PARG inhibitory compounds

Abstract

The present invention provides compounds of formula (I): JPEG2025535036000030.jpg57123 or a pharmaceutically acceptable salt thereof. The present invention further relates to a compound of formula (I) of the present invention for use in therapy. The compound is particularly useful as a PARG inhibitor and can be used in a method for treating a proliferative disorder, preferably cancer.

Description

[Technical Field]

[0001] The present invention provides compounds of formula (I):

[0002] [ka] or a pharmaceutically acceptable salt thereof. The present invention further relates to the compound of formula (I) of the present invention for use in therapy. The compound of the present invention is particularly useful as a PARG inhibitor and can be used in a method for treating a proliferative disorder, preferably cancer. [Background technology]

[0003] Cancer is a leading cause of death worldwide. Although progression-free survival and overall survival for cancer patients have improved over the past 20 years, millions of cancer patients still have few treatment options and poor survival outcomes (Jemal et al., J. Natl. Cancer Inst. 2017, 109, 1975).

[0004] DNA replication stress (DRS) is a hallmark of cancer cells and a major cause of genomic instability (a) Halazonetis et al., Science 2008, 319, 1352; b) Negrini et al., Nat. Rev. Mol. Cell Biol. 2010, 11, 220). Broadly defined, DRS refers to the deregulation of DNA replication and cell cycle progression. DRS can be induced by endogenous or exogenous causes, such as oncogene activation and chemotherapeutic agents, respectively (Zeman and Cimprich, Nat. Cell Biol. 2013, 16, 2). At the replication fork level, DRS leads to replication fork stalling, replisome disengagement, and eventual collapse. Several DNA repair proteins are involved in the stability, protection, and restart of replication forks under DRS conditions (a) Costantino et al., Science 2014, 343, 88; b) Scully et al., Curr. Opin. Genet. Dev. 2021 71, 154).

[0005] Poly(ADP) ribosylation (PARylation) is a transient and reversible post-translational modification that occurs at DNA damage sites and is catalyzed by the poly(ADP-ribose) polymerase (PARP) family of proteins (Cohen and Chang, Nat. Chem. Biol. 2018, 14, 236). PARylation of various DNA repair proteins leads to their activation. Degradation of poly(ADP) ribose chains is primarily mediated by poly(ADP-ribose) glycohydrolase (PARG) proteins. DNA damage-dependent PARylation / dePARylation is a rapid and dynamic process that must be well regulated, as an imbalance between the two processes can lead to DNA damage.

[0006] Human PARG encodes a 976-amino acid, 111-kDa protein. It contains an N-terminal regulatory domain, a catalytic domain, and an ADP-ribose-binding macrodomain. Five human PARG transcripts have been identified. Full-length PARG is predominantly nuclear; smaller isoforms are primarily localized in the cytoplasm. PARG functions primarily as an exohydrolase, hydrolyzing the α-O-glycosidic ribose-ribose bond in PAR, releasing primarily mono(ADP-ribose). PARG can also act as an endohydrolase. PARG preferentially degrades long, linear PAR chains, whereas its activity on small, branched PAR chains is significantly reduced (O'Sullivan et al., Nat. Commun. 2019, 10, 1182).

[0007] PARG is the major cellular PAR degrading enzyme, but it cannot act on terminal protein-ribose bonds. Additional hydrolases, such as terminal ADP-ribose protein glycohydrolase (TARG1) and ADP-ribosylhydrolase 3 (ARH3), are also known to catalyze PAR degradation. TARG1 and ARH3 complete the reversal of PARylation by removing the protein-bound mono(ADP-ribose) moiety (a) Fontana et al., Elife 2017, doi:10.7554 / eLife.28533; b) Rack et al., Genes Dev. 2020, 34, 263). TARG1 is located in the nucleus and cytoplasm. ARH3 is found primarily in the cytoplasm but can also be found in mitochondria and the nucleus (Rack et al., Genes Dev. 2020, 34, 263).

[0008] Genomic aberrations targeting tumor suppressor genes or oncogenes often render cancer cells dependent on specific DNA repair pathways. For example, PARP inhibitors are known to be particularly effective against tumors with mutations in the BRCA1 and BRCA2 genes (a) Bryant et al., Nature 2005, 434, 913; b) Farmer et al., Nature 2005, 434, 917). Targeting synthetic lethal interactions, such as those between PARP and BRCA, is an attractive novel therapeutic approach for cancer treatment.

[0009] PARG is involved in various DNA repair mechanisms, including DNA replication and single-strand break (SSB) repair and replication fork restart. PARG inhibitors demonstrated synthetic lethality in cells with high levels of DRS, which is caused by underexpression of genes involved in DNA replication and / or replication fork stability (Pillay et al., Cancer Cell. 2019, 35, 519). Furthermore, PARG inactivation, depletion, or inhibition sensitizes cells to DNA-damaging agents such as irradiation and alkylating agents (e.g., temozolomide and methyl methanesulfonate) (a) Fujihara et al., Curr. Cancer Drug Targets 2009, 9, 953; b) Gogola et al., Cancer Cell 2018, 33, 1078; c) Houl et al., Nat Commun. 2019, 10, 5654).

[0010] Considering the therapeutic potential of PARG inhibitors in cancer treatment, there is an increasing need for the development of highly potent and selective PARG inhibitors beyond those already described (a) James et al., ACS Chem. Biol. 2016, 11, 3179; b) Waszkowycz et al., J. Med. Chem. 2018, 61, 10767).

[0011] Certain compounds useful as PARG inhibitors are further disclosed in WO 2016 / 092326, WO 2016 / 097749 and WO 2021 / 055744.

[0012] US Patent Application Publication No. 2019 / 233411 discloses certain Gcn2 inhibitors and their uses.

[0013] WO 2009 / 050183 discloses certain imidazo[1,2-a]pyridine derivatives that are useful for treating diseases mediated by the ALK-5 and / or ALK-4 receptors. Summary of the Invention

[0014] The objective technical problem of the present invention was to provide compounds that are cell-permeable inhibitors of PARG. The technical problem of the present invention is solved by the embodiments described herein and characterized in the claims.

[0015] Thus, in a first embodiment, the present invention provides a compound of formula (I):

[0016] [ka] or a tautomer, a pharmaceutically acceptable solvate, a pharmaceutically acceptable crystal form, a pharmaceutically acceptable salt or a prodrug thereof.

[0017] Preferably, in this first embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0018] A further embodiment of the present invention relates to a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier.

[0019] In a further embodiment, the present invention relates to a compound of formula (I) of the present invention or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition of the present invention, for use in therapy.

[0020] The compounds of formula (I) are useful for treating diseases or disorders in which PARG activity is implicated.

[0021] The compounds of formula (I) are useful in methods for treating proliferative disorders. In a preferred embodiment of the invention, the proliferative disorder is cancer, preferably human cancer.

[0022] definition The following definitions apply throughout the specification and claims, unless stated otherwise.

[0023] The term "hydrogen" is used herein to refer to protium, deuterium and / or tritium, preferably protium. Thus, the term "non-hydrogen atom" refers to any atom that is not hydrogen, i.e., not protium, deuterium or tritium.

[0024] The term "hydrocarbon group" refers to a group consisting of carbon and hydrogen atoms.

[0025] The term "alicyclic" is used in reference to a cyclic group to indicate that the corresponding cyclic group is non-aromatic.

[0026] As used herein, the term "alkyl" refers to a monovalent saturated acyclic (i.e., non-cyclic) hydrocarbon group which may be straight-chain or branched. Thus, an "alkyl" group does not contain any carbon-carbon double bonds or any carbon-carbon triple bonds. 1~5 "Alkyl" means an alkyl group having 1 to 5 carbon atoms. Preferred exemplary alkyl groups are methyl, ethyl, propyl (e.g., n-propyl or isopropyl), or butyl (e.g., n-butyl, isobutyl, sec-butyl, or tert-butyl). Unless otherwise defined, the term "alkyl" preferably refers to a C 1~4 It refers to alkyl, more preferably methyl or ethyl, even more preferably methyl.

[0027] As used herein, the term "carbocyclyl" refers to hydrocarbon ring groups, including monocyclic rings as well as bridged, spiro, and / or fused ring systems (which may, for example, be composed of two or three rings), and the ring groups may be saturated, partially unsaturated (i.e., unsaturated but not aromatic), or aromatic. Unless otherwise defined, "carbocyclyl" preferably refers to aryl, cycloalkyl, or cycloalkenyl.

[0028] As used herein, the term "heterocyclyl" refers to ring groups including monocyclic rings as well as bridged, spiro, and / or fused ring systems (which may, for example, be composed of two or three rings), which contain one or more (e.g., one, two, three, or four, etc.) ring heteroatoms independently selected from O, S, and N, and the remaining ring atoms are carbon atoms, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) are optionally oxidized, one or more carbon ring atoms are optionally oxidized (i.e., to form an oxo group), and further wherein the ring group may be saturated, partially unsaturated (i.e., unsaturated but not aromatic), or aromatic. For example, each heteroatom-containing ring contained in a ring group may contain one or two O atoms and / or one or two S atoms (which may be optionally oxidized) and / or one, two, three, or four N atoms (which may be optionally oxidized), provided that the total number of heteroatoms in the corresponding heteroatom-containing ring is 1 to 4 and that there is at least one carbon ring atom (which may be optionally oxidized) in the corresponding heteroatom-containing ring. Unless otherwise defined, "heterocyclyl" preferably refers to heteroaryl, heterocycloalkyl, or heterocycloalkenyl.

[0029] As used herein, the term "aryl" refers to an aromatic hydrocarbon ring group, including monocyclic aromatic rings and bridged and / or fused ring systems containing at least one aromatic ring (e.g., a ring system composed of two or three fused rings, where at least one of the fused rings is aromatic; or a bridged ring system composed of two or three rings, where at least one of the bridged rings is aromatic). "Aryl" can refer to, for example, phenyl, naphthyl, diarylnyl (i.e., 1,2-dihydronaphthyl), tetralinyl (i.e., 1,2,3,4-tetrahydronaphthyl), indanyl, indenyl (e.g., 1H-indenyl), anthracenyl, phenanthrenyl, 9H-fluorenyl, or azulenyl. Unless otherwise defined, "aryl" preferably has 6 to 14 ring atoms, more preferably 6 to 10 ring atoms, and even more preferably refers to phenyl or naphthyl, and most preferably refers to phenyl.

[0030] As used herein, the term "heteroaryl" refers to aromatic ring groups, including monocyclic aromatic rings and bridged and / or fused ring systems containing at least one aromatic ring (e.g., a ring system composed of two or three fused rings, where at least one of the fused rings is aromatic; or a bridged ring system composed of two or three rings, where at least one of the bridged rings is aromatic), wherein the aromatic ring groups contain one or more (e.g., one, two, three, or four) ring heteroatoms independently selected from O, S, and N, the remaining ring atoms being carbon atoms, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) are optionally oxidized, and wherein one or more carbon ring atoms are optionally oxidized (i.e., to form an oxo group). For example, each heteroatom-containing ring within an aromatic ring group may contain one or two O atoms and / or one or two S atoms (which may be optionally oxidized) and / or one, two, three, or four N atoms (which may be optionally oxidized), provided that the total number of heteroatoms in the corresponding heteroatom-containing ring is one to four and that there is at least one carbon ring atom (which may be optionally oxidized) in the corresponding heteroatom-containing ring. "Heteroaryl" includes, for example, thienyl (i.e., thiophenyl), benzo[b]thienyl, naphtho[2,3-b]thienyl, thianthrenyl, furyl (i.e., furanyl), benzofuranyl, isobenzofuranyl, chromanyl, chromenyl (e.g., 2H-1-benzopyranyl or 4H-1-benzopyranyl), isochromenyl (e.g., 1H-2-benzopyranyl), chromonyl, xanthenyl, phenoxathiinyl, pyrrolyl (e.g., 1H-pyrrolyl), imidazolyl, pyrazolyl, pyridyl (i.e., pyridinyl; e.g., 2-pyridyl, 3-pyridyl, or 4-pyridyl), pyrazinyl, pyrimidinyl, pyridazinyl, indolyl (e.g., 3H-indolyl), isoindolyl, indazolyl, indolizinyl, purinyl, quinolyl, isoquinolyl, phthalazinyl, naphthyridinyl, quinoxalinyl, cinnolinyl, pteridinyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl, perimidinyl, phenanthrolinyl (e.g., [1,10]phenanthrolinyl, [1,7]phenanthrolinyl, or [4,7]phenanthrolinyl), phenazinyl, thiazolyl, isothiazolyl, phenothiazinyl, oxazolyl, isoxazolyl, oxadiazolyl (e.g., 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl (i.e., furazanyl), or 1,3,4-oxadiazolyl), thiadiazolyl (e.g., 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, or 1,3,4-thiadiazolyl), phenoxazinyl, pyrazolo[1,5-a]pyrimidinyl (e.g., pyrazolo[1,5-a]pyrimidin-3-yl), 1,2-benzisoxazol-3-yl, benzothiazolyl, benzothiadiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzo[b]thiazolyl, phenyl (i.e., benzothienyl), triazolyl (e.g., 1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl, or 4H-1,2,4-triazolyl), benzotriazolyl, 1H-tetrazolyl, 2H-tetrazolyl, triazinyl (e.g., 1,2,3-triazinyl, 1,2,4-triazinyl, or 1,3,5-triazinyl), furo[2,3-c]pyridinyl, dihydrofuropyridinyl (e.g., 2,3-dihydrofuro[2,3-c]pyridinyl or 1,3-dihydrofuro[3,4-c]pyridinyl), imidazopyridinyl (e.g., imidazo[1,2-a]pyridinyl or imidazo[3,2-a]pyridinyl), quinazolinyl, thienopyridinyl, tetrahydrothienopyridinyl (e.g., 4,5,6,7-tetrahydrothieno[3,2-c]pyridinyl), dibenzofuranyl, 1,3-benzodioxolyl, benzodioxanyl (e.g., 1,3-benzodioxanyl or 1,4-benzodioxanyl) or coumarinyl. Unless otherwise defined, the term "heteroaryl" preferably refers to a 5- to 14-membered (more preferably 5- to 10-membered) monocyclic ring or fused ring system containing one or more (e.g., 1, 2, 3, or 4) ring heteroatoms independently selected from O, S, and N, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) are optionally oxidized, and one or more carbon ring atoms are optionally oxidized; even more preferably, the term "heteroaryl" refers to a 5- or 6-membered monocyclic ring containing one or more (e.g., 1, 2, or 3) ring heteroatoms independently selected from O, S, and N, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) are optionally oxidized, and one or more carbon ring atoms are optionally oxidized.

[0031] As used herein, the term "cycloalkyl" refers to a saturated hydrocarbon ring group, which includes monocyclic rings as well as bridged rings, spiro rings, and / or fused ring systems (e.g., may be composed of two or three rings; e.g., fused ring systems composed of two or three fused rings, etc.). "Cycloalkyl" can refer to, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, decalinyl (i.e., decahydronaphthyl), or adamantyl. Unless otherwise defined, "cycloalkyl" preferably refers to C 3~11 It refers to cycloalkyl, more preferably C 3~7 Particularly preferred "cycloalkyl" is a monocyclic saturated hydrocarbon ring having 3 to 7 ring members (e.g., cyclopropyl or cyclohexyl).

[0032] As used herein, the term "cycloalkenyl" refers to an unsaturated alicyclic (non-aromatic) hydrocarbon ring group, which includes monocyclic rings as well as bridged rings, spiro rings, and / or fused ring systems (e.g., may be composed of two or three rings; e.g., a fused ring system composed of two or three fused rings, etc.), wherein the hydrocarbon ring group contains one or more (e.g., one or two) carbon-carbon double bonds and does not contain a carbon-carbon triple bond. "Cycloalkenyl" can refer to, for example, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, or cycloheptadienyl. Unless otherwise defined, "cycloalkenyl" preferably refers to C 3~11 It refers to cycloalkenyl, more preferably C 3~7 A particularly preferred "cycloalkenyl" is a monocyclic unsaturated alicyclic hydrocarbon ring having 3 to 7 ring members, which contains one or more (e.g., one or two; preferably one) carbon-carbon double bonds.

[0033] As used herein, the term "heterocycloalkyl" refers to saturated ring groups, including monocyclic rings as well as bridged, spiro, and / or fused ring systems (e.g., which may be composed of two or three rings; such as, for example, a fused ring system composed of two or three fused rings), which ring groups contain one or more (e.g., one, two, three, or four, etc.) ring heteroatoms independently selected from O, S, and N, the remaining ring atoms being carbon atoms, with one or more S ring atoms (if present) and / or one or more N ring atoms (if present) optionally being oxidized, and further one or more carbon ring atoms optionally being oxidized (i.e., to form an oxo group). For example, each heteroatom-containing ring contained in a saturated ring group may contain one or two O atoms and / or one or two S atoms (which may be optionally oxidized) and / or one, two, three, or four N atoms (which may be optionally oxidized), provided that the total number of heteroatoms in the corresponding heteroatom-containing ring is one to four and there is at least one carbon ring atom (which may be optionally oxidized) in the corresponding heteroatom-containing ring. "Heterocycloalkyl" includes, for example, aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl, azepanyl, diazepanyl (e.g., 1,4-diazepanyl), oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, morpholinyl (e.g., morpholin-4-yl), thiomorpholinyl (e.g., thiomorpholin-4-yl), oxazepanyl, oxazolidin ... It may refer to silanyl, oxetanyl, tetrahydrofuranyl, 1,3-dioxolanyl, tetrahydropyranyl, 1,4-dioxanyl, oxepanyl, thiiranyl, thietanyl, tetrahydrothiophenyl (i.e., thiolanyl), 1,3-dithiolanyl, thianyl, 1,1-dioxothianyl, thiepanyl, decahydroquinolinyl, decahydroisoquinolinyl, or 2-oxa-5-aza-bicyclo[2.2.1]hept-5-yl.Unless otherwise defined, the term "heterocycloalkyl" preferably refers to a 3- to 11-membered saturated ring group, which may be a monocyclic ring or a fused ring system (e.g., a fused ring system composed of two fused rings), wherein the ring group contains one or more (e.g., one, two, three, or four) ring heteroatoms independently selected from O, S, and N, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) are optionally oxidized, and one or more carbon ring atoms are optionally oxidized; more preferably, the term "heterocycloalkyl" refers to a 5- to 7-membered saturated monocyclic ring group containing one or more (e.g., one, two, or three) ring heteroatoms independently selected from O, S, and N, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) are optionally oxidized, and one or more carbon ring atoms are optionally oxidized.

[0034] As used herein, the term "heterocycloalkenyl" refers to an unsaturated alicyclic (non-aromatic) ring group, which includes monocyclic rings as well as bridged rings, spiro rings, and / or fused ring systems (e.g., which may be composed of two or three rings; e.g., a fused ring system composed of two or three fused rings, etc.), wherein the ring group contains one or more (e.g., one, two, three, or four, etc.) ring heteroatoms independently selected from O, S, and N, the remaining ring atoms are carbon atoms, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) are optionally oxidized, wherein one or more carbon ring atoms are optionally oxidized (i.e., to form an oxo group), and further wherein the ring group contains at least one double bond between adjacent ring atoms and does not contain any triple bonds between adjacent ring atoms. For example, each heteroatom-containing ring contained in an unsaturated alicyclic ring group may contain one or two O atoms and / or one or two S atoms (which may be optionally oxidized) and / or one, two, three, or four N atoms (which may be optionally oxidized), provided that the total number of heteroatoms in the corresponding heteroatom-containing ring is one to four and that there is at least one carbon ring atom (which may be optionally oxidized) in the corresponding heteroatom-containing ring. "Heterocycloalkenyl" includes, for example, imidazolinyl (e.g., 2-imidazolinyl (i.e., 4,5-dihydro-1H-imidazolyl), 3-imidazolinyl, or 4-imidazolinyl), tetrahydropyridinyl (e.g., 1,2,3,6-tetrahydropyridinyl), dihydropyridinyl (e.g., 1,2-dihydropyridinyl or 2,3-dihydropyridinyl), pyranyl (e.g., 2H-pyranyl or 4H- pyranyl), thiopyranyl (e.g., 2H-thiopyranyl or 4H-thiopyranyl), dihydropyranyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrazinyl, dihydroisoindolyl, octahydroquinolinyl (e.g., 1,2,3,4,4a,5,6,7-octahydroquinolinyl), or octahydroisoquinolinyl (e.g., 1,2,3,4,5,6,7,8-octahydroisoquinolinyl).Unless otherwise defined, "heterocycloalkenyl" preferably refers to a 3- to 11-membered unsaturated alicyclic ring group, which may be a monocyclic ring or a fused ring system (e.g., a fused ring system composed of two fused rings), wherein the ring group contains one or more (e.g., 1, 2, 3, or 4) ring heteroatoms independently selected from O, S, and N, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) are optionally oxidized, one or more carbon ring atoms are optionally oxidized, and the ring group has at least one carbon atom between adjacent ring atoms. and no triple bonds between adjacent ring atoms; more preferably, "heterocycloalkenyl" refers to a 5-7 membered monocyclic unsaturated non-aromatic ring group containing one or more (e.g., 1, 2, or 3) ring heteroatoms independently selected from O, S, and N, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) are optionally oxidized, and one or more carbon ring atoms are optionally oxidized, and the ring group contains at least one double bond between adjacent ring atoms and no triple bonds between adjacent ring atoms.

[0035] As used herein, the term "halogen" refers to fluoro (-F), chloro (-Cl), bromo (-Br), or iodo (-I).

[0036] As used herein, the term "haloalkyl" refers to an alkyl group substituted with one or more (preferably 1 to 6, more preferably 1 to 3) halogen atoms independently selected from fluoro, chloro, bromo, and iodo, preferably all fluoro atoms. It will be understood that the maximum number of halogen atoms is limited by the number of available bonding sites and, therefore, depends on the number of carbon atoms included in the alkyl portion of the haloalkyl group. "Haloalkyl" may refer to, for example, -CF3, -CHF2, -CH2F, -CF2-CH3, -CH2-CF3, -CH2-CHF2, -CH2-CF2-CH3-CH2-CF2-CF3, or -CH(CF3)2. A particularly preferred "haloalkyl" group is -CF3.

[0037] The terms "bond" and "covalently bonded" are used interchangeably herein unless expressly indicated otherwise or contradicted by context.

[0038] As used herein, unless expressly indicated otherwise or contradicted by context, the terms "a," "an," and "the" are used interchangeably with "one or more" and "at least one." Thus, for example, a composition that includes "a" compound of Formula (I) can be interpreted as referring to a composition that includes "one or more" compounds of Formula (I).

[0039] Whenever numerical ranges are provided / disclosed herein, it is to be understood that all values ​​and subranges encompassed within each numerical range are meant to be encompassed within the scope of the invention. Accordingly, the present invention specifically and individually relates to each value that falls within the numerical ranges disclosed herein, and each subrange encompassed by the numerical ranges disclosed herein.

[0040] As used herein, the term "about" preferably refers to ±10% of the stated numerical value, more preferably ±5% of the stated numerical value, and particularly the exact numerical value stated. When the term "about" is used in connection with the endpoints of a range, it preferably refers to a range from the lower endpoint of the stated numerical value minus 10% to the upper endpoint of the stated numerical value plus 10%, more preferably a range from the lower endpoint minus 5% to the upper endpoint plus 5%, and even more preferably a range defined by the exact numerical values ​​of the lower and upper endpoints.

[0041] As used herein, the term "comprising" (or "comprise," "comprises," "contain," "contains," or "containing") means "containing, among other things," i.e., "containing, among other optional elements," unless expressly indicated otherwise or contradicted by context. In addition, the term also includes the narrower meanings of "consisting essentially of" and "consisting of." For example, the term "A comprising B and C" means "A contains, among other things, B and C," and although A may contain additional optional elements (e.g., "A containing B, C, and D" is also included), the term also includes the meanings "A consisting essentially of B and C" and "A consisting of B and C" (i.e., A does not contain any components other than B and C). [Brief explanation of the drawings]

[0042] The present invention will be explained using the accompanying drawings, which are for illustrative purposes only and should not be construed as limiting the scope of the invention in any way. [Figure 1] 1 shows the results of testing the compound of Example 1 in an MDA-MB-436 xenograft model. DETAILED DESCRIPTION OF THE INVENTION

[0043] The present invention is described in detail below, with the understanding that the present invention relates specifically to each and every combination of the features and embodiments described herein, including any combination of the general and / or preferred features / embodiments.

[0044] In a first embodiment, the present invention provides a compound of formula (I):

[0045] [ka] or a tautomer, a pharmaceutically acceptable solvate, a pharmaceutically acceptable crystalline form, a pharmaceutically acceptable salt or prodrug thereof, or a pharmaceutically acceptable salt thereof. The compound may also be referred to as 3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-8-((3S,5S)-3,5-dimethylpiperazin-1-yl)-N-(1-methylcyclopropyl)imidazo[1,5-a]pyridine-6-sulfonamide.

[0046] As understood herein, a reference to a tautomer, pharmaceutically acceptable solvate, pharmaceutically acceptable crystal form, pharmaceutically acceptable salt or prodrug of a compound is preferably a reference to a pharmaceutically acceptable solvate (such as a hydrate), pharmaceutically acceptable crystalline form or pharmaceutically acceptable salt thereof, more preferably a pharmaceutically acceptable salt thereof.

[0047] Preferably, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0048] The present invention more specifically relates to the non-salt form of the compound of formula (I). The non-salt form may also be referred to as the free base form. Therefore, within the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof may be the free base form of the compound of formula (I).

[0049] In one embodiment, the present application relates to a formate salt of the compound of formula (I). Thus, within the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof may be a formate salt of the compound of formula (I).

[0050] The present invention also relates to each of the intermediates further described below in the Examples section of this specification, including any one of these intermediates in non-salt form or in the form of a salt (e.g., a pharmaceutically acceptable salt) of the respective compound. Such intermediates can be used, inter alia, in the synthesis of compounds of formula (I).

[0051] The scope of the present invention includes all pharmaceutically acceptable salt forms of compounds of formula (I), which may be formed, for example, by protonation of an atom having a lone pair of electrons susceptible to protonation, such as an amino group, with an inorganic or organic acid, or as a salt of an acid group (such as a carboxylic acid group) with a physiologically acceptable cation. Exemplary base addition salts include alkali metal salts such as, for example, sodium or potassium salts; alkaline earth metal salts, for example, calcium or magnesium salts; zinc salts; ammonium salts; aliphatic amine salts such as trimethylamine, triethylamine, dicyclohexylamine, ethanolamine, diethanolamine, triethanolamine, procaine salts, meglumine salts, ethylenediamine salts, choline salts, and the like; aralkylamine salts such as N,N-dibenzylethylenediamine salts, benzathine salts, benethamine salts, and the like; heterocyclic aromatic amine salts such as pyridine salts, picoline salts, quinoline salts, isoquinoline salts, and the like; quaternary ammonium salts such as tetramethylammonium salts, tetraethylammonium salts, benzyltrimethylammonium salts, benzyltriethylammonium salts, benzyltributylammonium salts, methyltrioctylammonium salts, tetrabutylammonium salts, and the like; and basic amino acid salts, for example, arginine salts, lysine salts, or histidine salts. Exemplary acid addition salts include, for example, mineral acid salts such as hydrochloride, hydrobromide, hydroiodide, sulfate (such as sulfate or hydrogen sulfate), nitrate, phosphate (such as phosphate, hydrogen phosphate, or dihydrogen phosphate), carbonate, bicarbonate, perchlorate, borate, or thiocyanate.Acetate, propionate, butyrate, pentanoate, hexanoate, heptanoate, octanoate, cyclopentanepropionate, decanoate, undecanoate, oleate, stearate, lactate, maleate, oxalate, fumarate, tartrate, malate, citrate, succinate, adipate, gluconate, glycolate, nicotinate, benzoate, salicylate, ascorbate, pamoate (embonate), camphorate, glucohepta Organic acid salts such as phosphates and pivalates; sulfonates such as methanesulfonate (mesylate), ethanesulfonate (esylate), 2-hydroxyethanesulfonate (isethionate), benzenesulfonate (besylate), p-toluenesulfonate (tosylate), 2-naphthalenesulfonate (naphsylate), 3-phenylsulfonate, and camphorsulfonate; glycerophosphate; and acidic amino acid salts such as aspartate or glutamate. Preferred pharmaceutically acceptable salts of the compound of formula (I) include hydrochloride, hydrobromide, mesylate, sulfate, tartrate, fumarate, acetate, citrate, and phosphate. A particularly preferred pharmaceutically acceptable salt of the compound of formula (I) is hydrochloride. Thus, the compound of formula (I), including any one of the specific compounds of formula (I) described herein, is preferably in the form of a hydrochloride, hydrobromide, mesylate, sulfate, tartrate, fumarate, acetate, citrate, or phosphate salt, with the compound of formula (I) being particularly preferably in the form of a hydrochloride salt. In one embodiment, the compound of formula (I) may be in the form of a formate salt.

[0052] The present invention also specifically relates to compounds of formula (I), including any one of the specific compounds of formula (I) described herein in non-salt form.

[0053] Furthermore, the scope of the present invention includes the compounds of formula (I) in any solvated form, including, for example, solvates with water (i.e., as hydrates) or solvates with organic solvents such as methanol, ethanol, isopropanol, acetic acid, ethyl acetate, ethanolamine, DMSO, or acetonitrile. All physical forms of the compounds of formula (I), including any amorphous or crystalline forms (i.e., polymorphs), are also included within the scope of the present invention. It is to be understood that such solvates and physical forms of pharmaceutically acceptable salts of the compounds of formula (I) are also encompassed by the present invention.

[0054] Furthermore, compounds of formula (I) may exist in different isomeric, particularly stereoisomers (including, for example, geometric isomers (or cis / trans isomers), enantiomers and diastereomers) or tautomers (including, in particular, prototropic tautomers such as keto / enol tautomers or thione / thiol tautomers). All such isomers of compounds of formula (I) are considered to be part of the present invention, either in admixture or in pure or substantially pure form. With respect to stereoisomers, the present invention encompasses isolated optical isomers of compounds according to the present invention, as well as any mixtures thereof (including, in particular, racemic mixtures / racemates). Racemates can be resolved by physical methods, such as fractional crystallization, separation or crystallization of diastereomeric derivatives, or separation by chiral column chromatography. Individual optical isomers can also be obtained from racemates by salt formation with an optically active acid followed by crystallization. The present invention further encompasses any tautomers of compounds of formula (I). It will be understood that some compounds may exhibit tautomerism. In such cases, the formulas provided herein explicitly represent only one of the possible tautomeric forms. The formulas and chemical names provided herein are intended to encompass any tautomeric form of the corresponding compounds, and are not limited to only the specific tautomeric form shown by the drawings or identified by the compound name.

[0055] The scope of the present invention also includes compounds of formula (I) in which one or more atoms have been replaced with a specific isotope of the corresponding atom. For example, the present invention includes compounds of formula (I) in which one or more hydrogen atoms (or, for example, all hydrogen atoms) have been replaced with a deuterium atom (i.e., 2 H; also referred to as "D"). Thus, the present invention also encompasses compounds of formula (I) that are enriched with deuterium. Naturally occurring hydrogen is approximately 99.98 mole % hydrogen-1 ( 1 H) and about 0.0156 mole % deuterium ( 2 The deuterium content at one or more hydrogen positions in a compound of formula (I) can be increased using deuteration techniques known in the art. For example, a compound of formula (I) or a reactant or precursor used in the synthesis of a compound of formula (I) can be subjected to an H / D exchange reaction using, for example, heavy water (DO). Further suitable deuteration techniques are described in: Atzrodt J et al., Bioorg Med Chem, 20(18), 5658-5667, 2012; William JS et al., Journal of Labeled Compounds and Radiopharmaceuticals, 53(11-12), 635-644, 2010; Modvig A et al., J Org Chem, 79, 5861-5868, 2014. The deuterium content can be determined, for example, using mass spectrometry or NMR spectroscopy. Unless otherwise specified, it is preferred that the compounds of formula (I) are not deuterium enriched. Thus, the hydrogen atoms or 1 Preferably, H hydrogen atoms are present.

[0056] The present invention also provides a method for preparing a compound in which one or more atoms are positron-emitting isotopes of the corresponding atom, e.g. 18 F, 11 C. 13 N, 15 O. 76 Br, 77 Br, 120 I and / or 124Such compounds can be used as tracers, trackers or imaging probes in positron emission tomography (PET). Accordingly, the present invention provides compounds of formula (I) in which (i) one or more fluorine atoms (or, for example, all fluorine atoms) are replaced by fluorine, such as fluorine. 18 (ii) a compound of formula (I) in which one or more carbon atoms (or, for example, all carbon atoms) are replaced with a F atom; 11 (iii) compounds of formula (I) in which one or more nitrogen atoms (or, for example, all nitrogen atoms) are replaced by a C atom; 13 (iv) compounds of formula (I) in which one or more oxygen atoms (or, for example, all oxygen atoms) are replaced by N atoms; 15 (v) one or more bromine atoms (or, for example, all bromine atoms) are replaced by an O atom; 76 (vi) compounds of formula (I) in which one or more bromine atoms (or, for example, all bromine atoms) are replaced by Br atoms; 77 (vii) compounds of formula (I) in which one or more iodine atoms (or, for example, all iodine atoms) are replaced by Br atoms; 120 (viii) compounds of formula (I) wherein one or more iodine atoms (or, for example, all iodine atoms) are replaced by: 124 I. Generally, it is preferred that none of the atoms in the compounds of formula (I) are replaced by a specific isotope.

[0057] The present invention further encompasses prodrugs of the compound of formula (I). As preferably understood herein, the term "prodrug" of the compound of formula (I) refers to a derivative of the compound of formula (I) that is metabolized to the compound of formula (I) upon administration to a subject. The prodrug of the compound of formula (I) may include modifications of the -OH, -NH2, or -COOH group, when present in the compound of formula (I), which can preferably be hydrolyzed to the -OH, -NH2, or -COOH group, respectively, for example, upon administration to a subject. For example, as known to those skilled in the art, such prodrugs preferably include modifications of the -OH moiety to -OR xand R x preferably comprises a moiety selected from -CO-, -CH2-O-CO, -CH2-O-CO-O-, and -CH(CH3)-O-COO-, more preferably R x is -CO-R y , -CH2-O-CO-R y , -CH2-O-CO-OR y , and -CH(CH3)-O-COO-R y Selected from R y is preferably carbocyclyl, heterocyclyl, C 1~5 Alkyl, -NH-(C 1~5 alkyl) or -S-(C 1~5 alkyl), wherein alkyl is optionally halogen, —CN, —OH, C 1~5 Alkyl, C 1~5 Haloalkyl, -O(C 1~5 alkyl), -O(C 1~5 haloalkyl), -SH, -S(C 1~5 alkyl), -S(C 1~5 haloalkyl), -NH2, -NH(C 1~5 alkyl), -NH(C 1~5 haloalkyl), -N(C 1~5 Alkyl)(C 1~5 alkyl), -N(C 1~5 haloalkyl)(C 1~5 alkyl), -CONH2, -CONH(C 1~5 alkyl), and -CON(C 1~5 Alkyl)(C 1~5 alkyl), and carbocyclyl and heterocyclyl are each optionally substituted by a group selected from halogen, —CN, —OH, C 1~5 Alkyl, C 1~5 Haloalkyl, -O(C 1~5 alkyl), -O(C 1~5 haloalkyl), -SH, -S(C 1~5 alkyl), -S(C 1~5 haloalkyl), -NH2, -NH(C 1~5 alkyl), -NH(C 1~5 haloalkyl), -N(C1~5 Alkyl)(C 1~5 alkyl), -N(C 1~5 haloalkyl)(C 1~5 alkyl), -CONH2, -CONH(C 1~5 alkyl), and -CON(C 1~5 Alkyl)(C 1~5 Further, for example, as known to those skilled in the art, such prodrugs preferably include those compounds of formula (I) that contain an -NH moiety derivative, where the -NH moiety is -NHCOO-R y and derivatives thereof, wherein R y is as defined above. Further, for example, as known to those skilled in the art, such prodrugs preferably include those compounds of formula (I) that contain a -COOH moiety, where the -COOH group is replaced by -COOR. y and derivatives thereof, wherein R y is as defined above. Further examples of groups that can be derivatized to give prodrugs will be known to those skilled in the art.

[0058] Pharmaceutical Composition The compounds provided herein may be administered as the compound per se or may be formulated as a medicament. The medicament / pharmaceutical composition may optionally include one or more pharmaceutically acceptable excipients such as carriers, diluents, fillers, disintegrants, lubricants, binders, colorants, pigments, stabilizers, preservatives, antioxidants, and / or solubility enhancers.

[0059] The pharmaceutical compositions may also include one or more solubility enhancers, such as poly(ethylene glycols) (e.g., PEG 200, PEG 300, PEG 400, or PEG 600), including poly(ethylene glycols) having a molecular weight ranging from about 200 to about 5,000 Da (e.g., PEG 200, PEG 300, PEG 400, or PEG 600), ethylene glycol, propylene glycol, glycerol, non-ionic surfactants, tyloxapol, polysorbate 80, macrogol-15-hydroxystearate (e.g., Kolliphor® HS), and the like. 15, CAS70142-34-6), phospholipids, lecithin, dimyristoylphosphatidylcholine, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, cyclodextrin, α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, hydroxyethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, hydroxyethyl-γ-cyclodextrin, hydroxypropyl-γ-cyclodextrin, dihydroxypropyl-β-cyclodextrin, sulfobutylether-β-cyclodextrin, sulfobutylether-γ-cyclodextrin, glucosyl-α-dextrin, The surfactant may include glucosyl-β-cyclodextrin, diglucosyl-β-cyclodextrin, maltosyl-α-cyclodextrin, maltosyl-β-cyclodextrin, maltosyl-γ-cyclodextrin, maltotriosyl-β-cyclodextrin, maltotriosyl-γ-cyclodextrin, dimaltosyl-β-cyclodextrin, methyl-β-cyclodextrin, carboxyalkyl thioether, hydroxypropyl methylcellulose, hydroxypropyl cellulose, polyvinylpyrrolidone, vinyl acetate copolymer, vinylpyrrolidone, sodium lauryl sulfate, sodium dioctyl sulfosuccinate, or any combination thereof.

[0060] The pharmaceutical compositions may also include one or more preservatives, particularly one or more antimicrobial preservatives, such as benzyl alcohol, chlorobutanol, 2-ethoxyethanol, m-cresol, chlorocresol (e.g., 2-chloro-3-methyl-phenol or 4-chloro-3-methyl-phenol), benzalkonium chloride, benzethonium chloride, benzoic acid (or a pharmaceutically acceptable salt thereof), sorbic acid (or a pharmaceutically acceptable salt thereof), chlorhexidine, thimerosal, or any combination thereof.

[0061] Pharmaceutical compositions are described in Remington: The Science and Practice of Pharmacy, Pharmaceutical Press, 22 nd Pharmaceutical compositions can be formulated by techniques known to those skilled in the art, such as those published in the "Publications of the United States of America" ​​edition. Pharmaceutical compositions can be formulated for oral, parenteral, e.g., intramuscular, intravenous, subcutaneous, intradermal, intraarterial, intracardiac, rectal, nasal, topical, aerosol, or vaginal administration. Dosage forms for oral administration include coated and uncoated tablets, soft and hard gelatin capsules, lozenges, troches, solutions, emulsions, suspensions, syrups, elixirs, powders and granules for reconstitution, dispersible powders and granules, medicated gums, chewable tablets, and effervescent tablets. Dosage forms for parenteral administration include solutions, emulsions, suspensions, dispersions, and powders and granules for reconstitution. Emulsions are preferred dosage forms for parenteral administration. Dosage forms for rectal and vaginal administration include suppositories and vaginal tablets. Dosage forms for nasal administration can be administered via inhalation and insufflation, for example, by metered-dose inhalers. Dosage forms for topical administration include creams, gels, ointments, salves, patches and transdermal delivery systems.

[0062] The compounds of formula (I) or the pharmaceutical compositions described above comprising compounds of formula (I) may be administered to a subject by any convenient route of administration, whether systemic / peripheral or at the desired site of action, including, but not limited to, one or more of oral (e.g., as a tablet, capsule, or as an ingestible solution), topical (e.g., transdermal, intranasal, ocular, buccal and sublingual), parenteral (e.g., using injection or infusion techniques, e.g., by implantation of a depot, e.g., subcutaneous or intramuscular, e.g., by injection, e.g., subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, subarachnoid, or intrasternal), pulmonary (e.g., by inhalation or insufflation therapy, e.g., via the mouth or nose, e.g., using an aerosol), gastrointestinal, intrauterine, intraocular, subcutaneous, intraocular (including intravitreal or intracameral), rectal, or intravaginal administration.

[0063] When a compound or pharmaceutical composition is administered parenterally, examples of such administration include one or more of intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, ​​intrasternal, intracardiac, intracranial, intramuscular, or subcutaneous administration of the compound or pharmaceutical composition, and / or using infusion techniques. For parenteral administration, the compound is best used in the form of a sterile aqueous solution which may contain other substances, for example, enough salts or glucose to make the solution isotonic with blood. The aqueous solution should be suitably buffered (preferably to a pH of 3 to 9), if necessary. The preparation of suitable parenteral formulations under sterile conditions is readily accomplished by standard pharmaceutical techniques well-known to those skilled in the art.

[0064] The compound or pharmaceutical composition may also be administered orally in the form of a tablet, capsule, ovoid, elixir, solution or suspension, which may contain flavoring or coloring agents for immediate, delayed, modified, sustained, pulsed, or controlled release applications.

[0065] Tablets may contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate, and glycine; disintegrants such as starch (preferably corn, potato, or tapioca starch), sodium starch glycolate, croscarmellose sodium, and certain complex silicates; and granulating binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin, and acacia. Additionally, lubricants such as magnesium stearate, stearic acid, glyceryl behenate, and talc may be included. Solid compositions of a similar type may also be employed as fillers in gelatin capsules. Preferred excipients in this regard include lactose, starch, cellulose, or high molecular weight polyethylene glycols. In the case of aqueous suspensions and / or elixirs, the drugs may be combined with various sweeteners or flavoring agents, pigments or dyes, emulsifying and / or suspending agents, and diluents such as water, ethanol, propylene glycol, and glycerin, as well as combinations thereof.

[0066] For oral administration, the compound or pharmaceutical composition is preferably administered by ingestion, particularly by swallowing. Thus, the compound or pharmaceutical composition can be administered so that it passes through the mouth and enters the gastrointestinal tract, which can also be referred to as "oral-gastrointestinal" administration.

[0067] Alternatively, the compounds or pharmaceutical compositions can be administered in the form of a suppository or pessary, or may be applied topically in the form of a gel, hydrogel, lotion, solution, cream, ointment or dusting powder. The compounds of the invention may also be administered dermally or transdermally, for example, by the use of a skin patch.

[0068] The compound or pharmaceutical composition can also be administered by a sustained-release system. Suitable examples of sustained-release compositions include semipermeable polymer matrices in the form of shaped articles, such as films or microcapsules. Examples of sustained-release matrices include polylactide, copolymers of L-glutamic acid and gamma-ethyl-L-glutamate, poly(2-hydroxyethyl methacrylate), ethylene vinyl acetate, or poly-D-(-)-3-hydroxybutyric acid. The sustained-release pharmaceutical composition also includes compounds encapsulated in liposomes. Therefore, the present invention also relates to liposomes containing the compounds of the present invention.

[0069] Compounds or pharmaceutical compositions can also be administered via pulmonary, rectal, or ocular routes.For ophthalmic use, they can be formulated as a micronized suspension in isotonic, pH-adjusted, sterile saline, or preferably as a solution in isotonic, pH-adjusted, sterile saline, optionally combined with a preservative such as benzalkonium chloride.Alternatively, they can be formulated into an ointment such as petrolatum.

[0070] It is also envisioned to prepare a dry powder formulation of the compound of formula (I) for pulmonary administration, particularly for inhalation.Such dry powder can be prepared by spray drying under conditions that produce a substantially amorphous glassy or substantially crystalline bioactive powder.Therefore, the dry powder of the compound of the present invention can be prepared according to the emulsification / spray drying process.

[0071] For topical application to the skin, the compounds or pharmaceutical compositions can be formulated into a suitable ointment containing the active compound suspended or dissolved in a mixture of, for example, one or more of mineral oil, liquid petrolatum, white petrolatum, propylene glycol, emulsifying wax, and water. Alternatively, they can be formulated into a suitable lotion or cream containing the active compound suspended or dissolved in a mixture of, for example, one or more of mineral oil, sorbitan monostearate, polyethylene glycol, liquid paraffin, polysorbate 60, cetyl esters wax, 2-octyldodecanol, benzyl alcohol, and water.

[0072] Therefore, the present invention relates to the compounds or pharmaceutical compositions provided herein, wherein the corresponding compounds or pharmaceutical compositions are administered by any one of the following routes: oral; topical route, including transdermal, intranasal, intraocular, buccal, or sublingual; parenteral route, including subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, intrathecal, intrasternal, intraventricular, intraurethral, ​​or intracranial; pulmonary route, including by inhalation therapy or insufflation therapy; gastrointestinal route; intrauterine route; intraocular route; subcutaneous route; ocular route, including by intravitreal or intracameral route; rectal route; or vaginal route. Preferred administration routes are oral or parenteral administration. For each of the compounds or pharmaceutical compositions provided herein, it is particularly preferred that the respective compounds or pharmaceutical compositions are administered orally (particularly by oral ingestion).

[0073] Typically, a physician will determine the actual dosage that will be most suitable for an individual subject. The specific dose level and frequency of administration for any particular individual subject may vary and will depend on a variety of factors, including the activity of the particular compound used, the metabolic stability and length of action of that compound, age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the individual subject being treated.

[0074] A suggested, but non-limiting, dosage of a compound according to the present invention for oral administration to humans (body weight approximately 70 kg) can be 0.05 to 2000 mg of active ingredient per unit dose, preferably 0.1 mg to 1500 mg, more preferably 0.1 mg to 1000 mg. The unit dose can be administered, for example, 1 to 3 times per day. The unit dose can also be administered 1 to 7 times per week, for example, no more than once a day. It will be understood that it may be necessary to vary the dosage routinely depending on the age and weight of the patient / subject and the severity of the condition being treated. The exact dosage and route of administration are ultimately at the discretion of the attending physician or veterinarian.

[0075] therapeutic use In one embodiment, the invention relates to a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition, for use in therapy.

[0076] As understood herein, a reference to a compound of formula (I), or a pharmaceutically acceptable salt, hydrate or solvate thereof, is preferably a reference to a compound of formula (I), or a pharmaceutically acceptable salt thereof.

[0077] The present invention provides compounds that function as inhibitors of PARG. Accordingly, the present invention provides a method of inhibiting PARG enzyme activity in vitro or in vivo, which method comprises contacting a cell with an effective amount of a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

[0078] The present invention also provides a method for selectively inhibiting PARG enzyme activity over PARP1 or ARH3 enzyme activity in vitro or in vivo, comprising contacting a cell with an effective amount of a compound defined herein, or a pharmaceutically acceptable salt, hydrate, or solvate thereof.

[0079] In a further embodiment, the present invention relates to a compound of formula (I) disclosed herein for use in a method of treating a disease or disorder in which PARG activity is implicated in a subject or patient in need of such treatment. The method of treatment comprises administering to the subject / patient a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein. In other words, in one embodiment, the present invention relates to a compound of formula (I) disclosed herein for use in treating a disease or disorder in which PARG activity is implicated.

[0080] In a further embodiment, the present invention relates to a method of inhibiting cell proliferation in vitro or in vivo, which method comprises contacting a cell with an effective amount of a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof.Accordingly, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in inhibiting cell proliferation in vitro or in vivo.

[0081] Therefore, in a further embodiment, the present invention relates to a method of treating a proliferative disorder in a subject or patient in need of such treatment. The method of treating a proliferative disorder in a subject or patient in need of such treatment comprises administering to the subject / patient a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein. Preferably, as disclosed herein, the proliferative disorder is cancer. Thus, the present invention relates to a method of treating cancer in a subject or patient in need thereof. The method of treating cancer in a subject or patient in need of such treatment comprises administering to the subject / patient a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition as defined herein. In a particular embodiment, the cancer is a human cancer.

[0082] In one embodiment, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt, hydrate, or solvate thereof for use in the treatment of a proliferative disorder. Preferably, as disclosed herein, the proliferative disorder is cancer. Accordingly, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt, hydrate, or solvate thereof for use in the treatment of cancer. In a particular embodiment, the cancer is a human cancer.

[0083] In a further embodiment, the present invention relates to a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, for use in the manufacture of a medicament for the treatment of a proliferative condition. In a preferred embodiment, the proliferative condition is cancer, more preferably human cancer. Thus, preferably, the present invention relates to a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, for use in the manufacture of a medicament for the treatment of cancer, preferably human cancer.

[0084] In a further embodiment, the present invention relates to a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, for use in the manufacture of a medicament for the inhibition of PARG enzyme activity. Preferably, the inhibition of PARG enzyme activity is selective inhibition of PARG enzyme activity over PARP1 or ARH3 enzyme activity. Accordingly, the present invention relates to a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, for use in the manufacture of a medicament for selectively inhibiting PARG enzyme activity over PARP1 or ARH3 enzyme activity.

[0085] The present invention further relates to a compound of formula (I) as defined herein, or a pharmaceutically acceptable salt, hydrate or solvate thereof, for use in the manufacture of a medicament for the treatment of a disease or disorder in which PARG activity is implicated, as defined herein.

[0086] As understood herein, the term "proliferative disorder" is used interchangeably herein and relates to unwanted or uncontrolled cell proliferation of unwanted, excessive, or abnormal cells, such as neoplastic or hyperplastic growth, whether in vitro or in vivo. Examples of proliferative conditions include premalignant and malignant cell proliferation, including, but not limited to, malignant neoplasms and tumors, cancer, leukemia, psoriasis, bone disease, fibroproliferative disorders (e.g., of connective tissue), and atherosclerosis. Any type of cell can be treated, including, but not limited to, lung, colon, breast, ovary, prostate, liver, pancreas, brain, and skin.

[0087] The anti-proliferative effects of the compounds of formula (I) of the present invention (through their inhibition of PARG enzyme activity) have particular utility in the treatment of human cancers. The anti-cancer effect may occur through one or more mechanisms, including, but not limited to, modulation of cell proliferation, inhibition of angiogenesis (the formation of new blood vessels), inhibition of metastasis (the spread of tumors from their origin), inhibition of invasion (the spread of tumor cells into adjacent normal structures), or promotion of apoptosis (programmed cell death).

[0088] The antiproliferative treatment with the compounds of formula (I) as defined above or their pharmaceutically acceptable salts, hydrates or solvates may be applied as a monotherapy or may involve, in addition to the compounds of the invention, conventional surgery or radiotherapy or chemotherapy. Such chemotherapy may include one or more of the following categories of antitumor agents: (i) other antiproliferative / antineoplastic agents and combinations thereof used in medical oncology, such as alkylating agents (e.g., cisplatin, oxaliplatin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulfan, temozolomide, and nitrosoureas); antimetabolites (e.g., gemcitabine and antifolates, e.g., fluoropyrimidines such as 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, and hydroxyurea); antitumor antibiotics (e.g., anthracyclines such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, and mithramycin; antimitotic agents (e.g., vinca alkaloids such as vincristine, vinblastine, vindesine, and vinorelbine, and taxoids such as taxol and taxotere, and polo kinase inhibitors); topoisomerase inhibitors (e.g., epipodophyllotoxins such as etoposide and teniposide, amsacrine, topotecan, and camptothecin); (ii) cytostatics, such as antiestrogens (e.g., tamoxifen, fluorefene, toremifene, raloxifene, droloxifene, and iodoxifene), antiandrogens (e.g., bicalutamide, flutamide, nilutamide, and cyproterone acetate), LHRH antagonists or agonists (e.g., goserelin, leuprorelin, and buserelin), progestogens (e.g., megestrol acetate), aromatase inhibitors (e.g., anastrozole, letrozole, vorazole, and exemestane), and inhibitors of 5oc-reductase, such as finasteride; (iii) anti-invasive agents [e.g., c-Src kinase family inhibitors such as 4-(6-chloro-2,3-methylenedioxyanilino)-7-[2-(4-methylpiperazin-1-yl)ethoxy]-5-tetrahydropyran-4-yloxyquinazoline (AZD0530; WO 01 / 94341), N-(2-chloro-6-methylphenyl)-2-{6-[4-(2-hydroxyethyl)piperazin-1-yl]ethoxy]-5-tetrahydropyran-4-yloxyquinazoline, perazin-1-yl]-2-methylpyrimidin-4-ylamino}thiazole 5-carboxamide (dasatinib, BMS-354825; J. Med. Chem., 2004, 47, 6658-6661) and bosutinib (SKI-606), as well as metalloproteinase inhibitors such as marimastat, inhibitors of urokinase plasminogen activator receptor function or antibodies against heparanase]; (iv) inhibitors of growth factor function: For example, such inhibitors include growth factor antibodies and growth factor receptor antibodies (e.g., the anti-erbB2 antibody trastuzumab [Herceptin®], the anti-EGFR antibody panitumumab, the anti-erbB1 antibody cetuximab [Erbitux, C225], and any growth factor or growth factor receptor antibody disclosed by Stern et al. (Critical reviews in oncology / haematology, 2005, Vol. 54, pp. 1 1-29; such inhibitors include tyrosine kinase inhibitors, such as inhibitors of the epidermal growth factor family (e.g., N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine (gefitinib, ZD1839), N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine (erlotinib, OSI-774), and 6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)-quinazolin-4-amine (CI EGFR family tyrosine kinase inhibitors such as EGFR-1 family tyrosine kinase inhibitors, e.g., EGFR-1 ... 1152, PH739358, VX-680, MLN8054, R763, MP235, MP529, VX-528 and AX39459) and cyclin-dependent kinase inhibitors, for example, CDK2 and / or CDK4 inhibitors; (v) antiangiogenic agents, such as those that inhibit the effects of vascular endothelial growth factor [e.g., the anti-vascular endothelial growth factor antibody bevacizumab (Avastin™) and, for example, VEGF receptor tyrosine kinase inhibitors, such as vandetanib (ZD6474), vatalanib (PTK787), sunitinib (SU1 1248), axitinib (AG-013736), pazopanib (GW 786034) and 4-(4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-1-ylpropoxy)quinazoline (AZD2171; Example 240 in WO 00 / 47212), compounds such as those disclosed in WO 97 / 22596, WO 97 / 30035, WO 97 / 32856 and WO 98 / 13354, as well as compounds acting by other mechanisms (e.g., linomide, an inhibitor of integrin ανβ3 function and angiostatin)]; (vi) vascular damaging agents such as combretastatin A4 and compounds disclosed in WO 99 / 02166, WO 00 / 40529, WO 00 / 41669, WO 01 / 92224, WO 02 / 04434 and WO 02 / 08213; (vii) endothelin receptor antagonists, such as zibotentan (ZD4054) or atrasentan; (viii) antisense therapy, e.g., directed to the targets listed above, such as the anti-ras antisense ISIS 2503; (ix) gene therapy approaches, including approaches to replace abnormal genes such as abnormal p53 or abnormal BRCA1 or BRCA2, GDEPT (gene-directed enzyme pro-drug therapy) approaches such as those using cytosine deaminase, thymidine kinase, or bacterial nitroreductase enzymes, and approaches to increase a patient's resistance to chemotherapy or radiation therapy, such as multidrug resistance gene therapy; and (x) Immunotherapeutic approaches, such as ex vivo and in vivo approaches to increase the immunogenicity of patient tumor cells, e.g., transfection with cytokines such as interleukin 2, interleukin 4, or granulocyte-macrophage colony-stimulating factor, approaches to reduce T-cell anergy, approaches using transfected immune cells, e.g., cytokine-transfected dendritic cells, approaches using cytokine-transfected tumor cell lines, and approaches using anti-idiotypic antibodies.

[0089] In certain embodiments, the antiproliferative treatment defined above may include, in addition to the compound of formula (I) of the present invention, conventional surgery or radiation therapy or chemotherapy. Such combined treatment can be achieved by simultaneous, separate or sequential administration of the individual components of the treatment. Such combination products use the compound of the present invention within the dosage ranges described hereinabove and other pharmaceutically active agents within their approved dosage ranges.

[0090] According to this aspect, the present invention further relates to a compound of formula (I) as defined herein or a pharmaceutically acceptable salt, hydrate or solvate thereof, for use in the treatment of cancer (e.g. cancer involving a solid tumor) in combination with another anti-tumor agent, which is preferably selected from the anti-tumor agents listed above.

[0091] As understood herein, the term "combination" refers to simultaneous, separate or sequential administration. In one embodiment of the present invention, "combination" refers to simultaneous administration. In another embodiment of the present invention, "combination" refers to separate administration. In a further embodiment of the present invention, "combination" refers to sequential administration. If the administration is sequential or separate, the delay in administration of the second component should not be such that the beneficial effect of the combination is lost. [Example]

[0092] The following examples are merely illustrative of the present invention and should not be construed as limiting the scope of the invention, which is defined by the appended claims.

[0093] Synthesis of Compounds of Formula (I) The synthesis of compounds of formula (I) according to the present invention is preferably carried out as described in, or analogously to, the specific synthetic procedures set out in the synthetic examples below.

[0094] Preparation Examples General Considerations Abbreviations used herein are as follows: AcOH (acetic acid); aq. (aqueous solution); Ar (argon); Atm (atmospheric pressure); BH3.THF (borane tetrahydrofuran complex); br. (broad, 1 H NMR signals; Boc2O (di-tert-butyl dicarbonate); (Cataxium APdG3 (mesylate [(di(1-adamantyl)-n-butylphosphine)-2-(2'-amino-1,1'-biphenyl)]palladium(II)); (CDCl3 (deuterated chloroform); cHex (cyclohexane); CMPB (cyanomethylenetrimethylphosphorane); Cs2CO3 (cesium carbonate); CuI (copper iodide); DABCO ((1,4-diazabicyclo[2.2.2]octane)); DAST (diethylaminosulfur trifluoride); DBU (1,8-diazabicyclo(5.4.0)undec-7-ene); DCE (dichloroethane); d (doublet, 1H NMR signal); DCM (dichloromethane); DIBAL-H (diisobutylaluminum hydride); DIPEA or DIEA (di-isopropylethylamine); DMAP (4-NN-dimethylaminopyridine), DME (1,2-dimethoxyethane), DMEDA (dimethylethylenediamine); DMF (NN-dimethylformamide); DMSO (dimethyl sulfoxide); DPPA (diphenylphosphoriazide); dtbbpy (bis(1,1-dimethylethyl)-2,2'-bipyridine); ES (electrospray); EtOAc or EA (ethyl acetate); EtOH (ethanol); h (hour); FA (formic acid); HATU (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate); HFIP (hexafluoroisopropanol); 1 H NMR (proton nuclear magnetic resonance spectroscopy); HPLC (high performance liquid chromatography), iPrOH (iso-propanol); K3PO4 (tripotassium phosphate); Ir[dF(CF3)(dtbbpy)PF6 ((4,4'-di-t-butyl-2,2'-bipyridine)bis[3,5-difluoro-2-[5-trifluoromethyl-2-pyridinyl-kN)phenyl-kC]iridium(III) hexafluorophosphate); LiOH (lithium hydroxide); m (multiplet, 1H NMR signal); mCPBA (meta-chloroperoxybenzoic acid), MeCN (acetonitrile), MeOH (methanol); min (minutes); MnO2 (manganese(IV) oxide); MS (mass spectrometry); MTBE (methyl tert-butyl ether); NaBH4 (sodium borohydride); NaHCO3 (sodium bicarbonate); Na2S2O3 (sodium thiosulfate); NCS (N-chlorosuccinimide); NH3 (ammonia); NH4Cl (ammonium fluoride); NiCl 2 (nickel dichloride); NIS (N-iodosuccinimide); NMP (N-methylpyrrolidone); NMR (nuclear magnetic resonance); Pd / C (palladium on charcoal); Pd2dba3 (tris(dibenzylideneacetone)dipalladium); Pd(dppf)Cl2 (1,1-bis(diphenylphosphino)ferrocenedichloropalladium); Pd(Ph3)2Cl2 (bis(triphenylphosphine)palladium(II) dichloride); PE (petroleum ether); Pd-PEPPSI-IPentCl o-Picoline ([1,3-bis[2,6-bis(1-ethylpropyl)phenyl]-4,5-dichloro-imidazol-2-ylidene]-dichloro-(2-methylpyridin-1-ium-1-yl)palladium; Pd(OH)2 (palladium hydroxide); Pd(Ph3)4 (palladium-tetrakis(triphenylphosphine); PhI(OAc)2 ((diacetoxyiodo)benzene)); P(tBu)3 (tri-tert-butylphosphine); Py (pyridine); q (quartet, 1H NMR signal); quin (quintet, 1H NMR signal); rac (racemic); RT (retention time); s (singlet, 1 H NMR signal); sat. (saturation); t (triplet, 1H NMR signal; TBAF (tetrabutylammonium fluoride); tert-BuBrettPhos-Pd-G3 ([(2-di-tert-butylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate); tBuXPhos Pd G3 (Methanesulfonato(2-di-t-butylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II)) TBDMSCl or TBSCl (tert-butyldimethylsilyl chloride); tBuOH (tert-butanol); TEA (triethylamine); TFA (trifluoroacetic acid); TFAA (trifluoroacetic anhydride), THF (tetrahydrofuran); TLC (thin layer chromatography); TMSCHN2 (trimethylsilyldiazomethane); TMSCN (trimethylsilyl cyanide); TMSOTf (trimethylsilyl trifluoromethanesulfonate); TTMSS (trimethylsilane); UPLC (ultra performance liquid chromatography), UV (ultraviolet), wt% (weight percent); Xantphos (4,5-bis(diphenylphosphino)-9,9-dimethylxanthene); Xantphos Pd G4 (methanesulfonato[9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene](2'-methylamino-1,1'-biphenyl-2-yl)palladium(II)).

[0095] General Procedures: All starting materials and solvents were obtained from commercial sources or prepared according to literature references. Commercially available reagents and anhydrous solvents were used as supplied without further purification. All reactions were stirred unless otherwise noted. Organic solutions were routinely dried over anhydrous sodium sulfate. Column chromatography was performed on pre-packed silica (100-1000 mesh, 40-63 μm) cartridges using the amounts indicated. All air- and moisture-sensitive reactions were performed in oven-dried (120 °C) glassware under an inert atmosphere of nitrogen or argon gas. Compound names were generated using ChemDraw Prime (Perkin Elmer). In some cases, commonly accepted names of commercially available reagents were used in place of ChemDraw-generated names.

[0096] Reversed phase HPLC conditions for LCMS analysis of final compounds: Method 1: SHIMADZU LCMS-2020 Kinetex EVO C18 2.1 x 30 mm, 5 μm column, 50 °C; Mobile phase: A: 0.0375% TFA (v / v) in water; B: 0.01875% TFA (v / v) in MeCN; Flow rate maintained at 1.5 mL / min; UV detection at 220 nm and 254 nm. Gradient information: 0-0.80 min: Ramp from 95% A-5% B to 5% A-95% B; 0.80-1.20 min: Hold at 5% A-95% B; 1.20-1.21 min: Return to 95% A-5% B; 1.21-1.55 min: Hold at 95% A-5% B.

[0097] Method 2: Shimadzu LCMS-2020 Kinetex EVO C18 2.1 x 30 mm, 5 μm column, 40 °C; Mobile phase: A: 0.025% NH₃·H₂O (v / v) in water, B: MeCN; Flow rate maintained at 1.5 mL / min; UV detection at 220 nm and 254 nm. Elution with the mobile phase over 1.55 min. Gradient information: 0–0.80 min: ramp from 95% A–5% B to 5% A–95% B; 0.80–1.20 min: hold at 5% A–95% B; 1.20–1.21 min: return to 95% A–5% B; 1.21–1.55 min: hold at 95% A–5% B.

[0098] Method 3: SHIMADZU LCMS-2020 Kinetex EVO C18 2.1 x 30 mm, 5 μm column, 50 °C; Mobile phase: A: 0.0375% TFA (v / v) in water; B: 0.01875% TFA (v / v) in MeCN; Flow rate maintained at 2.0 mL / min; UV detection at 220 nm and 254 nm. Elution with mobile phase over 0.80 min. Gradient information: 0-0.80 min: ramp from 95% A-5% B to 5% A-95% B; 0.80-1.20 min: hold at 5% A-95% B; 1.20-1.21 min: return to 95% A-5% B; 1.21-1.55 min: hold at 95% A-5% B.

[0099] Method 4: SHIMADZU LCMS-2020 Kinetex™ EVO C18 2.1 x 20 mm, 2.6 μm, 50 °C; Mobile phase: A: 0.0375% TFA (v / v) in water; B: 0.01875% TFA (v / v) in MeCN; Flow rate maintained at 2.0 mL / min; Elution with mobile phase over 1.00 min with UV detection at 220 nm and 254 nm. Gradient information: 0.01-0.60 min: Ramp from 95% A-5% B to 5% A-95% B; 0.61-0.78 min: Hold at 5% A-95% B; 0.78-0.79 min: Return to 95% A-5% B; 0.79-0.80 min: Hold at 95% A-5% B.

[0100] 1 H NMR spectrum: 1 H NMR spectra were acquired on a Bruker Avance III spectrometer at 400 MHz using residual non-deuterated solvent as the reference. 1 H NMR signals are specified with their multiplicity / bond multiplicity as evident from the spectrum. Possible higher order effects are not taken into account. The chemical shifts (δ) of the signals are specified in ppm (parts per million).

[0101] Salt stoichiometry: When compounds are referred to herein, particularly in the experimental section, in the synthesis of intermediates and examples of the present invention, as salts with the corresponding base or acid, the exact stoichiometric composition of the salt obtained by the respective preparation and / or purification process is unknown in most cases. Unless otherwise specified, suffixes to chemical names or structural formulas, such as "hydrochloride," "trifluoroacetate," "sodium salt," or "xHO," "xCF3COOH," "xNa+," etc., should be understood simply as salt forms, not as stoichiometric designations. This also applies when synthetic intermediates or example compounds or salts thereof are obtained by the described preparation and / or purification process as solvates, hydrates with unknown stoichiometric composition (if defined), etc.

[0102] Preparation of Intermediate 1.1 (5-Bromo-3-chloropyridin-2-yl)methanamine

[0103] [ka] To a mixture of 5-bromo-3-chloropicolinonitrile (2.0 g, 9.20 mmol) in THF (10 mL) under ice-water cooling, BH3 . THF (1M, 11.04 mL) was added over 5 min. The mixture was stirred at 0 °C for 30 min, then warmed to 20 °C and stirred at this temperature for an additional 30 min. The mixture was cooled to 0 °C and quenched by the dropwise addition of MeOH (10 mL) over 5 min. The mixture was heated to 70 °C and stirred at this temperature for 30 min. The reaction was concentrated in vacuo to give the crude product (2.2 g) as a light brown solid. The crude product was dissolved in HCl (aq. 2M, 20 mL) and washed with DCM (20 mL; 2x), and the aqueous phase was finally concentrated in vacuo to give the product (5-bromo-3-chloro-2-pyridyl)methanamine (1.5 g, 4.07 mmol, 44.26% yield, 70% purity, HCl salt) as a light brown solid. RT0.18 min (method 2); m / z222.9 (M+H) + (ESI + ), 1H NMR (400MHz, DMSO-d6) δ = 8.78 (d, J = 2.0 Hz, 1H), 8.69 (br, 3H), 8.47 (d, J = 2.0 Hz, 1H), 4.24 (d, J = 6.2 Hz, 2H).

[0104] Preparation of Intermediate 1.2 Ethyl 2-(((5-bromo-3-chloropyridin-2-yl)methyl)amino)-2-oxoacetate

[0105] [ka] To a mixture of (5-bromo-3-chloro-2-pyridyl)methanamine (1.5 g, 5.82 mmol, HCl salt) in DCM (30 mL) under ice-water cooling, DIPEA (2.25 g, 17.45 mmol) was added. Ethyl 2-chloro-2-oxoacetate (952.77 mg, 6.98 mmol) was then added over 5 minutes, and the mixture was stirred at 0°C for 30 minutes. The mixture was warmed to 20°C and stirred at this temperature for 30 minutes. The mixture was quenched with aqueous NaHCO3 (50 mL) and extracted with DCM (50 mL). The organic phase was separated, dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel (PE: EtOAc = 10:1 to 1:1) to give the product ethyl 2-(((5-bromo-3-chloropyridin-2-yl)methyl)amino)-2-oxoacetate (1300 mg, 3.64 mmol, 62.57% yield, 65.6% purity) as a white solid. RT 0.61 min (Method 1); m / z 322.8 (M+H) + (ESI + The product was used in the next step without further purification.

[0106] Preparation of Intermediate 1.3 Ethyl 6-bromo-8-chloroimidazo[1,5-a]pyridine-3-carboxylate

[0107] [ka] To a mixture of ethyl 2-(((5-bromo-3-chloropyridin-2-yl)methyl)amino)-2-oxoacetate (1300 mg, 4.04 mmol) in POCl (15 mL) under ice-water cooling was added phosphorus pentoxide (2.87 g, 20.21 mmol). The mixture was heated to 110 °C and stirred at this temperature for 5 h. The mixture was cooled to 25 °C and concentrated in vacuo to give a residue. The residue was dissolved in EtOAc (50 mL) and washed with water (30 mL) and aqueous NaHCO (30 mL). It was then finally concentrated in vacuo to give a residue. The residue was purified by column chromatography on silica gel (PE: EtOAc = 10: 1 to 3: 1) to give the product ethyl 6-bromo-8-chloroimidazo[1,5-a]pyridine-3-carboxylate (900 mg, 2.97 mmol, yield 73.34%) as a white solid. RT0.718 min (method 1), m / z 304.8 (M+H) + (ESI + ), 1 H NMR(400MHz,CHLOROFORM-d)δ=9.47(s,1H),7.77(s,1H),7.20(s,1H),4.65-4.42(m,2H),1.57-1.42(m,3H)

[0108] Preparation of Intermediate 1.4 6-Bromo-8-chloroimidazo[1,5-a]pyridine-3-carbohydrazide

[0109] [ka] To a mixture of ethyl 6-bromo-8-chloroimidazo[1,5-a]pyridine-3-carboxylate (900 mg, 2.97 mmol) in EtOH (20 mL), .HO (1.48 g, 29.65 mmol, 98%) was added. The mixture was heated to 80 °C and stirred at this temperature for 2 h. The reaction was cooled to 25 °C and the precipitated solid was separated. The crude product was triturated with EtOH (5 mL) to give 6-bromo-8-chloroimidazo[1,5-a]pyridine-3-carbohydrazide (650 mg, 2.25 mmol, 75.72% yield) as a white solid. RT0.56 min (method 1); m / z 290.8 (M+H) + (ESI + ); 1 H NMR (400MHz, DMSO-d6) δ = 10.02 (s, 1H), 9.50 (s, 1H), 7.72 (s, 1H), 7.51 (s, 1H), 4.58 (d, J = 4.0Hz, 2H).

[0110] Preparation of Intermediate 1.5 6-Bromo-8-chloro-N'-(2,2-difluoroacetyl)imidazo[1,5-a]pyridine-3-carbohydrazide

[0111] [ka] To a mixture of 6-bromo-8-chloroimidazo[1,5-a]pyridine-3-carbohydrazide (650 mg, 2.25 mmol) in EtOH (20 mL) was added ethyl 2,2-difluoroacetate (3.10 g, 22.45 mmol) and DBU (683.58 mg, 4.49 mmol). The mixture was heated to 100 °C and stirred at this temperature for 16 h. The mixture was cooled to 25 °C and concentrated in vacuo. The residue was dissolved in DCM (50 mL), washed with aqueous NH4Cl (30 mL; 2x), and concentrated in vacuo to give the crude product. The crude product was purified by column chromatography on silica gel (PE / EtOAc=1:1 to MeOH:EtOAc=1:10) to give the product 6-bromo-8-chloro-N′-(2,2-difluoroacetyl)imidazo[1,5-a]pyridine-3-carbohydrazide (650 mg, 1.56 mmol, 69.32% yield, 88% purity) as a white solid. RT0.62 min (method 1); m / z 368.8 (M+H) + (ESI + ); 1 H NMR (400MHz, DMSO-d6) δ = 10.95 (br, 2H), 9.44 (s, 1H), 7.81 (s, 1H), 7.59 (s, 1H), 6.38 (t, J = 53.2, 1H).

[0112] Preparation of Intermediate 1.6 2-(6-bromo-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(difluoromethyl)-1,3,4-thiadiazole

[0113] [ka] To a mixture of 6-bromo-8-chloro-N'-(2,2-difluoroacetyl)imidazo[1,5-a]pyridine-3-carbohydrazide (550 mg, 1.50 mmol) in toluene (20 mL) was added Lawesson's reagent (665.80 mg, 1.65 mmol) under a N atmosphere. The reaction was heated to 120 °C and stirred at this temperature for 2 hours. The reaction was cooled to 25 °C and concentrated in vacuo. The residue was triturated with MeOH (10 mL) at 70 °C for 1 hour, filtered, and the cake was collected and dried in vacuo to give the product, 2-(6-bromo-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(difluoromethyl)-1,3,4-thiadiazole (530 mg, 1.45 mmol, 96.88% yield) as a pale yellow solid. RT0.806 min (method 1); m / z 366.8 (M+H) + (ESI + ); 1 H NMR (400MHz, DMSO-d6) δ = 9.62 (s, 1H), 8.64 (s, 1H), 8.09 (s, 1H), 7.70 (t, J = 53.2, 1H).

[0114] Preparation of intermediate 1.7 2-(6-(benzylthio)-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(difluoromethyl)-1,3,4-thiadiazole

[0115] [ka] To a mixture of 2-(6-bromo-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(difluoromethyl)-1,3,4-thiadiazole (450 mg, 1.23 mmol) and phenylmethanethiol (168.17 mg, 1.35 mmol) in dioxane (10 mL) that had been degassed with nitrogen for 2 minutes, Pd(dba) (112.72 mg, 123.09 μmol), Xantphos (71.22 mg, 123.09 μmol), and DIEA (477.26 mg, 3.69 mmol) were added under nitrogen. The mixture was heated to 90 °C and stirred at this temperature for 16 h. The mixture was filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel (PE: EtOAc = 20: 1 to 5: 1) to give the product 2-(6-(benzylthio)-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(difluoromethyl)-1,3,4-thiadiazole (250 mg, 489.15 μmol, yield 39.74%, purity 80%) as a pale yellow solid. RT0.99min (method 1); m / z 409.0(M+H) + (ESI + ); 1 H NMR(400MHz,CHLOROFORM-d)δ=9.35(s,1H),7.69-7.67(m,1H),7.39-7.28(m,2H ),7.25-7.12(m,3H),7.05(t,J=53.2,1H),7.00(s,1H),6.90(s,1H),4.10(s,2H)

[0116] Preparation of Intermediate 1.8 2-(6-(benzylthio)-8-chloro-1-iodoimidazo[1,5-a]pyridin-3-yl)-5-(difluoromethyl)-1,3,4-thiadiazole

[0117] [ka] To a mixture of 2-(6-(benzylthio)-8-chloroimidazo[1,5-a]pyridin-3-yl)-5-(difluoromethyl)-1,3,4-thiadiazole (130 mg, 317.95 μmol) in MeCN (5 mL) at 0° C. was added NIS (78.68 mg, 349.74 μmol). The mixture was stirred at 25° C. for 5 h. The reaction mixture was used directly in the next step. RT0.99min (method 1); m / z 535.0(M+H) + (ESI + )

[0118] Preparation of Intermediate 1.9 8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-1-iodoimidazo[1,5-a]pyridine-6-sulfonyl chloride

[0119] [ka] A mixture of 2-(6-(benzylthio)-8-chloro-1-iodoimidazo[1,5-a]pyridin-3-yl)-5-(difluoromethyl)-1,3,4-thiadiazole (170 mg, 317.95 μmol) in MeCN (5 mL) was cooled to 0° C., and then HO (5.73 mg, 317.95 μmol), AcOH (38.19 mg, 635.89 μmol), and 1,3-dichloro-5,5-dimethylimidazolidine-2,4-dione (125.28 mg, 635.89 μmol) were added. The mixture was stirred at 0° C. for 2 h. The mixture was diluted with THF (8 mL), dried over NaSO, filtered, and concentrated in vacuo to give the crude product 8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-1-iodoimidazo[1,5-a]pyridine-6-sulfonyl chloride (160 mg, 219.14 μmol, 68.92% yield, 70% purity) as a light brown oil.

[0120] It should be noted that it cannot be excluded that the dichloro compound 1,8-dichloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)imidazo[1,5-a]pyridine-6-sulfonyl chloride was also formed in this process.

[0121] Preparation of Intermediate 1.10 8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-1-iodo-N-(1-methylcyclopropyl)imidazo[1,5-a]pyridine-6-sulfonamide

[0122] [ka] To a mixture of 1-methylcyclopropan-1-amine (37.80 mg, 531.49 μmol) in pyridine (1 mL) and NMP (N-methyl-2-pyrrolidone) (1 mL) at 0° C. was added 8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-1-iodoimidazo[1,5-a]pyridine-6-sulfonyl chloride (90 mg, 176.09 μmol) in MeCN (2 mL). The reaction was stirred at 0° C. for 50 min. The reaction mixture was quenched with water (10 mL) and extracted with EtOAc (10 mL; 2×). The organic phase was collected, dried over Na2SO4, filtered, and concentrated in vacuo to give a residue, which was purified by preparative TLC (PE: EtOAc = 3:1) to give the product 8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-1-iodo-N-(1-methylcyclopropyl) imidazo[1,5-a]pyridine-6-sulfonamide (25 mg, 45.81 μmol, 26.01% yield) as a light yellow solid.

[0123] It should be noted that it cannot be excluded that the dichloro compound 1,8-dichloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(1-methylcyclopropyl)imidazo[1,5-a]pyridine-6-sulfonamide was also formed in this process. RT0.510min (method 3); m / z 545.8(M+H) + (ESI + )

[0124] Preparation of intermediate 1.11 8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(1-methylcyclopropyl)imidazo[1,5-a]pyridine-6-sulfonamide

[0125] [ka] To a solution of 8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-1-iodo-N-(1-methylcyclopropyl)imidazo[1,5-a]pyridine-6-sulfonamide (25 mg, 45.81 μmol) in tetrahydrofuran (3 mL) was added Pd / C (5 mg, 10% purity). The reaction was degassed three times with H (15 Psi), and then the reaction was stirred at 20° C. for 3 hours. The reaction mixture was filtered, and the filtrate was concentrated in vacuo to give the product, 8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(1-methylcyclopropyl)imidazo[1,5-a]pyridine-6-sulfonamide (20 mg, 30.96 μmol, 67.59% yield, 65% purity) as a brown solid.

[0126] It should be noted that it cannot be excluded that the dichloro compound 1,8-dichloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(1-methylcyclopropyl)imidazo[1,5-a]pyridine-6-sulfonamide was also formed in this process. RT0.468 min (method 3); m / z 420.0 (M+H) + (ESI + )

[0127] Preparation of Intermediate 1.12 tert-Butyl (2S,6S)-4-(3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-6-(N-(1-methylcyclopropyl)sulfamoyl)imidazo[1,5-a]pyridin-8-yl)-2,6-dimethylpiperazine-1-carboxylate

[0128] [ka] To a mixture of 8-chloro-3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-N-(1-methylcyclopropyl)imidazo[1,5-a]pyridine-6-sulfonamide (30 mg, 0.0715 mmol) in dioxane (0.5 mL) was added tert-butyl (2S,6S)-2,6-dimethylpiperazine-1-carboxylate (15 mg, 0.0715 mmol), CsCO (70 mg, 0.214 mmol), and Pd-PEPPSI-IPentCl o-picoline (7.0 mg, 0.00715 mmol). The reaction mixture was degassed with N (3×) and then stirred at 98 °C for 1 h. The mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by preparative TLC (petroleum ether:ethyl acetate=1:2) to give the product tert-butyl (2S,6S)-4-(3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-6-(N-(1-methylcyclopropyl)sulfamoyl)imidazo[1,5-a]pyridin-8-yl)-2,6-dimethylpiperazine-1-carboxylate (16 mg, 0.0238 mmol, 33.34% yield) as a yellow solid. RT0.573 min (method 4); m / z 598.1 (M+H + )(ESI + ); 1H NMR(CDCl3,400MHz):9.74(s,1H),7.83(s,1H),7.08(t,J=53.6Hz,1H),6.36(s,1H),5.06(s,1H),4.20-4.37(m,2H),4.1 4-4.11(m,2H),3.67-3.48(m,2H),1.52(s,9H),1.40(s,3H),1.34(d,J=6.8Hz,6H),0.98-0.96(m,2H),0.63-0.58(m,2H)

[0129] Preparation of Example 1 3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-8-((3S,5S)-3,5-dimethylpiperazin-1-yl)-N-(1-methylcyclopropyl)imidazo[1,5-a]pyridine-6-sulfonamide formate

[0130] [ka] A solution of tert-butyl (2S,6S)-4-(3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-6-(N-(1-methylcyclopropyl)sulfamoyl)imidazo[1,5-a]pyridin-8-yl)-2,6-dimethylpiperazine-1-carboxylate (16 mg, 0.0322 mmol) in DCM (0.5 mL) and TFA (0.1 mL) was stirred at 25° C. for 1 h. The mixture was concentrated in vacuo to give a residue which was purified by preparative HPLC (column: Phenomenex luna C18 150 * 25mm * The product was purified by HPLC using a 10 μm column; mobile phase: A: 0.225% formic acid in water, B: MeCN; B%: 14%-44%, 10 min) and directly lyophilized to give the product 3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-8-((3S,5S)-3,5-dimethylpiperazin-1-yl)-N-(1-methylcyclopropyl)imidazo[1,5-a]pyridine-6-sulfonamide formate (4.0 mg, 0.00725 mmol, 22.56% yield, FA salt) as a yellow solid. RT0.362 min (method 4); m / z 498.2 (M+H) + (ESI + ); 1 H NMR (DMSO-d6,400MHz):9.57(s,1H),8.43(s,1H),8.22(s,1H),7.89(s,1H),7.67(t,J=53.2Hz,1H),6.66(s,1H),3.30-3. 27(m,2H),3.26-3.22(m,2H),3.06-3.01(m,2H),1.23(d,J=6.4Hz,6H),1.15(s,3H),0.77-0.67(m,2H),0.45-0.47(m,2H).

[0131] Preparation of Example 1b 3-(5-(difluoromethyl)-1,3,4-thiadiazol-2-yl)-8-((3S,5S)-3,5-dimethylpiperazin-1-yl)-N-(1-methylcyclopropyl)imidazo[1,5-a]pyridine-6-sulfonamide

[0132] [ka] The reaction was carried out twice on 29 g of tert-butyl (2S,6S)-4-[3-[5-(difluoromethyl)-1,3,4-thiadiazol-2-yl]-6-[(1-methylcyclopropyl)sulfamoyl]imidazo[1,5-a]pyridin-8-yl]-2,6-dimethyl-piperazine-1-carboxylate, and the reaction mixtures were then combined for workup.

[0133] A mixture of tert-butyl (2S,6S)-4-[3-[5-(difluoromethyl)-1,3,4-thiadiazol-2-yl]-6-[(1-methylcyclopropyl)sulfamoyl]imidazo[1,5-a]pyridin-8-yl]-2,6-dimethyl-piperazine-1-carboxylate (29.00 g, 48.5 mmol) was stirred at 20 °C for 2 hours. Both reaction mixtures were combined, and the formic acid (290 mL, 7686 mmol, 158 equiv.) was concentrated in vacuo to give a residue that was diluted with water (500 mL). The pH was adjusted to 9 with a saturated solution of NaHCO3, and the product was extracted with DCM (500 mL × 3). The combined organic layers were washed with brine (500 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The resulting yellow solid was dissolved in EtOH (1.4 L) and stirred at 110 °C until complete dissolution. The resulting clear solution was stirred for 12 h while cooling to room temperature. The precipitate was filtered off and dried under vacuum to give Fraction A of 3-[5-(difluoromethyl)-1,3,4-thiadiazol-2-yl]-N-(1-methylcyclopropyl)-8-[(3S,5S)-3,5-dimethylpiperazin-1-yl]imidazo[1,5-a]pyridine-6-sulfonamide as a yellow solid (33.00 g, 66.3 mmol, 68.6% yield).

[0134] The filtrate was concentrated under vacuum to give a residue (15 g) that was recrystallized in EtOH according to the same procedure as illustrated to give Fraction A. The precipitate was filtered off and dried under vacuum to give Fraction B of 3-[5-(difluoromethyl)-1,3,4-thiadiazol-2-yl]-N-(1-methylcyclopropyl)-8-[3S,5S)-3,5-dimethylpiperazin-1-yl]imidazo[1,5-a]pyridine-6-sulfonamide as a yellow solid (7.6 g, 15.3 mmol, 15.7% yield).

[0135] Fraction A 1H NMR(400MHz,DMSO-d6)δppm9.56(s,1H),8.43(s,1H),7.86(s,1H),7.67(t,J=53. 2Hz,1H),6.65(s,1H),3.25-3.31(m,2H),3.17-3.25(m,2H),3.00(m,2H),2.15(br s,1H),1.13-1.22(m,9H),0.66-0.77(m,2H),0.41-0.49(m,2H)

[0136] Fraction B 1 H NMR(400MHz,DMSO-d6)δppm9.56(s,1H),8.43(s,1H),7.86(s,1H),7.67(t,J=53.2Hz ,1H),6.65(s,1H),3.25-3.31(m,2H),3.17-3.25(m,2H),2.95-3.07(m,2H),2.15(br s,1H),1.13-1.22(m,9H),0.66-0.77(m,2H),0.41-0.49(m,2H)

[0137] Table 1 below provides a summary of the compounds described in the Examples section.

[0138] [Table 1]

[0139] Biological evaluation of example compounds Compounds of formula (I) were tested one or more times in selected biological and / or physicochemical assays. When tested multiple times, the data were reported as either the mean or median, where the mean, also called the arithmetic mean, represents the sum of the obtained values ​​divided by the number of times tested, and the median represents the middle number of a group of values ​​when ranked in ascending or descending order. If the number of values ​​in the data set is odd, the median is the middle value. If the number of values ​​in the data set is even, the median is the arithmetic mean of the two middle values. The in vitro pharmacological, pharmacokinetic, and physicochemical properties of the compounds can be determined according to the following assays and methods.

[0140] PARG protein expression and purification A codon-optimized gene encoding human PARG (448-976 [H446G, L447S, L473S, N479S, S802A, R811K, M841I, S858P, I916T, T924D, D927K, C963S, A967T]) was synthesized by Genscript and cloned into pET15b (NcoI / BamHI) with an N-terminal thrombin protease-cleavable 6His-TwinStrep tag. Protein expression in E. coli BL21(DE3) was induced by adding 0.2 mM IPTG to shake-flask cultures grown at 37°C to an OD600 of 0.8. Growth was continued at 30°C for an additional 20 hours before harvesting by centrifugation, and the cell pellets were stored at -80°C.

[0141] Proteins were purified by IMAC and SEC: Frozen cell pellets (typically 40 g wet weight) were resuspended by homogenization in 5 volumes of Buffer A (25 mM Tris / HCl pH 8.0, 200 mM NaCl, 2 mM DTT) supplemented with 1 mg of DNase I (Sigma-Aldrich) from bovine pancreas and protease inhibitors (Roche Complete™ EDTA-free protease inhibitor tablets), and lysed by passage through a Constant Systems Basic Z homogenizer. The lysate was clarified by centrifugation at 25,000 g for 60 min at 4°C, and the lysate supernatant was applied to a 5 mL StrepTrap HP (Cytiva) column pre-equilibrated with Buffer A. The column was washed with Buffer A (-10 CV) and then with Buffer B containing 1 M KCl (-5 CV), and the protein was then eluted with Buffer A containing 2.5 mM d-desthiobiotin. Pooled fractions containing 6HisTwinStrep-TEV-hPARG were incubated with TEV protease overnight at 4°C. hPARG was separated from uncleaved material and thrombobin protease by gel filtration using a Superdex 75 sizing column (GE Healthcare) pre-equilibrated with SEC buffer (15 mM Tris / HCl pH 8.5, 100 mM NaCl, 2 mM DTT). Pooled fractions containing pure hPARG were concentrated to 10 mg / mL using a 10k MWCO spin concentrator (VivaSpin) and then either used immediately for crystallization or flash-frozen in liquid nitrogen for storage at -80°C.

[0142] PARG enzyme IC 50 Assay PARG enzyme was incubated with compound or vehicle (DMSO) in a 384-well plate for 15 minutes or 2 hours. After addition of the PARG substrate ADP-ribose-pNP, the plate was read for absorbance intensity at 405 nm. Vehicle (DMSO) with high absorbance intensity indicates no inhibition of the enzyme reaction, while the low control (no enzyme) with low absorbance intensity indicates complete inhibition of the enzyme reaction.

[0143] material: hPARG: peak protein, 30 nM Substrate: ADP-pNP, 800 μM, Jena Bioscience Catalog No. NU-955 Reaction time: 60 minutes Assay buffer: 50 mM Tris-HCl pH 8.0, 100 mM NaCl, 2 mM DTT Temperature: 30℃ Total volume: 30 μL Control: 0% inhibitor control: DMSO 100% inhibition control: no enzyme

[0144] The protocol used for the enzymatic reaction and detection was as follows. 1. Transfer 100 nL of the final concentration of test compound or vehicle (DMSO) to the appropriate wells of the microtiter plate. 2. Centrifuge the plate at 1000 rpm for 1 minute. 3. 14.6 µL of 2x final concentration of enzyme in assay buffer or assay buffer alone Transfer to appropriate wells. 4. Centrifuge the plate at 1000 rpm for 1 minute. 5. Incubate the plate at room temperature for 15 minutes or 2 hours. 6. Transfer 15.4 μL of 2× substrate in assay buffer to all test wells. 7. Centrifuge the plate at 1000 rpm for 1 minute. 8. Read the plate on a plate reader (e.g., Spark Tecan). Absorbance IC of the compound of formula (I) in Example 1 50 The value is Provided in Table 2 below.

[0145] Cellular PAR chain assay The ability of compounds to inhibit PARG in response to DNA damage was evaluated using U2OS cells pretreated with compound for 1 hour after 1 hour of treatment with or without the DNA alkylating agent temozolomide (TMZ). Cells were harvested, fixed in 70% ethanol, rehydrated with glucose and EDTA in PBS, and subsequently blocked for 1 hour with PBS 1% BSA and 0.01% Tween-20 (PBT). Cells were incubated with a mouse monoclonal antibody against poly(ADP) ribose (PAR) polymer for 2 hours at room temperature. Cells were washed and incubated with an anti-mouse Alexa-488-conjugated secondary antibody for 1 hour at room temperature. DNA content in cells was determined using propidium iodide staining (overnight staining at 4°C). Cell fluorescence intensity was assessed by flow cytometry (Beckman Cytoflex) to determine the percentage of PAR chain-positive cells (gated against the TMZ + DMSO-treated control). The % PAR chain positive cells were fit to the compound concentration using a four-parameter log-logistic function to obtain the PAR chain EC 50 Generated the value:

[0146]

number

[0147] PAR chain EC of the compound of formula (I) in Example 1 50 The values ​​are provided in Table 2 below.

[0148] Cell viability assay NCI-H460 and MDA-MB-436, which are PARG inhibition-sensitive cell lines, and U2OS, which is a PARG inhibition-insensitive cell line, were seeded at 1,000, 5,000, and 2,000 cells / well, respectively, in a 96-well, clear-bottom, white plate. After 24 hours, compounds were added in duplicate using a Tecan digital dispenser (D300e). The outer wells of the plate were excluded. After 96 hours of incubation, 150 μl of growth medium was removed, and 50 μl of Cell Titer-Glo (Promega) was added per well. After 10 minutes of incubation, luminescence was read using a plate reader (Tecan) for NCI-H460 and U2OS cell lines, and a 2104 EnVision plate reader for MDA-MB-436. The mean values ​​of samples were normalized to the DMSO-treated control sample. Curves were fitted as % of control vs. log of compound concentration using a 4-parameter log-logistic function:

[0149]

number

[0150] PARGi (NCI-H460, MDA-MB-436 and U2OS) cell viability EC 50 The values ​​are provided in Table 2 below.

[0151] Table 2: Inhibition of PARG and cellular activity of compounds according to the invention. I C 50 (Inhibitory concentration at 50% of the maximal effect) Values ​​are given in μM, blank means that the corresponding compound was not tested in the respective assay.

[0152] [Table 2]

[0153] Further assays Kinetic solubility assay The kinetic solubility assay uses a shake flask method followed by HPLC-UV analysis. For exemplary compounds, kinetic solubility was measured according to the following protocol. 1. Sample was weighed and dissolved in 100% DMSO to make a 10 mM stock solution. Approximately 100 μL of stock solution is needed to cover this assay. 2. Test compounds and controls (10 mM in DMSO, 10 μL / tube) were added to buffer (490 μL / well) which was placed into Mini-UniPrep filters. Buffers were prepared according to customer requirements. 3. The kinetic solubility samples were vortexed for 2 minutes. 4. The solubility solution was shaken on an orbital shaker at room temperature for 24 hours. 5. If each sample was directly filtered through a syringeless filter device, transfer 200 μL of each soluble solution to a 96-deep well for analysis. 6. The test compound concentration of the filtrate was determined using HPLC-UV. 7. Three UV standard solutions were injected into the HPLC from low to high concentration, followed by testing the KS supernatant. Test samples were injected in duplicate.

[0154] Bidirectional permeability in Caco2 A bidirectional permeability assay in Caco-2 cells was performed for exemplary compounds of formula (I) according to the following protocol. 1. Caco-2 cells purchased from ATCC were plated at 1 x 105 cells / cm2 on polyethylene membrane (PET) in a 96-well BD insert plate. 2 21 for confluent cell monolayer formation. 日目 ~28 日目 The medium was refreshed every 4 to 5 days. 2. Monolayer integrity was verified by performing a Lucifer Yellow rejection assay. 3. The quality of the monolayer was verified by measuring the unidirectional (A → B) permeability of fenoterol / nadolol (low permeability markers), propranolol / metopronolol (high permeability markers) and the bidirectional permeability of digoxin (P-glycoprotein substrate marker) in duplicate wells. 4. Standard assay conditions for test compounds: - Test concentration: 2 μM (DMSO ≦ 1%); - Repetition: n=2; - Direction: Two-way transport including A → B and B → A; - Incubation time: single time point, 2 hours; - Transport buffer: HBSS containing 10 mM HEPES, pH 7.40 ± 0.05; - Incubation conditions: 37±1°C, 5% CO2, relative saturated humidity. 5. The dosing solution was spiked and mixed with transport buffer and stop solution (containing the appropriate internal standard (IS)) as the TO sample. 6. At the end of the incubation, the sample solutions from both the donor and receiver wells were immediately mixed with the stop solution. 7. All samples, including TO samples, donor samples, and receiver samples, were analyzed using LC / MS / MS. The concentrations of test compounds are expressed as the peak area ratio of the analyte to IS without a standard curve.

[0155] Liver microsome stability assay

[0156] 1.Material

[0157] 1.1 Liver microsomes Animal or human liver microsomes were purchased from Xenotech or Corning and stored in a freezer (below -60°C) before use.

[0158] 1.2 β-Nicotinamide adenine dinucleotide phosphate reduced form, tetrasodium salt, Supplier: Chem-Impex International, Cat. No. 00616

[0159] 1.3 Control compounds: testosterone, diclofenac and propafenone.

[0160] 2. Preparation of Working Solutions Stock solution: 10 mM test compound in DMSO. Working solution: 100 μM test or control compound in 100% acetonitrile (concentration of organic solvents: 1% (v / v) DMSO and 99% (v / v) acetonitrile)

[0161] 3. Assay Procedure A total of two sample plates with a 96-well format were prepared for incubation and labeled "Incubation" T60 and "Incubation" NCF60. Empty "Incubation" T60 and NCF60 plates were pre-warmed for 10 minutes. Liver microsomes were diluted to 0.56 mg / mL in 100 mM phosphate buffer. Microsome working solution (0.56 mg / mL) was transferred (445 μL) to the pre-warmed "Incubation" T60 and NCF60 plates, followed by incubation at 37°C for 10 minutes with constant shaking.

[0162] Liver microsomes (54 μL) were transferred to a Blank60 plate, followed by the addition of 6 μL of NAPDH cofactor and 180 μL of stop solution (acetonitrile containing an internal standard) to each well.

[0163] Aliquots (5 μL) of compound working solutions (100 μM) were added to the "incubation" plates (T60 and NCF60) containing microsomes and mixed thoroughly three times.

[0164] For the "incubation" NCF60 plate, 50 uL of buffer was added and mixed thoroughly three times. The plate was incubated at 37°C for 60 minutes with shaking. Samples were mixed once and after the 60 minute incubation, 60 μL was transferred from the NCF60 incubation plate to the stop plate containing stop solution.

[0165] Stop solution (180 μL) and NAPDH cofactor (6 μL) were added to the "quench" plate TO. The plate was chilled to prevent evaporation.

[0166] For the "incubation" T60 plate: Mix thoroughly three times and immediately remove 54 μL of the mixture at 0 minutes to stop the plate ("quench" plate TO). NAPDH cofactor (44 μL) was added to the "incubation" T60 plate. The plate was incubated at 37°C for 60 minutes with shaking. At 5, 15, 30, 45, and 60 minutes, 180 μL of stop solution was added to the "quench" plate, the samples were mixed once, and 60 μL was serially transferred from the "incubation" T60 plate at each time point.

[0167] Consequently, for wells containing test or control compounds, the final concentrations were 1 μM for test compounds, testosterone, diclofenac, and propafenone, 0.5 mg / mL for animal or human liver microsomes, 0.01% (v / v) for DMSO, and 0.99% (v / v) for acetonitrile.

[0168] All sampling plates were shaken for 10 min and then centrifuged at 3220 × g for 20 min at 4 ° C. The supernatant (80 μL) was transferred to 240 μL of HPLC water and mixed on a plate shaker for 10 min. Each bioanalysis plate was sealed and shaken for 10 min before LC-MS / MS analysis.

[0169] 4. Bioanalytical analysis The concentrations of the test compounds and the positive controls, testosterone, diclofenac and propafenone, in the samples were measured using a liquid chromatography-tandem mass spectrometry (LC-MS / MS) method.

[0170] 5. Data Calculation In determining the in vitro elimination constants k e of test and control compounds, the analyte / internal standard peak area ratio (PAR) was used to calculate the percentage remaining (% remaining) using the following formula:

[0171]

number

[0172] The parameters in the formula are listed in the table below.

[0173] JPEG2025535036000024.jpg44170

[0174] If the % remaining at the maximum incubation time, which was 60 min in this study, was higher than 75%, it would be considered within acceptable experimental variation, i.e., CV = 25%. Therefore, the corresponding T value of >145 min would be reported. Consequently, the corresponding CLint(mic) value would be reported as <9.6 μL / min / mg protein.

[0175] 6.References [1]Brian Davies and Tim Morris,Physiological Parameters in Laboratory Animals and Human.Pharmaceutical Research,Vol.10 No.7,1993 [2]Journal of Pharmacology and Experimental Therapeutics,1997,283(1):46-58

[0176] Hepatocyte metabolic stability

[0177] 1.Material

[0178] 1.1 Hepatocytes Animal or human hepatocytes were purchased from Bioreclamation IVT or RILD.

[0179] 1.2 Control Compounds: 7-ethoxycoumarin and 7-hydroxycoumarin

[0180] 2. Preparation of Working Solutions Stock solutions: 10 mM test compound and 30 mM control compound in DMSO. Working solution: 100 μM test compound or 300 μM control compound in 100% acetonitrile (concentration of organic solvents: 1% (v / v) DMSO and 99% (v / v) acetonitrile)

[0181] 3. Assay Procedure Cryopreserved hepatocytes were thawed, isolated, suspended in Williams Medium E, and then diluted with pre-incubated Williams Medium E to a final concentration of 0.510 x 106 cells / mL.

[0182] 198 μL of cell suspension (0.510 × 10 cells / mL) was added to the appropriate wells. The incubation plates were pre-incubated in a 37.0°C incubator for approximately 10 minutes. 2 μL of test compound and positive control were then added to each plate except the blank plate. All plates were incubated at 37.0°C in a 95.0% humidified incubator with 5.0% CO2 and constant shaking to initiate the reaction.

[0183] For the TO plate, a corresponding quench plate was prepared by adding 125 μL / well of acetonitrile containing 200 ng / mL tolbutamide and 200 ng / mL labetalol as an internal standard (stop solution), and after shaking for 1 minute to ensure uniformity, 25 μL / well of the incubation sample was transferred to this plate.

[0184] At each time point, the corresponding plate was removed from the incubator and 25 μL / well of the corresponding sample was transferred to its corresponding quench plate containing 125 μL / well of stop solution. Medium control (MC) plates (T0-MC and T90-MC) were prepared by adding everything except Williams medium E at the corresponding time points.

[0185] The plates were then sealed and shaken for 10 minutes before being centrifuged for 20 minutes at 4000 rpm and 4° C. 80 μL / well of the resulting supernatant was diluted with 240 μL / well of pure water, sealed, and shaken for 10 minutes before LC-MS / MS analysis.

[0186] 4. Bioanalytical analysis The concentrations of the test compounds and the positive controls, 7-ethoxycoumarin and 7-hydroxycoumarin, in the samples were determined using a liquid chromatography-tandem mass spectrometry (LC-MS / MS) method.

[0187] 5. Data Calculation In determining the in vitro elimination constants k e of test and control compounds, the analyte / internal standard peak area ratio (PAR) was used to calculate the percentage remaining (% remaining) using the following formula:

[0188]

number

[0189] The parameters in the formula are listed in the table below.

[0190] JPEG2025535036000026.jpg44170

[0191] If the % remaining at the maximum incubation time, which was 90 min in this study, was higher than 75%, it would be considered within acceptable experimental variation, i.e., CV = 25%. Therefore, a corresponding T value of >216.8 min would be reported. Consequently, the corresponding CLint(hep) (µL / min / 106 cells) would be reported as <7.5.

[0192] 6.References [1]Anna-Karin Sohlenius-Sternbeck.Determination of the hepatocellularity number for human,dog,rabbit,rat and mouse livers from protein concentration measurements.Toxicology in Vitro,Vol.20 No.8,2006 [2]Brian Davies and Tim Morris,Physiological Parameters in Laboratory Animals and Human.Pharmaceutical Research,Vol.10 No.7,1993 [3]Obach RS, Baxter JG, Liston TE, et al.The prediction of human pharmacokinetic parameters from preclinical and in vitro metabolism data[J].Journal of Pharmacology and Experimental Therapeutics,1997,283(1):46

[0193] Cell-derived xenograft (CDX) testing material: animal Species:Mus musculus Lineage:BALB / c nude Age: 6-9 weeks Gender: Female Weight: 17-20g Animal supplier: Beijing Vital River Laboratory Animal Technology Co.,Ltd Certification number: 20230209Abzz0619000498

[0194] cell MDA-MB-436 tumor cells (ATCC, catalog number HTB-130) were maintained in vitro as monolayer cultures in L-15 medium supplemented with 10% fetal bovine serum, 10 μg / mL insulin, 16 μg / mL glutathione, and 1% antibiotic-antimycotic in an atmosphere of 0% CO in air at 37° C. Cells growing in exponential phase were harvested and counted for tumor inoculum.

[0195] In vivo CDX studies: For tumor development, BALB / c nude female mice were inoculated with MDA-MB-436 tumor cells (10 × 10) in 0.2 mL of PBS containing Matrigel (1:1) into the right flank. 6 Animals were randomized and the mean tumor volume was approximately 126 mm 3Treatment was initiated 25 days after tumor inoculation, when tumor size reached 10. Each group consisted of 10 tumor-bearing mice. The compound of formula (I) was administered to the mice according to a predetermined regimen as shown in Experimental Design Table 3.

[0196] [Table 3]

[0197] Tumor size was measured twice weekly in two dimensions using calipers and the volume was expressed in mm 3 The tumor size was then used to calculate the TGI value.

[0198] TGI was calculated for each group using the following formula: TGI (%) = [1 - (Ti - T0) / (Vi - V0)] × 100; Ti is the mean tumor volume of the treatment group on a given day, T0 is the mean tumor volume of the treatment group on the first day of treatment, Vi is the mean tumor volume of the vehicle control group on the same day as Ti, and V0 is the mean tumor volume of the vehicle group on the first day of treatment.

[0199] One-way ANOVA was performed to compare tumor volumes between groups.

[0200] The compound of formula (I) (i.e., Example 1 - its formate salt form) administered at 100 mg / kg produced a significant and robust anti-tumor response, with tumor regression observed in all mice (Table 4, Figure 1).

[0201] Tumor growth inhibition assay

[0202] [Table 4]

Claims

1. Compounds of formula (I): 【Chemical 1】 or a tautomer, a pharmaceutically acceptable solvate, a pharmaceutically acceptable crystalline form, a pharmaceutically acceptable salt, or a prodrug thereof.

2. 2. The compound of claim 1, wherein the compound is a compound of formula (I) or a pharmaceutically acceptable salt thereof:

3. The compound of claim 2 in a non-salt form.

4. 3. The compound of claim 2, which is a formic acid salt of the compound of formula (I).

5. A pharmaceutical composition comprising the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt, hydrate or solvate thereof, and a pharmaceutically acceptable carrier.

6. A compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition according to claim 5, for use in therapy.

7. 7. A compound for use or a pharmaceutical composition for use according to claim 6 for use in a method for treating a disease or disorder in which PARG activity is implicated.

8. 8. A compound for use or a pharmaceutical composition for use according to claim 7 for use in a method for treating a proliferative disorder.

9. 9. The compound for use or pharmaceutical composition for use according to claim 8, wherein the proliferative disorder is cancer.

10. Use of a compound according to any one of claims 1 to 4 in the manufacture of a medicament for the inhibition of PARG enzyme activity.

11. 10. Use of a compound according to any one of claims 1 to 4 in the manufacture of a medicament for the treatment of a proliferative condition.

12. 12. The use of claim 11, wherein the proliferative condition is cancer.

13. 10. A method of treating a disease or disorder associated with PARG activity in a subject or patient in need thereof, comprising administering to said subject / patient a therapeutically effective amount of a compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

14. 10. A method of treating a proliferative disorder in a subject or patient in need thereof, comprising administering to said subject / patient a therapeutically effective amount of a compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, hydrate or solvate thereof.

15. 15. The method of claim 14, wherein the proliferative disorder is cancer.

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