are hydrogen;

R₅ is hydrogen, substituted C1-C6 alkyl, or methoxy; and

R₆ is hydrogen or substituted C1-C6 alkyl.

[0022] In some aspects, Rs is substituted C1-C6 alkyl.

[0023] In some aspects, the substituted C1-C6 alkyl is trifluoromethane.

[0024] In some aspects, Rs is substituted C1-C6 alkyl.

[0025] In some aspects, the substituted C1-C6 alkyl is substituted with one or more of fluorine or hydroxyl.

[0026] In some aspects, the substituted C1-C6 alkyl is 2,2,2-trifluoroethan-1-ol or propan-2-ol.

[0027] In some aspects, B is aryl and the compound is substituted imidazole with Formula (I- c-1): wherein R₂ and R₃ are hydrogen.

[0028] In some aspects, A is substituted thiazole and with Formula (l-d), Formula (l-e), or Formula (l-f): wherein C is i-Ce alkyl, substituted C1-C6 alkyl, C3-C6 cycloalkyl, substituted C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, substituted C3-C6 heterocycloalkyl, C3-C6 heterocyclyl, substituted C3-C6 heterocyclyl, aryl, substituted aryl, amide, or substituted amide.

[0029] In some aspects, C is C3-C6 heterocyclyl or substituted C3-C6 heterocyclyl, with any one of Formula (l-d-1), Formula (l-d-2), Formula (l-d-3), Formula (l-d-4), Formula (l-d-5), Formula (l-d-6), or Formula (l-d-7):

wherein:

R10 and R12-R13 are hydrogen;

R11 is C1-C6 alkyl, substituted C1-C6 alkyl, or C3-C6 cycloalkyl;

R14-R18 are each independently hydrogen, fluorine, substituted methoxy, or substituted ethyl.

[0030] In some aspects, the C1-C6 alkyl is methyl.

[0031] In some aspects, the substituted C1-C6 alkyl is substituted with one or more of cyano, halogen, or alkoxy.

[0032] In some aspects, the substituted C1-C6 alkyl is trifluoromethyl.

[0033] In some aspects, the substituted C1-C6 alkyl is isobutyronitrile, tri-fluoroethyl, tri- fluoropropyl, or (trifluoromethoxy)ethyl.

[0034] In some aspects, C is aryl or substituted aryl with Formula (l-d-8), Formula (l-e-1 ), or Formula (l-f-1 ):

wherein:

R14-R15 are each independently hydrogen or fluorine;

R16 is hydrogen, fluorine, substituted methoxy, or substituted ethyl; and

R17-R18 are hydrogen.

[0035] In some aspects, C is C1-C6 alkyl, substituted C1-C6 alkyl, C3-C6 cycloalkyl, or C3-C6 substituted cycloalkyl. In some aspects, the C1-C6 alkyl is methyl. In some aspects, the substituted C1-C6 alkyl is trifluoromethyl. In some aspects, the C3-C6 substituted cycloalkyl is a bridged cyclobutyl.

[0036] In some aspects, A is pyridine or substituted pyridine and with Formula (l-g): wherein R19 is C1-C6 alkyl, aryl, C3-C6 heterocycloalkyl, substituted C3-C6 heterocycloalkyl, amide, or substituted amide. [0037] In some aspects, R19 is methyl. In some aspects, R19 is amide, methyl amide, or dimethyl amide. In some aspects, R19 is a nitrogen containing C3-C6 heterocycloalkyl.

[0038] In other aspects, A is oxazole with Formula (l-h) or Formula (l-i); wherein R24 is aryl.

[0039] In other aspects, A is oxadiazole and with Formula (l-j) or Formula (l-k): wherein R25 is aryl.

[0040] In other aspects, A is thiadiazole and with Formula (l-l): wherein R26 is aryl.

[0041] In other aspects, A is triazole and with Formula (l-m): wherein R27 is aryl.

DETAILED DESCRIPTION

[0042] Provided are compounds, pharmaceutically acceptable salts of the compounds, and pharmaceutical composition comprising the compounds or their salts for increasing the processivity of POLy.

(I) Definitions

[0043] The term “alkyl” as used herein refers to both branched- and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms in a specified range. For example, the term “C1-C6 alkyl” means linear or branched chain alkyl groups, including all possible isomers, having 1 , 2, 3, 4, 5, or 6 carbon atoms. Furthermore, alkyl groups allow for substituents to be located on any of the carbon atoms. For example, a substituted C3 alkyl group allows for the substituent to be located on any of the three carbon atoms.

[0044] The term “diazole” as used herein refers to a five-membered heterocyclic compound containing three carbon atoms and two nitrogen atoms. Examples of diazoles include imidazole, pyrazole, and the like.

[0045] The term “oxazole” as used herein refers to a five-membered heterocyclic compound containing an oxygen atom and a nitrogen atom separated by one carbon.

[0046] The term “thiazole” as used herein refers to a five-membered heterocyclic compound containing both a sulfur atom and a nitrogen atom.

[0047] The term “thiadiazole” as used herein refers to a five-membered heterocyclic compound containing both a sulfur atom and two nitrogen atoms.

[0048] The term “oxadiazole” as used herein refers to a five-membered heterocyclic compound containing a nitrogen atom and at least one other noncarbon atom in the ring. Examples of oxadiazoles include 1,2,3-oxadiazole, 1 ,2,4-oxadiazole, 1 ,2,5-oxadiazole, 1,3,4-oxadiazole, and the like.

[0049] The term “triazole” as used herein refers to a five-membered heterocyclic compound containing three nitrogen atoms and two double bonds. Four triazole isomers (1 H-1 ,2,3-, 2H- 1 ,2,3-, 1 H-1 , 2, 4-, and 2H-1 ,2,4-) differ by the arrangement of the nitrogen atoms and the locations of their three hydrogen atoms.

[0050] The term “oxetane” as used herein refers to a heterocyclic organic compound having a four-membered ring with three carbon atoms and one oxygen atom.

[0051] The term “alkoxy” or “alkoxyl” as used herein refers to an -O-alkyl group. For example, the term “C1-C4 alkoxyl” means -O-C1-C4 alkyl. Examples of alkoxyl include methoxyl, ethoxyl, propoxyl (e.g., n-propoxyl and isopropoxyl), and the like.

[0052] The term “haloalkoxy” or “haloalkoxyl” as used herein refers to an -O-alkyl group in which at least one of the hydrogen atoms of the alkyl group is replaced with a halogen atom. Examples of haloalkoxyl include trifluoromethoxyl, 2,2,2-trifluoroethoxyl, and the like.

[0053] The term “alkanoyl” or “acyl” as used herein refers to an -C(O)-alkyl group. For example, the term “C1-C6 alkanoyl” means -C(O)-C1-C6 alkyl. Examples of alkanoyl include acetyl, propionyl, butyryl, and the like.

[0054] The term “cycloalkyl” as used herein refers to a cyclized alkyl ring having the indicated number of carbon atoms in a specified range. Thus, for example, “C3-C6 cycloalkyl” encompasses each of cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. [0055] The term “aryl” as used herein refers to a monocyclic or fused bicyclic ring system having the characteristics of aromaticity, wherein at least one ring contains a completely conjugated pi-electron system. Typically, aryl groups contain 6 to 14 carbon atoms (“Ce- C14 aryl”) or preferably, 6 to 12 carbon atoms (“C6-C12 aryl”). Fused aryl groups may include an aryl ring (e.g., a phenyl ring) fused to another aryl ring, or fused to a saturated or partially unsaturated carbocyclic or heterocyclic ring. The point of attachment to the base molecule on such fused aryl ring systems may be a C atom of the aromatic portion or a C or N atom of the non-aromatic portion of the ring system. Examples, without limitation, of aryl groups include phenyl, biphenyl, naphthyl, anthracenyl, indanyl, indenyl, and tetrahydronaphthyl.

[0056] The term “heteroaryl” as used herein refers to (i) a 5- or 6-membered ring having the characteristics of aromaticity containing at least one heteroatom selected from N, O and S, wherein each N is optionally in the form of an oxide, and (ii) a 9- or 10- membered bicyclic fused ring system, wherein the fused ring system of (ii) contains at least one heteroatom independently selected from N, O and S, wherein each ring in the fused ring system contains zero, one or more than one heteroatoms, at least one ring is aromatic, each N is optionally in the form of an oxide, and each S in a ring which is not aromatic is optionally S(O) or S(O)2. Typically, heteroaryl groups contain 5 to 14 ring atoms (“5-14 membered heteroaryl”), and preferably 5 to 12 ring atoms (“5-12 membered heteroaryl”). Heteroaryl rings are attached to the base molecule via a ring atom of the heteroaromatic ring, such that aromaticity is maintained. Suitable 5- and 6- membered heteroaromatic rings include, for example, pyridyl, 3-fluroropyridyl, 4-fluoropyridyl, 3- methoxypyridyl, 4-methoxypyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, thienyl, furanyl, imidazolyl, pyrazolyl, triazolyl (i.e., 1 ,2,3-triazolyl or 1 ,2,4- triazolyl), tetrazolyl, oxazolyl, isooxazolyl, oxadiazolyl (i.e., the 1,2,3-, 1 ,2,4-, 1 ,2,5- (furazanyl), or 1 ,3,4-isomer), oxatriazolyl, thiazolyl, isothiazolyl, and thiadiazolyl. Suitable 9- and 10-membered heterobicyclic, fused ring systems include, for example, benzofuranyl, indolyl, indazolyl, naphthyridinyl, isobenzofuranyl, benzisoxazolyl, benzoxazolyl, benzothiazolyl, chromenyl, quinolinyl, isoquinolinyl, benzopiperidinyl, benzofuranyl, imidazo[1,2-a]pyridinyl, benzotriazolyl, indazolyl, indolinyl, and isoindolinyl.

[0057] The term “pyridine” as used herein refers to a heteroaryl compound in which a carbon atom of benzene is exchanged with nitrogen.

[0058] The term “heteroaryloxy” or “heteroaryloxyl” as used herein refers to an -O- heteroaryl group.

[0059] The term “heterocycle”, “heterocyclyl”, or “heterocyclic” as used herein represents a stable 3- to 10-membered monocyclic, non-aromatic ring that is either saturated or unsaturated, and that consists of carbon atoms and from one to two heteroatoms selected from the group consisting of N, O, and S. Examples include oxiranyl, aziridinyl, oxetanyl, azetidinyl, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl, piperidinyl, 1 ,4-dioxanyl, morpholinyl, piperazinyl, azepanyl, oxepanyl, and oxazepanyl.

[0060] The term “oxo” as used herein refers to a group which consists of oxygen which is double bonded to carbon or any other element.

[0061] The term “carboxyl” as used herein refers to a combination of two functional groups attached to a single carbon atom, namely, hydroxyl (OH) and carbonyl (O).

[0062] The term “substituted”, “substituents”, “optionally substituted”, or “optional substituents”, as used herein means that the groups are either unsubstituted or substituted with one or more of the substituents specified. When the groups are substituted with more than one substituent, the substituents may be the same or different. Furthermore, when using the terms “independently,” “independently are,” and “independently selected from” mean that the groups may be the same or different.

[0063] The term “deuterium” as used herein refers to an isotope of hydrogen that has one proton and one neutron in its nucleus and that has twice the mass of ordinary hydrogen. Deuterium herein is represented by the symbol “D”.

[0064] The term “deuterated” by itself or used to modify a compound or group as used herein refers to the presence of at least one deuterium atom attached to carbon. For example, the term “deuterated compound” refers to a compound which contains one or more carbon-bound deuterium(s). In a deuterated compound of the present invention, when a particular position is designated as having deuterium, it is understood that the abundance of deuterium at that position is substantially greater than the natural abundance of deuterium, which is about 0.015 %.

[0065] The term “undeuterated” or “non-deuterated” as used herein refers to the ratio of deuterium atoms of which is not more than the natural isotopic deuterium content, which is about 0.015 %; in other words, all hydrogen are present at their natural isotopic percentages. Unless otherwise stated, when a position is designated specifically as “H” or “hydrogen”, the position is understood to have hydrogen at its natural abundance isotopic composition.

[0066] The term “pharmaceutically acceptable salt” as used herein refers to a salt that is not biologically or otherwise undesirable (e.g., not toxic or otherwise harmful). A salt of a compound of the invention is formed between an acid and a basic group of the compound, or a base and an acidic group of the compound. For example, when the compounds of the invention contain at least one basic group (i.e., groups that may be protonated), the invention includes the compounds in the form of their acid addition salts with organic or inorganic acids such as, for example, but not limited to salts with hydrogen chloride, hydrogen bromide, phosphoric acid, sulfuric acid, nitric acid, benzenesulfonic acid, acetic acid, citric acid, glutamic acid, lactic acid, and methanesulfonic acid. When compounds of the invention contain one or more acidic groups (e.g., a carboxylic acid), the invention includes the pharmaceutically acceptable salts of the compounds formed with but not limited to alkali metal salts, alkaline earth metal salts or ammonium salts. Examples of such salts include, but are not limited to, sodium salts, potassium salts, calcium salts, magnesium salts or salts with ammonia or organic amines such as, for example, ethylamine, ethanolamine, triethanolamine or amino acids. Additional examples of such salts may be found in Stahl, P. H. et al. Pharmaceutical Salts: Properties, Selection, and Use, 2nd Revised Edition, Wiley, 2011.

(II) Compounds

[0067] In certain embodiments, the present disclosure is directed to a compound, or a pharmaceutically acceptable salt thereof, represented by Formula (I): wherein A is substituted diazole, oxazole, substituted oxazole, thiazole, substituted thiazole, thiadiazole, substituted thiadiazole, oxadiazole, substituted oxadiazole, triazole, substituted triazole, pyridine or substituted pyridine; and wherein when the substituted diazole is a pyrazole the substitution is not an unsubstituted phenyl.

[0068] In some aspects of Formula (I), A may be a substituted diazole. For example, A may be a substituted pyrazole or a substituted imidazole.

[0069] In some embodiments, A may be substituted pyrazole or a substituted imidazole with Formula (l-a) or Formula (l-b) respectively:

[0070] In at least one example, R1-R3 may each independently be hydrogen or C1-C6 alkyl. For example, R1-R3 may be methyl, ethyl, propyl, butyl, pentyl, or hexyl.

[0071] In at least one example, B may be hydrogen, substituted C1-C6 alkyl, C3-C6 cycloalkyl, substituted C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, substituted C3-C6 heterocycloalkyl, C3-C6 heterocyclyl, substituted C3-C6 heterocyclyl, substituted aryl, amide, or substituted amide.

[0072] In at least one example, B may be substituted C1-C6 alkyl. The substituted C1-C6 alkyl may be substituted with one or more of benzyl, fluorine, or dimethyl amine. In one embodiment, the substituted C1-C6 alkyl may be methylbenzene. In one embodiment, the substituted Ci- Ce alkyl may be methylbenzene. In another embodiment, the substituted C1-C6 alkyl may be ethylbenzene. In yet another embodiment, the substituted C1-C6 alkyl may be trifluoroethyl. In yet another embodiment, the substituted C1-C6 alkyl may be difluoroethyl. In yet another embodiment, the substituted C1-C6 alkyl may be N,N-dimethylethanamine.

[0073] In at least one example, B may be substituted aryl with Formula (l-a-1) or Formula (l-b- 1):

[0074] In at least one example, Ri-R₂ may each independently be hydrogen or methyl. In at least one example, R₃ may be hydrogen. In at least one example, R₄ may be hydrogen or halogen. In at least one example, R₅ and R₆ may each independently be hydrogen, halogen, C1-C6 alkyl, C1-C6 substituted alkyl, carboxylic acid, substituted carboxylic acid, sulfonyl, substituted sulfonyl, alkoxy, amide, substituted amide, amine, substituted amine, C3- Ce heterocycloalkyl, substituted C3-C6 heterocycloalkyl, C3-C6 cycloalkyl, or substituted C3- Ce cycloalkyl. In at least one example, R₇ and R_s may each independently be hydrogen or alkoxy. R₄-R₈ may not be hydrogen at the same time.

[0075] In one embodiment, R1 and R₂ may be hydrogen with Formula (l-a-2):

[0076] In at least one example, R₄ may be hydrogen or fluorine. In at least one example, R₅ may be hydrogen, fluorine, bromine, chlorine, methyl, trifluoromethyl, carboxylic acid, methyl substituted carboxylic acid, methoxy, substituted ethyl, amide, methyl amide, dimethyl amide, methyl sulfonyl, N-methyl sulfonyl amide, or substituted C1-C6 alkyl. In at least one example, R₆ may be hydrogen, fluorine, methoxy, methoxy methyl, methoxy ethyl, substituted ethyl, amide, methyl amide, amide, methyl amide, dimethyl amide, substituted alkyl, substituted sulfonyl, cyclobutyl, substituted cyclobutyl, oxetane, substituted oxetane. In at least one example, R? and Rs may each independently be hydrogen or methoxy. R4-R8 may not be hydrogen at the same time.

[0077] The substituted alkyl may be methyl or ethyl substituted with one or more of methyl, amine, methyl amine, dimethyl amide, hydroxyl, or fluorine.

[0078] In one embodiment, R₆ may be substituted oxetane. The substituted oxetane may be 3,3-difluorocyclobutan-1-ol.

[0079] In one embodiment, Re may be substituted cyclobutane. The substituted cyclobutane may be 3,3-difluorocyclobutan-1-ol.

[0080] The substituted sulfonyl may be represented by Formula (l-a-3):

[0081] In Formula (l-a-3), R9 may be methyl, ethyl, trifluoromethyl, cyclopropyl, or methyl amide.

[0082] In at least one example, B may be C3-C6 cycloalkyl or substituted C3-C6 cycloalkyl with Formula (l-a-4):

[0083] R' and R" may each independently be hydrogen or fluorine. [0084] In at least one example, B may be C3-C6 heterocyclyl or substituted C3-C6 heterocyclyl, and with any one of Formula (l-a-5), Formula (l-a-6), Formula (l-a-7), Formula (l-a-8), Formula (l-a-9), Formula (l-a-10), or Formula (l-a-11):

[0085] In some examples of Formulae (l-a-5) to (l-a-11), R₄ and R7R8 may be hydrogen. In some examples of Formulae (l-a-5) to (l-a-11) R5 may be hydrogen, substituted C1-C6 alkyl, or methoxy. In some examples of Formulae (l-a-5) to (l-a-11), R6 may be hydrogen or substituted C1-C6 alkyl.

[0086] In one embodiment, R₅ may be substituted C1-C6 alkyl. The substituted C1-C6 alkyl may be trifluoromethane.

[0087] In one embodiment, R₆ may be substituted C1-C6 alkyl. The substituted C1-C6 alkyl may be substituted with one or more of fluorine or hydroxyl. In one embodiment, R₆ may be 2,2,2- trifluoroethan-1-ol. In another embodiment, R₆ may be propan-2-ol.

[0088] In some aspects, B may be aryl and the compound may be a substituted imidazole with Formula (l-c-1):

[0089] In some embodiments, R₂ and R₃ are hydrogen.

[0090] In some aspects, A may be substituted thiazole with any one of Formula (l-d), Formula

(l-e), or Formula (l-f): [0091] C may be hydrogen, C1-C6 alkyl, substituted C1-C6 alkyl, C3-C6 cycloalkyl, substituted C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, substituted C3-C6 heterocycloalkyl, C3-C6 heterocyclyl, substituted C3-C6 heterocyclyl, aryl, substituted aryl, amide, or substituted amide.

[0092] In some embodiments, C may be C3-C6 heterocyclyl or substituted C3-C6 heterocyclyl, with any one of Formula (l-d-1), Formula (l-d-2), Formula (l-d-3), Formula (l-d-4), Formula (I- d-5), Formula (l-d-6), or Formula (l-d-7):

[0093] In at least one example, R10 and R12-R13 may be hydrogen.

[0094] In at least one example, R11 may be C1-C6 alkyl, substituted C1-C6 alkyl, or C3-C6 cycloalkyl.

[0095] In at least one example, R14-R18 may each independently be hydrogen, fluorine, substituted methoxy, or substituted ethyl.

[0096] In some embodiments, the C1-C6 alkyl may be methyl. [0097] In some embodiments, the substituted C1-C6 alkyl may be substituted with one or more of cyano, halogen, or alkoxy.

[0098] In some embodiments, the substituted C1-C6 alkyl may be trifluoromethyl.

[0099] In some embodiments, the substituted C1-C6 alkyl may be isobutyronitrile, trifluoroethyl, tri-fluoropropyl, or (trifluoromethoxy)ethyl.

[0100] In certain embodiments, C is aryl or substituted aryl with Formula (l-d-8), Formula (l-e- 1), or Formula (l-f-1):

[0101] In at least one example, R14-R15 may each independently be hydrogen or fluorine.

[0102] In at least one example, R16 may be hydrogen, fluorine, substituted methoxy, or substituted ethyl.

[0103] In at least one example, R17-R18 may be hydrogen.

[0104] In some embodiments, C may be C1-C6 alkyl, substituted C1-C6 alkyl, C3-C6 cycloalkyl, or C3-C6 substituted cycloalkyl. The C3-C6 substituted cycloalkyl may be a bridged cyclobutyl. The C1-C6 alkyl may be methyl, ethyl, propyl, butyl, pentyl, or hexyl. The substituted C1-C6 alkyl may be trifluoromethyl

[0105] In some aspects of Formula (I), A may be pyridine or substituted pyridine with Formula (l-g):

[0106] In at least one example, R19 may be C1-C6 alkyl, aryl, C3-C6 heterocycloalkyl, substituted C3-C6 heterocycloalkyl, amide, or substituted amide. In some embodiments, R19 may be methyl. In some embodiments, R19 may be amide, methyl amide, or dimethyl amide. In some embodiments, R19 is a nitrogen containing C3-C6 heterocycloalkyl.

[0107] In some aspects of Formula (I), A may be an oxazole with Formula (l-h) or Formula (I-

[0108] In at least one example, R24 may be aryl.

[0109] In some aspects of Formula (I), A may be an oxadiazole with Formula (l-j) or Formula

(l-k):

[0110] In at least one example, R25 may be aryl.

[0111] In some aspects of Formula (I), A may be a thiadiazole with Formula (l-l):

[0112] In at least one example, R26 may be aryl.

[0113] In some aspects of Formula (I), A may be a triazole with Formula (l-m):

[0114] In at least one example, R27 may be aryl.

[0115] In certain embodiments, the compounds inhibits POL y.

[0116] In certain embodiments, the compounds promote POL y.

[0117] In one embodiment, the compound is Compound 1 , or a pharmaceutically acceptable salt thereof:

[0118] In one embodiment, the compound is Compound 2, or a pharmaceutically acceptable salt thereof:

[0119] In one embodiment, the compound is Compound 3, or a pharmaceutically acceptable salt thereof:

[0120] In one embodiment, the compound is Compound 4, or a pharmaceutically acceptable salt thereof: [0121] In one embodiment, the compound is Compound 5, or a pharmaceutically acceptable salt thereof:

[0122] In one embodiment, the compound is Compound 6, or pharmaceutically acceptable salt thereof:

[0123] In one embodiment, the compound is Compound 7, or pharmaceutically acceptable salt thereof:

[0124] In one embodiment, the compound is Compound 8, or a pharmaceutically acceptable salt thereof:

[0125] In one embodiment, the compound is Compound 9 (Example 23), or a pharmaceutically acceptable salt thereof:

[0126] In one embodiment, the compound is Compound 10 (Example 21), or a pharmaceutically acceptable salt thereof:

[0127] In one embodiment, the compound is Compound 11 (Example 22), or a pharmaceutically acceptable salt thereof:

[0128] In one embodiment, the compound is Compound 12, or a pharmaceutically acceptable salt thereof:

[0129] In one embodiment, the compound is Compound 13, or a pharmaceutically acceptable salt thereof: [0130] In one embodiment, the compound is Compound 14, or a pharmaceutically acceptable salt thereof:

[0131] In one embodiment, the compound is Compound 15, or a pharmaceutically acceptable salt thereof:

[0132] In one embodiment, the compound is Compound 16, or a pharmaceutically acceptable salt thereof:

[0133] In one embodiment, the compound is Compound 17, or a pharmaceutically acceptable salt thereof:

[0134] In one embodiment, the compound is Compound 18, or a pharmaceutically acceptable salt thereof:

[0135] In one embodiment, the compound is Compound 19, or a pharmaceutically acceptable salt thereof:

[0136] In one embodiment, the compound is Compound 20, or a pharmaceutically acceptable salt thereof:

[0137] In one embodiment, the compound is Compound 21 , or a pharmaceutically acceptable salt thereof:

[0138] In one embodiment, the compound is Compound 22, or a pharmaceutically acceptable salt thereof:

[0139] In one embodiment, the compound is Compound 23, or a pharmaceutically acceptable salt thereof:

[0140] In one embodiment, the compound is Compound 24, or a pharmaceutically acceptable salt thereof:

[0141] In one embodiment, the compound is Compound 25, or a pharmaceutically acceptable salt thereof:

[0142] In one embodiment, the compound is Compound 26, or a pharmaceutically acceptable salt thereof:

[0143] In one embodiment, the compound is Compound 27 (Example 3), or a pharmaceutically acceptable salt thereof:

[0144] In one embodiment, the compound is Compound 28 (Example 2), or a pharmaceutically acceptable salt thereof:

[0145] In one embodiment, the compound is Compound 29 (Example 5), or a pharmaceutically acceptable salt thereof:

[0146] In one embodiment, the compound is Compound 30 (Example 4), or a pharmaceutically acceptable salt thereof:

[0147] In one embodiment, the compound is Compound 31 (Example 19), or a pharmaceutically acceptable salt thereof:

[0148] In one embodiment, the compound is Compound 32 (Example 20), or a pharmaceutically acceptable salt thereof: [0149] In one embodiment, the compound is Compound 33, or a pharmaceutically acceptable salt thereof:

[0150] In one embodiment, the compound is Compound 34, or a pharmaceutically acceptable salt thereof:

[0151] In one embodiment, the compound is Compound 35, or a pharmaceutically acceptable salt thereof:

[0152] In one embodiment, the compound is Compound 36, or a pharmaceutically acceptable salt thereof:

[0153] In one embodiment, the compound is Compound 37 (Example 1), or a pharmaceutically acceptable salt thereof: [0154] In one embodiment, the compound is Compound 38, or a pharmaceutically acceptable salt thereof:

[0155] In one embodiment, the compound is Compound 39, or a pharmaceutically acceptable salt thereof:

[0156] In one embodiment, the compound is Compound 40 (Example 17), or a pharmaceutically acceptable salt thereof:

[0157] In one embodiment, the compound is Compound 41 (Example 16), or a pharmaceutically acceptable salt thereof:

[0158] In one embodiment, the compound is Compound 42, or a pharmaceutically acceptable salt thereof: [0159] In one embodiment, the compound is Compound 43, or a pharmaceutically acceptable salt thereof:

[0160] In one embodiment, the compound is Compound 44, or a pharmaceutically acceptable salt thereof:

[0161] In one embodiment, the compound is Compound 45, or a pharmaceutically acceptable salt thereof:

[0162] In one embodiment, the compound is Compound 46, or a pharmaceutically acceptable salt thereof:

[0163] In one embodiment, the compound is Compound 47 (Example 18), or a pharmaceutically acceptable salt thereof:

[0164] In one embodiment, the compound is Compound 48, or a pharmaceutically acceptable salt thereof:

[0165] In one embodiment, the compound is Compound 49, or a pharmaceutically acceptable salt thereof:

[0166] In one embodiment, the compound is Compound 50, or a pharmaceutically acceptable salt thereof:

[0167] In one embodiment, the compound is Compound 51 , or a pharmaceutically acceptable salt thereof: [0168] In one embodiment, the compound is Compound 52, or a pharmaceutically acceptable salt thereof:

[0169] In one embodiment, the compound is Compound 53, or a pharmaceutically acceptable salt thereof:

[0170] In one embodiment, the compound is Compound 54, or a pharmaceutically acceptable salt thereof:

[0171] In one embodiment, the compound is Compound 55, or a pharmaceutically acceptable salt thereof:

[0172] In one embodiment, the compound is Compound 56, or a pharmaceutically acceptable salt thereof:

[0173] In one embodiment, the compound is Compound 57, or a pharmaceutically acceptable salt thereof:

[0174] In one embodiment, the compound is Compound 58, or a pharmaceutically acceptable salt thereof:

[0175] In one embodiment, the compound is Compound 59, or a pharmaceutically acceptable salt thereof:

[0176] In one embodiment, the compound is Compound 60, or a pharmaceutically acceptable salt thereof:

[0177] In one embodiment, the compound is Compound 61 , or a pharmaceutically acceptable salt thereof:

[0178] In one embodiment, the compound is Compound 62, or a pharmaceutically acceptable salt thereof:

[0179] In one embodiment, the compound is Compound 63, or a pharmaceutically acceptable salt thereof:

[0180] In one embodiment, the compound is Compound 64, or a pharmaceutically acceptable salt thereof:

[0181] In one embodiment, the compound is Compound 65, or a pharmaceutically acceptable salt thereof:

[0182] In one embodiment, the compound is Compound 66, or a pharmaceutically acceptable salt thereof: [0183] In one embodiment, the compound is Compound 67, or a pharmaceutically acceptable salt thereof:

[0184] In one embodiment, the compound is Compound 68, or a pharmaceutically acceptable salt thereof:

[0185] In one embodiment, the compound is Compound 69, or a pharmaceutically acceptable salt thereof:

[0186] In one embodiment, the compound is Compound 70, or a pharmaceutically acceptable salt thereof:

[0187] In one embodiment, the compound is Compound 71 , or a pharmaceutically acceptable salt thereof:

[0188] In one embodiment, the compound is Compound 72, or a pharmaceutically acceptable salt thereof:

[0189] In one embodiment, the compound is Compound 73, or a pharmaceutically acceptable salt thereof:

[0190] In one embodiment, the compound is Compound 74, or a pharmaceutically acceptable salt thereof:

[0191] In one embodiment, the compound is Compound 75, or a pharmaceutically acceptable salt thereof:

[0192] In one embodiment, the compound is Compound 76, or a pharmaceutically acceptable salt thereof:

[0193] In one embodiment, the compound is Compound 77, or a pharmaceutically acceptable salt thereof:

[0194] In one embodiment, the compound is Compound 78, or a pharmaceutically acceptable salt thereof:

[0195] In one embodiment, the compound is Compound 79, or a pharmaceutically acceptable salt thereof:

[0196] In one embodiment, the compound is Compound 80, or a pharmaceutically acceptable salt thereof:

[0197] In one embodiment, the compound is Compound 81 , or a pharmaceutically acceptable salt thereof:

[0198] In one embodiment, the compound is Compound 82, or a pharmaceutically acceptable salt thereof:

[0199] In one embodiment, the compound is Compound 83, or a pharmaceutically acceptable salt thereof:

[0200] In one embodiment, the compound is Compound 84, or a pharmaceutically acceptable salt thereof:

[0201] In one embodiment, the compound is Compound 85, or a pharmaceutically acceptable salt thereof:

[0202] In one embodiment, the compound is Compound 86, or a pharmaceutically acceptable salt thereof:

[0203] In one embodiment, the compound is Compound 87, or a pharmaceutically acceptable salt thereof:

[0204] In one embodiment, the compound is Compound 88, or a pharmaceutically acceptable salt thereof:

[0205] In one embodiment, the compound is Compound 89, or a pharmaceutically acceptable salt thereof:

[0206] In one embodiment, the compound is Compound 90, or a pharmaceutically acceptable salt thereof:

[0207] In one embodiment, the compound is Compound 91 , or a pharmaceutically acceptable salt thereof:

[0208] In one embodiment, the compound is Compound 92, or a pharmaceutically acceptable salt thereof:

[0209] In one embodiment, the compound is Compound 93, or a pharmaceutically acceptable salt thereof:

[0210] In one embodiment, the compound is Compound 94, or a pharmaceutically acceptable salt thereof:

[0211] In one embodiment, the compound is Compound 95, or a pharmaceutically acceptable salt thereof:

[0212] In one embodiment, the compound is Compound 96, or a pharmaceutically acceptable salt thereof:

[0213] In one embodiment, the compound is Compound 97, or a pharmaceutically acceptable salt thereof:

[0214] In one embodiment, the compound is Compound 98, or a pharmaceutically acceptable salt thereof:

[0215] In one embodiment, the compound is Compound 99, or a pharmaceutically acceptable salt thereof:

[0216] In one embodiment, the compound is Compound 100, or a pharmaceutically acceptable salt thereof:

[0217] In one embodiment, the compound is Compound 101 , or a pharmaceutically acceptable salt thereof:

[0218] In one embodiment, the compound is Compound 102, or a pharmaceutically acceptable salt thereof: [0219] In one embodiment, the compound is Compound 103, or a pharmaceutically acceptable salt thereof:

[0220] In one embodiment, the compound is Compound 104, or a pharmaceutically acceptable salt thereof:

[0221] In one embodiment, the compound is Compound 105, or a pharmaceutically acceptable salt thereof:

[0222] In one embodiment, the compound is Compound 106, or a pharmaceutically acceptable salt thereof:

[0223] In one embodiment, the compound is Compound 107, or a pharmaceutically acceptable salt thereof: [0224] In one embodiment, the compound is Compound 108, or a pharmaceutically acceptable salt thereof:

[0225] In one embodiment, the compound is Compound 109, or a pharmaceutically acceptable salt thereof:

[0226] In one embodiment, the compound is Compound 110, or a pharmaceutically acceptable salt thereof:

[0227] In one embodiment, the compound is Compound 111 , or a pharmaceutically acceptable salt thereof:

[0228] In one embodiment, the compound is Compound 112, or a pharmaceutically acceptable salt thereof: [0229] In one embodiment, the compound is Compound 113, or a pharmaceutically acceptable salt thereof:

[0230] In one embodiment, the compound is Compound 114, or a pharmaceutically acceptable salt thereof:

[0231] In one embodiment, the compound is Compound 115a, or a pharmaceutically acceptable salt thereof:

[0232] In one embodiment, the compound is Compound 115b, or a pharmaceutically acceptable salt thereof:

[0233] In one embodiment, the compound is Compound 116, or a pharmaceutically acceptable salt thereof: [0234] In one embodiment, the compound is Compound 117, or a pharmaceutically acceptable salt thereof:

[0235] In one embodiment, the compound is Compound 118, or a pharmaceutically acceptable salt thereof:

[0236] In one embodiment, the compound is Compound 119, or a pharmaceutically acceptable salt thereof:

[0237] In one embodiment, the compound is Compound 120, or a pharmaceutically acceptable salt thereof:

[0238] In one embodiment, the compound is Compound 121 , or a pharmaceutically acceptable salt thereof:

[0239] In one embodiment, the compound is Compound 122, or a pharmaceutically acceptable salt thereof:

[0240] In one embodiment, the compound is Compound 123, or a pharmaceutically acceptable salt thereof:

[0241] In one embodiment, the compound is Compound 124, or a pharmaceutically acceptable salt thereof:

[0242] In one embodiment, the compound is Compound 125, or a pharmaceutically acceptable salt thereof:

[0243] In one embodiment, the compound is Compound 126, or a pharmaceutically acceptable salt thereof:

[0244] In one embodiment, the compound is Compound 127, or a pharmaceutically acceptable salt thereof:

[0245] In one embodiment, the compound is Compound 128, or a pharmaceutically acceptable salt thereof:

[0246] In one embodiment, the compound is Compound 129 (Example 14), or a pharmaceutically acceptable salt thereof:

[0247] In one embodiment, the compound is Compound 130 (Example 13), or a pharmaceutically acceptable salt thereof:

[0248] In one embodiment, the compound is Compound 131 (Example 15), or a pharmaceutically acceptable salt thereof:

[0249] In one embodiment, the compound is Compound 132, or a pharmaceutically acceptable salt thereof:

[0250] In one embodiment, the compound is Compound 133, or a pharmaceutically acceptable salt thereof:

[0251] In one embodiment, the compound is Compound 134 (Example 11), or a pharmaceutically acceptable salt thereof:

[0252] In one embodiment, the compound is Compound 135 (Example 10), or a pharmaceutically acceptable salt thereof:

[0253] In one embodiment, the compound is Compound 136 (Example 12), or a pharmaceutically acceptable salt thereof:

[0254] In one embodiment, the compound is Compound 137 (Example 9), or a pharmaceutically acceptable salt thereof:

[0255] In one embodiment, the compound is Compound 138 (Example 8), or a pharmaceutically acceptable salt thereof:

[0256] In one embodiment, the compound is Compound 139 (Example 7), or a pharmaceutically acceptable salt thereof:

[0257] In one embodiment, the compound is Compound 140 (Example 6), or a pharmaceutically acceptable salt thereof:

[0258] In one embodiment, the compound is Compound 141 , or a pharmaceutically acceptable salt thereof:

[0259] The compounds of the present invention may contain asymmetric carbon atoms (sometimes as the result of a deuterium atom) and thereby may exist as either individual stereoisomers or mixtures of the enantiomers or mixtures of diastereomers. Accordingly, a compound of the present invention may exist as either a racemic mixture, a mixture of diastereomers, or as individual stereoisomers that are substantially free of other stereoisomers. Synthetic, separation, or purification methods to be used to obtain an enantiomer of a given compound are known in the art and are applicable for obtaining the compounds identified herein. [0260] Unless otherwise indicated, when a disclosed compound is named or depicted by a structure without specifying the stereochemistry and has one or more chiral centers, it is understood to represent all possible stereoisomers of the compound. Carbon atoms labelled with * or ** refer to a compound that is chiral but the absolute stereochemistry has not been determined.

[0261] The compounds of the present invention may contain double bonds that may exist in more than one geometric isomer. Examples of such double bonds are carbon- carbon double bonds which form alkenes. In the case of carbon-carbon double bonds, the geometric isomers may be E or Z isomers.

[0262] Unless otherwise indicated, when a disclosed compound is named or depicted by a structure without specifying the geometric isomerism and has one or more possible geometric isomers, it is understood to represent all possible geometric isomers of the compound.

[0263] Certain compounds of the present invention may be able to exist as tautomers. All tautomeric forms of these compounds, whether isolated individually or in mixtures, are within the scope of the present invention. For example, in instances where an — OH substituent is permitted on a heteroaromatic ring and ketoenol tautomerism is possible, it is understood that the substituent might in fact be present, in whole or in part, in the oxo (=0) form.

[0264] Compounds of the present invention may exist in amorphous form and/or one or more crystalline forms. As such all amorphous and crystalline forms and mixtures thereof of the compounds of the invention are intended to be included within the scope of the present invention. In addition, some of the compounds of the present invention may form solvates with water (i.e. , a hydrate) or common organic solvents. Such solvates and hydrates, particularly the pharmaceutically acceptable solvates and hydrates, of the compounds of this invention are likewise encompassed within the scope of the compounds of the invention and the pharmaceutically acceptable salts thereof, along with un-solvated and anhydrous forms of such compounds.

[0265] In one embodiment, deuterium isotope content at the deuterium substituted position is greater than the natural isotopic deuterium content (0.015%), more preferably greater than 50%, more preferably greater than 60%, more preferably greater than 75%, more preferably greater than 90%, more preferably greater than 95%, more preferably greater than 97%, more preferably greater than 99%. It will be understood that some variation of natural isotopic abundance may occur in any compound depending upon the source of the reagents used in the synthesis. Thus, a preparation of undeuterated compounds may inherently contain small amounts of deuterated isotopologues, such amounts being insignificant as compared to the degree of stable isotopic substitution of the deuterated compounds of the invention. See, e.g., Gannes, L Z et al., Comp Biochem Physiol Mol Integr Physiol, 119, 725 (1998). Replacement of hydrogen with deuterium may affect the activity, toxicity, and pharmacokinetics (e.g., absorption, distribution, metabolism, and excretion (“ADME”)) of some drugs. For instance, such replacement may alter the chemical stability and biochemical reactivity of a compound through kinetic isotope effects. Because of the increased mass of deuterium relative to hydrogen, epimerization at stereogenic carbons may be slowed down when hydrogen is replaced with deuterium. See Pirali et al, J. Med. Chem.62, 5276-97 (2019). Additionally, the presence of deuterium may affect how a molecule interacts with enzymes, thereby impacting enzyme kinetics. While in certain cases the increased mass of deuterium as compared to hydrogen may stabilize a compound and thereby improve activity, toxicity, or half-life, such impact is not predictable. In other instances deuteration may have little to no impact on these properties, or may affect them in an undesirable manner. Whether and/or how such replacement will impact drug properties may only be determined if the drug is synthesized, evaluated, and compared to its non-deuterated counterpart. Because some drugs have multiple sites of metabolism or more than one active sites for binding to a target, it is unpredictable as to which sites may benefit by deuterium replacement or to what extent isotope enrichment is necessary to produce a beneficial effect.

(Ill) Methods of making compounds

[0266] The starting materials and reagents used in each step in the preparation are known and may be readily prepared or purchased from commercial sources.

[0267] The compound obtained in each step may also be used for the next reaction as a reaction mixture thereof or after obtaining a crude product thereof. Alternatively, the compound obtained in each step may be isolated and/or purified from the reaction mixture by a separation means such as concentration, crystallization, recrystallization, distillation, solvent extraction, fractionation, chromatography and the like according to a conventional method. [0268] In each reaction step, while the reaction time varies depending on the reagents and solvents to be used, unless otherwise specified, it is generally 1 min. to 48 h., preferably 10 min. to 8 h. In the reaction of each step, while the reaction temperature varies depending on the reagents and solvents to be used, unless otherwise specified, it is generally -78 °C to 300 °C, preferably -78 °C to 150 °C. In the reaction of each step, unless otherwise specified, a reagent is used in 0.5 equivalent to 20 equivalents, preferably 0.8 equivalent to 5 equivalents, relative to the substrate. When a reagent is used as a catalyst, the reagent is used in 0.001 equivalent to 1 equivalent, preferably 0.01 equivalent to 0.2 equivalent, relative to the substrate. When the reagent is also a reaction solvent, the reagent is used in a solvent amount.

[0269] In the reaction of each step, unless otherwise specified, it is performed without solvent or by dissolving or suspending in a suitable solvent. Specific examples of the solvent include the following. Alcohols: methanol, ethanol, tert-butyl alcohol, 2- methoxyethanol and the like; ethers: diethyl ether, diphenyl ether, tetrahydrofuran, 1,2- dimethoxyethane and the like; aromatic hydrocarbons: chlorobenzene, toluene, xylene and the like; saturated hydrocarbons: cyclohexane, hexane and the like; amides: N,N- dimethylformamide, N-methylpyrrolidone and the like; halogenated hydrocarbons: dichloromethane, carbon tetrachloride and the like; nitriles: acetonitrile and the like; sulfoxides: dimethyl sulfoxide and the like; aromatic organic bases: pyridine and the like; acid anhydrides: acetic anhydride and the like; organic acids: formic acid, acetic acid, trifluoroacetic acid and the like; inorganic acids: hydrochloric acid, sulfuric acid and the like; esters: ethyl acetate and the like; ketones: acetone, methyl ethyl ketone and the like; and water.

[0270] Two or more kinds of the above-mentioned solvents may be used by mixing at an appropriate ratio.

[0271] Unless otherwise specified, the reaction of each step is performed according to a known method, for example, the methods described in “Reactions and Syntheses: In the Organic Chemistry Laboratory 2nd Edition” (Lutz F. Tietze, Theophil Eicher, Ulf Diederichsen, Andreas Speicher, Nina Schutzenmeister) Wiley, 2015; “Organic Syntheses Collective Volumes 1 - 12” (John Wiley & Sons Inc); “Comprehensive Organic Transformations, Third Edition” (Richard C. Larock) Wiley, 2018 and the like.

[0272] In each step, protection or deprotection of a functional group is performed by a known method, for example, the methods described in “Protective Groups in Organic Synthesis, 4^th Ed.” (Theodora W. Greene, Peter G. M. Wuts) Wiley-lnterscience, 2007; “Protecting Groups 3rd Ed.” (P. J. Kocienski) Thieme, 2004 and the like. [0273] Deuterated POLy modulators of the present invention may be prepared using chemical reactions known to a person of ordinary skill in the art using deuterated starting materials or reagents. Deuterium-containing reagents are well known in the art and may be prepared using known procedures or purchased from commercial sources. The deuterated compounds obtained may be characterized by analytical techniques known to persons of ordinary skill in the art. For example, nuclear magnetic resonance (“NMR”) may be used to determine a compound’s structure while mass spectroscopy (“MS”) may be used to determine the amount of deuterium atom in the compound by comparison to its non-deuterated form.

(IV) Compositions

[0274] The present invention further includes pharmaceutical compositions of the compounds or a pharmaceutically acceptable salt of said compounds. In addition to the compound of the invention or a salt thereof, the pharmaceutical compositions comprise one or more pharmaceutically acceptable excipients, such excipients being compatible with other ingredients in the composition and also being not toxic or otherwise harmful. Examples of excipients include carriers, lubricants, binders, disintegrants, solvents, solubilizing agents, suspending agents, isotonic agents, buffers, soothing agents, preservatives, antioxidants, colorants, taste-modifying agents, absorbents, and/or wetting agents.

[0275] The pharmaceutical compositions of the invention include those suitable for oral, rectal, nasal, topical, buccal, sublingual, vaginal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration. Such compositions may be prepared by any methods well known in the art of pharmaceutical formulations and pharmacy. See, e.g., Remington: The Science and Practice of Pharmacy, Elsevier Science, 23^rd ed. (2020). Formulations for parenteral administration include sterile aqueous or non- aqueous solutions, suspensions, or emulsions. A variety of aqueous carriers may be used, e.g., water, buffered water, saline, and the like. Examples of other suitable vehicles include polypropylene glycol, polyethylene glycol, vegetable oils, hydrogels, gelatin, hydrogenated naphthalenes, and injectable organic esters, such as ethyl oleate. Such formulations may also contain auxiliary substances, such as preserving, wetting, buffering, emulsifying, and/or dispersing agents. Biocompatible, biodegradable lactide polymer, lactide/glycolide copolymer, or polyoxyethylenepolyoxypropylene copolymers may be used to control the release of the active ingredients. Alternatively, the compositions may be administered by oral ingestion. Compositions intended for oral use may be prepared in solid or liquid forms, according to any method known to a person of ordinary skill in the art for the manufacture of pharmaceutical compositions. Solid dosage forms for oral administration include capsules (both soft and hard gelatin capsules), tablets, powders, and granules. Generally, these pharmaceutical preparations contain active ingredients admixed with pharmaceutically acceptable excipients. These excipients include, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, sucrose, glucose, mannitol, cellulose, starch, calcium phosphate, sodium phosphate, kaolin and the like; binding agents, buffering agents, and/or lubricating agents (e.g., magnesium stearate) may also be used. Tablets and capsules may additionally be prepared with release-controlling coatings such as enteric coatings. The compositions may optionally contain sweetening, flavoring, coloring, perfuming, and preserving agents in order to provide a more palatable preparation.

[0276] In another embodiment, a pharmaceutical composition of this invention further comprises a second therapeutic agent. The second therapeutic agent may be selected from any pharmaceutically active compound; preferably the second therapeutic agent is known to correct mitochondrial dysfunction. Alternatively, the compounds of the invention and second therapeutic agent may be administered together (within less than 24 hours of one another, consecutively or simultaneously) but in separate pharmaceutical compositions. In certain embodiments, the compounds on the invention and second therapeutic agent may be administered separately (e.g., more than 24 hours of one another.) If the second therapeutic agent acts synergistically with the compounds of this invention, the therapeutically effective amount of such compounds and/or the second therapeutic agent may be less that such amount required when either is administered alone.

[0277] For the treatment of cancer, the compounds described herein may be administered in combination with a chemotherapeutic agent. Therapeutically effective amounts of the additional chemotherapeutic agent(s) are well known to those skilled in the art. However, it is well within the attending physician to determine the amount of other chemotherapeutic agent(s) to be delivered. Examples of these chemotherapeutic agents include, but are not limited to, Abitrexate (Methotrexate Injection), Abraxane (Paclitaxel Injection), Actemra (Tocilizumab), Adcetris (Brentuximab Vedotin Injection), Adriamycin (Doxorubicin), Adrucil Injection (5-FU (fluorouracil)), Afinitor (Everolimus), Afinitor Disperz (Everolimus), Aldara (Imiquimod), Alimta (PEMET EXED), Alkeran Injection (Melphalan Injection), Alkeran Tablets (Melphalan), Aredia (Pamidronate), Arimidex (Anastrozole), Aromasin (Exemestane), Arranon (Nelarabine), Arzerra (Ofatumumab Injection), Avastin (Bevacizumab), Avelumab, Bexxar (Tositumomab), BiCNU (Carmustine), Blenoxane (Bleomycin), Blincyto (Blinatumomab), Bosulif (Bosutinib), Busulfex Injection (Busulfan Injection), Campath (Alemtuzumab), Camptosar (Irinotecan), Caprelsa (Vandetanib), Casodex (Bicalutamide), CeeNU (Lomustine), CeeNU Dose Pack (Lomustine), Cerubidine (Daunorubicin), Clolar (Clofarabine Injection), Cometriq (Cabozantinib), Cosmegen (Dactinomycin), CytosarU (Cytarabine), Cytoxan (Cytoxan), Cytoxan Injection (Cyclophosphamide Injection), Cyramza (Ramucirumab), Dacogen (Decitabine), Darzalex (Daratumumab), DaunoXome (Daunorubicin Lipid Complex Injection), Decadron (Dexamethasone), DepoCyt (Cytarabine Lipid Complex Injection), Dexamethasone Intensol (Dexamethasone), Dexpak Taperpak (Dexamethasone), Docefrez (Docetaxel), Doxil (Doxorubicin Lipid Complex Injection), Droxia (Hydroxyurea), DTIC (Decarbazine), Durvalumab, Eligard (Leuprolide), Ellence (Ellence (epirubicin)), Eloxatin (Eloxatin (oxaliplatin)), Elspar (Asparaginase), Emcyt (Estramustine), Empliciti (Elotuzumab), Enhertu (fam- trastuzumab deruxtecan-nxki), Erbitux (Cetuximab), Erivedge (Vismodegib), Erwinaze (Asparaginase Erwinia chrysanthemi), Ethyol (Amifostine), Etopophos (Etoposide Injection), Eulexin (Flutamide), Fareston (Toremifene), Faslodex (Fulvestrant), Femara (Letrozole), Firmagon (Degarelix Injection), Fludara (Fludarabine), Folex (Methotrexate Injection), Folotyn (Pralatrexate Injection), FUDR (FUDR (floxuridine)), Gazyva (Obinutuzumab), Gemzar (Gemcitabine), Gilotrif (Afatinib), Gleevec (Imatinib Mesylate), Gliadel Wafer (Carmustine wafer), Halaven (Eribulin Injection), Herceptin (Trastuzumab), Hexalen (Altretamine), Hycamtin (Topotecan), Hycamtin (Topotecan), Hydrea (Hydroxyurea), Iclusig (Ponatinib), Idamycin PFS (Idarubicin), Ifex (Ifosfamide), Inlyta (Axitinib), Intron A alfab (Interferon alfa-2a), Iressa (Gefitinib), Istodax (Romidepsin Injection), Ixempra (Ixabepilone Injection), Jakafi (Ruxolitinib), Jevtana (Cabazitaxel Injection), Kadcyla (Ado-trastuzumab Emtansine), Kyprolis (Carfilzomib), Leflunomide (SU101), Lartruvo (Olaratumab), Leukeran (Chlorambucil), Leukine (Sargramostim), Leustatin (Cladribine), Libtayo (Cemiplimab), Lupron (Leuprolide), Lupron Depot (Leuprolide), Lupron DepotPED (Leuprolide), Lysodren (Mitotane), Marqibo Kit (Vincristine Lipid Complex Injection), Matulane (Procarbazine), Megace (Megestrol), Mekinist 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Removab (Catumaxomab), Rheumatrex (Methotrexate), Rituxan (Rituximab), RoferonA alfaa (Interferon alfa-2a), Rubex (Doxorubicin), Sandostatin (Octreotide), Sandostatin LAR Depot (Octreotide), Sarclisa (Isatuximab-irfc), Soltamox (Tamoxifen), Sprycel (Dasatinib), Sterapred (Prednisone), Sterapred DS (Prednisone), Stivarga (Regorafenib), Supprelin LA (Histrelin Implant), Sutent (Sunitinib), Sylatron (Peginterferon Alfa-2b Injection (Sylatron)), Synribo (Omacetaxine Injection), Tabloid (Thioguanine), Taflinar (Dabrafenib), Tarceva (Erlotinib), Targretin Capsules (Bexarotene), Tasigna (Decarbazine), Taxol (Paclitaxel Injection), Taxotere (Docetaxel), Tecentriq (Atezolizumab), Temodar (Temozolomide), Temodar (Temozolomide Injection), Tepadina (Thiotepa), Thalomid (Thalidomide), TheraCys BCG (BCG), Thioplex (Thiotepa), TICE BCG (BCG), Toposar (Etoposide Injection), Torisel (Temsirolimus), Treanda (Bendamustine hydrochloride), Tremelimumab, Trelstar (Triptorelin Injection), Trexall (Methotrexate), Trisenox (Arsenic trioxide), Tykerb (lapatinib), Unituxin (Dinutuximab), Valstar (Valrubicin Intravesical), Vantas (Histrelin Implant), Vectibix (Panitumumab), Velban (Vinblastine), Velcade (Bortezomib), Vepesid (Etoposide), Vepesid (Etoposide Injection), Vesanoid (Tretinoin), Vidaza (Azacitidine), Vincasar PFS (Vincristine), Vincrex (Vincristine), Votrient (Pazopanib), Vumon (Teniposide), Wellcovorin IV (Leucovorin Injection), Xalkori (Crizotinib), Xeloda (Capecitabine), Xtandi (Enzalutamide), Yervoy (Ipilimumab Injection), Zaltrap (Ziv-aflibercept Injection), Zanosar (Streptozocin), Zelboraf (Vemurafenib), Zevalin (Ibritumomab Tiuxetan), Zoladex (Goserelin), Zolinza (Vorinostat), Zometa (Zoledronic acid), Zortress (Everolimus), Zytiga (Abiraterone), Nimotuzumab and immune checkpoint inhibitors such as nivolumab, pembrolizumab/MK-3475, pidilizumab and AMP-224 targeting PD-1 ; and BMS-935559, MEDI4736, MPDL3280A and MSB0010718C targeting.

[0278] The examples and preparations provided below further illustrate and exemplify the compounds of the present invention and methods of preparing such compounds. It is to be understood that the scope of the present invention is not limited in any way by the scope of the following examples and preparations.

[0279] The structures of the compounds are confirmed by either mass spectrometry or nuclear magnetic resonance spectroscopy (NMR), where peaks assigned to the characteristic protons in the title compound are presented where appropriate. ¹H NMR shift (5H) are given in parts per million (ppm) down field from an internal reference standard. Acceptable ¹H NMR shifts and masses are within 5% of the reported values (i.e. , all reported measurement values herein also include ± 5%, ± 4%, ± 3%, ± 2%, and/or ± 1 %). All reported LCMS values herein also include ± 5%, ± 4%, ± 3%, ± 2%, and/or ± 1%. [0280] The following examples illustrate various non-limiting embodiments of the present disclosure.

EXAMPLES

EXAMPLE 1

Synthesis of 1-((S)-1-cyclopropylethyl)-3-(1-(4-((S)-2,2,2-trifluoro-1-hydroxyethyl)phenyl)- 1 H-pyrazol-3-yl)urea, Example 1 (Compound 37)

[0281] Synthesis of tert-butyl (S)-3-(3-(1-cyclopropylethyl)ureido)-1 H-pyrazole-1 - carboxylate, 1 -3 [Step 1]: To a stirred solution of tert-butyl 3-amino-1 H-pyrazole-1-carboxylate (1-1 , 10 g, 54.6 mmol) in dichloromethane (800 mL) was added triphosgene (9.7 g, 32.7 mmol) followed by triethylamine (29 mL, 218 mmol) at -15 °C and stirred for 30 min at same condition. (S)-1-cyclopropylethan-1-amine (1-2, 5.1 mL, 54.6 mmol) was added under same condition and allowed to stir for 3 h at ambient temperature. The reaction mixture was diluted with dichloromethane and washed with water followed by brine solution. The organic layer was separated, dried over anhydrous Na2SC>4, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to afford tert-butyl (S)-3-(3-(1- cyclopropylethyl)ureido)-1 H-pyrazole-1-carboxylate (1-3, 11 g). LCMS (ESI) Calcd. For C14H22N4O3: 294.2, found [M+H]⁺= 295.3. ¹H NMR (400 MHz, DMSO-d6) δ_H: 9.19 (s, 1 H), 8.06 (d, 1 H), 6.72 (d, 1 H), 6.46 (d, 1 H), 3.31-3.22 (m, 1 H), 1.54 (s, 9H), 1.12 (d, 3H), 0.88-0.83 (m, 1 H), 0.43-0.36 (m, 2H), 0.30-0.25 (m, 1 H), 0.10 (m, 1 H). [0282] Synthesis of (S)-1-(1-cyclopropylethyl)-3-(1H-pyrazol-3-yl)urea, 1-4 [Step 2]: To a solution of tert-butyl (S)-3-(3-(1-cyclopropylethyl)ureido)-1 H-pyrazole-1-carboxylate (1-3, 16 g, 54.4 mmol) in dichloromethane (150 ml_) was added trifluoroacetic acid (42 mL, 544 mmol) drop wise at ice cold condition and stirred at ambient temperature for 4h. Volatiles were evaporated under reduced pressure. The crude product was dissolved in Ethyl acetate and washed with saturated aqueous NaHCO3 solution (twice). The organic layer was dried over anhydrous Na₂SC>4 and concentrated under reduced pressure to afford (S)-1-(1-cyclopropylethyl)-3-(1 H- pyrazol-3-yl) urea (1-4, 9.5 g). LCMS (ESI) Calcd. For C9H14N4O: 194.12, found [M+H]⁺= 195.2. ¹H NMR (400 MHz, DMSO-d6) δ_H: 12.08 (bs, 1 H), 8.59 (s, 1 H), 7.51 (s, 1 H), 6.9 (bs, 1 H), 6.0 (s, 1 H), 3.27-3.21 (m, 1 H), 1.18 (d, 3H), 0.87-0.81 (m, 1 H), 0.46-0.36 (m, 2H), 0.25 (m, 1 H), 0.10 (m, 1 H).

[0283] Synthesis of 1-((S)-1-cyclopropylethyl)-3-(1-(4-((S)-2,2,2-trifluoro-1- hydroxyethyl)phenyl)-1H-pyrazol-3-yl)urea, Example 1 (Compound 37) [Step 3]: To a solution of (S)-1-(1-cyclopropylethyl)-3-(1 H-pyrazol-3-yl)urea (1-4, 3.56 g, 18.3 mmol) and (S)- 1-(4-bromophenyl)-2,2,2-trifluoroethan-1-ol (1-5, 5.14 g, 20.2 mmol) in acetonitrile (140 mL) was added dried potassium carbonate (6.3 g, 45.8 mmol) at ambient temperature. The reaction mixture was degassed with argon for 10 min followed by the addition of trans-A/,A/’- dimethylcyclohexane-1,2-diamine (2.9 mL, 18.3 mmol) and Cui (1.7 g, 9.2 mmol). The reaction mixture was heated in a sealed tube at 90 °C for 16 h. The reaction mixture was cooled to ambient temperature, filtered through celite bed and the bed was washed with ethyl acetate (twice). The combined filtrate was concentrated under reduced pressure. The crude product was purified by combi flash chromatography followed by trituration with 20% EtOAc in n-pentane to afford 1-((S)-1-cyclopropylethyl)-3-(1-(4-((S)-2,2,2-trifluoro-1-hydroxyethyl)phenyl)-1H-pyrazol- 3-yl)urea (Example 1 (Compound 37), 2.5 g). LCMS (ESI) Calcd. For C17H19F3N4O2: 368.1 , found [M+H]⁺: 369.2. ¹H NMR (400 MHz, DMSO-d6) δ_H: 8.96 (s, 1 H), 8.34-8.33 (m, 1 H), 7.75- 7.73 (m, 2H), 7.57-7.55 (m, 2H), 6.86-6.85 (m, 1 H), 6.68-6.66 (m, 1 H), 6.44 (s, 1 H), 5.20-5.15 (m, 1 H), 3.28-3.23 (m, 1 H), 1.16-1.14 (m, 3H), 0.92-0.86 (m, 1H), 0.47-0.36 (m, 2H), 0.29-0.25 (m, 2H).

EXAMPLES 2 & 3

Synthesis of 1 -((S)-1 -cyclopropylethyl)-3-(1 -(4-( 1 , 1 ,1 -trifluoro-2-hydroxypropan-2- yl)phenyl)-1H-pyrazol-3-yl)urea, Example 2 (Compound 28) & 1-((S)-1-cyclopropylethyl)-3- (1-(4-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)-1H-pyrazol-3-yl)urea, Example 3

(Compound 27):

[0284] Synthesis of 1-((S)-1-cyclopropylethyl)-3-(1-(4-(1,1,1-trifluoro-2-hydroxypropan- 2-yl)phenyl)-1H-pyrazol-3-yl)urea, 2-3 [Step 1]: To a stirred solution of (S)-1-(1 - cyclopropylethyl)-3-(1/7-pyrazol-3-yl)urea (2-1 , 200 mg, 1 .0 mmol) and 2-(4-bromophenyl)-1 ,1 ,1- trifluoropropan-2-ol (2-2, 305 mg, 1.1 mmol) in acetonitrile (8 mL) was added oven dried K2CO3 (294 mg, 2.1 mmol) at ambient temperature and the reaction mixture was degassed with argon for 10 min. Copper(l) iodide (81 mg, 0.4 mmol) followed by trans-N,N'-dimethylcyclohexane-1 ,2- diamine (0.13 mL, 0.8 mmol) were added into the reaction mixture. The reaction mixture was allowed to stir at 90°C for 16 h. The reaction mixture was cooled to ambient temperature, filtered through celite and the filtrate was concentrated under reduced pressure. Product was purified by reverse phase Prep HPLC and lyophilized to afford 1-((S)-1-cyclopropylethyl)-3-(1-(4-(1 ,1 ,1- trifluoro-2-hydroxypropan-2-yl)phenyl)-1/7-pyrazol-3-yl)urea (2-3, 160 mg). LCMS (ESI) Calcd. For C18H21F3N4O2: 382.2, found [M+H]⁺: 383.2.

[0285] Synthesis of 1-((S)-1-cyclopropylethyl)-3-(1-(4-(1,1,1-trifluoro-2-hydroxypropan-

2-yl)phenyl)-1H-pyrazol-3-yl)urea, Example 2 (Compound 28) & 1-((S)-1-cyclopropylethyl)-

3-(1-(4-(1,1,1-trifluoro-2-hydroxypropan-2-yl)phenyl)-1H-pyrazol-3-yl)urea, Example 3 (Compound 27) [Step 2]:The racemic Compound 1-((S)-1-cyclopropylethyl)-3-(1-(4-(1 ,1 ,1- trifluoro-2-hydroxypropan-2-yl)phenyl)-1 /7-pyrazol-3-yl)urea (2-3, 120 mg, 0.3 mmol) was submitted for HPLC SFC prep purification. The fractions obtained are lyophilized to yield peak 1 as 1-((S)-1-cyclopropylethyl)-3-(1-(4-(®-1 ,1 ,1-trifluoro-2-hydroxypropan-2-yl)phenyl)-1/7- pyrazol-3-yl)urea (Example 2 (Compound 28) , 50 mg) and peak 2 as 1-((S)-1- cyclopropylethyl)-3-(1-(4-((S)-1 ,1 , 1-trifluoro-2-hydroxypropan-2-yl)phenyl)-1H-pyrazol-3-yl)urea (Example 3 (Compound 27), 48 mg). Stereochemistry of both the compounds are arbitrarily assigned.

[0286] SFC method: SFC PREP PURIFICATION of sample has been completed on Waters SFC 80 instrument equipped with Waters 2489 UV/Visible Detector by using CHIRALPAK IG(30 mm x 250mm ), 5μ Column operating at 35 °C temperature, maintaining flow rate of 70 ml/min .using 60% CO2 in super critical state & 40% of [100% MeOH] as Mobile phase, Run this isocratic mixture up to 8.0 minutes and also maintained the isobaric condition of 100 bar at 282nm wavelength.

[0287] 1 -((S)-1-cyclopropylethyl)-3-(1-(4-(1,1 ,1-trifluoro-2-hydroxypropan-2-yl)phenyl)- 1H-pyrazol-3-yl)urea, Example 2 (Compound 28) (Peak 1): LCMS (ESI): Calcd. For C18H21 F3N4O2: 382.2, found [M+H]⁺: 383.2. 1 H NMR (400 MHz, DMSO-d6 at 25°C) δ_H: 8.98 (s, 1 H), 8.35-8.34 (m, 1 H), 7.74-7.72 (m, 2H), 7.66-7.64 (m, 2H), 6.69-6.68 (m, 1 H), 6.63 (s, 1 H), 6.44-6.43 (m, 1 H), 3.28-3.23 (m, 1 H), 1.70 (s, 3H), 1.16-1.14 (m, 3H), 0.92-0.87 (m, 1 H), 0.44- 0.38 (m, 2H), 0.30-0.22 (m, 2H).

[0288] 1 -((S)-1-cyclopropylethyl)-3-(1-(4-(1,1 ,1-trifluoro-2-hydroxypropan-2-yl)phenyl)- 1H-pyrazol-3-yl)urea, Example 3 (Compound 27) (Peak 2): LCMS (ESI): Calcd. For C18H21 F3N4O2: 382.2, found [M+H]⁺: 383.2. 1 H NMR (400 MHz, DMSO-d6 at 25°C) δ_H: 8.99 (s, 1 H), 8.35-8.34 (m, 1 H), 7.74-7.72 (m, 2H), 7.66-7.64 (m, 2H), 6.71-6.69 (m, 1 H), 6.64 (s, 1 H), 6.44-6.43 (m, 1 H), 3.28-3.23 (m, 1 H), 1.70 (s, 3H), 1.16-1.14 (m, 3H), 0.91-0.88 (m, 1 H), 0.45- 0.38 (m, 2H), 0.30-0.20 (m, 2H).

EXAMPLES 4 & 5

Synthesis of 1 -((S)-1-cyclopropylethyl)-3-(1 -(6-(2,2,2-trifluoro-1-hydroxyethyl)69yridine-3- yl)-1 H-pyrazol-3-yl)urea, Example 4 (Compound 30) and 1-((S)-1-cyclopropylethyl)-3-(1 -(6- (2, 2, 2-trifluoro-1 -hydroxyethyl) 69 yridine-3-yl)-1H-pyrazol-3-yl)urea, Example 5 (Compound 29):

[0289] Synthesis of (S)-1-(1-cyclopropylethyl)-3-(1H-pyrazol-3-yl)urea, 3-2 [Step 1]: To a solution of tert-butyl 3-[(1 S)-1-cyclopropylethyl]carbamoylamino]pyrazole-1-carboxylate (3-1 , 300 mg, 1.02 mmol) in dichloromethane (10 mL) at 0 °C, was added trifluoroacetic acid (0.78 mL, 10.2 mmol) and stirred for 4 h at ambient temperature. The reaction mixture was then concentrated under reduced pressure, crude residue was dissolved in ethyl acetate. Organic extract was washed with saturated NaHCOs solution, dried over anhydrous Na2SO4 and then concentrated under reduced pressure to afford 1-(1H-pyrazol-3-yl)-3-[(1 S)-1- cyclopropylethyl]urea (3-2, 200 mg). LCMS (ESI) Calcd. For C9H14N4O: 194.1 , found [M+H]⁺: 195.02.

[0290] Synthesis of 1-((S)-1-cyclopropylethyl)-3-(1-(6-(2,2,2-trifluoro-1- hydroxyethyl)70yridine-3-yl)-1H-pyrazol-3-yl)urea, 3-4 [Step 2]: To a stirred solution of 1-(5- bromo-2-pyridyl)-2,2,2-trifluoro-ethanol (3-3, 377 mg, 1.47 mmol) and 1-(1 H-pyrazol-3-yl)-3- [(1 S)-1-cyclopropylethyl]urea (3-2, 220 mg, 1.13 mmol) in MeCN (5 mL), was added anhydrous K2CO3 (469 mg, 3.40 mmol) at ambient temperature and the reaction mixture was degassed with argon balloon for 10 min, followed by the addition of N1,N1-dimethylcyclohexane-1 ,2- diamine (0.18 mL, 1.13 mmol) and Cui (108 mg, 0.56 mmol). The reaction mixture was heated in a sealed tube at 90 °C for 16 h. The reaction mixture was cooled to ambient temperature, diluted with ethyl acetate, filtered through celite bed and the filtrate was concentrated under reduced pressure to obtain crude compound. The crude was purified by combi flash column chromatography to afford 1-[(1 S)-1-cyclopropylethyl]-3-[1-[6-(2,2,2-trifluoro-1-hydroxy-ethyl)-3- pyridyl]pyrazol-3-yl]urea (3-4, 110 mg). LCMS (ESI) Cacld. For C16H18F3N5O2: 369.1 , found [M+H]⁺: 370.0

[0291] Synthesis of 1-((S)-1-cyclopropylethyl)-3-(1-(6-(2,2,2-trifluoro-1- hydroxyethyl)70yridine-3-yl)-1H-pyrazol-3-yl)urea, Example 4 (Compound 30) and 1-((S)- 1 -cyclopropylethyl)-3-(1 -(6-(2, 2, 2-trifluoro-1 -hydroxyethyl) 71 yridine-3-yl)-1 H-pyrazol-3- yljurea, Example 5 (Compound 29) [Step 3]: 1-[(1 S)-1-cyclopropylethyl]-3-[1-[6-(2,2,2- trifluoro-1-hydroxy-ethyl)-3-pyridyl]pyrazol-3-yl]urea (3-4, 101 mg) was purified on Waters SFC 80 instrument equipped with Waters 2489 UV/Visible Detector by using CHIRALPAK IG (30.0 mm x 250 mm ), 5μ Column operating at 35 °C temperature, maintaining flow rate of 70 ml/min .using 50% CO2 in super critical state & 50% [100% methanol] as Mobile phase, Run this isocratic mixture up to 12.0 minutes and also maintained the isobaric condition of 100 bar at 291 nm wavelength to afford the first fraction 1-((S)-1-cyclopropylethyl)-3-(1-(6-((S)-2,2,2-trifluoro- 1-hydroxyethyl)71yridine-3-yl)-1 H-pyrazol-3-yl)urea (Example 4 (Compound 30), 40 mg) as Peak 1 and the second fraction 1-((S)-1-cyclopropylethyl)-3-(1-(6-(®-2,2,2-trifluoro-1- hydroxyethyl)71yridine-3-yl)-1H-pyrazol-3-yl)urea (Example 5 (Compound 29), 40 mg) as Peak 2.

[0292] 1 -((S)-1 -cyclopropylethyl)-3-(1 -(6-(2, 2, 2-trifluoro-1 -hydroxyethyl) 71 yridine-3-yl)- 1H-pyrazol-3-yl)urea, Example 4 (Compound 30): LCMS (ESI) Calcd. For C16H18F3N5O2: 369.1 , found [M+H]⁺: 370.2, HPLC: Rt (min) = 6.86 (96.26 %). ¹H NMR (400 MHz, DMSO-d₆): δ_H 9.06 (s, 1 H), 8.98 (d, 1 H), 8.45 (d, 1 H), 8.19 (dd, 1 H), 7.69 (d, 1 H), 7.08 (brs, 1 H), 6.60 (d, 1 H), 6.53 (d, 1H), 5.18-5.12 (m, 1 H), 3.27-3.22 (m, 1 H), 1.15 (d, 3H), 0.92-0.87 (m, 1 H), 0.44- 0.37 (m, 2H), 0.29-0.15 (m, 2H).

[0293] 1 -((S)-1 -cyclopropylethyl)-3-(1 -(6-(2, 2, 2-trifluoro-1 -hydroxyethyl) 71 yridine-3-yl)- 1H-pyrazol-3-yl)urea, Example 5 (Compound 29): LCMS (ESI) Cacld. For C16H18F3N5O2: 369.1 , found [M+H]⁺: 370.2, HPLC: Rt (min) = 6.86 (99.80 %). ¹H NMR (400 MHz, DMSO-d₆): δH 9.06 (s, 1 H), 8.98 (d, 1 H), 8.45 (d, 1 H), 8.19 (dd, 1 H), 7.69 (d, 1 H), 7.07 (brs, 1 H), 6.60 (d, 1 H), 6.53 (d, 1H), 5.18-5.13 (m, 1 H), 3.27-3.22 (m, 1 H), 1.15 (d, 3H), 0.93-0.87 (m, 1 H), 0.46- 0.35 (m, 2H), 0.30-0.15 (m, 2H). Note: absolute stereochemistry of the compounds are unknown.

EXAMPLES 6 & 7

1 -[(1 S)-1 -cyclopropylethyl]-3-[1 -[5-[2,2,2-trifluoro-1 -hydroxy-1 -methyl-ethyl]-2- pyridyl]pyrazol-3-yl]urea, Example 6 (Compound 140) and 1-[(1S)-1-cyclopropylethyl]-3-[1-[5- [2,2,2-trifluoro-1-hydroxy-1-methyl-ethyl]-2-pyridyl]pyrazol-3-yl]urea, Example 7 (Compound 139):

[0294] To an ice-cold solution of 1-(6-bromo-3-pyridyl)72yridine (4-1 , 1.00 g, 5.00 mmol) in THF (20 ml_) was added trimethyl(trifluoromethyl)silane (1.07 g, 7.50 mmol) followed by TBAF (1M in THF) (13 mL, 12.5 mmol). The mixture was stirred at ambient temperature for 16 h. It was quenched with water and extracted with ethyl acetate. Organic extract was washed with brine, dried over anhydrous Na₂SO4, concentrated under reduced pressure to afford crude which was purified by combiflash column chromatography to afford 2-(6-bromo-3-pyridyl)- 1 ,1 ,1 - trifluoro-propan-2-ol (4-2, 1 .00 g, merged with SM). LCMS (ESI) calcd. For CsH7BrF3NO: 268.97, found [M+H]⁺: 370.0.

[0295] To a stirred solution of 2-(6-bromo-3-pyridyl)-1,1,1-trifluoro-propan-2-ol (4-2, 500 mg, 1.85 mmol) and 1-[(1 S)-1-cyclopropylethyl]-3-(1 H-pyrazol-3-yl)urea (4-3, 360 mg, 1.85 mmol) in MeCN (30 mL) was degassed with argon balloon for 20 min. Then added Cui (0.26 g, 0.926 mmol) followed by /V,/V-frans-dimethyl-cy-diamine (263.36 mg, 1.85 mmol) at ambient temperature. The reaction mixture was heated in a sealed tube at 90 °C for 16 h. Reaction mixture was cooled to ambient temperature, filtered through celite bed and the filtrate was concentrated under reduced pressure to obtain crude compound, which was purified by combi flash column chromatography followed by reverse phase preparative HPLC to afford 1-[(1 S)-1 - cyclopropylethyl]-3-[1-[5-(2,2,2-trifluoro-1-hydroxy-1-methyl-ethyl)-2-pyridyl]pyrazol-3-yl]urea (4-4, 130 mg). LCMS (ESI) calcd for C18H21F3N4O2: 382.16, found [M+H]⁺: 383.3.

[0296] 1-[(1 S)-1-cyclopropylethyl]-3-[1-[5-(2,2,2-trifluoro-1-hydroxy-1-methyl-ethyl)-2- pyridyl]pyrazol-3-yl]urea (4-4, 120 mg) was purified by SFC HPLC on Pic Solutions- 175 instrument equipped with Knauer 40D UV/Visible Detector by using Chiralpak-IG (30.0 mm x 250 mm), 5μ Column operating at 35°C temperature, maintaining flow rate of 90 ml/min, using 75% CO₂ in super critical state & 25% of 0.2% (7N ammonia in methanol) in methanol as mobile phase. Run this isocratic mixture up to 10.0 min and also maintained the isobaric condition of 100 bar at 299 nm wavelength to afford the first fraction 1-[(1 S)-1 -cyclopropylethyl]- 3-[1-[5-[(1 S)-2, 2, 2-trifluoro-1 -hydroxy-1 -methyl-ethyl]-2-pyridyl]pyrazol-3-yl]urea (Example 6 (Compound 140), 58 mg) as peak 1 and 1-[(1 S)-1-cyclopropylethyl]-3-[1-[5-[(1R)-2,2,2- trifluoro-1 -hydroxy-1 -methyl-ethyl]-2-pyridyl]pyrazol-3-yl]urea (Example 7 (Compound 139), 52 mg) as peak 2.

[0297] 1-[(1 S)-1-cyclopropylethyl]-3-[1-[5-[2,2,2-trifluoro-1-hydroxy-1-methyl-ethyl]-2- pyridyl]pyrazol-3-yl]urea, Example 6 (Compound 140) [Peak 1]: LCMS (ESI) calcd. For Ci7H2oF₃N₅02:383.16 ; found [M+H]⁺: 384.2; HPLC :Rt(min)=7.319 (99.71 %); Chiral purity=98.94% de., ¹H NMR (400 MHz, DMSO-d₆): δH 9.07 (s, 1 H), 8.58 (d, 1 H), 8.44 (d, 1H), 8.13-8.11 (m, 1 H), 7.70 (d, 1 H), 6.86 (s, 1 H), 6.67-6.65 (m, 1H), 6.52 (d, 1 H), 3.27-3.21 (m, 1 H), 1.74 (s, 3H), 1.16 (d, 3H), 0.94-0.86 (m, 1 H), 0.46-0.36 (m, 2H), 0.30-0.27 (m, 2H).

[0298] 1-[(1 S)-1-cyclopropylethyl]-3-[1-[5-[2, 2, 2-trifluoro-1 -hydroxy-1 -methyl-ethyl]-2- pyridyl]pyrazol-3-yl]urea, Example 7 (Compound 139) [Peak 2]: LCMS (ESI) calcd. For C17H20F3N5O2: 383.16; found [M+H]⁺:384.2, HPLC: Rt(min)= 7.693 (98.57%). Chiral purity= 97.18% de; ¹H NMR (400 MHz, DMSO-d₆): δH 9.07 (s, 1 H), 8.58 (d, 1H), 8.44 (d, 1 H), 8.13- 8.11 (m, 1 H), 7.70 (d, 1 H), 6.87 (s, 1H), 6.67-6.65 (m, 1 H), 6.52 (d, 1 H), 3.27-3.21 (m, 1H), 1.74 (s, 3H), 1.16 (d, 3H), 0.91-0.86 (m, 1 H), 0.44-0.38 (m, 2H), 0.30-0.18 (m, 2H). Note: Both isomers are single diastereomer with unknown absolute configuration.

EXAMPLES 8 & 9

Synthesis of 1-((S)-1-cyclopropylethyl)-3-(1-(6-(1,1,1-trifluoro-2-hydroxypropan-2- yl)73yridine-3-yl)-1H-pyrazol-3-yl)urea, Example 8 (Compound 138) and 1-((S)-1- cyclopropylethyl)-3-(1-(6-(1,1,1-trifluoro-2-hydroxypropan-2-yl)73yridine-3-yl)-1H-pyrazol-3- yl)urea, Example 9 (Compound 137): [0299] Synthesis of 1-(5-bromopyridin-2-yl)ethan-1-ol, 2 [Step 1]: To a solution of 5- bromopyridine-2-carbaldehyde (5-1 , 3.00 g, 16.1 mmol) in 2-Me-THF (20 mL) under nitrogen atmosphere, was added MeMgBr (3M in THF) (7.0 mL, 21.0 mmol) at 0 °C, and stirred for 2 h at the ambient temperature. Reaction mixture was quenched with saturated NH4CI solution, then extracted with ethyl acetate, dried over anhydrous Na2SC>4 and concentrated under reduced pressure to obtain crude compound, which was purified by combi flash column chromatography to afford 1-(5-bromo-2-pyridyl)ethanol (5-2, 3g). ¹H NMR (400 MHz, DMSO-d₆): δH 8.58 (d, 1 H), 8.03-8.00 (m, 1 H), 7.48 (d, 1 H), 5.45 (d, 1H), 4.72-4.66 (m, 1 H), 1.34 (d, 3H).

[0300] Synthesis of 1-(5-bromopyridin-2-yl)ethan-1-one, 5-3 [Step 2]: To a solution of 1-(5- bromo-2-pyridyl)ethanol (2, 1.50 g, 7.42 mmol) in dichloromethane (20 mL), was added dessmartin periodinane (4.09 g, 9.65 mmol) and stirred at the ambient temperature for 16 h. The reaction mixture was filtered, washed with NaHCO3 solution, dried over anhydrous Na₂SC>4 and concentrated under reduced pressure to obtain crude compound, which was purified by combi flash column chromatography to afford 1-(5-bromo-2-pyridyl)74yridine (5-3, 900 mg) ¹H NMR (400 MHz, CDCI3): δH 8.72 (s, 1H), 7.96-7.90 (m, 2H),2.68 (s, 3H).

[0301] Synthesis of 2-(5-bromopyridin-2-yl)-1,1,1-trifluoropropan-2-ol, 4 [Step 3]: To an ice-cold solution of 1-(5-bromo-2-pyridyl)74yridine (5-3, 1.10 g, 5.50 mmol) in THF (10 mL) was added trimethyl(trifluoromethyl)silane (1.56 g, 11.0 mmol) followed by TBAF (1M in THF) (14 mL, 13.7 mmol) . The mixture was stirred at ambient temperature for 16 h. Water (30 mL) was added followed by extraction with ethyl acetate. The combined organic was washed with brine, dried over anhydrous Na₂SC>4, filtered and concentrated in vacuo to get crude product. The product was purified by combi flash column chromatography to afford 2-(5-bromo-2-pyridyl)- 1 ,1 ,1-trifluoro-propan-2-ol (5-4, 1.10 g). LCMS (ESI) calcd. For C₈H₇BrF₃NO: 268.97, [M+H]⁺ = 270.05.

[0302] Synthesis of 1-((S)-1-cyclopropylethyl)-3-(1-(6-(1,1,1-trifluoro-2-hydroxypropan- 2-yl)74yridine-3-yl)-1H-pyrazol-3-yl)urea, 5-6 [Step 4]: To a solution of 1-(1 H-pyrazol-3-yl)-3- [(1 S)-1-cyclopropylethyl]urea (5-4, 280 mg, 1.44 mmol), 2-(5-bromo-2-pyridyl)-1 ,1 ,1-trifluoro- propan-2-ol (5-5, 584 mg, 2.16 mmol) and potassium carbonate (598 mg, 4.32 mmol) in MeCN (3 mL), was degassed by argon for 15 min, then /V,/V-dimethyl-trans-cy-1,2-diamine (205 mg, 1.44 mmol) and Cui (137 mg, 0.721 mmol) was added and then stirred at 90 °C for 16 h. The reaction mixture was then filtered through celite bed and washed with ethyl acetate. Organic extract was washed with water, dried over anhydrous Na₂SO4 and concentrated under reduced pressure to afford 1-((S)-1-cyclopropylethyl)-3-(1-(6-(1,1,1-trifluoro-2-hydroxypropan-2- yl) 75yridine-3-yl)-1 H-pyrazol-3-yl)urea (5-6, 140 mg) LCMS (ESI) calcd. For C17H20F3N5O2: 383.1 , [M+H]⁺ = 384.1.

[0303] Synthesis of 1-((S)-1-cyclopropylethyl)-3-(1 -(6-(1,1 ,1-trifluoro-2-hydroxypropan-

2-yl) 75 yridine-3-yl)-1 H-pyrazol-3-yl)urea, Example 8 (Compound 138) and 1-((S)-1- cyclopropylethyl)-3-(1-(6-(1,1,1-trifluoro-2-hydroxypropan-2-yl) 75 yridine-3-yl)-1 H- pyrazol-3-yl)urea, Example 9 (Compound 137) [Step 5]: Purification of 1-((S)-1- cyclopropylethyl)-3-(1 -(6-(1 ,1 ,1 -trifluoro-2-hydroxypropan-2-yl) 75 y rid i ne-3-y I) - 1 H-pyrazol-3- yl)urea was done by SFC prep purification on waters SFC 150 instrument equipped with waters 2489 UV/Visible detector by using C amylose A (30 mm x 250mm), 5μ column operating at 35°C temperature, maintaining flow rate of 100 ml/min, using 60% CO2 in super critical state & 40% of [100% methanol] as mobile phase, run this isocratic mixture up to 10.0 min and also maintained the isobaric condition of 100 bar at 290 nm wavelength to afford the first fraction 1- ((S)-1-cyclopropylethyl)-3-(1-(6-((S)-1 ,1 ,1-trifluoro-2-hydroxypropan-2-yl) 75 yridine-3-yl)-1 H- pyrazol-3-yl)urea (Example 8 (Compound 138), 48 mg) as peak 1 and the second fraction 1- ((S)-1-cyclopropylethyl)-3-(1-(6-(®-1 ,1 , 1-trifluoro-2-hydroxypropan-2-yl) 75 yridine-3-yl)-1 H- pyrazol-3-yl)urea (Example 9 (Compound 137), 48 mg) as peak 2.

[0304] 1 -((S)-1-cyclopropylethyl)-3-(1-(6-(1,1 ,1-trifluoro-2-hydroxypropan-2-yl)75yridine-

3-yl)-1 H-pyrazol-3-yl)urea, Example 8 (Compound 138) [Peak 1]: LCMS (ESI) calcd. For C17H20F3N5O2: 383.1 , found [M+H]⁺: 384.3, HPLC: Rt(min) = 7.71 (99.55%). ¹H NMR (400 MHz, DMSO-d₆): δ_H 9.04 (s, 1 H), 8.97 (d, 1 H), 8.43 (d, 1 H), 8.18-8.15 (m, 1 H), 7.85 (d, 1 H), 6.79 (s, 1 H), 6.59 (d, 1 H), 6.53 (d, 1 H), 3.27-3.21 (m, 1 H), 1.72 (s, 3H), 1.15 (d, 3H), 0.90-0.87 (m, 1 H), 0.44-0.37 (m, 2H), 0.28-0.20 (m, 2H). Note: absolute stereochemistry of both compounds are unknown.

[0305] 1 -((S)-1-cyclopropylethyl)-3-(1-(6-(1,1 ,1-trifluoro-2-hydroxypropan-2-yl)75yridine- 3-yl)-1 H-pyrazol-3-yl)urea, Example 9 (Compound 137) [Peak 2]: LCMS (ESI) calcd. For C17H20F3N5O2: 383.1 , found [M+H]⁺: 384.3, HPLC: Rt(min) = 7.71 (99.49%). ¹H NMR (400 MHz, DMSO-d₆): δ_H 9.04 (s, 1 H), 8.97 (d, 1 H), 8.43 (d, 1 H), 8.18-8.15 (m, 1 H), 7.85 (d, 1 H), 6.79 (s, 1 H), 6.59 (d, 1 H), 6.53 (d, 1 H), 3.27-3.23 (m, 1 H), 1.72 (s, 3H), 1.15 (d, 3H), 0.90-0.87 (m, 1 H), 0.43-0.38 (m, 2H), 0.28-0.17 (m, 2H). Note: absolute stereochemistry of both compounds are unknown.

EXAMPLES 10 & 11

Synthesis of 1 -((S)-1 -cyclopropylethyl)-3-(1 -(2-1 , 1 , 1 -trifluoro-2-hydroxypropan-2-yl)75yridine-

4-yl)-1 H-pyrazol-3-yl)urea, Example 10 (Compound 135) and 1-((S)-1-cyclopropylethyl)-3-(1-(2- 1,1,1-trifluoro-2-hydroxypropan-2-yl) 76 yridine-4-yl)-1H-pyrazol-3-yl)urea, Example 11

(Compound 134):

[0306] Synthesis of 2-(4-bromo-2-pyridyl)-1,1,1-trifluoro-propan-2-ol, 6-2 [Step 1]: To a stirred solution of 1-(4-bromo-2-pyridyl)76yridine (6-1 , 200 mg, 1.00 mmol) in THF (2 mL) was added trimethyl(trifluoromethyl)silane (213 mg, 1 .50 mmol) at 0°C followed by dropwise addition of TBAF (1M in THF) (2.50 mL, 2.50 mmol). The reaction mixture was stirred at ambient temperature for 16 h. Reaction mixture was quenched with ice cold water and extracted with ethyl acetate. Organic extract was washed with brine, dried over anhydrous Na₂SO4, and concentrated under reduced pressure to get crude product. The product was purified by combi- flash to afford 2-(4-bromo-2-pyridyl)-1 ,1 ,1-trifluoro-propan-2-ol (6-2, 1.14 g). LCMS (ESI) calcd. For C8H₇BrF₃NO: 268.97, found [M+H]⁺=269.8.

[0307] Synthesis of 1-[(1S)-1-cyclopropylethyl]-3-[1-[2-(2,2,2-trifluoro-1-hydroxy-1- methyl-ethyl)-4-pyridyl]pyrazol-3-yl]urea, 6-4 [Step 2]: To a stirred solution of 2-(4-bromo-2- pyridyl)-1,1 ,1-trifluoro-propan-2-ol (6-2, 600 mg, 2.22 mmol) and 1-[(1 S)-1-cyclopropylethyl]-3- (1 H-pyrazol-3-yl)urea (6-3, 475 mg, 2.44 mmol) and anhydrous K2CO3 (920 mg, 6.67 mmol) in MeCN (10 mL), was degassed with argon balloon for 10 min. Then was added N,N-trans- dimethyl-cy-diamine (0.35 mL, 2.22 mmol) followed by Cui (126 mg, 0.444 mmol) at ambient temperature. The reaction mixture was heated in a sealed tube at 90°C for 16 h. Reaction mixture was cooled to ambient temperature, filtered through celite bed and the filtrate was concentrated under reduced pressure to obtain crude compound, which was purified by reverse phase preparative HPLC to afford 1-[(1 S)-1-cyclopropylethyl]-3-[1-[2-(2,2,2-trifluoro-1-hydroxy- 1-methyl-ethyl)-4-pyridyl]pyrazol-3-yl]urea (6-4, 150 mg, 0.383 mmol). LCMS (ESI) calcd. For C17H20F3N5O2: 383.16, found [M+H]⁺ =384.0. [0308] Synthesis of 1-((S)-1-cyclopropylethyl)-3-(1-(2-1,1,1-trifluoro-2-hydroxypropan-2- yl) 77 yridine-4-yl)-1H-pyrazol-3-yl)urea, Example 10 (Compound 135) and 1-((S)-1- cyclopropylethyl)-3-(1-(2-((S)-1,1,1-trifluoro-2-hydroxypropan-2-yl) 77 yridine-4-yl)-1H- pyrazol-3-yl)urea, Example 11 (Compound 134) [Step 3]: The racemic compound 1-[(1 S)-1- cyclopropylethyl]-3-[1-[2-(2,2,2-trifluoro-1-hydroxy-1-methyl-ethyl)-4-pyridyl]pyrazol-3-yl]urea (99 mg) was purified by SFC prep HPLC on Pic Solutions-175 instrument equipped with Knauer 40D UV/Visible Detector by using Unichiral CND-5H (21.2 mm x 250 mm ), 5μ Column operating at 35 °C, maintaining flow rate of 50 ml/min, using 80 % CO2 in super critical state and 20% of 0.1% Ipamine in Acetonitrile/lsopropanol (70:30) as mobile phase. Run this isocratic mixture was run up to 15.0 minutes and also maintained the isobaric condition of 100 bar at 295 nm wavelength. The fractions obtained are lyophilized to afford the first fraction 1-[(1 S)-1- cyclopropylethyl]-3-[1-[2-[(1R)-2,2,2-trifluoro-1-hydroxy-1-methyl-ethyl]-4-pyridyl]pyrazol-3- yl]urea (Example 10 (Compound 135), 35 mg) as Peak 1 and 1-[(1 S)-1-cyclopropylethyl]-3-[1- [2-[(1 S)-2,2,2-trifluoro-1-hydroxy-1-methyl-ethyl]-4-pyridyl]pyrazol-3-yl]urea (Example 11 (Compound 134), 34 mg) as Peak 2. Note: Peak 1 and Peak 2 are arbitrarily assigned.

[0309] 1-((S)-1-cyclopropylethyl)-3-(1-(2-1,1,1-trifluoro-2-hydroxypropan-2-yl) 77 yridine- 4-yl)-1H-pyrazol-3-yl)urea, Example 10 (Compound 135) [Peak 1]: LCMS (ESI): calcd. For C17H20F3N5O2: 383.16, found [M+H]⁺:384.3; ¹H NMR (400 MHz, DMSO-d₆): δH 9.17 (s, 1H), 8.58- 8.57 (m, 2H), 8.09 (d, 1H), 7.74-7.72 (m, 1 H), 6.83 (s, 1 H), 6.67 (d, 1 H), 6.57 (d, 1 H), 3.28-3.24 (m, 1 H), 1.72 (s, 3H), 1.16 (d, 3H), 0.90-0.88 (m, 1 H), 0.45-0.40 (m, 2H), 0.29-0.27 (m, 1 H), 0.21-0.19 (m, 1 H); HPLC :Rt(min)=7.418 (99.49 %).

[0310] 1-((S)-1-cyclopropylethyl)-3-(1-(2-1,1,1-trifluoro-2-hydroxypropan-2-yl) 77 yridine- 4-yl)-1H-pyrazol-3-yl)urea, Example 11 (Compound 134) [Peak 2]: LCMS (ESI): calcd. For C17H20F3N5O2: 383.16, found [M+H]⁺:384.3; ¹H NMR (400 MHz, DMSO-d₆): δH 9.17 (s, 1H), 8.58- 8.57 (m, 2H), 8.09 (d, 1H), 7.74-7.72 (m, 1 H), 6.83 (s, 1 H), 6.69 (d, 1 H), 6.56 (d, 1 H), 3.28-3.23 (m, 1 H), 1.72 (s, 3H), 1.16 (d, 3H), 0.91-0.88 (m, 1 H), 0.46-0.40 (m, 2H), 0.29-0.27 (m, 1 H), 0.27-0.21 (m, 1 H); HPLC :Rt(min)=7.411 (99.60 %). EXAMPLE 12

Synthesis of (S)-1 -(1 -cyclopropylethyl)-3-(1 -(4-(3-hydroxyoxetan-3-yl)phenyl)-1 H-pyrazol-3- yl)urea, Example 12 (Compound 136):

[0311] Synthesis of (S)-1-(1-(4mmol) dropwise1H-pyrazol-3-yl)-3-(1- cyclopropylethyl)urea, 7-3 [Step 1]: To a stirred solution of 1-bromo-4-iodobenzene (7-1 , 2.36 g, 8.33 mmol) in THF (10 mL) was added n-BuLi, 2M in hexane (4.2 mL, 8.33 mmol) dropwise under nitrogen atmosphere at -78 °C and stirred for 1h. To the reaction mixture was added oxetan-3-one (7-2, 500 mg, 6.94 mmol) and stirred for 3h at -78°C. The reaction mixture was gradually warmed to ambient temperature and stirred for 16 h. The reaction mixture was quenched with saturated aqueous NH4CI solution and extracted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous Na2SO4 and concentrated under reduced pressure. The crude product was purified by combi flash column chromatography to afford 3-(4- bromophenyl)oxetan-3-ol (7-3, 450 mg). ¹H NMR (400 MHz, CDCI3): δ_H 7.55-7.52 (m, 2H), 7.49- 7.47 (m, 2H), 4.94-4.88 (m, 2H), 4.85-4.83 (m, 2H), 2.59 (bs, 1H).

[0312] Synthesis of (S)-1-(1-cyclopropylethyl)-3-(1-(4-(3-hydroxyoxetan-3-yl)phenyl)-1H- pyrazol-3-yl)urea, Example 12 (Compound 136) [Step 2]: To a stirred solution of 3-(4- bromophenyl)oxetan-3-ol (7-3, 153 mg, 0.67 mmol) in MeCN (5 mL) was added (S)-1-(1- cyclopropylethyl)-3-(1 H-pyrazol-3-yl)urea (7-4, 100 mg, 0.51 mmol) followed by K2CO3 (178 mg, 1.29 mmol) and degassed with argon for 10 min. trans N,N-dimethyl cyclohexanedimine (0.08 mL, 0.51 mmol) and Cui ( 49 mg, 0.26 mmol) were added to it and stirred at 90 °C for 16 h. The reaction mixture was filtered through celite bed and washed with ethyl acetate. Filtrate was concentrated under reduced pressure and purified by reverse phase PREP HPLC and lyophilized to afford (S)-1-(1-cyclopropylethyl)-3-(1-(4-(3-hydroxyoxetan-3-yl)phenyl)-1 H- pyrazol-3-yl)urea (Example 12 (Compound 136), 45 mg). LCMS (ESI) Calcd. for C18H22N4O3: 342.4, found [M+H]⁺= 343.3. ¹H NMR (400 MHz, DMSO-d₆): δH 8.95 (s, 1 H), 8.34-8.33 (m, 1 H), 7.74-7.72 (m, 2H), 7.67-7.65 (m, 2H), 6.72 (bs, 1 H), 6.42-6.38 (m, 2H), 4.78-4.76 (m, 2H), 4.70- 4.68 (m, 2H), 3.27-3.25 (m, 1 H), 1.15 (d, 3H), 0.89-0.87 (m, 1H), 0.44-0.38 (m, 3H), 0.27-0.16 (m, 2H).

EXAMPLES 13 & 14

Synthesis of (S)-1 -(1 -cyclopropylethyl)-3-(1 -(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1 H- pyrazol-3-yl)urea, Example 13 (Compound 130) a and (S)-1-(1-cyclopropylethyl)-3-(1-(5-(2- hydroxypropan-2-yl)pyridin-2-yl)-1H-pyrazol-3-yl)urea, Example 14 (Compound 129):

[0313] Synthesis of (S)-1-(1-cyclopropylethyl)-3-(1-(6-(2-hydroxypropan-2-yl)pyridin-3- yl)-1H-pyrazol-3-yl)urea, Example 13 (Compound 130) [Step 1]: A stirred solution of 2-(5- bromopyridin-2-yl)propan-2-ol (8-2A, 275 mg, 1.27 mmol) and (S)-1-(1-cyclopropylethyl)-3-(1 H- pyrazol-3-yl)urea (8-1, 190 mg, 0.98 mmol) and potassium carbonate (406 mg, 2.93 mmol) in MeCN (5 mL) was degassed by argon gas for 10 min. Then A/,A/-dimethylcyclohexane- 1 ,2-diamine (139 mg, 0.98 mmol) followed by cuprous iodide (93 mg, 0.49 mmol) were added and the resultant reaction mixture was stirred for 16 h at 80 °C. The crude was filtered through celite bed, washed with ethyl acetate, solvent was concentrated under reduced pressure to afford crude product. The product was purified by reverse phase preparative HPLC to afford (S)- 1-(1-cyclopropylethyl)-3-(1-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)-1 H-pyrazol-3-yl)urea (Example 13 (Compound 130), 90.0 mg). LCMS (ESI) calcd for C17H23N5O2: 329.19, found [M+H]⁺ = 330.2; HPLC: Rt(min) = 5.92 (99.79%). ¹H NMR (400 MHz, DMSO-d₆): δH 9.00 (s, 1 H), 8.86 (d, 1 H), 8.36 (d, 1 H), 8.07-8.04 (m, 1 H), 7.72 (d, 1 H), 6.61 (d, 1 H), 6.47 (d, 1 H), 5.25 (s, 1 H), 3.34-3.21 (m, 1 H), 1.41-1.14 (m, 6H), 0.93-0.90 (m, 3H), 0.89-0.85 ( s, 1H), 0.46-0.19 (m, 4H). [0314] Synthesis of (S)-1 -(1-cyclopropylethyl)-3-(1 -(5-(2-hydroxypropan-2-yl)pyridin-2- yl)-1 H-pyrazol-3-yl)urea, Example 14 (Compound 129) [Step 2]: To a stirred solution of 2-(6- bromopyridin-3-yl)propan-2-ol (8-2B, 275 mg, 1.27 mmol) and (S)-1-(1-cyclopropylethyl)-3-(1 H- pyrazol-3-yl)urea (8-1 , 190 mg, 0.98 mmol) and potassium carbonate (406 mg, 2.93 mmol) in MeCN (5 mL) was degassed by argon gas for 10 min. Then A/,/\/-dimethylcyclohexane- 1 ,2-diamine (139 mg, 0.98 mmol) followed by cuprous iodide (93 mg, 0.49 mmol) were added and the resultant reaction mixture was stirred for 16 h at 80 °C. The crude was filtered through celite bed, washed with ethyl acetate. Solvent was concentrated under reduced pressure to afford crude product. The crude was purified by reverse phase preparative HPLC to afford (S)- 1-(1-cyclopropylethyl)-3-(1-(5-(2-hydroxypropan-2-yl)pyridin-2-yl)-1 H-pyrazol-3-yl)urea (Example 14 (Compound 129), 45 mg). LCMS (ESI) calcd for C17H23N5O2: 329.19, found [M+H]⁺ = 330.3; HPLC: Rt(min) = 7.03 (99.26%). ¹ H NMR (400 MHz, DMSO-d₆): δ_H 9.01 (s, 1 H), 8.48 (s, 1 H), 8.41 (s, 1 H), 7.99 (d, 1 H), 7.60 (d, 1 H), 6.68 (d, 1 H), 6.47 (s, 1 H), 5.24 (s, 1 H), 3.31-3.23 (m, 1 H), 1.47 (s, 6H), 1.15 (d, 3H), 0.90 ( d, 1 H), 0.42 (d, 2H), 0.28 (d, 1 H), 0.20 (d, 1 H).

Example 15

Synthesis of (S)-1-(1-cyclopropylethyl)-3-(2-(6-(2-hydroxypropan-2-yl)pyridin-3-yl)thiazol-4- yl)urea, Example 15 (Compound 131):

[0315] Synthesis of 1 -[2-(6-acetyl-3-pyridyl)thiazol-4-yl]-3-[(1 S)-1 -cyclopropylethyl]urea, 9-3 [Step 1]: To a solution of 1-(2-bromothiazol-4-yl)-3-[(1 S)-1-cyclopropylethyl]urea (9-1 , 212 mg, 0.72 mmol), 1-[5-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)-2-pyridyl]ethanone (9-2, 200 mg, 0.80 mmol) and potassium phosphate (516 mg, 2.42 mmol) in 1 ,4-dioxane (10 ml_) and water (1 ml_), was purged by argon for 15 min, then Pd-118 (52 mg, 0.08 mmol) was added and stirred at 90 °C for 3 h. The reaction mixture was cooled to ambient temperature, diluted with water and extracted with ethyl acetate. Organic extract was dried over anhydrous Na₂SO4 and concentrated under reduced pressure to afford crude compound. The product was purified by combiflash column chromatography to afford 1-[2-(6-acetyl-3-pyridyl)thiazol-4-yl]-3-[(1 S)-1- cyclopropylethyl]urea (9-3, 200 mg). LCMS (ESI) calcd. for C16H18N4O2S: 330.1 , found [M+H]⁺= 331.0.

[0316] Synthesis of (S)-1 -(1-cyclopropylethyl)-3-(2-(6-(2-hydroxypropan-2-yl)pyridin-3- yl)thiazol-4-yl)urea, Example 15 (Compound 131 ) [Step 2]: To a solution of (S)-1-(2-(6- acetylpyridin-3-yl)thiazol-4-yl)-3-(1-cyclopropylethyl)urea (9-2, 200 mg, 0.60 mmol) in 2-Me-THF (5 mL), was added MeMgBr (3 M in THF) (0.80 mL, 2.42 mmol) and stirred for 16 h at 0 °C to ambient temperature. Reaction mixture was quenched with saturated NH₄CI solution and extracted with ethyl acetate. Organic extract was dried over anhydrous Na₂SO4 and concentrated under reduced pressure to afford crude compound. The product was purified by reverse phase preparative HPLC to afford (S)-1-(1-cyclopropylethyl)-3-(2-(6-(2-hydroxypropan- 2-yl)pyridin-3-yl)thiazol-4-yl)urea (Example 15 (Compound 131), 78 mg). LCMS (ESI) calcd. for CI₇H₂₂N₄O₂S: 346.1 , found [M + H]⁺= 347.2. HPLC: Rt(min) = 7.43 (99.87%).¹H NMR (400 MHz, DMSO-d₆): δH 9.28 (s, 1 H), 8.97 (s, 1 H), 8.18 (d, 1 H), 7.77 (d, 1 H), 7.23 (s, 1 H), 6.30 (d, 1 H), 5.32 (s, 1 H), 3.27-3.21 (m, 1 H), 1.46 (s, 6H), 1.14 (d, 3H), 0.89-0.85 (m, 1 H), 0.42-0.17 (m, 4H).

EXAMPLE 16

Synthesis of (S)-1-(1-cyclopropylethyl)-3-(6-phenylpyridin-2-yl)urea, Example 16 (Compound 41):

[0317] Synthesis of (S)-1-(1-cyclopropylethyl)-3-(6-phenylpyridin-2-yl)urea, Example 16 (Compound 41) [Step 1]: To a stirred solution of 6-phenylpyridin-2-amine (10-1 , 100 mg, 0.58 mmol) in dichloromethane (10 ml) was added triphosgene (174 mg, 0.58 mmol) and triethylamine (0.25 mL, 1.76 mmol) at -15 °C and stirred for 10 min. (S)-1-cyclopropylethan-1- amine (10-2, 100 mg, 1.18 mmol) was added to the reaction mixture at -15 °C. The reaction mixture was allowed to stir for 6 h at ambient temperature. Volatiles were evaporated under reduced pressure. The reaction mixture was diluted with ice cold water and extracted with ethyl acetate. The organic extract was washed with brine, dried over anhydrous Na₂SC>4, filtered and concentrated under reduced pressure. The crude product was purified by reverse phase prep HPLC and lyophilized to afford (S)-1-(1-cyclopropylethyl)-3-(6-phenylpyridin-2-yl)urea (Example 16 (Compound 41 ), 70 mg). LCMS (ESI) Calcd. for C17H19N3O: 281.35, found [M+H]⁺= 282.2. ¹H NMR (400 MHz, d₆-DMSO) b_H 9.30 (s, 1 H), 8.73 (bs, 1 H), 7.97-7.95 (m, 1 H), 7.76 (t, 1 H), 7.52-7.44 (m, 4H), 7.19 (d, 1 H), 3.34-3.30 (m, 1 H), 1.20 (d, 3H), 0.93-0.91 (m, 1 H), 0.48-0.40 (m, 2H), 0.31-0.22 (m, 2H).

EXAMPLE 17

Synthesis of (S)-6-(3-(1-cyclopropylethyl)ureido)-N,N/-dimethylpicolinamide, Example 17

(Compound 40):

[0318] Synthesis of N,N-dimethyl-6-nitropicolinamide, 11 -2 [Step 1]: To a stirred solution of 6-nitropicolinic acid (11-1 , 2.5 g, 14.9 mmol) in DMF (25 mL) were added HATU (6.8 g, 17.8 mmol) and N,N-diisopropylethylamine (6.5 mL, 37.2 mmol) at 0 °C and stirred for 10 min. Dimethylamine, 2M in THF (0.8 g, 17.8 mmol) was added to this reaction mixture and stirred at ambient temperature for 2 h. The reaction mixture was diluted with ice cold water and extracted with ethyl acetate. The organic extract was dried over anhydrous Na₂SC>4, filtered and concentrated under reduced pressure. The crude product was purified by combi flash chromatography to afford A/,A/-dimethyl-6-nitropicolinamide (11-2, 1.4 g). LCMS (ESI) Calcd. for C8H9N3O3: 195.06, found [M+H]⁺= 195.9.

[0319] Synthesis of 6-amino-/V,A/-dimethylpicolinamide, 11-3 [Step 2]: To a stirred solution of A/,A/-dimethyl-6-nitropicolinamide (11 -2, 1.4 g, 7.2 mmol) in methanol (20 mL) was added 10% Pd/C (140 mg) and hydrogenated under hydrogen balloon pressure. The reaction mixture was stirred at ambient temperature for 16 h. The reaction mixture was filtered through celite bed and washed with ethyl acetate. The filtrate was concentrated under reduced pressure to afford 6- amino-A/,A/-dimethylpicolinamide (11-3, 1 g). LCMS (ESI) Calcd. for C8H11N3O: 165.09, found [M+H]⁺= 166.2.

[0320] Synthesis of (S)-6-(3-(1-cyclopropylethyl)ureido)-A/,/V-dimethylpicolinamide, Example 17 (Compound 40) [Step 3]: To a stirred solution of 6-amino-/V,A/- dimethylpicolinamide (11 -3, 80 mg, 0.5 mmol) in dichloromethane (10 mL) was added triethylamine (0.3 mL, 1.9 mmol) followed by a solution of triphosgene (144 mg, 0.5 mmol) in DCM (1 mL) at -10 °C and stirred the reaction for 40 min. (S)-1-cyclopropylethan-1-amine (11 - 4, 82 mg, 0.1 mmol) was added and stirred at same temperature for 2 h. Volatiles were removed under reduced pressure, diluted with water and extracted with ethyl acetate. The organic extract was dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure. The crude product was purified by reverse phase preparative HPLC and lyophilized to afford (S)-6-(3-(1- cyclopropylethyl)ureido)-A/,A/-dimethylpicolinamide (Example 17 (Compound 40), 54 mg,). LCMS (ESI) Calcd. for C14H20N4O2: 276.16, found [M+H]⁺= 277.2. ¹H NMR (400 MHz, DMSO- d₆): 6H 9.22 (s, 1 H), 7.78-7.71 (m, 2H), 7.47 (d, 1 H), 7.02 (d, 1 H), 2.98 (s, 3H), 2.91 (s, 3H), 1.14 (d, 3H), 0.88-0.85 (m, 1 H), 0.44-0.37 (m, 2H), 0.28-0.25 (m, 1 H), 0.21-0.20 (m, 1 H). One proton merged with solvent residual peak at 3.33.

Example 18

Synthesis of 1-(6-methyl-2-pyridyl)-3-[(1 S)-1-cyclopropylethyl] urea, Example 18 (Compound 47):

[0321] Synthesis of 1-(6-methyl-2-pyridyl)-3-[(1 S)-1-cyclopropylethyl] urea, 9m [Step 1]: To a stirred solution of 6-methylpyridin-2-amine (12-1 , 100 mg, 0.93 mmol) in dichloromethane (2 mL) was added (1 R)-1-cyclopropylethanamine (12-2, 87 mg, 1.02 mmol), followed by addition of triethylamine (0.39 mL, 2.77 mmol) and triphosgene (165 mg, 0.56 mmol) at -5°C. The reaction mixture was allowed to stir for 40 min at the same temperature. The reaction mixture was diluted with water and extracted with dichloromethane. Combined organic extracts were dried over anhydrous Na2SO4 and concentrated under reduced pressure to afford crude product. The product was purified by reverse phase preparative HPLC to afford 1-(6-methyl-2-pyridyl)-3- [(1 S)-1-cyclopropylethyl] urea (Example 18 (Compound 47), 65 mg). LCMS (ESI) Calcd. for C12H17N3O: 219.1 , found [M+H]⁺ = 220.1 , ¹H NMR (400 MHz, DMSO-d₆): <δ_H 9.05 (s, 1 H), 8.40 (s, 1 H), 7.53 (t, 1 H), 7.05 (d, 1 H), 6.76 (d,1 H), 3.37-3.31 (m, 1 H), 2.36 (s, 3H), 1.15 (d, 3H), 0.94-0.86 (m, 1 H), 0.42-0.36 (m, 2H), 0.28-0.18 (m, 2H). Examples 19 & 20

Synthesis of 1-((S)-1-cyclopropylethyl)-3-(6-((S)-3-hydroxypyrrolidin-1-yl)pyridin-2-yl)urea

Example 19 (Compound 31):

[0322] Synthesis of (S)-1-(6-bromopyridin-2-yl)-3-(1-cyclopropylethyl)urea, 13-3 [Step 1]: (S)-1-(6-bromopyridin-2-yl)-3-(1-cyclopropylethyl)urea was synthesized following scheme 1 , Step-1 .

[0323] Synthesis of 1-((S)-1-cyclopropylethyl)-3-(6-((S)-3-hydroxypyrrolidin-1-yl)pyridin- 2-yl)urea, Example 19 (Compound 31 ) [Step 2]: In a sealed tube, to a stirred solution of (S)-1- (6-bromopyridin-2-yl)-3-(1-cyclopropylethyl)urea (13-3, 100 mg, 0.352 mmol) in THF (0.6 ml_) was added DBU (0.60 mL, 4.2 mmol), (S)-pyrrolidin-S-ol (13-4, 6.00 eq, 180 mg, 2.1 mmol) and continued stirring at 70 °C for 16 h. The reaction mixture was diluted with ethyl acetate, washed with water and brine. The combined organic extracts were dried over anhydrous sodium sulphate and concentrated under reduced pressure. The product was purified by reverse phase preparative HPLC to afford 1-((S)-1-cyclopropylethyl)-3-(6-((S)-3-hydroxypyrrolidin-1-yl)pyridin-

2-yl)urea (Example 19 (Compound 31), 50 mg). LCMS (ESI) Calcd. for C15H22N4O2: 290.2, found [M + H]⁺= 291.2. ¹H NMR (400 MHz, DMSO-d₆): <δ_H 9.20 (s, 1 H), 8.90 (s, 1 H), 7.33 (t, 1 H), 6.16 (d, 1 H), 5.92 (d, 1 H), 4.98 (d, 1 H), 4.38 (s, 1 H), 3.49-3.40 (m, 3H), 3.28-3.22 (m, 2H), 2.03- 1.98 (m, 1 H), 1.90 (brs, 1 H), 1.18 (d, 3H), 0.88-0.86 (m, 1 H), 0.45-0.36 (m, 2H), 0.29-0.25 (m, 1 H), 0.21-0.19 (m, 1 H).

[0324] Synthesis of 1-((S)-1-cyclopropylethyl)-3-(6-((/?)-3-hydroxypyrrolidin-1-yl) pyridin-2-yl) urea (Example 20 (Compound 32)) [Step 1]: 1-((S)-1-cyclopropylethyl)-3-(6-((R)-

3-hydroxypyrrolidin-1-yl) pyridin-2-yl) urea was synthesized following Scheme 35, Step 2.

LCMS (ESI) Calcd. for Ci5H₂2N₄O2:290.36, found [M+H] ⁺ = 291.2. ¹H NMR (400 MHz, DMSO-d₆): <δ_H9.21(d, 1H), 8.90 (s, 1 H), 7.33 (t, 1 H), 6.15 (d, 1 H), 5.91 (d, 1 H), 4.98 (d, 1 H), 4.38 (brs, 1H), 3.49- 3.39 (m, 3H), 3.30-3.23 (m, 2H), 2.07-1.98 (m, 1 H), 1.91-1.87 (m, 1 H), 1.17-1.16 (m, 3H), 0.90-0.84 (m, 1 H), 0.46-0.31 (m, 4H).

Examples 21 & 22

Synthesis of 1-((S)-1-cyclopropylethyl)-3-(2-(4-(2,2,2-trifluoro-1-hydroxyethyl)phenyl)thiazol- 4-yl)urea, Example 22 (Compound 11) and 1-((S)-1-cyclopropylethyl)-3-(2-(4-(2,2,2-trifluoro-1- hydroxyethyl)phenyl)thiazol-4-yl)urea, Example 21 (Compound 10):

[0325] Synthesis of (S)-1-(2-bromothiazol-4-yl)-3-(1-cyclopropylethyl)urea, 14-3 [Step 1]: To a stirred solution of tert-butyl-A/-(2-bromothiazol-4-yl)carbamate (14-1 , 0.98 g, 3.52 mmol) in dichloromethane (30 mL) were added 2-chloropyridine (2.0 mL, 21.1 mmol) and trifluoromethanesulfonic anhydride (1.8 mL, 10.6 mmol) at ice cold condition and stirred at ambient temperature for 50 min. A solution of triethylamine (4.9 mL, 35.2 mmol), and (1 S)-1- cyclopropylethanamine (14-2, 300 mg, 3.52 mmol) in dichloromethane (15 mL) was added at ice cold condition and the reaction mixture was stirred for 16 h at ambient temperature. Reaction mixture was diluted with dichloromethane and washed with water. Organic layer was separated, dried over anhydrous Na₂SO4 and concentrated under reduced pressure to afford crude product. The product was purified by combiflash column chromatography to afford 1-(2-bromothiazol-4- yl)-3-(1 S)-1-cyclopropylethyl]urea (14-3, 350 mg). LCMS (ESI) Cacld. for C₉Hi₂BrN₃OS: 288.99, found [M+H]⁺: 290.05 & [M+2+H]+: 292.05

[0326] Synthesis of 1 -((S)-1-cyclopropylethyl)-3-(2-(4-(2,2,2-trifluoro-1- hydroxyethyl)phenyl)thiazol-4-yl)urea, 14-5 [Step 2]: To a solution of 1-(2-bromothiazol-4-yl)- 3-(1 S)-1-cyclopropylethyl]urea (14-3, 350 mg, 1.21 mmol), 2,2,2-trifluoro-1-[4-(4,4,5,5- tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl]ethanol (14-4, 547 mg, 1.81 mmol) and potassium phosphate (768 mg, 3.62 mmol) in 1 ,4-dioxane (10 mL) and water (1 mL), was degassed by argon for 15 min, then Pd-118 (79 mg, 0.121 mmol) was added and stirred at 90 °C for 4 h. The reaction mixture was diluted with water and extracted with ethyl acetate. Organic extract was dried over anhydrous Na2SO4 and concentrated under reduced pressure to afford 1-(1 S)-1- cyclopropylethyl]-3-[2-[4-(2,2,2-trifluoro-1-hydroxy-ethyl)phenyl]thiazol-4-yl)urea (14-5, 230 mg). LCMS (ESI) Cacld. for C17H18F3N3O2S: 385.1 , found [M+H]⁺: 386.2.

[0327] Separation of 1-((S)-1-cyclopropylethyl)-3-(2-(4-((S)-2,2,2-trifluoro-1- hydroxyethyl)phenyl)thiazol-4-yl)urea, Example 22 (Compound 11) and 1 -((S)-1- cyclopropylethyl)-3-(2-(4-((R)-2,2,2-trifluoro-1-hydroxyethyl)phenyl)thiazol-4-yl)urea, Example 21 (Compound 10) [Step 3]: Purification of 1-(1 S)-1-cyclopropylethyl]-3-[2-[4-(2,2,2- trifluoro-1-hydroxy-ethyl)phenyl]thiazol-4-yl)urea (110.0 mg) was done by SFC prep purification on Pic Solutions-175 instrument equipped with Knauer 40D UV/Visible Detector by using I- Cellulose-J (30.0 mm x 250 mm ), 5p Column operating at 35°C temperature, maintaining flow rate of 100 ml/min, using 70% CO2 in super critical state & 30% of 100% Isopropanol as Mobile phase. Run this isocratic mixture up to 10.0 minutes and also maintained the isobaric condition of 100 bar at 240 nm wavelength to afford the first fraction 1-((S)-1-cyclopropylethyl)-3-(2-(4- ((S)-2,2,2-trifluoro-1-hydroxyethyl)phenyl)thiazol-4-yl)urea (Example 22 (Compound 11), 49 mg) as peak 1 and the second fraction 1-((S)-1-cyclopropylethyl)-3-(2-(4-((R)-2,2,2-trifluoro-1- hydroxyethyl)phenyl)thiazol-4-yl)urea (Example 21 (Compound 10), 49 mg) as peak 2. Note: Absolute stereochemistry of the compounds are unknown, single pure unknown diastereomers.

[0328] 1 -((S)-1-cyclopropylethyl)-3-(2-(4-(2,2,2-trifluoro-1-hydroxyethyl)phenyl)thiazol-4- yl)urea, Example 22 (Compound 11): LCMS (ESI) Cacld. for C17H18F3N3O2S: 385.1 , found [M+H]⁺: 386.2, HPLC: Rt(min) = 7.42 (99.55%). ¹H NMR (400 MHz, DMSO-d₆): δ_H 9.24 (s, 1 H), 7.90 (d, 2H), 7.60 (d, 2H), 7.19 (s, 1 H), 6.94 (d, 1 H), 6.31 (d, 1 H), 5.27-5.23 (m, 1 H), 3.25-3.23 (m, 1 H), 1.14 (d, 3H), 0.90-0.85 (m, 1 H), 0.44-0.37 (m, 2H), 0.28-0.19 (m, 2H).

[0329] 1 -((S)-1-cyclopropylethyl)-3-(2-(4-(2,2,2-trifluoro-1-hydroxyethyl)phenyl)thiazol-4- yl)urea, Example 21 (Compound 10): LCMS (ESI) Cacld. for C17H18F3N3O2S: 385.1 , found [M+H]⁺: 386.2, HPLC: Rt(min) = 7.44 (99.84%). ¹H NMR (400 MHz, DMSO-d₆): δH 9.24 (s, 1 H), 7.90 (d, 2H), 7.60 (d, 2H), 7.19 (s, 1 H), 6.94 (d, 1 H), 6.31 (d, 1 H), 5.27-5.23 (m, 1 H), 3.25-3.21 (m, 1 H), 1.14 (d, 3H), 0.89-0.84 (m, 1 H), 0.44-0.37 (m, 2H), 0.29-0.19 (m, 2H).

Example 23 Synthesis of (S)-1-(2-(1-cyclopropyl-1H-imidazol-4-yl)thiazol-4-yl)-3-(1-cyclopropylethyl)urea,

Example 23 (Compound 9):

[0330] Synthesis of (S)-1-(2-(1-cyclopropyl-1 H-imidazol-4-yl)thiazol-4-yl)-3-(1- cyclopropylethyl)urea, Example 23 (Compound 9) [Step 1]: To a solution of 1-(2- bromothiazol-4-yl)-3-[(1 S)-1-cyclopropylethyl]urea (14-3, 100 mg, 0.345 mmol), 1-cyclopropyl- 4-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)imidazole (15-1 , 121 mg, 0.517 mmol) and potassium phosphate (219 mg, 1.03 mmol) in 1 ,4-dioxane (5 mL) and water (0.5 mL), was degassed by argon for 15 min, then Pd-118 (22 mg, 0.0345 mmol) was added and stirred at 90 °C for 4 h. The reaction mixture was diluted with water and extracted with ethyl acetate. Organic extract was dried over anhydrous Na2SO4 and concentrated under reduced pressure to afford crude compound, which was purified by reverse phase preparative HPLC to afford 1-[2-(1- cyclopropylimidazol-4-yl)thiazol-4-yl]-3-[(1 S)-1-cyclopropylethyl]urea (Example 23 (Compound 9), 26 mg). LCMS (ESI) Calcd. for C15H19N5OS: 317.1 , found [M+H]⁺: 318.2, HPLC: Rt(min) = 7.13 (98.47%). ¹H NMR (400 MHz, DMSO-d₆): δH 9.00 (s, 1 H), 7.80 (s, 1 H), 7.60 (s, 1 H), 6.94 (s, 1 H), 6.32 (d, 1 H), 3.60-3.54 (m, 1 H), 3.26-3.19 (m, 1 H), 1.13 (d, 3H), 1.05-1.02 (m, 2H), 0.99- 0.96 (m, 2H), 0.88-0.83 (m, 1 H), 0.43-0.36 (m, 2H), 0.28-0.19 (m, 2H).

Biological/ Biochemical Evaluation

DNA POLYMERIZATION ASSAY TO MONITOR EFFECTS ON HUMAN WILDTYPE POLI" AND FIVE DISEASE-CAUSING MUTANT VARIANTS OF HUMAN POLI".

[0331] The ability of small molecules to stimulate the polymerase activity of POLy was analyzed in a quantitative fluorescence SYBR Green I assay with fluorescence intensity (Fl) readout.

[0332] In the presence of dNTPs, wild type human POLy and mutant derivatives thereof were incubated with single-stranded, circular M13mp18 ssDNA hybridized to a short DNAoligonucleotide that functions as a primer for initiation of DNA synthesis. POLy and mutantderivatives thereof will extend the primer and use the M13mp18 molecule as a template tosynthesize long stretches of double-stranded DNA. SYBR Green I, which is a double-stranded DNA-binding dye, was used to quantify formation of double-stranded DNA in the reaction. When bound to double-stranded DNA, SYBR Green I fluorescence increases by up to 100-fold.

[0333] The assay was performed in the 384-well plate format. Upon addition of compounds, changes in double-stranded DNA synthesis activity can be monitored by following effects on fluorescent intensity.

[0334] Proteins used in the polymerization assays were wild type POLyA (POLyA:WT) and mutant derivatives thereof in which an Alanine in position 467 of the amino acids sequence had been changed to Threonine (POLyA:A467T), a Glycine in position 848 of the amino acids sequence had been changed to Serine (POLyA:G848S), an Arginine in position 309 of the amino acids sequence had been changed to Cysteine (POLyA:R309C), or a Tyrosine in position 955 of the amino acid sequence had been changed to Cysteine (POLyA:Y955C). W748S substitution arises because of a G to C mutation at position 2243 in exon 13. The W748S mutation is generally found in cis with the E1143G polymorphism, which is caused by an A to G transition at nucleotide 3428 in exon 21 of POLy. The reactions also contained the accessory POLyB subunit, and the human mitochondrial single stranded DNA binding protein (mtSSB).

[0335] The protein mixture used in the assays contained POLyA or a mutant derivative (1 nM), POLyB (1.3 nM, concentration calculated as a dimer), 25 mM Tris-HCI (pH 8.0), 0.1 mg/mL bovine serum albumin, 1 mM Bond-Breaker TCEP solution (pH 7) [Thermo Fisher], 25 mM NaCI, and 0.02% Triton X-100.

[0336] The primed circular ssDNA template used in the polymerization assay was generated by hybridizing circular, single-stranded DNA from M13mp18 with a 20 nucleotides longoligonucleotide (5'-GTA AAA CGA CGG CCA GTG CC-3') using a Bio-Rad T100 Thermal Cycler.

[0337] The DNA template mixture used in the assay contained 0.5 nM primed M13 mp18ssDNA, 400 nM mtSSB, 0.1 mM Tris-HCI (pH 8.0), 0.1 mM dNTP, 10 mM MgCI2, 0.1 mg/mL BSA, 1 mM Bond-Breaker TCEP solution [Thermo Fisher], and 0.02% Triton X-100. The DNA template mixture for the wild type protein contained either 100 pM dNTP or 0.1 pM dNTP.

[0338] Microplates with compounds (0.1 pL in each well) to be tested in the assay were prepared from 10 mM compound stocks in 100% DMSO, and equal amounts of DMSO without any compound were added to positive and negative control wells. [0339] The protein mixture was dispensed (5 pL into each well) into compound plates and incubated at 37 °C for 15 minutes. After incubation, 5 pL of the DNA template mixture was dispensed into each well. The plates were then incubated at 37 °C for 2 hours.

[0340] During incubation, a combined stop buffer and detection reagent solution was prepared. The buffer contained 25 mM EDTA (pH 8.0), 0.02% Triton X-100, and SYBR Green I diluted 10,000 X (for fluorescent readout). EDTA will stop the enzymatic reaction by chelating magnesium ions.

[0341] After the incubation period, SYBR Green I solution was added (10 pL) to the screening plates and the plates were incubated at room temperature in the dark for 20 minutes before the fluorescent signal was read between 485-520 nM on a microtiter plate reader. The compound concentrations for half maximum activity (AC50) are provided in Table 1.

[0342] Table 1 : AC50 Values on Human POLy and Human Mutant POLy.

Claims

CLAIMS What is claimed is:

1. A compound, or a pharmaceutically acceptable salt thereof, according to Formula (I): wherein:

A is substituted diazole, oxazole, substituted oxazole, thiazole, substituted thiazole, thiadiazole, substituted thiadiazole, oxadiazole, substituted oxadiazole, triazole, substituted triazole, pyridine, or substituted pyridine; wherein when the substituted diazole is a pyrazole the substitution is not an unsubstituted phenyl.

2. The compound of claim 1 , wherein A is substituted diazole and with any one of Formula (I- a) or Formula (l-b):

wherein:

R1-R3 are each independently hydrogen or C1-C6 alkyl; and

B is substituted C1-C6 alkyl, C3-C6 cycloalkyl, substituted C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, substituted C3-C6 heterocycloalkyl, C3-C6 heterocyclyl, substituted C3-C6 heterocyclyl, substituted aryl, amide, or substituted amide.

3. The compound of claim 2, wherein B is substituted C1-C6 alkyl.

4. The compound of claim 3, wherein the substituted C1-C6 alkyl is substituted with one or more of benzyl, fluorine, or methyl.

5. The compound of claim 4, the substituted C1-C6 alkyl is methylbenzene, ethylbenzene, trifluoroethyl, difluoroethyl, or N,N-dimethylethanamine.

6. The compound of claim 2, wherein B is substituted aryl and the compound is substituted pyrazole with Formula (l-a-1): wherein:

R1-R2 are each independently hydrogen or methyl; R₃ is hydrogen;

R₄ is hydrogen or halogen;

R₅ and R₆ are each independently hydrogen, halogen, C1-C6 alkyl, C1-C6 substituted alkyl, carboxylic acid, substituted carboxylic acid, sulfonyl, substituted sulfonyl, alkoxy, amide, substituted amide, amine, substituted amine, C3-C6 heterocycloalkyl, substituted C3-C6 heterocycloalkyl, C3-C6 cycloalkyl, or substituted C3-C6 cycloalkyl;

R₇ and R₈ is are each independently hydrogen or alkoxy; and

R₄- R₈ can not be hydrogen at the same time.

7. The compound of claim 6, wherein R1 and R2 are hydrogen, and with Formula (l-a-2): wherein:

R₄ is hydrogen or fluorine;

R5 is hydrogen, fluorine, bromine, chlorine, methyl, trifluoromethyl, carboxylic acid, methyl substituted carboxylic acid, methoxy, substituted ethyl, amide, methyl amide, dimethyl amide, methyl sulfonyl, N-methyl sulfonyl amide, or substituted Ci-C₈ alkyl;

Re is hydrogen, fluorine, methoxy, methoxy methyl, methoxy ethyl, substituted ethyl, amide, methyl amide, amide, methyl amide, dimethyl amide, substituted alkyl, substituted sulfonyl, cyclobutyl, substituted cyclobutyl, oxetane, substituted oxetane;

R₇ and R₈ are each independently hydrogen or methoxy; and

R₄-R₈ can not be hydrogen at the same time.

8. The compound of claim 7, wherein R₆ is substituted oxetane.

9. The compound of claim 8, wherein the substituted oxetane is oxetan-3-ol.

10. The compound of claim 7, wherein R₆ is substituted cyclobutane.

11. The compound of claim 10, wherein the substituted cyclobutane is 3,3-difluorocyclobutan- 1-ol.

12. The compound of claim 7, wherein the substituted sulfonyl is with Formula (l-a-3): wherein R₉ is methyl, ethyl, trifluoromethyl, cyclopropyl, or methyl amide.

13. The compound of claim 7, wherein the substituted alkyl is methyl or ethyl substituted with one or more of methyl, amine, methyl amine, dimethyl amide, hydroxyl, or fluorine.

14. The compound of claim 2, wherein B is C3-C6 cycloalkyl or substituted C3-C6 cycloalkyl, with Formula (l-a-4): wherein R' and R" are each independently hydrogen or fluorine.

15. The compound of claim 2, wherein B is C3-C6 heterocyclyl or substituted C3-C6 heterocyclyl, and with any one of Formula (l-a-5), Formula (l-a-6), Formula (l-a-7), Formula (I- a-8), Formula (l-a-9), Formula (l-a-10), or Formula (l-a-11):

wherein:

R4 and R7-R8 are hydrogen;

R5 is hydrogen, substituted C1-C6 alkyl, or methoxy; and R6 is hydrogen or substituted C1-C6 alkyl.

16. The compound of claim 15, wherein R5 is substituted C1-C6 alkyl.

17. The compound of claim 15, wherein the substituted C1-C6 alkyl is trifluoromethane.

18. The compound of claim 15, wherein Re is substituted C1-C6 alkyl.

19. The compound of claim 18, wherein the substituted C1-C6 alkyl is substituted with one or more of fluorine or hydroxyl.

20. The compound of claim 18, wherein the substituted C1-C6 alkyl is 2,2,2-trifluoroethan-1-ol or propan-2-ol.

21. The compound of claim 2, wherein B is aryl and the compound is substituted imidazole with Formula (l-c-1): wherein R₂ and R₃ are hydrogen.

22. The compound of claim 1 , wherein A is substituted thiazole and with Formula (l-d), Formula (l-e), or Formula (l-f):

wherein C is 1-C6 alkyl, substituted C1-C6 alkyl, C3-C6 cycloalkyl, substituted C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, substituted C3-C6 heterocycloalkyl, C3-C6 heterocyclyl, substituted C3-C6 heterocyclyl, aryl, substituted aryl, amide, or substituted amide.

23. The compound of claim 22, wherein C is C3-C6 heterocyclyl or substituted C3-C6 heterocyclyl, with any one of Formula (l-d-1), Formula (l-d-2), Formula (l-d-3), Formula (l-d- 4), Formula (l-d-5), Formula (l-d-6), or Formula (l-d-7):

wherein:

R10 and R12-R13 are hydrogen;

R11 is C1-C6 alkyl, substituted C1-C6 alkyl, or C3-C6 cycloalkyl; R14-R18 are each independently hydrogen, fluorine, substituted methoxy, or substituted ethyl.

24. The compound of claim 23, wherein the C1-C6 alkyl is methyl.

25. The compound of claim 23, wherein the substituted C1-C6 alkyl is substituted with one or more of cyano, halogen, or alkoxy.

26. The compound of claim 25, wherein the substituted C1-C6 alkyl is trifluoromethyl.

27. The compound of claim 25, wherein the substituted C1-C6 alkyl is isobutyronitrile, trifluoroethyl, tri-fluoropropyl, or (trifluoromethoxy)ethyl.

28. The compound of claim 22, wherein C is aryl or substituted aryl with Formula (l-d-8), Formula (l-e-1) , or Formula (l-f-1 ): wherein:

R14-R15 are each independently hydrogen or fluorine;

R16 is hydrogen, fluorine, substituted methoxy, or substituted ethyl; and R17-R18 are hydrogen.

29. The compound of claim 22, wherein C is C1-C6 alkyl, substituted C1-C6 alkyl, C3-C6 cycloalkyl, or C3-C6 substituted cycloalkyl.

30. The compound of claim 29, wherein the C1-C6 alkyl is methyl.

31. The compound of claim 29, wherein the substituted C1-C6 alkyl is trifluoromethyl.

32. The compound of claim 29, wherein the C3-C6 substituted cycloalkyl is a bridged cyclobutyl.

33. The compound of claim 1 , wherein A is pyridine or substituted pyridine and with Formula (i-g): wherein R19 is C1-C6 alkyl, aryl, C3-C6 heterocycloalkyl, substituted C3-C6 heterocycloalkyl, amide, or substituted amide.

34. The compound of claim 33, wherein R19 is methyl.

35. The compound of claim 33, wherein R19 is amide, methyl amide, or dimethyl amide.

36. The compound of claim 33, wherein R19 is a nitrogen containing C3-C6 heterocycloalkyl.

37. The compound of claim 1 , wherein A is oxazole with Formula (l-h) or Formula (l-i);

wherein R24 is aryl.

38. The compound of claim 1 , wherein A is oxadiazole and with Formula (l-j) or Formula (l-k): wherein R 25 is aryl.

39. The compound of claim 1 , wherein A is thiadiazole and with Formula (l-l):

wherein R26 is aryl.

40. The compound of claim 1 , wherein A is triazole and with Formula (l-m): wherein R27 is aryl.

41. A compound, or a pharmaceutically acceptable salt thereof, according to one of the following formulas: