WO2025212360A1 - Isoquinolones as pi3k inhibitors - Google Patents

Abstract

Novel PI3K inhibitors of the general Formula (1) are described along with methods of their preparation and their use in the treatment of diseases associated with the elevation or activation of the PI3K pathway, where R₁ to R₈, X₁, X₂, X₃ and X₄ are defined as described.

Description

ISOQUINOLONES AS PI3K INHIBITORS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63/572,657 , filed April 1, 2024, the disclosure of which is incorporated by reference in its entirety for all purposes.

BACKGROUND OF THE INVENTION

[0002] Phosphatidylinositol lipids (Pls) and their various phosphorylated subspecies are second messengers involved in a wide array of cellular vesicle trafficking and signal transduction processes. Phosphoinositide 3' kinases (PI3Ks) are a family of enzymes responsible for phosphorylation of the 3' hydroxyl position of the inositol ring of Pls. PI3Ks are subdivided into 3 classes according to their structure and substrates. Class II PI3Ks (PI3K-C2a, PI3K-C2P, PI3K-C2y) and Class III PI3Ks (vps34) are monomeric enzymes primarily associated with endocytosis and autophagy (Posor et al., Biochim Biophys Acta 2015, 1851, 794; Backer, Biochem J. 2016, 473, 2251). The Class I PI3Ks are heterodimeric, consisting of a catalytic kinase subunit (pllOa, p, y, 6) and one of several regulatory subunits that determine binding partners and subcellular localization. Class I PI3Ks are activated upon interaction with receptor tyrosine kinases (RTKs), Ras-related GTPases, G-protein coupled receptors, and/or related adaptor proteins, and in their active form convert phosphatidylinositol 4,5-diphosphate (PIP2) to phosphatidyl 3,4,5- triphosphate (PIP3) (Fruman et al., Cell 2017, 170, 605).

[0003] High local concentrations of PIP3 promote the recruitment and activation of downstream signaling partners, including AKT and mTOR. Activation of the AKT/mTOR pathways are implicated in several growth-related roles and pathologies including glucose regulation, cell survival, angiogenesis, and proliferation (Porta et al., Front Oncol. 2014, 4, 1), indicating a role for Class I PI3Ks as a critical upstream regulator of these functions.

[0004] Class I PI3Ks are further subdivided into 4 isoforms (a, p, y, and 6) based on the identity of their catalytic (pllOa, pliop, pllOy, or pll06) and regulatory (p85a or its various splice variants, p85P, p55y, or plOl) subunits, giving rise to distinct roles in cellular physiology (Vanhaesebroeck et al., J Mol Med (Berl). 2016, 94, 5). PI3Ky and PI3K6 are mostly expressed in leukocytes and play an important role in pro-inflammatory pathways (Hawkins et. al., Biochimica et Biophysica Acta 2015, 1851, 882; Okkenhaug et al., Science 2002, 297, 1031; Ali et al., Nature 2004, 431, 1007). PI3Ka and p are more ubiquitously expressed and share similar but not identical roles. For example, PI3Ka has a nonredundant role in angiogenesis (Soler et al., J Exp Med. 2013, 210, 1937), while PI3KP is known to serve a specific function in platelet aggregation (Liu et. al., Nat Rev Drug Discov. 2009, 8, 627; Jackson et al., Nat Med. 2005, 11, 507).

[0005] Elevation or constitutive activation of the PI3K pathway is one of the most frequent events in human cancers. The PI3K pathway is overactivated through a variety of mechanisms, including activating mutation of PI3K isoforms, up-regulation of PI3K isoforms, loss or inactivation of the tumor suppressor PTEN, or hyperactivation of tyrosine kinase growth factor receptors or other upstream signaling partners (Yang et al., Mol Cancer 2019, 18, 1). Mutations in the gene coding for PI3Ka or mutations which lead to upregulation of PI3Ka have been found to occur in many human cancers such as lung, stomach, endometrial, ovarian, bladder, breast, colon, brain, prostate, and skin cancers (Goncalves et al., N Eng J Med. 2018, 379,2052). In particular, PIK3CA, the gene encoding the pllOa subunit of PI3Ka, is frequently mutated or amplified in a variety of tumor types. Missense mutations occur in all domains of pllOa, but cluster in two 'hot spots', the most common being E542K and E545K in the helical domain, and H1047R in the kinase domain. Helical domain mutations reduce inhibition of pllOa by p85 or facilitate direct interaction of pllOa with insulin receptor substrate 1 (IRS1)37, whereas kinase domain mutations increase interaction of pllOa with lipid membranes, concomitantly upregulating signaling events. (Thorpe et al., Nat Rev Cancer 2015, 15, 7).

[0006] The development of inhibitors for the PI3K pathway has been challenging due to the inability to achieve dosing sufficient for tumor suppression without adverse events. To date PI3K inhibitors in the clinic (alpelisib, buparlisib, copanlisib, duvelisib, idelal isib, pictilisib, taselisib, and others) have caused dose-dependent adverse events such as hyperglycemia, rash, fatigue, diarrhea, etc. (Jiang et al., Mol Biol Rep. 2020, 47, 4587) which are known on-target toxicities. Hyperglycemia is a result of the body not producing enough insulin or aberrant utilization. The pancreas regulates insulin release in response to changes in blood glucose levels, resulting in either glucose uptake by muscle and fat cells when insulin levels are high or gluconeogenesis by the liver when insulin levels are low. Tissue cellular response to insulin requires PI3K signaling through the ubiquitously expressed pllOa sub-unit. As a result, pan-PI3K inhibition of the target disrupts glucose metabolism in tissues, leading to insulin resistance (Hopkins et al., Nature 2018, 560, 499). To mitigate adverse events, selective PI3K isoform inhibitors were developed. The severity of the adverse event is dependent on the select isoform, for example PI3Ka inhibitors are associated with hyperglycemia and rash due to the pllOa sub-unit role in insulin response (Rugo et al., The Breast 2022, 61, 156). Similarly, use of a selective PI3K6 inhibitor (idelalisib), where the pll06 sub-unit is highly expressed in immune cells, causes severe diarrhea and colitis. Inhibition with a dual inhibitor (taselisib), a potent PI3K6 inhibitor possessing modest PI3Ka inhibition led to gastrointestinal (Gl) side effects, but a highly selective and potent PI3K6 inhibitor (umbralisib) reported no Gl related adverse events (Gadkar et al., CPT Pharmacometrics Syst Pharmacol. 2021, 11, 616). Such amelioration of adverse events with highly isoform selective and potent inhibitors demonstrates that a strategy to mitigate toxicity by developing mutant selective isoform inhibitors is promising for decreasing the severity of toxicity.

Furthermore, selective inhibition of the mutant PI3Ka isoform over wild type may suppress cancer signaling while having minimal effect on PI3K signaling in healthy cells bearing just wild type PI3Ka, leading to a reduction in the toxicities associated with nonselective PI3K inhibition (Castel et al., Nat Cancer 2021 2, 587).

[0007] There is currently an interest in developing PI3K inhibitors for cancer therapy (WO 2023/081209, WO 2023/078401, WO 2023/060262, WO 2023/056407, WO 2021/202964, WO 2023/159155). However, there is a continued need for novel potent and selective PI3K inhibitors, either as single agents or as combination therapies, in the treatment of cancer.

SUMMARY OF THE INVENTION

[0008] An aspect of the invention is a compound of Formula (1) or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein:

Ri is heteroaryl, aryl, heterocyclyl or cycloalkyl, where each of the heteroaryl, aryl, heterocyclyl and cycloalkyl is substituted or unsubstituted; each of Xi, X2, X3 and X₄ is independently N, CH or substituted C (where C may be substituted with, but not limited to, C1-C4 alkyl, C3-C7 cycloalkyl, halogen, CN, CF3, OCF3, CFH2 or CF2H, where the C1-C4 alkyl and C3-C7 cycloalkyl are unsubstituted or substituted), with the proviso that X₄ cannot be a carbon atom substituted with a carboxylic acid or ester thereof, or Xi and Ri together with the ring carbon atom attached to Ri form a 5- or 6- membered heteroaryl or heterocyclyl ring containing 1-3 nitrogen atoms, where the heteroaryl or heterocyclyl ring is substituted or unsubstituted;

R2 is H, C1-C4 alkyl, C3-C7 cycloalkyl, CF3, CFH2 or CF2H, where the C1-C4 alkyl and C3-C7 cycloalkyl is unsubstituted or substituted, and where when R₂ is not H, the carbon atom attached to R2 is a chiral center and exists as a (R)- and (S)-racemic mixture or as either the (R)- or (S)- enantiomer;

R3 is H or C1-C4 alkyl, where the C1-C4 alkyl is unsubstituted or substituted;

R₅ is H, C1-C4 alkyl, C3-C7 cycloalkyl, heteroaryl, CF3, CH2F or CF2H, where each of the C1-C4 alkyl, C3-C7 cycloalkyl and heteroaryl is unsubstituted or substituted;

Rs is H, halogen, C1-C4 alkyl, C3-C7 cycloalkyl, CN, CF3, OCF3, CH2F or CF2H, where the C1-C4 alkyl and C3-C7 cycloalkyl is unsubstituted or substituted; each R₇ is independently H, halogen, C1-C4 alkyl, C3-C7 cycloalkyl, CN, CF3, OCF3, CH2F or CF2H, where the C1-C4 alkyl and C3-C7 cycloalkyl is unsubstituted or substituted;

Rs is H, C1-C4 alkyl, C3-C7 cycloalkyl, halogen, CN, CF3, OCF3, CFH2 or CF2H, where the C1-C4 alkyl and C3-C7 cycloalkyl is unsubstituted or substituted; and

R₄ is heteroaryl, C3-C7 cycloalkyl (such as C5-C7 cycloalkyl) or heterocyclyl, where the heterocyclic is optionally part of a bridged, fused or spiro ring system, and where each of the heteroaryl, C3-C7 cycloalkyl, and heterocyclyl is unsubstituted or substituted.

[0009] In an exemplary embodiment of the compound of Formula (1), R₄ is unsubstituted or unsubstituted heteroaryl.

[00010] In an exemplary embodiment of the compound of Formula (1), R₄ is unsubstituted or unsubstituted C3-C7 cycloalkyl (such as C5-C7 cycloalkyl). [00011] In an exemplary embodiment of the compound of Formula (1), R₄ is unsubstituted or unsubstituted heterocyclyl.

[00012] In an exemplary embodiment of the compound of Formula (1), R₄ is selected from wherein the above rings may be further substituted and Rg is selected from H, halogen, Ci-C₄ alkyl, C3-C7 cycloalkyl, aryl, heteroaryl, heterocyclyl, CN, CF3, OCF3, CFH? or CF?H, wherein each of the Ci-C₄ alkyl, C3-C7 cycloalkyl, aryl, heteroaryl and heterocyclyl is unsubstituted or substituted.

[00013] In an exemplary embodiment of the compound of Formula (1), R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2- oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2-azaadamantane bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane.

[00014] In an exemplary embodiment of the compound of Formula (1), R₄ is substituted or unsubstituted heterocyclyl and is not part of a bridged, fused or spiro ring system.

[00015] In an exemplary embodiment of the compound of Formula (1), R₄ is substituted or unsubstituted heterocyclyl and is part of a bridged, fused or spiro ring system.

[00016] In an exemplary embodiment of the compound of Formula (1), R₄ is substituted or unsubstituted heterocyclyl containing at least one ring nitrogen atom and is not part of a bridged, fused or spiro ring system.

[00017] In an exemplary embodiment of the compound of Formula (1), R₄ is substituted or unsubstituted heterocyclyl containing at least one ring nitrogen atom and is part of a bridged, fused or spiro ring system.

[00018] In an exemplary embodiment of the compound of Formula (1), R₄ is substituted or unsubstituted heterocyclyl containing at least two ring nitrogen atoms and is not part of a bridged, fused or spiro ring system. [00019] In an exemplary embodiment of the compound of Formula (1), R₄ is substituted or unsubstituted heterocyclyl containing at least two ring nitrogen atoms and is part of a bridged, fused or spiro ring system.

[00020] In an exemplary embodiment of the compound of Formula (1), R₄ is substituted or unsubstituted C3-C7 cycloalkyl and is not part of a bridged, fused or spiro ring system.

[00021] In an exemplary embodiment of the compound of Formula (1), R₄ is substituted or unsubstituted C5-C7 cycloalkyl and is part of a bridged, fused or spiro ring system.

[00022] In an exemplary embodiment of the compound of Formula (1), Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7- azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4- thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4- b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or triazolo-pyridine.

[00023] In an exemplary embodiment of the compound of Formula (1), R? is CH3 or CH?F.

[00024] In an exemplary embodiment of the compound of Formula (1), R3 is H.

[00025] In an exemplary embodiment of the compound of Formula (1), R₅ is CH3.

[00026] In an exemplary embodiment of the compound of Formula (1), Rg is CH3.

[00027] In an exemplary embodiment of the compound of Formula (1), each R₇ is independently H or F.

[00028] In an exemplary embodiment of the compound of Formula (1), Rg is H or F.

[00029] In an exemplary embodiment of the compound of Formula (1), R?, R₅ and Rg are CH3.

[00030] In an exemplary embodiment of the compound of Formula (1), R? is CH3 or CH?F; and R3 is H.

[00031] In an exemplary embodiment of the compound of Formula (1), R? is CH3 or CH?F; R3 is H; and each R₇ is independently H or F.

[00032] In an exemplary embodiment of the compound of Formula (1), R? is CH3 or CH?F; R3 is H; each R₇ is independently H or F; and R₅ is CH3. [00033] In an exemplary embodiment of the compound of Formula (1), R₂ is CH3 or CH₂F; R3 is H; and R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2- azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane.

[00034] In an exemplary embodiment of the compound of Formula (1), Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7- azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4- thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4- b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or triazolo-pyridine; R₃ is H; and R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2- oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2-azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane.

[00035] In an exemplary embodiment of the compound of Formula (1), Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7- azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4- thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4- b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or triazolo-pyridine; R₂ is CH3 or CH₂F; R3 is H; and R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2- azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane.

[00036] In an exemplary embodiment of the compound of Formula (1), Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7- azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4- thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4- b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or triazolo-pyridine; R2 is CH3 or CH?F; R3 is H; R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2- azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane; and each R₇ is independently H or F.

[00037] In an exemplary embodiment of the compound of Formula (1), Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7- azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4- thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4- b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or triazolo-pyridine; R2 is CH3 or CH2F; R3 is H; R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2- azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane; each R₇ is independently H or F; and R₅ is CH3.

[00038] In an exemplary embodiment of the compound of Formula (1), Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7- azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4- thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4- b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or triazolo-pyridine; R2 is CH3 or CH2F; R3 is H; R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2- azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane; R₇ is H; R₅ is CH3; and Rg is CH₃.

[00039] In an exemplary embodiment of the compound of Formula (1), Ri is heteroaryl, where the heteroaryl is unsubstituted or substituted, R₂ is CH3 or CH₂F; R3 is H; R₄ is heterocyclyl, where the heterocyclyl is unsubstituted or substituted; each R₇ is independently H or F; R₅ is CH3; and Rg is CH₃.

[00040] In an exemplary embodiment of the compound of Formula (1), Ri is heteroaryl, where the heteroaryl is unsubstituted or substituted, R₂ is CH3 or CH₂F; R3 is H; R₄ is C₃-C₇ cycloalkyl, where the C₃-C₇ cycloalkyl is unsubstituted or substituted; each R₇ is independently H or F; R₅ is CH3; and Rg is CH₃.

[00041] In an exemplary embodiment of the compound of Formula (1), at least one of Xi, X₂, X3 and X₄ is N.

[00042] In an exemplary embodiment of the compound of Formula (1), at least two of Xi, X₂, X3 and X₄ is N.

[00043] In an exemplary embodiment of the compound of Formula (1), at least three of Xi, X₂, X3 and X₄ is N.

[00044] In an exemplary embodiment of the compound of Formula (1), Xi is N.

[00045] In an exemplary embodiment of the compound of Formula (1), X₂ is N.

[00046] In an exemplary embodiment of the compound of Formula (1), X3 is N.

[00047] In an exemplary embodiment of the compound of Formula (1), X₄ is N.

[00048] In an exemplary embodiment of the compound of Formula (1), none of Xi, X₂, X3 and X₄ is N.

[00049] In an exemplary embodiment, the compound of Formula (1) is a compound of Formula (2)

or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein:

Ri, R₄, XI, X2 X3 and X₄ are defined as described in the compound of Formula (1), the carbon marked with * is a chiral center and exists as a (R)- and (S)-racemic mixture or as either the (R)- or (S)- enantiomer, and the listing of substituents within brackets for a given compound indicates individual compounds containing one of each of the substituents.

[00050] In an exemplary embodiment of the compound of Formula (2), Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7- azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4- thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4- b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or a triazolo-pyridine.

[00051] In an exemplary embodiment of the compound of Formula (2), R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2- oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2-azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane.

[00052] In an exemplary embodiment of the compound of Formula (2), Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7- azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4- thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4- b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or a triazolo-pyridine; and R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2- oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2-azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane.

[00053] In an exemplary embodiment of the compound of Formula (2), at least one of Xi, X₂, X3 and X₄ is N.

[00054] In an exemplary embodiment of the compound of Formula (2), at least two of Xi, X₂, X3 and X₄ is N.

[00055] In an exemplary embodiment of the compound of Formula (2), at least three of Xi, X₂, X3 and X₄ is N.

[00056] In an exemplary embodiment of the compound of Formula (2), Xi is N.

[00057] In an exemplary embodiment of the compound of Formula (2), X₂ is N.

[00058] In an exemplary embodiment of the compound of Formula (2), X3 is N.

[00059] In an exemplary embodiment of the compound of Formula (2), X₄ is N.

[00060] In an exemplary embodiment of the compound of Formula (2), none of Xi, X₂, X3 and X₄ is N.

[00061] In an exemplary embodiment, the compound of Formula (1) is a compound of Formula (3) or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein:

Ri is substituted or unsubstituted heteroaryl,

Xi, X2 X3 and X₄ are defined as described in the compound of Formula (1), and the listing of substituents within brackets for a given compound indicates individual compounds containing one of each of the substituents.

[00062] In an exemplary embodiment of the compound of Formula (3), Ri is a 5- to 6- membered heteroaryl ring containing from 1-4 nitrogen atoms. In a further embodiment, the 5-membered heteroaryl ring additionally contains a sulfur atom or an oxygen atom. In every case, the 5- to 6- membered heteroaryl ring may be further substituted.

[00063] In an exemplary embodiment of the compound of Formula (3), Ri is a substituted or unsubstituted pyrrole, pyridine, oxazole, isoxazole, pyrazole, imidazole, thiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4-thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine or 1,3,5-triazine.

[00064] In an exemplary embodiment of the compound of Formula (3), at least one of Xi, X2, X3 and X₄ is N.

[00065] In an exemplary embodiment of the compound of Formula (3), at least two of Xi, X2, X3 and X₄ is N. [00066] In an exemplary embodiment of the compound of Formula (3), at least three of Xi, X₂, X3 and X₄ is N.

[00067] In an exemplary embodiment of the compound of Formula (3), Xi is N.

[00068] In an exemplary embodiment of the compound of Formula (3), X₂ is N.

[00069] In an exemplary embodiment of the compound of Formula (3), X3 is N.

[00070] In an exemplary embodiment of the compound of Formula (3), X₄ is N.

[00071] In an exemplary embodiment of the compound of Formula (3), none of Xi, X₂, X3 and X₄ is N.

[00072] In an exemplary embodiment of the [heteroaryl or heterocyclyl]-[heteroaryl or heterocyclyl] covalent linkage depicted in Formula (3), each heteroaryl is independently a 5- to 6-membered heteroaryl ring containing from 1-3 nitrogen atoms. In a further embodiment, the 5-membered heteroaryl ring additionally contains a sulfur atom or an oxygen atom. In every case, the 5- to 6- membered heteroaryl ring may be further substituted.

[00073] In an exemplary embodiment of the [heteroaryl or heterocyclyl]-[heteroaryl or heterocyclyl] covalent linkage depicted in Formula (3), each heterocyclyl is independently a non-aromatic 4- to 7-membered ring containing from 1 to 3 nitrogen atoms. In a further embodiment, the 4- to 7- membered heterocyclic ring additionally contains from 1 to 2 oxygen or sulfur atoms, with the proviso that if the ring size is 4 or 5, the total number of ring heteroatoms is 1, 2 or 3, and if the ring size is 6 or 7, the total number of ring heteroatoms is 1, 2, 3 or 4. In every case, the 4- to 7- membered heterocyclic ring may be further substituted.

[00074] In an exemplary embodiment of the [heteroaryl or heterocyclyl]-[heteroaryl or heterocyclyl] covalent linkage depicted in Formula (3), each heteroaryl is independently a pyridine, pyridine oxide, pyridone, oxazole, isoxazole, thiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4-thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4- triazine or 1,3,5-triazine, where the heteroaryl ring may be further substituted.

[00075] In an exemplary embodiment of the [heteroaryl or heterocyclyl]-[heteroaryl or heterocyclyl] covalent linkage depicted in Formula (3), each heterocyclyl is independently an azetidine, morpholine, thiomorpholine, pyrrolidinone, pyrrolidinine, 2-pyrroline, 3-pyrroline, pyrazolidine, 2-pyrazoline, 2-imidazoline, imidazolidine, piperidine or piperazine, where the heterocyclyl ring may be further substituted.

[00076] An aspect of the invention is a pharmaceutical composition comprising any compound of the invention as described herein (such as a compound of Formula (1), Formula (2) or Formula (3)) or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[00077] In an exemplary embodiment, the pharmaceutical composition comprising any compound of the invention as described herein or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof further comprises one or more anti-cancer agents.

[00078] Another aspect of the invention is a method of treating a disease in which PI3K activity is implicated in a subject in need of such treatment, the method comprising administering to the subject a therapeutically effective amount of any compound of the invention as described herein (such as a compound of Formula (1), Formula (2) or Formula (3)) or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof.

[00079] In an exemplary embodiment, the disease to be treated is cancer. In a particular embodiment, the disease is a cancer bearing a PI3Ka H1047 mutation (such as H1047R, H1047L, or H1047Y), a PI3Ka E545 mutation (such as E545K), or a PI3Ka E542 mutation (such as E542K).

DETAILED DESCRIPTION OF THE INVENTION

[00080] The term "at risk for" as used herein, refers to a medical condition or set of medical conditions exhibited by a patient which may predispose the patient to a particular disease or affliction. For example, these conditions may result from influences that include, but are not limited to, behavioral, emotional, chemical, biochemical, or environmental influences.

[00081] The term "effective amount" as used herein, refers to a particular amount of a pharmaceutical composition comprising a therapeutic agent that achieves a clinically beneficial result (/.e., for example, a reduction of symptoms). Toxicity and therapeutic efficacy of such compositions can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD₅₀ (the dose lethal to 50% of the population) and the ED₅₀ (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index, which can be expressed as the ratio LD₅o/ ED50. Compounds that exhibit large therapeutic indices are preferred. The data obtained from these cell culture assays and additional animal studies can be used in formulating a range of dosages for human use. The dosages of such compounds lie preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage varies within this range depending upon the dosage form employed, the sensitivity of the patient, and the route of administration.

[00082] The term "symptom" as used herein, refers to any subjective or objective evidence of disease or physical disturbance observed by the patient. For example, subjective evidence is usually based upon patient self-reporting and may include, but is not limited to, pain, headache, visual disturbances, nausea and/or vomiting. Alternatively, objective evidence is usually a result of medical testing including, but not limited to, body temperature, complete blood count, lipid panels, thyroid panels, blood pressure, heart rate, electrocardiogram, tissue body imaging scans and other medical testing results.

[00083] The term "disease" as used herein, refers to any impairment of the normal state of the living animal or one of its parts that interrupts or modifies the performance of the vital functions. Typically manifested by distinguishing signs and symptoms, a disease is usually a response to i) environmental factors (such as malnutrition, industrial hazards, or climate); ii) specific infective agents (such as worms, bacteria, or viruses); iii) inherent defects of the organism (such as genetic anomalies); and/or iv) combinations of these factors.

[00084] The terms "reduce", "inhibit", "diminish", "suppress", "decrease", "prevent" and grammatical equivalents thereof (including "lower", "smaller", etc.) when used in reference to the expression of any symptom in an untreated subject relative to a treated subject, indicate that the quantity and/or magnitude of the symptoms in the treated subject is lower than in the untreated subject by any amount that is recognized as clinically relevant by any medically trained personnel. In one embodiment, the quantity and/or magnitude of the symptoms in the treated subject is at least 10% lower than, at least 25% lower than, at least 50% lower than, at least 75% lower than, and/or at least 90% lower than the quantity and/or magnitude of the symptoms in the untreated subject.

[00085]The term "inhibitory compound" as used herein, refers to any compound capable of interacting with (/.e., for example, attaching, binding, etc.) to a binding partner under conditions such that the binding partner becomes unresponsive to its natural ligands. Inhibitory compounds may include, but are not limited to, small organic molecules, antibodies, and proteins/peptides.

[00086] The term "attached" as used herein, refers to any interaction between a medium (or carrier) and a drug. Attachment may be reversible or irreversible. Such attachment includes, but is not limited to, covalent bonding, ionic bonding, Van der Waals forces or friction, and the like. A drug is attached to a medium (or carrier) if it is impregnated, incorporated, coated, in suspension with, in solution with, mixed with, etc.

[00087] The term "drug" or "compound" as used herein, refers to any pharmacologically active substance capable of being administered which achieves a desired effect. Drugs or compounds can be synthetic or naturally occurring, non-peptide, proteins or peptides, oligonucleotides or nucleotides, polysaccharides, or sugars.

[00088]The term "administered" or "administering" as used herein, refers to any method of providing a composition to a patient such that the composition has its intended effect on the patient. An exemplary method of administering is by a direct mechanism such as, local tissue administration (/.e., for example, extravascular administration, such as subcutaneous, intramuscular, or intraperitoneal), intravenous, oral ingestion, transdermal patch, topical, inhalation, suppository, etc.

[00089]The term "patient" as used herein, is a human or animal and needs not be hospitalized. For example, out-patients and persons in nursing homes are "patients." A patient may be a human or non-human animal of any age and therefore includes both adults and juveniles (/.e., children). It is not intended that the term "patient" connote a need for medical treatment. Therefore, a patient may voluntarily be subject to experimentation, whether clinical or in support of basic science studies.

[00090] The term "subject" as used herein, refers to, but is not limited to, humans (e.g., a male or female of any age group, e.g., a pediatric subject (e.g., infant, child, adolescent) or adult subject (e.g., young adult, middle-aged adult or senior adult)) and/or other primates (e.g., monkeys); non-human mammals, such as cows, pigs, horses, sheep, mice, goats, cats, dogs; and/or birds, such as chickens, ducks and/or geese.

[00091] The term "affinity" as used herein, refers to any attractive force between substances or particles that causes them to enter into and remain in chemical combination. For example, an inhibitor compound that has a high affinity for a receptor will provide greater efficacy in preventing the receptor from interacting with its natural ligands, than an inhibitor with a low affinity.

[00092] The term "derived from" as used herein, refers to the source of a compound or sequence. In one respect, a compound or sequence may be derived from an organism or particular species. In another respect, a compound or sequence may be derived from a larger complex or sequence. [00093]The term "test compound" as used herein, refers to any compound or molecule considered a candidate as an inhibitory compound.

[00094] The term "combination therapy" as used herein refers to refers to a dosing regimen of two or more different therapeutically active agents during a period of time, wherein the therapeutically active agents are administered together or separately. In one embodiment the combination therapy is a non-fixed combination.

[00095] The term "non-fixed combination" as used herein refers to two or more different therapeutic agents that are formulated as separate compositions or dosages such that they may be administered separately to a subject in need thereof either simultaneously or sequentially with variable intervening time limits.

[00096] The term "synergy" or "synergistic" as used herein refers to the phenomenon where the combination of two therapeutic agents of a combination therapy is greater in terms of measured results than the sum of the effect of each agent when administered alone.

[00097] The term "in vivo" as used herein refers to an event that takes place in a subject's body.

[00098] The term "in vitro" as used herein refers to an event that takes places outside of a subject's body.

[00099] The term "protein" as used herein, refers to any of numerous naturally occurring extremely complex substances (such as an enzyme or antibody) that contain amino acid residues joined by peptide bonds, and which include carbon, hydrogen, nitrogen, oxygen, and typically sulfur. In general, a protein comprises amino acids having an order of magnitude within the hundreds.

[000100] The term "peptide" as used herein, refers to any of various amides that are derived from two or more amino acids by combination of the amino group of one acid with the carboxyl group of another and are usually obtained by partial hydrolysis of proteins. In general, a peptide comprises amino acids having an order of magnitude with the tens.

[000101] The term "pharmaceutically acceptable" or "pharmacologically acceptable" as used herein, refers to molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered to an animal or a human.

[000102] The term, "pharmaceutically acceptable carrier" as used herein, includes any and all solvents, or a dispersion medium including, but not limited to, water, ethanol, a polyol (such as, for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, vegetable oils, coatings, isotonic and absorption delaying agents, liposome, commercially available cleansers, and the like. Supplementary bioactive ingredients also can be incorporated into such carriers.

[000103] The term "pharmaceutically acceptable salt" as used herein, refers to a salt that does not adversely impact the biological activity and properties of the compound and is suitable for use in contact with the tissues of subjects without undue toxicity, irritation and/or allergic response and the like. Pharmaceutically acceptable salts include those derived from suitable inorganic acids, organic acids and bases, and include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, nitric acid, acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, malonic acid, ascorbic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, benzoic acid, naphthalene sulfonic acid, lactic acid, succinic acid, oxalic acid, stearic acid, and the like. In some instances, pharmaceutically acceptable salts are obtained by reacting a compound having acidic group described herein with a base to form a salt such as an ammonium salt, an alkali metal salt (e.g., a sodium or a potassium salt), an alkaline earth metal salt (e.g., a calcium or a magnesium salt), a salt formed from an organic base, and an amino acid salt. Pharmaceutically acceptable salts derived from appropriate bases include alkali metals, alkaline earth metals, and ammonium and quaternary ammonium compounds. Specific metals include, but are not limited to, sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Organic bases from which salts may be prepared include, for example, primary, secondary, and tertiary amines.

[000104] The term "prodrug" as used herein, refers to a compound that is transformed in vivo to yield a disclosed compound or a pharmaceutically acceptable form of the compound. A prodrug may be inactive when administered to a subject, but is converted in vivo to an active compound. In various instances, a prodrug has improved physicochemical properties (such as bioavailability) and/or delivery properties over the parent compound. Prodrugs are typically designed to enhance pharmaceutically and/or pharmacokinetically based properties associated with the parent compound. The prodrug compound often offers advantages of solubility, tissue compatibility or delayed release in subject. Prodrugs include compounds wherein a hydroxy, amino, or mercapto group is bonded to any group that, when the prodrug is administered to a subject, cleaves to form a free hydroxy, free amino, or free mercapto group, respectively. Prodrugs are well known to be prepared from carboxylic acids in the form of, for example, carboxylate esters or thioesters. [000105] The term, "purified" or "isolated" as used herein, may refer to a composition (such as, for example, a peptide composition) that has been subjected to treatment (e.g., fractionation) to remove various other components, and which composition substantially retains its expressed biological activity.

[000106] The term "sample" as used herein, includes, for example, environmental and biological samples. Environmental samples include material from the environment such as soil and water. Biological samples include animal (e.g., human), fluids (e.g., blood, plasma, and serum), solids (e.g., stool), tissue, liquid foods (e.g., milk), and solid foods (e.g., vegetables). For example, a pulmonary sample may be collected by bronchoalveolar lavage (BAL) which comprises fluid and cells derived from lung tissues. A biological sample may comprise a cell, tissue extract, body fluid, chromosomes or extrachromosomal elements isolated from a cell, genomic DNA (in solution or bound to a solid support such as for Southern blot analysis), RNA (in solution or bound to a solid support such as for Northern blot analysis), cDNA (in solution or bound to a solid support) and the like.

[000107] The term "biologically active" as used herein, refers to any molecule having structural, regulatory or biochemical functions. For example, biological activity may be determined, for example, by restoration of wild-type growth in cells lacking protein activity. Cells lacking protein activity may be produced by many methods (i.e., for example, point mutation and frame-shift mutation). Complementation is achieved by transfecting cells which lack protein activity with an expression vector which expresses the protein, a derivative thereof, or a portion thereof.

[000108] The term "label" or "detectable label" as used herein, refers to any composition detectable by spectroscopic, photochemical, biochemical, immunochemical, electrical, optical or chemical means. Such labels include biotin for staining with labeled streptavidin conjugate, magnetic beads (e.g., Dynabeads*), fluorescent dyes (e.g., fluorescein, Texas Red*, rhodamine, green fluorescent protein, and the like), radiolabels (e.g., ³H, ¹²⁵l, ³⁵S, ¹⁴C, or ³²P), enzymes (e.g., horse radish peroxidase, alkaline phosphatase and others commonly used in an ELISA), and calorimetric labels such as colloidal gold or colored glass or plastic (e.g., polystyrene, polypropylene, latex, etc.) beads. Patents teaching the use of such labels include, but are not limited to, U.S. Patent Nos. 3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149; and 4,366,241 (all herein incorporated by reference in their entireties). The labels contemplated in the present invention may be detected by conventional methods. For example, radiolabels may be detected using photographic film or scintillation counters, fluorescent markers may be detected using a photodetector to detect emitted light. Enzymatic labels are typically detected by providing the enzyme with a substrate and detecting, the reaction product produced by the action of the enzyme on the substrate, and calorimetric labels are detected by simply visualizing the colored label.

[000109] The term "conjugate" as used herein, refers to any compound that has been formed by the joining of two or more moieties.

[000110] A "moiety" or "group" as used herein, is any type of molecular arrangement designated by formula, chemical name, or structure. Within the context of certain embodiments, a conjugate comprises one or more moieties or chemical groups. This means that the formula of the moiety is substituted at some position in order to be joined and be a part of the molecular arrangement of the conjugate. Although moieties may be directly covalently joined, it is not intended that the joining of two or more moieties must be directly to each other. A linking group, a crosslinking group, or a joining group refers to any molecular arrangement that will connect moieties by covalent bonds such as, but not limited to, one or more amide group(s). Additionally, although the conjugate may be unsubstituted, the conjugate may have a variety of additional substituents connected to the linking groups and/or connected to the moieties.

[000111] A "polymer" or "polymer group" as used herein, refers to a chemical species or group composed of repeatedly linked moieties. Within certain embodiments, it is preferred that the number of repeating moieties is 3 or more or greater than 10. The linked moieties may be identical in structure or may vary in their moiety structures. A "monomeric polymer" or "homopolymer" is a polymer that contains the same repeating, asymmetric subunit. A "copolymer" is a polymer derived from two or more types of monomeric species (/.e., two or more different chemical asymmetric subunits). "Block copolymers" are polymers comprised of two or more species of polymer subunits linked by covalent bonds.

[000112] The term "substituted" as used herein, refers to at least one hydrogen atom of a molecular arrangement that is replaced with a non-hydrogen substituent. The number of substituents present depends on the number of hydrogen atoms available for replacement and includes replacement of more than one hydrogen atom bound to a single atom (such as in the case of a carbon atom or a silicon atom which may be available for mono-, di- or tri-substitution or in the case of a nitrogen atom which may be available for mono-, di- or tri-substitution or in the case of an oxygen atom or a sulfur atom which may be available for mono-substitution). In the case of an oxo substituent ("=O"), two hydrogen atoms are replaced (which provides, for example, -(CH2)-C(=O)-CH3 as a substituent when the two hydrogen atoms of the middle carbon atom of -CH2-CH2-CH3 are replaced). When substituted, one or more of the groups below are "substituents." Substituents include, but are not limited to, halogen (e.g., F, Cl, Br, I), hydroxy (OH), hydroxyalkyl (e.g., CH2-OH, CH(CH3)OH, CfCHshOH), oxo, cyano (CN), cyanoalkyl (e.g., CH2- CN, CH(CH3)CN, CfCHshCN), nitro (NO2), amino, alkylamino, dialkylamino, branched or unbranched alkyl (e.g., methyl, ethyl, propyl, isopropyl, sec-butyl, etc.), cycloalkyl (e.g., cyclopropyl), fluoroalkyl (e.g., CF₃, CF₂H, CH₂F, CH2CF3, CH2CF2H, CHFCHF2, CF2CH2F, CF2CF3, CF2CH3, CF(CH₃)2, CH2CH2CF3, CF2CH2CF3, CF2CF2CF3, etc.) or more generally, haloalkyl (e.g., CH₂CI, CH(CH₃)Br, etc.), O-alkyl (alkoxy) (e.g., OCH3, OCH2CH3, OCHfCHsh, etc.), O-cycloalkyl (e.g., O- cyclopropyl), O-haloalkyl (e.g., OCF₂H, OCFH₂, OCF₃, OCH2CF3, OCH2CF2H, OCHFCHF2, OCF2CH2F, OCF2CF3, OCF2CH3, OCF(CH₃)2, OCH2CH2CF3, OCF2CH2CF3, OCF2CF2CF3 or OCH2CI), O-aryl (e.g., O- phenyl), O-heteroaryl, O-heterocyclyl, (CH2)i-3-cycloalkyl, (CH2)i-3-haloalkyl, (CH2)i-3-heterocyclyl, (CH2)i-3-aryl, (CH2)i-3-heteroaryl, thioalkyl (e.g., S-CH3), hydroxyalkyl (e.g., CH2OH), alkyl ether (e.g., CH2OCH3), alkynyl (e.g., -C = CRr), alkenyl (e.g., -CRf=CRfR_g), aryl (e.g., phenyl), arylalkyl (e.g., CH2Ph), heteroaryl (e.g., pyridyl or any 5- or 6- membered heteroaryl ring), heteroarylalkyl (e.g., CH₂-pyridine), heterocyclyl, heterocycloalkyl and as well as

-NRfRg, -NRfC(=O)R_g, -NR_fC(=O)NR_fNRg, -NR_fC(=O)OR_fSO₂Rg, -C(=O)R_f, -C(=O)OR_f, -C(=O)(CH₂)i-₃Rf, -C(=0)0(CH2)i-3Rf, -C(=0)(CH(CH3))(CH2)o-3Rf, -C(=0)0(CH(CH₃))(CH2)o-3Rf, -C(=0)(C(CH₃)2)(CH2)o-3Rf, -C(=0)0(C(CH₃)2)(CH2)o-3Rf, -OR_f, -C(=O)NR_fR_g, -OC(=O)NR_fR_g, -SR_f, -SOR_f, -S(=O)₂R_f, -OS(=O)₂R_f, -S( =O)ORf, and P(O)RfR_g, where each Rf and R_g may be the same or different and are independently, hydrogen, alkyl (e.g., CH3), substituted alkyl, cycloalkyl, substituted cycloalkyl, haloalkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocyclyl, substituted heterocyclyl, heterocycloalkyl, substituted heterocycloalkyl, heteroaryl or substituted heteroaryl. In addition, the above substituents may be further substituted with one or more of the above substituents, such that the substituent may constitute, for example, a substituted alkyl, a substituted aryl, a substituted heteroaryl, a substituted arylalkyl, a substituted heterocyclyl, or a substituted heterocycloalkyl.

[000113] The term "unsubstituted" as used herein, refers to any compound that does not contain extra substituents attached to the compound. For example, an unsubstituted compound refers to the chemical makeup of the compound without added substituents (e.g., no non-hydrogen substituents). For example, unsubstituted proline is a proline amino acid even though the amino group of proline may be considered as disubstituted with alkyl groups. [000114] The term "bond" as used herein in describing a substituent with atoms on both sides, refers to the absence of that substituent. For example, in the 4-atom sequence A-B-C-D, when B and C are both listed as being bonds, the result is the 2-atom sequence A-D. If only B is listed as being a bond, the result is the 3-atom sequence A-C-D.

[000115] The term "alkyl" as used herein, refers to any straight chain or branched, non-cyclic or cyclic, unsaturated or saturated aliphatic hydrocarbon containing from 1 to 10 carbon atoms, while the term "lower alkyl" has the same meaning as alkyl but contains from 1 to 3 carbon atoms. The term "higher alkyl" has the same meaning as alkyl but contains from 4 to 10 carbon atoms. Representative saturated straight chain alkyls include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, and the like, while saturated branched alkyls include, but are not limited to, isopropyl, sec-butyl, isobutyl, tertbutyl, isopentyl, and the like. As used herein, a methyl substituent may be depicted as "CH₃" or "Me" or as a terminal bond with no indication of specific atoms.

[000116] The term "cycloalkyl" as used herein, refers to saturated and unsaturated cyclic alkyls. Representative saturated cyclic alkyls include, but are not limited to, C3-C14 (such as C3-C7) cycloalkyls, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclododecyl, and the like; while unsaturated cyclic alkyls include, but are not limited to, cyclobutenyl, cyclopentenyl and cyclohexenyl, cyclohexadiene, and the like. Cyclic alkyls are also referred to herein as "homocycles" or "homocyclic rings".

[000117] The term "bicyclic compounds" as used herein, encompasses "bridged" compounds, "fused" compounds and "spiro" compounds as described.

[000118] The term "spiro" or "spirocyclic" as used herein, refers to chemical structures having at least two rings sharing one common atom. The rings may be cycloalkyl, heterocyclyl or a combination thereof, and may include one or more aryl or heteroaryl rings. Examples include, but are not limited to, spirocyclic cyclopropanes, spirocyclic aziridines, spirocyclic cyclobutanes, spirocyclic azetidines, spirocyclic oxetanes, spirocyclic cyclopentanes, spirocyclic pyrrolidines, spirocyclic 1,3-dioxolanes, spirocyclic dioxanes, spirocyclic oxathiolanes, spirocyclic thiazolidines, spirocyclic cyclohexanes, spirocyclic piperidines and spirocyclic piperizines, where the other ring is cycloalkyl (e.g., cyclobutane, cyclopentane or cyclohexane) or heterocyclyl (e.g., piperidine, tetrahydropyran, tetrahydrofuran, azetidine or pyrrolidine). Exemplary embodiments include, but are not limited to, l,4-dioxaspiro[4.5]decane, l,4-dioxa-8-azaspiro[4.5]decane, 2- azaspiro[4.4]nonane, 2-azaspiro[4.4]nonane, 2,7-diazaspiro[4.4]nonane, 3- azaspiro[5.5]undecane, 3,9-diazaspiro[5.5]undecane, 6-azaspiro[3.4]octane, 6- azaspiro[2.5]octane, l,3-dihydrospiro[indene-2,3'-pyrrolidine] and 3,4-dihydro-2H- spiro[naphthalene-l,4'-piperidine].

[000119] The term "bridged" as used herein, refers to a compound containing two nonadjacent atoms common to two rings. Exemplary embodiments include, but are not limited to, norbornane, bicyclo[l.l.l]pentane, bicyclo[2.2.1]heptane, l,4-diazabicyclo[2.2.2]octane, 3,8- diazabicyclo[3.2.1]octane, 3-azabicyclo[3.2.1]octane, bicyclo[3.2.1]octane, 3,6- diazabicyclo[3.1.1]heptane, 3,6-diazabicyclo[2.2.1]heptane, 3-azabicylo[3.1.0]hexane and other bridged piperazines and bridged piperidines.

[000120] The term "fused" as used herein, refers to polycyclic ring systems in which any two adjacent rings have two, and only two, adjacent atoms in common (ortho-fused) and polycyclic ring systems in which a ring contains two, and only two, adjacent atoms in common with each of two or more rings of a contiguous series of ortho-fused rings (ortho- and peri-fused). An exemplary embodiment is pentalene and dibenzoxepine (ortho-fused) and pyrene (ortho- and peri-fused). Ortho-fused systems have "n" common sides and "2n" common atoms while perifused systems have "n" common sides and less than "2n" atoms in common. Other exemplary fused systems include, but are not limited to, fused cyclopropyl rings, fused aziridine rings, fused cyclobutane rings, fused azetidine rings, fused cyclopentane rings, fused pyrrolidine rings, fused cyclohexane rings, fused piperidine rings, fused tetrahydropyran rings and fused piperazine rings, where each of these rings may be fused to an identical or different ring, such a pyrrolidine ring fused to another pyrrolidine ring (e.g., octahydropyrrolo[3,4-c]pyrrole) or to a cyclohexane ring (e.g., octahydro-lH-indole or octahydro-lH-isoindole). Other examples include fused aryl or heteroaryl rings, such as a pyridine ring fused with a cycloalkyl ring (e.g., cyclopentane or cyclohexane) or with a heterocyclyl ring (e.g., tetrahydrofuran or tetrahydropyran).

[000121] The term "aromatic" or "aryl" as used herein, refers to any aromatic carbocyclic (i.e., all of the ring atoms are carbon) substituent such as, but not limited to, phenyl (from benzene), tolyl (from toluene), xylyl (from xylene) or multi-ring systems (e.g., naphthyl (from naphthalene) and anthracenyl (from anthracene).

[000122] The term "arylalkyl" or "aralkyl" as used herein, refers to any alkyl having at least one alkyl hydrogen atom replaced with an aryl moiety such as, but not limited to, benzyl, - (d- phenyl, -(CHzhphenyl, -CH(phenyl)2, and the like. [000123] The term "halogen" as used herein, refers to any fluoro, chloro, bromo, or iodo moiety.

[000124] The term "haloalkyl" as used herein, refers to any alkyl where at least one hydrogen atom (and including all hydrogen atoms) has been replaced with a halogen atom, such as, for example, trifluoromethyl, dichloromethyl, difluoromethyl, monofluoromethyl, monobromomethyl, 1,1,1-trifluoroethyl and the like.

[000125] The term "aminoalkyl" as used herein, refers to any alkyl where at least one hydrogen atom has been replaced with a nitrogen atom, such as, for example, -(CH₂)I.₅-NH₂, (CH₂)I.5-NHCH₃, -(CH₂)I-₅-N(CH₃)₂, -(CH₂)I-5-NH-(CH₂)I.₅-N(CH3)₂, and the like.

[000126] The term "heteroaromatic" or "heteroaryl" as used herein, refers to any aromatic heterocyclic ring of 5 to 10 or more members and having at least one heteroatom selected from nitrogen, oxygen or sulfur, and containing at least 1 carbon atom, including, but not limited to, both mono- and bicyclic- ring systems, and where the nitrogen atom may be in an oxidized state. The heteroaryl ring may be attached as a substituent via a ring heteroatom or a carbon atom. Representative heteroaromatics include, but are not limited to, furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, indazole, 7-azaindole, 4-azaindole, 5-azaindole, 6- azaindole, 7-azaindazole, pyridine, pyridone (e.g., 2-pyridone, 3-pyridone or 4-pyridone), pyrimidinone, oxopyrazine, pyridine oxide, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, imidazopyrimidine, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, oxadiazole (e.g., 1,2,3-oxadiazole, 1,2,4- oxadiazole, 1,2,5-oxadiazole, 1,3,4-oxadiazole), thiadiazole (e.g., 1,2,3-thiadiazole, 1,2,4- thiadiazole, 1,2,5-thiadiazole, 1,3,4-thiadiazole), pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, triazolopyrazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4-b]pyrazine, pyrido[2,3-b]pyrazine, pteridine, triazolopyridines (e.g., [l,2,4]triazolo[4,3-a]pyridine) and the like.

[000127] The term "heteroarylalkyl" as used herein, means any alkyl having at least one alkyl hydrogen atom replaced with a heteroaryl moiety, such as -CH₂pyridinyl, -CH₂pyrimidinyl, and the like.

[000128] The term "heterocycle" or "heterocyclyl" or "heterocyclic ring" as used herein, refers to a nonaromatic ring which is either saturated or unsaturated and which contains 1 or more heteroatoms independently selected from nitrogen, oxygen, sulfur, phosphorus and silicon, wherein each of the nitrogen, phosphorus and sulfur heteroatoms may be in an oxidized state, and each of the nitrogen and silicon heteroatoms is substituted or unsubstituted and the nitrogen heteroatoms may be optionally quaternized, and includes bicyclic rings in which any of the above heterocycles are fused to an aryl or heteroaryl ring. The heterocyclic ring may be attached as a substituent via a ring heteroatom or a carbon atom. In various embodiments, heterocycles may contain 3 to 14 or more ring atoms (such as 3- to 7-membered monocyclic rings or 7- to 10-membered bicyclic rings) and include, but are not limited to, 2H-azirine, azetidine, 2,3-dihydroazete, 1,3-diazetidine, 2H-oxete, thietane, 2H-thiete, azetidin-2-one, morpholine, thiomorpholine, pyrrolidinone, pyrrolidinine, 2-pyrroline, 3-pyrroline, pyrazolidine, 2-pyrazoline, pyridazinone, pyrazinone, oxazolidin-2-one, 2-imidazoline, imidazolidine, piperidine, oxopiperidine, tetrahydropyrimidinone, piperazine, oxopiperazine, diazepane, ethylene oxide (oxirane), ethylene imine (aziridine), 1,1-dioxoisothiazolidine, ethylene sulfide (thiirane), oxetane, propylene oxide, 1,3-dioxolane, 1,2-oxathiolane, 1,3-oxathiolane, sulfolane, 2,4-thiazolidinedione, succinimide, 4-methyl-l,4-azaphosphinane 4-oxide, oxadiazoIone, dioxane (e.g. 1,4-dioxane and 1,3-dioxane), hydantoin, valerolactam, tetrahydrofuran, tetrahydropyran, 2H-pyran, 4H-pyran, thiane, 2H-thiopyran, 1,3-dithiane, 1,4-dithiane, 1,3,5-trithiane, pyrrolizidine, l,4,5,6-tetrahydrocyclopenta[b]pyrrole, tetrahydropyridine, tetrahydropyrimidine, dihydropyridazine, 6-oxo-l,6-dihydropyridazine, 6-oxo-l,4-dihydropyridazine, 6-oxo-l,6- dihydropyrazine, 6-oxo-l,4-dihydropyrazine, tetrahydrothiophene, tetrahydrothiopyran, tetrahydrotriazolopyrazine, tetrahydropyrazolopyridine, dihydrotriazolopyrazine, dihydropyrazolopyrazine, dihydroimidazopyrazine, indoline, isoindoline, decahydroisoquinoline, decahydroquinoline, 1,2,3,4-tetrahydroquinoline, 1,2-dihydroquinoline, 2H- benzo[e][l,3]oxazine, 2H-benzo[b][l,4]oxazine, quinolin-2(lH)-one, isoquinolin-l(2H)-one, quinuclidine, triethylenediamine, 1-azaadamantane, 2-azaadamantane, 2,3-dihydroazepine, 2,5- dihydroazepine, oxepane, azonane, spiro[cyclobutane-l,3'-indole], l-oxaspiro[4,5]decane, 1,6- dioxaspiro[3,4]octane, 2-oxa-7-azaspiro[3,5]nonane, l,4-dioxa-7-azaspiro[4,4]nonane, 1,3- diazaspiro[4,4]non-2-en-4-one, 2,9-diazaspiro[5,5]undecan-l-one, oxa- diazabicyclo[3.3.1]nonane, 8-azaspiro[4,5]decane-7, 9-dione, l,4-dithia-7-azaspiro[4,4]nonane, and the like.

[000129] The term "heterocycloalkyl" as used herein, refers to any alkyl having at least one alkyl hydrogen atom replaced with a heterocycle, such as -CHjmorpholinyl, and the like.

[000130] The term "alkylamino" as used herein, means at least one alkyl moiety attached through a nitrogen bridge (/.e., -N-(al kyl)_n), where n = 1 or 2, such as alkylamino or dialkylamino) including, but not limited to, methylamino, ethylamino, dimethylamino, diethylamino, and the like.

[000131] The term "alkyloxy" or "alkoxy", as used herein, means any alkyl moiety attached through an oxygen bridge (/.e., -O-alkyl) such as, but not limited to, methoxy, ethoxy, and the like.

[000132] The term "thioalkyl" as used herein, means any alkyl moiety attached through a sulfur bridge (/.e., -S-alkyl) such as, but not limited to, methylthio, ethylthio, and the like.

[000133] The term "alkenyl" as used herein, refers to an unbranched or branched hydrocarbon chain having one or more carbon-carbon double bonds therein and may also be referred to as an "unsaturated alkyl". The double bond of an alkenyl group can be unconjugated or conjugated to another unsaturated group. Suitable alkenyl groups include, but are not limited to vinyl, allyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, 2-ethylhexenyl, 2- propyl-2-butenyl, 4-(2-methyl-3-butene)-pentenyl. An alkenyl group can be unsubstituted or substituted with one or two suitable substituents.

[000134] The term "alkynyl" as used herein, refers to unbranched or branched hydrocarbon chain having one or more carbon-carbon triple bonds therein and may also be referred to as an "unsaturated alkyl". The triple bond of an alkynyl group can be unconjugated or conjugated to another unsaturated group. Suitable alkynyl groups include, but are not limited to ethynyl, propynyl, butynyl, pentynyl, hexynyl, methylpropynyl, 4-methyl-l-butynyl, 4-propyl-2-pentynyl-, and 4-butyl-2-hexynyl. An alkynyl group can be unsubstituted or substituted with one or two suitable substituents.

[000135] As used herein, "reactive groups" refer to nucleophiles, electrophiles, or radically active groups, i.e., groups that react in the presence of radicals. A nucleophile is a moiety that forms a chemical bond to its reaction partner (the electrophile) by donating both bonding electrons. Electrophiles accept these electrons. Nucleophiles may take part in nucleophilic substitution, whereby a nucleophile becomes attracted to a full or partial positive charge on an element and displaces the group it is bonded to. Alternatively, nucleophiles may take part in substitution of carbonyl group. Carboxylic acids are often made electrophilic by creating succinyl esters and reacting these esters with aminoalkyls to form amides. Other common nucleophilic groups are thiolalkyls, hydroxylalkyls, primary and secondary amines, and carbon nucleophiles such as enols and alkyl metal complexes. Other preferred methods of ligating proteins, oligosaccharides and cells using reactive groups are disclosed (Lemieux et al., Trends in Biotechnology 1998, 16, 506, incorporated herein by reference in its entirety). In yet another preferred method, one provides reactive groups for the Staudinger ligation, i.e., "click chemistry" with an azide comprising moiety and alkynyl reactive groups to form triazoles. Michael additions of a carbon nucleophile enolate with an electrophilic carbonyl, or the Schiff base formation of a nucleophilic primary or secondary amine with an aldehyde or ketone may also be utilized. Other methods of bioconjugation are provided (Hang et al. Accounts of Chemical Research 2001, 34, 1 , and Kiick et al. Proc Natl Acad Sci US.A. 2002, 99, 19, both of which are incorporated by reference in its entirety).

[000136] The term "biocompatible" as used herein, refers to any material that does not illicit a substantial detrimental response in the host. There is always concern when a foreign object is introduced into a living body that the object will induce an immune reaction, such as an inflammatory response that will have negative effects on the host. In the context of this invention, biocompatibility is evaluated according to the application for which it was designed: for example, a bandage is regarded as biocompatible with the skin, whereas an implanted medical device is regarded as biocompatible with the internal tissues of the body. Preferably, biocompatible materials include, but are not limited to, biodegradable and biostable materials. A substantial detrimental response has not occurred if an implant comprising the material is in close association to its implant site within the host animal and the response is better than a tissue response recognized and established as suitable from materials provided in an ASTM. ASTM subcommittee F04.16 on Biocompatibility Test Methods has developed biocompatibility standards for medical and surgical materials and devices which includes E1262-88, F612-20, F719-20el, F720-17, F748-16, F749-20, F750-20, F756-17; F763-04, F813-20, F895-11, F981-04, F1027-86, F1408-20a, F1439-03, F1877-16, F1903-18, F1904-14, F1983-14, F1984-99, F2147-01, F2148-18, F2382-18, F2808-17, F1288-19 and F2909-19, each of which is incorporated herein by reference. For example, materials that are to be used in contact with the blood stream must be composed of materials that meet hemocompatibility standards. One of these tests is for damage to red blood cells, which can result in hemolysis that is, rupturing of the cells, as described in F756-17 Standard Practice for Assessment of Hemolytic Properties of Materials.

[000137] As used herein, a "bioactive substance" refers to any of a variety of chemical moieties and that binds with a biomolecule such as, but not limited to, peptides, proteins, enzymes, receptors, substrates, lipids, antibodies, antigens, and nucleic acids. In certain preferred embodiments, the bioactive substance is a biomolecule but it is not intended that the bioactive substance be limited to biomolecules. In other preferred embodiments, the bioactive substances provide hydrophobic, hydrophilic, or electrostatic interactions, such as polycarboxylic acids that are anionic at physiological pH. In other preferred embodiment, the alkaline growth factors (with isoelectric point above 7) are retained via favorable electrostatic interactions by the polycarboxylates, and subsequently released in a controlled and sustained manner.

[000138] "Cancer" is a term used for a physiological condition in mammals that is typically characterized by unregulated cell growth. Examples of cancer include, but are not limited to, carcinoma, lymphoma, leukemia, blastoma, and sarcoma. More particular examples of such cancers include squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer (NSCLC), glioma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, acute myeloid leukemia (AML), multiple myeloma, gastrointestinal cancer, renal cell carcinoma, renal cancer (e.g., advanced renal cell carcinoma), ovarian cancer, liver cancer, lymphoblastic leukemia, lymphocytic leukemia, colorectal cancer, endometrial cancer, kidney cancer, prostate cancer, thyroid cancer, melanoma, chondrosarcoma, neuroblastoma, pancreatic cancer, glioblastoma multiforme, cervical cancer, brain cancer, stomach cancer, urothelial carcinoma (including local advanced or metastatic urothelial carcinoma), bladder cancer, hepatoma, breast cancer and head and neck cancer.

[000139] The term "stereoisomer" refers to compounds that have the same atomic connectivity but different atomic arrangement in space. Stereoisomers include cis-trans isomers, E and Z isomers, enantiomers, diastereomers and atropisomers. In the context of the present invention, the term "enantiomerically pure" is understood to mean that the compound in question with respect to the absolute configuration of the chiral center is present in an enantiomeric excess of more than 95%, preferably more than 97%.

[000140] The present disclosure contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers isomers, (D)-isomers, (L)-isomers, atropisomers, tautomers and racemic and other mixtures thereof, such as enantiomers or diastereomeric enriched mixtures, all of which are within the scope of the present disclosure. Insofar as compounds of the invention as defined herein may exist in optically active or racemic forms by virtue of one or more asymmetric carbon atoms, the invention includes in its definition any such optically active or racemic form. The synthesis of optically active compounds may be carried out by standard techniques of organic chemistry well known in the art such as, for example, by synthesis from optically active starting materials or by resolution of a racemic compound. Similarly, the enantiomeric or diastereomeric purity of a compound may be evaluated using standard laboratory techniques. [000141] In experimental procedures described herein where mixtures of stereoisomers (including but not limited to diastereomers, enantiomers, and geometric isomers) were generated during synthesis, their separation into stereochemically-enriched components was achieved by conventional techniques, such as chiral SFC, prep-HPLC, or other appropriate methods. Where appropriate, the absolute stereochemistry of the separated components was assigned by comparing the observed biological activity of the target compounds or appropriate intermediates thereof to similar components of known absolute stereochemistry and biological activity described in the literature. An exemplary publication used for reference in this manner includes, but is not limited to, Ketcham et al., J Med Chem (2024), 67, 4936-4949.

[000142] The pharmaceutical compositions of the invention can take any suitable form for the desired route of administration. Where the composition is to be administered orally, any suitable orally deliverable dosage form can be used, including without limitation water, glycols, oils, alcohols, and the like in the case of oral liquid preparations such as suspensions, syrups, elixirs, emulsions, and solutions; or solid carriers such as starches, sugars, kaolin, diluents, lubricants, binders, disintegrating agents, and the like in the case of powders, pills, capsules, and tablets. Because of their ease in administration, tablets and capsules represent the most advantageous oral dosage unit forms. Injectable compositions or intravenous infusions are also provided in the form of solutions, suspensions, and emulsions. For parenteral compositions, the carrier usually comprises sterile water and possibly other ingredients to aid solubility. Injectable solutions may be prepared in which the carrier comprises a saline solution, a glucose solution, or a mixture of a saline and a glucose solution. Suitable oils include, for example, peanut oil, sesame oil, cottonseed oil, corn oil, soybean oil, synthetic glycerol esters of long chain fatty acids, and mixtures of these and other oils. In compositions suitable for percutaneous administration, the carrier optionally comprises a penetration enhancing agent and/or a suitable wetting agent, optionally combined with suitable additives as needed, where the additives may facilitate administration of the composition to the skin and/or may facilitate preparation of the compositions to be delivered. These compositions may be administered in various ways, e.g., as a transdermal patch or as an ointment. Acid or base addition salts of the compounds of the invention are typically more suitable in the preparation of aqueous compositions due to their increased water solubility over the corresponding neutral form of the compounds.

[000143] The pharmaceutical compositions of the invention may comprise one or more of a filler, diluent, adjuvant, vehicle, or other excipient to facilitate storage and/or administration of the active ingredients contained therein. [000144] In an exemplary embodiment, a pharmaceutical composition according to the present invention may contain one or more additional therapeutic agents, for example, to increase efficacy or to decrease undesired side effects. In a particular embodiment, the pharmaceutical composition further contains one or more additional therapeutic agents useful to treat or inhibit a disease mediated directly or indirectly by PI3K. Examples of such agents include, without limitation, agents to treat or inhibit cancer, Huntington's disease, cystic fibrosis, liver fibrosis, renal fibrosis, pulmonary fibrosis, skin fibrosis, rheumatoid arthritis, diabetes, or heart failure.

[000145] In a specific embodiment, the additional therapeutic agent to be included is an anticancer agent. Examples of an anti-cancer agent include, but are not limited to, DNA-damaging cytotoxic drugs, alkylating agents such as cyclophosphamide, dacarbazine, and cisplatin; antimetabolites such as methotrexate, mercaptopurine, thioguanine, fluorouracil, and cytarabine; plant alkaloids such as vinblastine and paclitaxel; antitumor antibiotics such as doxorubicin, bleomycin and mitomycin; hormones/antihormones such as prednisone, tamoxifen, and flutamide; other types of anticancer agents such as asparaginase, rituximab, trastuzumab, imatinib, retinoic acid, and derivatives, colony stimulating factors, amifostine, camptothecin, topotecan, thalidomide analogs such as lenalidomide, and proteasome inhibitors such as Velcade.

[000146] In another embodiment, the present invention provides a method of inhibiting or treating diseases arising from abnormal cell proliferation and/or differentiation in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of one or more compounds according to the present invention. In one embodiment, the method of inhibiting or treating disease comprises administering to a subject in need thereof, a composition comprising an effective amount of one or more compounds of the invention and a pharmaceutically acceptable carrier. The composition to be administered may further contain a therapeutic agent such as an anti-cancer agent.

[000147] The compounds of the invention are defined herein by their chemical structures and/or chemical names and are generally listed according to the IUPAC or CAS nomenclature system. Abbreviations that are well known to one of ordinary skill in the art may be used. When a compound is referred to by both a chemical structure and a chemical name, and the chemical structure and chemical name conflict, the chemical structure is intended to be determinative of the compound's identity, except in cases where reproduction through publication of the chemical structure is compromised. [000148] The present invention includes compounds labeled with various radioactive or nonradioactive isotopes. Examples of atomic isotopes may include, but are not limited to, deuterium (²H), tritium (³H), iodine-125 (¹²⁵l), carbon-14 (¹⁴C), nitrogen-15 (¹⁵N), sulfur-35 (³⁵S) and chlorine-36 (³⁶CI). In an exemplary embodiment, one or more hydrogen atoms in a compound of the invention can be replaced by deuterium. In various embodiments, a compound of the invention includes at least one deuterium atom, or two or more deuterium atoms, or three or more deuterium atoms, etc. As described herein, compounds of the invention may also be radiolabeled with a radioactive isotope such as tritium (³H), iodine-125 (¹²⁵l), and carbon-14 (¹⁴C). A radiolabeled compound is useful as a therapeutic or prophylactic agent, provides a reagent for research such as for an assay, and/or provides a diagnostic agent for techniques such as in vivo imaging. Synthetic methods for incorporating isotopes into organic compounds are well known in the art.

[000149] In an embodiment of the invention, a compound of the invention as defined herein (such as a compound of Formula (1), Formula (2) or Formula (3)) or a pharmaceutically- acceptable salt thereof, exists as a single enantiomer being in an enantiomeric excess (% ee) of > 95%, such as > 98%, such as > 99%.

[000150] In an embodiment of the invention, a pharmaceutical composition comprises a compound of the invention as defined herein (such as a compound of Formula (1), Formula (2) or Formula (3)) or a pharmaceutically-acceptable salt thereof, where the compound exists as a single enantiomer being in an enantiomeric excess (% ee) of > 95%, such as > 98%, such as > 99%.

[000151] In an exemplary embodiment of the invention, the disease or disorder to be treated by the compounds of the invention is selected from congenital lipomatous overgrowth, vascular malformations, epidermal naevi, scoliosis/skeletal and spinal syndrome (CLOVES), mosaic tissue overgrowth syndromes, venous malformations and brain malformations associated with severe epilepsy or PIK3CA-related overgrowth syndrome (PROS) (Keppler-Noreuil et at., Am J Med Genet A. 2015, 167A, 287; Kurek et al. Am. J. Hum. Genet. 2012, 90, 1108).

[000152] In an exemplary embodiment of the invention, the cancer to be treated is a cancer bearing a PI3K H1047 mutation (such as H1047R) (Thorpe et al., Nat Rev Cancer 2015, 15, 7).

[000153] The compounds of the invention (such as defined by Formula (1), Formula (2), or Formula (3)) are typically PI3Ka mutant-selective inhibitors that exhibit greater selectivity for any of the H1047R, H1047L, H1047Y, E542K, or E545K mutations over the wild-type. As such, the compounds may decrease the amount of phosphorylated AKT (pAKT) and decrease proliferation selectively in PI3Ka mutant cell lines, across several tumor types.

[000154] A PI3Ka mutant selective inhibitor of the invention (such as defined by Formula (1), Formula (2) or Formula (3)) dosed in combination with an aromatase inhibitor (Al) such as, but not limited to letrozole or anastrozole, a selective estrogen receptor modulator (SERM) such as, but not limited to tamoxifen, or a selective estrogen receptor degrader (SERD) such as, but not limited to, fulvestrant, elacestrant, or camizestrant may exhibit a combination benefit leading to tumor regression in ER+/PI3Ka mutant tumors such as, but not limited to, the breast cancer xenograft model T47D, the breast cancer xenograft model MCF7, or the breast cancer xenograft model BT483, at doses where little or no regression would be observed with either single agent. Similarly, triple combinations of a PI3Ka mutant selective inhibitor of the invention (such as defined by Formula (1), Formula (2) or Formula (3)) dosed in combination with an aromatase inhibitor (Al), selective estrogen receptor modulator (SERM), or selective estrogen receptor degrader (SERD) in addition to a CDK4 or CDK4/6 inhibitor such as, but not limited to atirmociclib, ribociclib, abemaciclib, or palbociclib, may exhibit a combination benefit leading to tumor regression in ER+/PI3Ka mutant tumors such as, but not limited to, the breast cancer xenograft model T47D, the breast cancer xenograft model MCF7, or the breast cancer xenograft model BT483, at doses where little or no regression would be observed with either single agent and greater regressions that would be observed with doublet combinations.

[000155] A PI3K H1047R mutant selective inhibitor of the invention (such as defined by Formula (1), Formula (2) or Formula (3)) dosed in combination with a HER2 inhibitor such as, but not limited to, tucatinib or trastuzumab may exhibit a combination benefit leading to tumor regression in ER-/HER2+/PI3K H1047R mutant tumors such as, but not limited to, the breast cancer xenograft model HCC1954, at doses where little or no regression would be observed with either single agent.

[000156] Compounds of Formula (1), Formula (2) and Formula (3) of the present invention were generally prepared according to the synthetic route identified in Schemes 1-3.

[000157] In Scheme 1, the synthesis of isoquinolone cores may begin with an appropriately substituted 2,3-dihydro-lH-inden-l-one. In the case of 1 where R₆ is methyl, this is commercially available. In other cases, the starting material may be prepared via established methods known to those skilled in the art. Nitrosation of indenones 1 to convert to the oxime derivatives 2 may be accomplished using established methods (for examples see Touster, O.; Org. Reactions, VII, 1953, 327). A Beckmann-type rearrangement mediated by phosphorus pentachloride can convert oximes 2 into chloroisoquinolones 3 (Cushman, M.; Dekow, F.W. Tetrahedron 1978, 34(10), 1435-9). Alkylation of isoquinolone intermediates 3 with an appropriate electrophile and base can give nitrogen-substituted isoquinolones 4. In the case of where R₅ is methyl this may be accomplished with methyl iodide and an appropriate base (such as sodium hydride). Other electrophiles and alkylating agents, which would be known to those skilled in the art, may also be employed. In order to convert the bromide of isoquinolones 4 to methyl ketones 5, a Stille coupling reaction may be employed using an appropriate tin reagent, such as (a-ethoxyvinyl)- tributyl tin, followed by acid hydrolysis (Kosugi, M. et al., Bull. Chem. Soc. Jpn. 1987, 60(2), 767- 768). Alternatively, conversion of 4 to 5 may be accomplished by other established methods (for example, using a Heck coupling reaction with an appropriate enol-ether followed by acid hydrolysis (Mingcui, L. et al., Org. Biomol. Chem., 2010, 8, 2012-2015). Ketones 5 may be converted to chiral sulfinyl-imines 6 via known procedures, which may then in turn be reduced to sulfinyl-amines 7 in a stereo-controlled fashion using a suitable reducing agent (Datta and Ellman, J. Org. Chem. 2010, 75, 6283-6285; Ellman et al., Acc. Chem. Res. 2002, 35, 984-995; Ellman et al., J. Org. Chem. 2007, 72, 626-629; Colyer et al., Journal of Organic Chemistry 2006, 71(18), 6859-6862). With use of the R isomer of the sulfinyl group within 6, generally, the R, R - isomer of the product 7 predominantly results when (for example) the reducing agent used is a mixture of sodium borohydride and cerium chloride-heptahydrate. The use of this particular reducing system has been shown to be effective at reducing imines and may often give enhanced stereo-control in similar reductions (Hua, D.H., et al., Synthesis 1991, (11), 970-4; Zhu, X., et al., Journal of Chemical Research 2015, 39(7), 390-393). The major isomer may be separated from the other minor isomer via standard chromatographic means. As has been demonstrated in the preceding literature references, a judicious choice of the antipode of the sulfinyl-imine and the reducing agent may give access to either antipode of the sulfinyl-amine. The sulfinyl-amines can be cleaved to the single enantiomer of the chiral amine 8 using standard conditions (such as hydrogen chloride in dioxane). Scheme 1

[000158] Intermediates 8 may be further elaborated upon to afford the compounds in the present invention as depicted in Scheme 2. Standard coupling reactions of amines 8 with aryl halides 9 (such as an Ullman coupling or Buchwald-Hartwig coupling) may then give anthranilic acid derivatives 10 (see references in the review by Yang, Q. et al., Organic Process Research & Development 2022, 26(6), 1690-1750; Surry, D.S. and Buchwald ,S.L., Chemical Science 2011, 2(1), 27-50). Intermediates 10 may be reacted under suitable coupling conditions with an amine, wherein R¹ and R" each may be either alkyl or aryl or one of R¹ and R" may be hydrogen. R¹ and R" may also be joined to form a ring. In some cases, the R¹ and/or R" groups may be further elaborated prior to subsequent steps. Also, halogen-substituted isoquinolones 10 could be reacted under suitable coupling conditions with alkyl or aromatic boronates or boronic acids to give carbon-linked versions of 11.

Scheme 2

[000159] Alternatively, the order of reactions may be adjusted as shown in Scheme 3. Final compounds 11 can be prepared from the enantiomerically pure intermediates 7 using synthetic methods analogous to those shown in Scheme 1, but the order in which the positions of the isoquinolone cores are elaborated upon is altered.

Scheme 3

Experimental

[000160] All commercially available solvents and reagents were used as received. All ¹H NMR spectra were recorded using a Bruker Avance III HD 300 MHz or Bruker Avance III HD 400 MHz. MS samples were analyzed on a Shimadzu LCMS-2O2O mass spectrometer with electrospray ionization operating in positive and negative ion mode. Samples were introduced into the mass spectrometer using chromatography. All final products had a purity of > 90 %, unless specified otherwise in the experimental details. HPLC purity was measured on a Shimadzu Acquity HPLC system.

[000161] The following represents acronyms used in the experimental section for well-known chemical solvents, reagents, parameters and techniques:

¹H NMR: proton nuclear magnetic resonance spectroscopy

AcjO: acetic anhydride

ACN: acetonitrile

AcOH: acetic acid c-Bu: cyclobutyl c-Pr: cyclopropyl

CeCI₃: cerium (III) chloride

CH2CI2: dichloromethane

CHCI3: chloroform

CS2CO3: cesium carbonate

Cui: copper iodide

CU2O: copper oxide

DBAD: di-tert-butyl azodicarboxylate

DCM: dichloromethane

DIBAL: diisobutylaluminum hydride

DIEA: /V,/V-diisopropylethylamine

DIPEA: /V,/V-diisopropylethylamine

DMF: /V,/V-dimethylformamide

DMSO: dimethyl sulfoxide

DTAD: di-tert-butyl azodicarboxylate EA: ethyl acetate ee: enantiomeric excess

EtjO: diethyl ether

EtaN: triethylamine

EtOAc: ethyl acetate

EtOH: ethanol

FA: formic acid

H2O: water h: hours

HATU: l-[Bis(dimethylamino)methylene]-lH-l,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

HCI: hydrochloric acid

Hex: hexanes

HPLC: high-performance liquid chromatography

IPA: isopropanol

K2CO3: potassium carbonate

K3PO4: postassium phosphate

KOAc: potassium acetate

LiOH: lithium hydroxide mCPBA: meto-chloroperoxybenzoic acid

Me: methyl

MeCN: acetonitrile

MeOH: methanol mg: milligram min: minutes mL: milliliter Ms2O: methanesulfonic anhydride

MsCI: methanesulfonyl chloride

NaBH₄: sodium borohydride

N?: nitrogen

Na2CO₃: sodium carbonate

Na2SCO₄: sodium sulfate

NaCI: sodium chloride

NaH: sodium hydride

NaOH: sodium hydroxide

NaHCO₃: sodium bicarbonate

NaH2PO₄: monosodium phosphate

NH3: ammonia

NH4HCO3: ammonium bicarbonate

NMP: /V-methylpyrrolidone

Oxetane: 4-membered ring containing 3 carbon ring atoms and 1 oxygen ring atom.

PBr3: phosphorous tribromide

PCI₅: phosphorous pentachloride

Pd/C: palladium on carbon

Pd-PEPPSI-IHeptCI 3-chloropyridine: dichloro[l,3-bis(2,6-di-4-heptylphenyl)imidazol-2- ylidene](3-chloropyridyl)palladium(ll)

Pd(dppf)Cl₂: (l,r-bis(diphenylphosphino)ferrocene)palladium(ll) dichloride

Pd(PPh3)₄: tetrakis(triphenylphosphine)palladium(0)

Pd₂(dba)₃: tris(dibenzylideneacetone)dipalladium(0)

PdfPPh3)2Cl₂: bis(triphenylphosphine)palladium(ll) dichloride

PE: petroleum ether

POCI3: phosphorus oxychloride PPha: triphenylphosphine

Prep: preparative

RuPhos: 2-dicyclohexylphosphino-2',6'-diisopropoxybiphenyl

TEA: triethylamine

TFA: trifluoroacetic acid

THF: tetrahydrofuran

Ti(Oi-Pr)₄: Titanium(IV) isopropoxide

TLC: thin-layer chromatography

Xantphos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene

EXAMPLES

Intermediates

[000162] Preparation of Intermediate 1: 5-(2-iodophenyl)pyrimidine

[000163] To a stirred solution of 1,2-diiodobenzene (1.0 g, 3.0 mmol) in dioxane (10 mL) and

H₂O (3 mL) were added pyrimidin-5-ylboronic acid (751 mg, 6.1 mmol), K2CO3 (1.26 g, 9.1 mmol) and Pd(PPha)4 (350 mg, 0.3 mmol) at room temperature. The resulting mixture was stirred overnight at 100 °C under a nitrogen atmosphere. The reaction was quenched by the addition of water (20 mL) at room temperature. The resulting mixture was extracted with DCM (3 x 50 mL). The combined organic layers were washed with brine (3 x 50 mL) and then dried over anhydrous Na2SO₄. The filtrate was concentrated under reduced pressure. The residue was purified by prep-TLC (PE:EA = 1:1) to afford 5-(2-iodophenyl) pyrimidine (300 mg, 35% yield) as a light yellow solid. MS: (ES⁺) m/z = 282.9 [M+H]⁺.

[000164] Preparation of Intermediate 2: 4-fluoro-l-(2-iodophenyl)imidazole [000165] To a stirred mixture of 4-fluoro-lH-imidazole (200 mg, 2.3 mmol) and 1,2- diiodobenzene (3.8 g, 11.6 mmol) in DMF (4 mL) were added Cui (44 mg, 0.23 mmol), CS2CO3 (1.14 g, 3.5 mmol) and methyl[2-(methylamino)ethyl]amine (41 mg, 0.47 mmol) in portions at room temperature. The resulting mixture was stirred overnight at 60 °C under an argon atmosphere. The reaction was quenched with water (20 mL) at room temperature. The resulting mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SC The filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography to give 4-fluoro-l- (2-iodophenyl)imidazole (100 mg, 15% yield) as a yellow solid. ¹H NMR (400 MHz, DMSO-dg) 6 8.05-8.03 (m, 1H), 7.60 - 7.52 (m, 2H), 7.51-7.49 (m, 1H), 7.31-7.29 (m, 1H), 7.17-7.19 (m, 1H).

[000166] Preparation of Intermediate 3: 4-chloro-l-(2-iodophenyl)-lH-imidazole

[000167] A solution of 4-chloro-lH-imidazole (200 mg, 2.0 mmol) in DMSO (5 mL) was treated with 1,2-diiodobenzene (3.21 g, 9.8 mmol), Cui (37 mg, 0.2 mmol), L-proline (45 mg, 0.39 mmol) and K3PO4 (621 mg, 2.9 mmol) at 25 °C under an argon atmosphere. The resulting mixture was stirred overnight at 60 “C. The reaction was then quenched with water (30 mL) at 0 °C. The aqueous layer was extracted with EtOAc (3 x 30 mL), and the combined organics were dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography to give the crude product (400 mg, 67% yield) as a white solid. ^TH NMR (400 MHz, DMSO-d₆) 6 8.05 - 8.03 (m, 1H), 7.83 (d, J = 1.5 Hz, 1H), 7.60 - 7.52 (m, 2H), 7.51-7.49 (m, 1H), 7.32 - 7.27 (m, 1H).

[000168] Preparation of Intermediate 4: l-(difluoromethyl)-4-(2-iodophenyl)pyrazole

[000169] To a stirred mixture of l-(difluoromethyl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-

2-yl)pyrazole (1 g, 4.1 mmol) and 1,2-diiodobenzene (1.62 g, 4.9 mmol) in THF (8 mL) and water

(2 mL) was added K2CO3 (1.70 g, 12.3 mmol) and Pd PPhshCl₂ (1.44 g, 2.0 mmol) at room temperature under an argon atmosphere. The resulting mixture was stirred overnight at 70 °C. The mixture was allowed to cool down to room temperature and the reaction was quenched with water (40 mL) at room temperature. The resulting mixture was extracted with EtOAc (3 x 40 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SCU. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography to give l-(difluoromethyl)-4-(2-iodophenyl)pyrazole (350 mg, 27% yield) as a light yellow oil. ^TH NMR (400 MHz, DMSO-d₆) 68.50 (d, J = 0.7 Hz, 1H), 8.03 (d, J = 0.7 Hz, 1H), 8.01 - 7.97 (m, 1H), 7.80 (d, J = 59.0 Hz, 1H), 7.49 - 7.41 (m, 2H), 7.14- 7.07 (m, 1H).

[000170] Preparation of Intermediate 5: 4-(2-iodophenyl)-l,5-dimethyl-lH-pyrazole

[000171] To a stirred mixture of l,5-dimethyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)pyrazole (500 mg, 2.3 mmol) and 1,2-diiodobenzene (3.71 g, 11.3 mmol) in THF (20 mLj/HjO (5 mL) were added K2CO3 (933 mg, 6.8 mmol) and PdfPPhshCl₂ (158 mg, 0.23 mmol) in portions at room temperature under an argon atmosphere. The mixture was stirred overnight at 70 °C under an argon atmosphere. The mixture was allowed to cool down to room temperature and then quenched by the addition of water (80 mL). The resulting mixture was extracted with EtOAc (3 x 80 mL). The combined organic layers were washed with brine and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA (1:1), to afford 4-(2- iodophenyl)-l,5-dimethylpyrazole (250 mg, 37% yield) as a yellow solid. ^XH NMR (400 MHz, DMSO-dg) 67.99 - 7.92 (m, 1H), 7.46 - 7.40 (m, 1H), 7.38 (s, 1H), 7.27 - 7.20 (m, 1H), 7.11 - 7.02 (m, 1H), 3.78 (s, 3H), 2.13 (s, 3H). LCMS (ESI) m/z = 299.0 (M+H).

[000172] Preparation of Intermediate 6: 4-(2-bromophenyl)-l,3-dimethylpyrazole

[000173] To a stirred solution/mixture of l,3-dimethyl-4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)pyrazole (500 mg, 2.3 mmol) and l-bromo-2-iodobenzene (637 mg, 2.3 mmol) and PdfPPhshCl₂ (158 mg, 0.23 mmol) and K2CO3 (933 mg, 6.8 mmol) in THF (4 mL) was added H2O (2 mL) at room temperature under an argon atmosphere. The resulting mixture was stirred overnight at 50 °C and then quenched by the addition of water/ice (20 mL) at 0 °C. The resulting mixture was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA (1:1), to afford 4-(2-bromophenyl)-l,3-dimethylpyrazole (350 mg, 62% yield) as an off-white oil. LCMS (ESI) m/z = 251.1 (M+H).

[000174] Preparation of Intermediate 7: 4-(2-bromophenyl)-3-fluoro-l-methylpyrazole)

[000175] A mixture of 4-bromo-3-fluoro-l-methylpyrazole (300 mg, 1.68 mmol) and 4, 4, 5, 5- tetramethyl-2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (638 mg, 2.5 mmol), KOAc (493 mg, 5 mmol) and Pd(dppf)Ck (123 mg, 0.17 mmol) in 1,4-dioxane (4 mL) was stirred for 2 h at 100 °C under an argon atmosphere. The crude product/ resulting mixture was used in the next step directly without further purification. To a stirred mixture of 3-fluoro-l- methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazole (300 mg, 1.3 mmol) in benzene (15 mL), 2-bromoiodobenzene (375 mg, 1.3 mmol), Pd PPhshCh (93 mg, 0.13 mmol), and K2CO3 (550 mg, 4 mmol) in THF (4 mL) was added H₂O (1 mL) portions at room temperature under an argon atmosphere. The resulting mixture was stirred for 2 h at 50 °C under an argon atmosphere. The reaction was quenched by the addition of water/ice (30 mL) at 0 °C. The resulting mixture was extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine and dried over anhydrous Na₂SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA (1:1), to afford 4-(2-bromophenyl)-3-fluoro-l- methylpyrazole) (100 mg, 24% yield) as a light yellow oil. LCMS (ESI) m/z = 255.1.

[000176] Preparation of Intermediate 8: 3-(2-bromophenyl)pyridine [000177] To a solution of l-bromo-2-iodo-benzene (2.0 g, 7.1 mmol) and 3-pyridylboronic acid (1.3 g, 10.6 mmol) in dioxane (20 mL) and H2O (2 mL) was added K2CO3 (2.93 g, 21.2 mmol) and Pd(PPh₃)₂CI₂ (496 mg, 0.71 mmol). The mixture was stirred at 80 °C for 12 h under a nitrogen atmosphere. The reaction mixture was cooled to room temperature and water (50 mL) was added. The mixture was then extracted with ethyl acetate (2 x 50 mL), and the combined organic layers were washed with brine (50 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to afford 3-(2- bromophenyl)pyridine (0.54 g, 27% yield) as a yellow oil. LCMS (ESI) m/z = 234.1 and 236.1 (M+H). ^TH NMR (400 MHz, CDCI3): <58.59 (d, J = 1.6 Hz, 1H), 8.55 (dd, J = 1.2, 4.8 Hz, 1H), 7.71- 7.65 (m, 1H), 7.62 (dd, J = 1.2, 8.0 Hz, 1H), 7.36-7.21 (m, 4H).

[000178] Preparation of Intermediate 9: l-(2-bromophenyl)-lH-l,2,3-triazole

[000179] To a solution of lH-triazole (5 g, 72.4 mmol) and (2-bromophenyl)boronic acid (20 g, 99.6 mmol) in CH3CN (200 mL) was added CU2O (2.07 g, 14.5 mmol), 4A molecular sieves (20 g), 1,10-phenanthroline (2.61 g, 14.5 mmol), and DIPEA (51.9 g, 401 mmol). The resulting mixture was stirred at 80 °C for 12 h under an oxygen atmosphere (15 psi). The mixture was then filtered through a celite pad and water (300 mL) was added. It was then extracted with ethyl acetate (3 x 200 mL), and the combined organic layers were washed with brine (300 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to afford l-(2-bromophenyl) triazole (2.6 g, 14% yield) as a brown oil. LCMS (ESI) m/z = 224.1 and 226.1 (M+H). ^XH NMR (400 MHz, CDCI3): <57.97 (d, J = 0.8 Hz, 1H), 7.85 (d, J = 1.2 Hz, 1H), 7.75 (dd, J = 1.2, 8.0 Hz, 1H), 7.56-7.51 (m, 1H), 7.48 (dt, J = 1.2, 7.6 Hz, 1H), 7.56-7.45 (m, 1H), 7.44-7.35 (m, 1H).

[000180] Preparation of Intermediate 10: 4-(2-bromophenyl)-5-fluoro-l-methylpyrazole

Step 1: Preparation of 5-fluoro-l-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazole [000181] To a stirred solution of 4-bromo-5-fluoro-l-methylpyrazole (1 g, 5.6 mmol) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2- dioxaborolane (2.13 g, 8.4 mmol) in dioxane (10 mL) were added Pd(dppf)CI₂ (409 mg, 0.56 mmol) and KOAc (1.37 g, 14 mmol) in portions at room temperature. The resulting mixture was stirred for 2 h at 100 °C under an argon atmosphere. The resulting mixture was used in the next step directly without further purification. LCMS (ESP) m/z = 227.1 (M+H).

Step 2: Preparation of 4-(2-bromophenyl)-5-fluoro-l-methylpyrazole

[000182] A solution of 5-fluoro-l-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)pyrazole (1 g, 4.4 mmol) in dioxane (10 mL) was treated with l-bromo-2-iodobenzene (292 mg, 1.0 mmol), Pd(PPh3)₂CI₂ (310 mg, 0.44 mmol), K2CO3 (1.83 g, 13.2 mmol) at room temperature followed by the addition of H₂O (2.5 mL) dropwise at room temperature. The resulting mixture was stirred overnight at 50 °C under a nitrogen atmosphere. The reaction was then quenched by the addition of water/ice (30 mL) at room temperature. The resulting mixture was extracted with EtOAc (3 x 30 mL). The combined organic phases were washed with brine and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography to give 4-(2- bromophenyl)-5-fluoro-l-methylpyrazole (120 mg, 11% yield) as a light-yellow oil. LCMS (ESP) m/z = 255.0 (M+H).

[000183] Preparation of Intermediate 11: (R)-5-(l-aminoethyl)-2,7-dimethyl-3-(4-(2- methylpyrimidin-5-yl)piperazin-l-yl)isoquinolin-l(2H)-one

Step 1: Preparation of benzyl 4-(2-methylpyrimidin-5-yl)piperazine-l-carboxylate

[000184] To a stirred mixture of 5-bromo-2-rnethylpyrirnidine (2.0 g, 11.5 mmol) and benzyl piperazine-l-carboxylate (2.0 g, 8.9 mmol) in toluene (30 mL) were added CS2CO3 (5.8 g, 17.8 mmol), tris(dibenzylideneacetone)dipalladium(0) (1.6 g, 1.8 mmol) and RuPhos (0.8 g, 1.8 mmol) at room temperature. The resulting mixture was stirred overnight at 100 °C under a nitrogen atmosphere. The reaction was then quenched with water (40 mL) at room temperature and the resulting mixture was extracted with EtOAc (2 x 40 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SCU. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CHjCl₂/MeOH to afford benzyl 4-(2-methylpyrimidin-5- yl)piperazine-l-carboxylate (2.0 g, 72% yield) as a yellow solid. ¹H NMR (400 MHz, DMSO-dg) 6 8.40 (br, 2H), 7.38 (d, J = 4.3 Hz, 4H), 7.37 - 7.28 (m, 1H), 5.11 (s, 2H), 3.55 (t, J = 4.9 Hz, 4H), 3.20 (t, J = 5.2 Hz, 4H), 2.49 (s, 3H).

Step 2: Preparation of 2-methyl-5-(piperazin-l-yl)pyrimidine

[000185] To a stirred mixture of benzyl 4-(2-methylpyrimidin-5-yl)piperazine-l-carboxylate (2.0 g, 6.4 mmol) in MeOH (30 mL) was added Pd/C (500 mg, 4.7 mmol) at room temperature. The resulting mixture was stirred for 2 h at room temperature under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with MeOH (3 x 10 mL). The filtrate was concentrated under reduced pressure. This resulted in 2-methyl-5-(piperazin-l- yl)pyrimidine (900 mg, 79% yield) as a yellow solid. ¹H NMR (400 MHz, DMSO-dg) 6 8.35 (s, 2H), 3.11 - 3.04 (m, 4H), 2.87 - 2.78 (m, 4H), 2.48 (s, 3H).

Step 3: Preparation of 4-bromo-2-(hydroxyimino)-6-methyl-3H-inden-l-one

[000186] To a stirred solution of 4-bromo-6-methyl-2,3-dihydroinden-l-one (2 g, 8.89 mmol) in 12 M aqueous HCI (10 mL) and EtjO (10 mL) was added 3-methylbutyl nitrite (1.25 g, 10.7 mmol) slowly dropwise at 0 °C. The resulting solution was stirred for 4 h at room temperature. The mixture was then cooled to 0 °C and the precipitated solids were collected by filtration and washed sequentially with H2O (3 x 50 mL) and EtjO (2 x 20 mL). The collected solids were concentrated and dried under vacuum. The crude product was used in the next step directly without further purification. This resulted in 4-bromo-2-(hydroxyimino)-6-methyl-3H-inden-l- one (1.5 g, 66%) as an off-white solid. MS: (ES⁺) m/z = 253.9 [M+H]⁺.

Step 4: Preparation of 5-bromo-3-chloro-7-methyl-2H-isoquinolin-l-one

[000187] To a stirred solution of 4-bromo-2-(hydroxyimino)-6-methyl-3H-inden-l-one (1.5 g, 5.90 mmol) in CHCI3 (30 mL) was slowly added PCI₅ (2.46 g, 11.81 mmol) in portions at 0 °C. The resulting mixture was stirred for 3 h at room temperature and then concentrated under reduced pressure. To the crude product was added 4 M HCI in 1,4-dioxane (30 mL). The resulting solution was stirred overnight at room temperature and then concentrated under reduced pressure. The residue was purified by trituration, PE:EA = 5:1, to afford 5-bromo-3- chloro-7-methyl-2H-isoquinolin-l-one (1 g, 61%) as a yellow solid. MS: (ES ) m/z = 269.9 [M-l]’.

Step 5: Preparation of 5-bromo-3-chloro-2,7-dimethylisoquinolin-l-one

[000188] To a stirred solution of 5-bromo-3-chloro-7-methyl-2H-isoquinolin-l-one (1.3 g, 4.77 mmol) in DMF (10 mL) was slowly added NaH (0.17 g, 7.16 mmol) in portions at 0 °C. The resulting solution was stirred for 20 min at 0 °C. lodomethane (0.81 g, 5.72 mmol) was slowly added dropwise at 0 °C and the resulting solution was stirred overnight at room temperature. The reaction was quenched with water (40 mL) and the resulting mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (3 x 40 mL), dried over anhydrous Na2SCU, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA = 4:1, to afford 5-bromo-3-chloro-2,7- dimethylisoquinolin-l-one (900 mg, 65%) as a reddish brown solid. MS: (ES⁺) m/z = 286.0 [M+H]⁺.

Step 6: Preparation of 5-acetyl-3-chloro-2,7-dimethylisoquinolin-l-one

[000189] A mixture of 5-bromo-3-chloro-2,7-dimethylisoquinolin-l-one (2.6 g, 9.07 mmol), tributyl(l-ethoxyethenyl)stannane (3.60 g, 9.98 mmol) and Pd(PPh3)₄ (1.05 g, 0.91 mmol) in anhydrous 1,4-dioxane (25 mL) was stirred overnight at 100 °C under a nitrogen atmosphere. The reaction mixture was cooled to room temperature, treated with aqueous 1 N HCI (10 mL), and stirred for 15 min. The resulting mixture was diluted with water (60 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organics were washed with water (3 x 60 mL) and brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford 5-acetyl-3-chloro-2,7-dimethylisoquinolin-l-one (1.9 g, 83%) as a yellow solid. MS: ES⁺ (m/z) =250.1 [M+H]⁺.

Step 7: Preparation of (R,E)-N-(l-(3-chloro-2,7-dimethyl-l-oxo-l,2-dihydroisoquinolin-5- yl)ethylidene)-2-methylpropane-2-sulfinamide

[000191] To a stirred solution of 5-acetyl-3-chloro-2,7-dimethylisoquinolin-l-one (9.8 g, 39.2 mmol) and (R)-2-methylpropane-2-sulfinamide (23.8 g, 196 mmol) in THF (200 mL) was added Ti(Oi-Pr)₄ (55.8 g, 196 mmol). The resulting mixture was stirred overnight at 80 °C under a nitrogen atmosphere. The reaction was quenched with saturated aqueous sodium chloride (200 mL). The resulting mixture was filtered, and the filter cake was washed with ethyl acetate (3 x 300 mL). The filtrate was extracted with ethyl acetate (3 x 300 mL). The combined organic layers were washed with H₂O (3 x 200 mL), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA = 3:2, to afford (R,E)-N-(l-(3- chloro-2,7-dimethyl-l-oxo-l,2-dihydroisoquinolin-5-yl)ethylidene)-2-methylpropane-2- sulfinamide (9 g, 64%) as a yellow solid. MS: (ES⁺) m/z = 353.1 [M+H]⁺.

Step 8: Preparation of (R)-N-((R)-l-(3-chloro-2,7-dimethyl-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)-2- methylpropane-2-sulfinamide

[000192] To a stirred solution of (R,E)-N-(l-(3-chloro-2,7-dimethyl-l-oxo-l,2- dihydroisoquinolin-5-yl)ethylidene)-2-methylpropane-2-sulfinamide (10.5 g, 29.8 mmol) and CeCk’TE O (16.6 g, 44.6 mmol) in MeOH (120 mL) was added NaBH₄ (2.81 g, 74.4 mmol) at -78 °C. The resulting solution was stirred for 2 h at room temperature. The reaction mixture was quenched with saturated aqueous ammonium chloride (150 mL) and extracted with ethyl acetate (3 x 200 mL). The combined organic layers were washed with brine (2 x 200 mL), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA = 1:2, and then purified further by HP-Flash chromatography (25%-55% ACN in H2O (0.1% FA) in 45 min) to afford (R)-N-((R)-l-(3-chloro-2,7-dimethyl-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)-2- methylpropane-2-sulfinamide (6.5 g, 61%) as an off-white solid. MS: (ES⁺) m/z = 355.0 [M+H]⁺.

Step 9: Preparation of (R)-N-((R)-l-(2,7-dimethyl-3-(4-(2-methylpyrimidin-5-yl)piperazin-l-yl)-l-oxo- l,2-dihydroisoquinolin-5-yl)ethyl)-2-methylpropane-2-sulfinamide

[000193] A mixture of (R)-N-((R)-l-(3-chloro-2,7-dimethyl-l-oxo-l,2-dihydroisoquinolin-5- yl)ethyl)-2-methylpropane-2-sulfinamide (2 g, 5.6 mmol) and 2-methyl-5-(piperazin-l- yl)pyrimidine (1.0 g, 5.6 mmol) and CS2CO3 (21 mg, 0.065 mmol) and Pd2(dba)s (1.55 g, 1.7 mmol) and RuPhos (1.58 g, 3.4 mmol) in 1,4-dioxane (20 mL) was stirred overnight at 100 °C under an argon atmosphere. The reaction was quenched with water (50 mL) at room temperature. The resulting mixture was extracted with EtOAc (2 x 50 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with C^Cl₂/MeOH = 5:1, to afford (R)-N-((R)-l-(2,7-dimethyl-3-(4-(2- methylpyrimidin-5-yl)piperazin-l-yl)-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)-2-methylpropane- 2-sulfinamide (1.5 g, 54% yield) as a brown solid. ¹H NMR (400 MHz, DMSO-dg) 6 8.46 (m, J = 7.0 Hz, 2H), 7.93 (s, 1H), 7.61 (d, J = 1.9 Hz, 1H), 6.36 (d, J = 12.3 Hz, 1H), 5.65 (d, J = 5.7 Hz, 1H), 4.93 - 4.86 (m, 1H), 4.1-3.9 (m, 2H), 3.52 (s, 3H), 3.4-2.7 (br, 6H), 2.52 (s, 3H), 2.42 (s, 3H), 1.52-1.47 (m, 3H), 1.07 (d, J = 9.3 Hz, 9H).

Step 10: Preparation of (R)-5-(l-aminoethyl)-2,7-dimethyl-3-(4-(2-methylpyrimidin-5-yl)piperazin-l- yl)isoquinolin-l(2H)-one

[000194] To a stirred mixture of (R)-N-((R)-l-(2,7-dimethyl-3-(4-(2-methylpyrimidin-5- yl)piperazin-l-yl)-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)-2-methylpropane-2-sulfinamide (1.5 g, 3.0 mmol) in DCM (15 mL) was added HCI in 1,4-dioxane (3 mL, 4 M) dropwise at 0 °C under an argon atmosphere. The resulting mixture was stirred for 3h at room temperature and then diluted via the addition of water/ice (50 mL) at 0 °C. The resulting mixture was extracted with EtOAc (2 x 50 mL). The aqueous layer was basified to pH 12 via the addition of aqueous ammonium hydroxide. The resulting mixture was then extracted with CH2CI2 (3 x 80 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SO₄. After filtration, the filtrate was concentrated under reduced pressure to afford (R)-5-(l- aminoethyl)-2,7-dimethyl-3-(4-(2-methylpyrimidin-5-yl)piperazin-l-yl)isoquinolin-l(2H)-one (950 mg, 80% yield) as a brown solid. ^TH NMR (400 MHz, DMSO-d₆) 6 8.46 (br, 2H), 7.87 (s, 1H), 7.69 (s, 1H), 6.34 (s, 1H), 4.56-4.50 (m, 1H), 3.53 (s, 3H), 3.30-2.88 (m, 8H), 2.41 (s, 3H), 2.52 (s, 3H), 1.30 (d, J = 6.5 Hz, 3H).

[000195] Preparation of Intermediate 12: (R)-5-(l-aminoethyl)-3-chloro-2,7- dimethylisoquinolin-l-one hydrochloride

[000197] A mixture of (R)-N-[(lR)-l-(3-chloro-2,7-dimethyl-l-oxoisoquinolin-5-yl)ethyl]-2- methylpropane-2-sulfinamide (240 mg, 0.68 mmol) in 1:1 4 M HCI in l,4-dioxane/CHzCl2 (10 mL) was stirred overnight at room temperature under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure to afford 5-[(lR)-l-aminoethyl]-3-chloro-2,7- dimethylisoquinolin-l-one hydrochloride (165 mg, 85%) as a white solid. The crude product was used in the next step directly without further purification. MS: (ES⁺) m/z = 250.0 [M+H]⁺.

[000198] Preparation of Intermediate 13: 2'-methyl-5-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-2,5'-bipyrimidine

Step 1: Preparation of 5-bromo-2'-methyl-2,5'-bipyrimidine [000199] To a stirred mixture of 2-methylpyrimidin-5-ylboronic acid (1 g, 7.25 mmol) and 5- bromo-2-iodopyrimidine (2.07 g, 7.25 mmol) in dioxane (15 mL) and H2O (3 mL) were added Pd(dppf)Ck (0.53 g, 0.73 mmol) and K2CO3 (3.01 g, 21.8 mmol) in portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred for 2 h at 80 °C and then was filtered and the filter cake was washed with ethyl acetate (3 x 10 mL). The filtrate was concentrated under reduced pressure and the resulting residue was purified by silica gel chromatography (eluted with PE:EA = 1:1) to afford 5-bromo-2'-methyl-2,5'-bipyrimidine (1.2 g, 67% yield) as a brown solid. LCMS: (ES ) m/z = 251.0 (M+H).

Step 2: Preparation of 2'-methyl-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-2,5'-bipyrimidine

[000200] To a stirred mixture of 5-bromo-2'-methyl-2,5'-bipyrimidine (400 mg, 1.59 mmol) and bis(pinacolato)diboron (607 mg, 2.39 mmol) in dioxane (10 mL) were added Pd(dppf)Ck (117 mg, 0.16 mmol) and KOAc (469 mg, 4.78 mmol) in portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred for 2 h at 100 °C. The resulting mixture was then filtered, and the filter cake was washed with CH2CI2 (3 x 20 mL). The filtrate was concentrated under reduced pressure to afford 2'-methyl-5-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-2,5'-bipyrimidine (400 mg, 84% crude yield) which was used directly in the next step without further purification. LCMS: (ESP) m/z = 299.1 (M+H).

[000201] Intermediate 14: (2-(l-methyl-6-oxo-l,6-dihydropyridin-2-yl)pyrimidin-5-yl)boronic acid Step 1: Preparation of 6-bromo-l-methylpyridin-2(lH)-one

[000202] To a mixture of 6-bromo-lH-pyridin-2-one (4.97 g, 28.5 mmol) in ACN (50 mL) were added K2CO3 (7.90 g, 57.2 mmol) and iodomethane (8.11 g, 57.2 mmol). The mixture was stirred at 25 °C for 12 hr and then poured into ice-water (20 mL) and stirred for 3 min. The aqueous phase was extracted with ethyl acetate (20 mL x 3). The combined organics were washed with brine (20 mL x 2), dried with anhydrous Na2SCU, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (PE:EA = 1:0 to 65:35) to afford 6-bromo-l-methylpyridin-2(lH)-one (4.55 g, 80% yield) as a white solid. LCMS: (ES ) m/z = 188.1 (M+H).

Step 2: Preparation of (l-methyl-6-oxo-l,6-dihydropyridin-2-yl)boronic acid

[000203] A mixture of 6-bromo-l-methylpyridin-2(lH)-one (2.5 g, 13.3 mmol), 4, 4, 5, 5- tetramethyl-2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (4.05 g, 16.0 mmol), potassium acetate (3.91 g, 39.9 mmol), Pd(dppf)Cl₂ (973 mg, 1.33 mmol) in dioxane (8 mL) was degassed and purged with nitrogen three times. The mixture was then stirred at 110 °C for 0.5 h, then cooled to ambient temperature, filtered, and concentrated under reduced pressure to afford (l-methyl-6-oxo-l,6-dihydropyridin-2-yl)boronic acid (2.5 g, crude yield) as a dark brown oil. The crude product was used in the next step without further purification. LCMS: (ESP) m/z = 154.0 (M+H).

Step 3: Preparation of 6-(5-bromopyrimidin-2-yl)-l-methylpyridin-2(lH)-one

[000204] A mixture of 5-bromo-2-iodo-pyrimidine (2.12 g, 7.44 mmol), (l-methyl-6-oxo-l,6- dihydropyridin-2-yl)boronic acid (2.5 g, 10.6 mmol), K2CO3 (4.41 g, 31.9 mmol), and Pd(dppf)Cl₂ (778 mg, 1.06 mmol) in dioxane (10 mL) and water (1 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 90 °C for 0.5 h, cooled to ambient temperature, and poured into water (10 mL). The mixture was extracted with ethyl acetate (10 mL x 5). The combined organics were washed with brine (10 mL x 3), dried over anhydrous Na2SCU, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (PE:EA = 1:0 to 49:51) to afford 6-(5-bromopyrimidin-2-yl)-l-methylpyridin- 2(lH)-one (1.2 g, 17% yield) as a yellow solid. LCMS: (ES ) m/z = 266.0 (M+H).

Step 4: Preparation of (2-(l-methyl-6-oxo-l,6-dihydropyridin-2-yl)pyrimidin-5-yl)boronic acid

[000205] A mixture of 6-(5-bromopyrimidin-2-yl)-l-methylpyridin-2(lH)-one (500 mg, 1.88 mmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (954 mg, 3.76 mmol), Pd(dppf)Cl₂ (137 mg, 0.18 mmol), potassium acetate (553 mg, 5.64 mmol) in dioxane (8 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 100 °C for 1 h, then cooled to ambient temperature, filtered, and the filtrate was concentrated under reduced pressure to afford (2-(l-methyl-6-oxo-l,6-dihydropyridin-2- yl)pyrimidin-5-yl)boronic acid (600 mg, crude yield) as a brown oil that was used directly. LCMS: (ESP) m/z = 232.1 (M+H).

[000206] Intermediate 15: 4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)aniline

Pd(dppf)CI₂,

Step 1: Preparation of (2-amino-5-fluorophenyl)boronic acid [000207] To a mixture of 2-bromo-4-fluoro-aniline (5 g, 26.3 mmol) and 4,4,5,5-tetramethyl-2- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (10.0 g₇ 39.5 mmol), potassium acetate (7.75 g, 78.9 mmol) in dioxane (50 mL) was added Pd(dppf)CI₂»CH₂CI₂ (2.15 g, 2.63 mmol) at 25 °C under a nitrogen atmosphere. The mixture was then heated to 90 °C for 1 hour. The reaction mixture was cooled to ambient temperature, filtered, and the filtrate was concentrated. The resulting residue was purified by silica gel chromatography (PE:EA = 10:1 to 5:1) to give (2-amino-5-fluorophenyl)boronic acid (3.98 g, 94% yield) as a colorless oil. LCMS: (ESP) m/z = 156.1 (M+H).

Step 2: Preparation of 4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)aniline

[000208] A mixture of 4-bromo-l-methyl-triazole (3.69 g, 22.8 mmol), (2-amino-5- fluorophenyl)boronic acid (3.6 g, 15.2 mmol), Pd(dppf)CI₂ (2.22 g, 3.04 mmol), Na₂CO₃ (4.83 g, 45.6 mmol) in dioxane (30 mL) and H₂O (3 mL) was degassed and purged with nitrogen three times. The mixture was then stirred at 90 °C for 6 hours, cooled to an ambient temperature, and poured into ice-water (100 mL). The mixture was extracted with DCM (100 mL x 3). The combined organics were washed with brine (100 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (PE:EA = 1:1) to afford 4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)aniline (460 mg, 14% yield) as a yellow solid. LCMS: (ESP) m/z = 193.1 (M+H).

[000209] Intermediate 16: 5-bromo-2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4- yl)pyrimidine 1-oxide

Step 1: Preparation of 3-chloro-4-iodopyridin-2-ol Cl

K .OH tr

[000210] To a mixture of 3-chloro-2-fluoro-4-iodo-pyridine (25 g, 97.1 mmol) was added HCI (6 M, 100 mL). The mixture was stirred at 100 °C for 0.5 h, then cooled and lyophilized to give 3- chloro-4-iodo-pyridin-2-ol (22 g, 79% yield) as a white solid. ¹H NMR (400 MHz, CDCU): 6 7.09 (d, J = 6.8 Hz, 1H), 6.83 (d, J = 6.8 Hz, 1H).

Step 2: Preparation of 3-chloro-4-iodo-l-methylpyridin-2(lH)-one

[000211] To a mixture of 3-chloro-4-iodo-pyridin-2-ol (22 g, 86.1 mmol) in MeOH (220 mL) was added K2CO3 (23.8 g, 172 mmol) and Mel (14.7 g, 103 mmol). The mixture was stirred at 60 °C for 2 h. The mixture was then poured into ice-water (300mL) and stirred for 3 min. The mixture was then extracted with ethyl acetate (250 mL x 3). The combined organics were washed with brine (100 mL), dried with anhydrous Na2SCU, filtered and concentrated under reduced pressure. The crude product was triturated with PE:EA = 10:1 at 25 °C for 10 min to afford 3-chloro-4-iodo-l-methyl-pyridin-2(lH)-one (22 g, 93% yield) as a white solid. ^TH NMR (400 MHz, DMSO-dg): 6 7.48 (d, J = 7.2 Hz, 1H), 6.73 (d, J = 7.2 Hz, 1H), 3.44 (s, 3H).

Step 3: Preparation of 3-chloro-l-methyl-2-oxo-l,2-dihydropyridine-4-carbonitrile

[000212] A mixture of 3-chloro-4-iodo-l-methyl-pyridin-2(lH)-one (22 g, 81.6 mmol), copper (I) cyanide (8.04 g, 89.8 mmol) in NMP (150 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 100 °C for 12 h under a nitrogen atmosphere. After the mixture was cooled to ambient temperature, 13% aqueous NH₄OH (500 mL) was added dropwise over 5 min. The resulting mixture was diluted with water (250 mL), stirred for 0.5 h, and then extracted with DCM (150 mL x 3). The combined organics were washed with brine (150 mL), dried over Na2SO₄, filtered, and concentrated under reduced pressure to give a residue. The crude product was purified by reversed-phase HPLC to afford 3-chloro-l-methyl-2-oxo-pyridine-4-carbonitrile (9 g, 64% yield) as a white solid. ^TH NMR (400 MHz, DMSO-d₆): 6 7.93 (d, J = 7.2 Hz, 1H), 6.66 (d, J = 7.2 Hz, 1H), 3.54 (s, 3H).

Step 4: Preparation of 3-chloro-N'-hydroxy-l-methyl-2-oxo-l,2-dihydropyridine-4-carboximidamide

[000213] To a mixture of 3-chloro-l-methyl-2-oxo-pyridine-4-carbonitrile (4 g, 23.7 mmol) in EtOH (60 mL) was added TEA (12.0 g, 119 mmol) and hydroxylamine hydrochloride (3.30 g, 47.5 mmol). The mixture was stirred at 70 °C for 0.5 h and then concentrated under reduced pressure. Water (20 mL) was then added and the resulting precipitate was collected by filtration to afford 3-chloro-N'-hydroxy-l-methyl-2-oxo-l,2-dihydropyridine-4-carboximidamide (7.5 g, 78% yield) as a white solid. ^XH NMR (400 MHz, DMSO-d₆): 6 9.68 (s, 1H), 7.72 (d, J = 6.8 Hz, 1H), 6.23 (d, J = 6.8 Hz, 1H), 5. 92 (br s, 2H), 3.51 (s, 3H).

Step 5: Preparation of 5-bromo-2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)pyrimidine 1- oxide

[000214] To a mixture of 3-chloro-N'-hydroxy-l-methyl-2-oxo-l,2-dihydropyridine-4- carboximidamide (7 g, 34.7 mmol) and 2-bromopropanedial (7.86 g, 52.1 mmol) in IPA (200 mL) was added TFA (4.75 g, 41.7 mmol). The mixture was stirred at 90 °C for 1 h and then concentrated under reduced pressure to afford a residue. The crude product was crystallized from PE:EA = 1:1 (60 mL) at 25 °C and then triturated with H2O (10 mL) at 25 °C for 5 min to afford 5-bromo-2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)pyrimidine 1-oxide (3.2 g, 30% yield) as a yellow solid. LCMS: (ES ) m/z = 316.0 (M+H). [000215] Intermediate 17: 4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4-yl)aniline

\

N I

Step 1: Preparation of 4-fluoro-2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)aniline

[000216] To a mixture of 2-bromo-4-fluoro-aniline (5 g, 26.3 mmol), 4,4,5,5-tetramethyl-2- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (10.0 g, 39.5 mmol), potassium acetate (7.75 g, 78.9 mmol) in dioxane (50 mL) was added P , 2.63 mmol) at 25 °C under a nitrogen atmosphere. The mixture was then heated to 90 °C for 1 h. The mixture was then cooled to ambient temperature, filtered, and the filtrate was concentrated to give a residue. The residue was purified by silica gel chromatography (PE:EA = 10:1 to 5:1) to afford 4-fluoro-2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)aniline (6.1 g, 93% yield) as a colorless oil. LCMS: (ESP) m/z = 156.1 (M+H, boronic acid observed).

Step 2: Preparation of 4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4-yl)aniline

[000217] To a mixture of 4-fluoro-2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)aniline (3.18 g, 13.4 mmol), 4-bromo-3-fluoro-l-methyl-pyrazole (1.5 g, 8.38 mmol), and K2CO3 (3.47 g, 25.1 mmol) in dioxane (20 mL) and water (2 mL) was added PdfdppfjCl₂’Cl-bCk (684 mg, 0.84 mmol) at 25 °C under a nitrogen atmosphere. The mixture was heated to 100 °C for 1 h, then cooled to ambient temperature and concentrated to afford a residue. The residue was purified by silica gel chromatography (PE:EA = 10:1 to 5:1) to afford 4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4- yl)aniline (0.95 g, 54% yield) as a black solid. LCMS: (ES ) m/z = 210.1 (M+H).

[000218] Intermediate 18: 2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5- iodopyridazin-3(2H)-one

Step 1: Preparation of 2-(3-chloro-2-methoxypyridin-4-yl)-5-iodopyridazin-3(2H)-one

[000219] In 4 parallel batches, a mixture of (3-chloro-2-methoxy-4-pyridyl)boronic acid (1.90 g, 10.1 mmol), 4-iodo-lH-pyridazin-6-one (1.5 g, 6.76 mmol), Cu(OAc)2 (245.47 mg, 1.35 mmol), isoquinolin-6-ol (196 mg, 1.35 mmol) and TEA (2.05 g, 20.3 mmol) in DCE (30 mL) was degassed and purged with O2 three times. The mixture was stirred at 50 °C for 16 h, then cooled to ambient temperature and filtered. All filtrates were combined, concentrated, and the residue was suspended in water (30 mL) and stirred for 2 min. The aqueous mixture was extracted with ethyl acetate (30 mL x 3) and the combined organics were dried with anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EA = 100:0 to 90:10) to afford 2-(3-chloro-2-methoxypyridin-4-yl)-5- iodopyridazin-3(2H)-one (1.7 g, 14% yield) as a yellow solid. LCMS: (ESP) m/z = 363.9 (M+H).

Step 2: Preparation of 2-(3-chloro-2-hydroxypyridin-4-yl)-5-iodopyridazin-3(2H)-one [000220] A mixture of 2-(3-chloro-2-methoxypyridin-4-yl)-5-iodopyridazin-3(2H)-one (1.65 g, 4.54 mmol) and trimethylsilyl iodide (4.54 g, 22.7 mmol) in ACN (15 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 80 °C for 2 h, then cooled to ambient temperature, filtered, and the filtrate was concentrated to afford 2-(3-chloro-2- hydroxypyridin-4-yl)-5-iodopyridazin-3(2H)-one (1.8 g, crude yield) as a brown solid. The crude product was used in the next step without further purification. LCMS: (ES ) m/z = 349.8 (M+H).

Step 3: Preparation of 2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-iodopyridazin-3(2H)- one

[000221] To a mixture of 2-(3-chloro-2-hydroxypyridin-4-yl)-5-iodopyridazin-3(2H)-one (1.75 g, 5.01 mmol) in ACN (20 mL) was added CS2CO3 (4.89 g, 15.0 mmol) and iodomethane (1.42 g, 10.0 mmol). The mixture was stirred at 25 °C for 12 h, then was poured into ice-water (20 mL) and stirred for 1 min. The aqueous mixture was extracted with DCM (50 mL x 3). The combined organics were washed with brine (50 mL x 2), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EA = 100:0 to 45:55) to afford 2-(3-chloro-l-methyl-2-oxo-l,2- dihydropyridin-4-yl)-5-iodopyridazin-3(2H)-one (1.42 g, 76% yield) as an off-brown solid. LCMS: (ESP) m/z = 363.9 (M+H).

[000222] Intermediate 19: 5-acetyl-3,7-dichloro-2-methylisoquinolin-l(2H)-one

Step 1: Preparation of 4-bromo-6-chloro-2-(hydroxyimino)-2,3-dihydro-lH-inden-l-one

[000223] To a stirred mixture of 4-bromo-6-chloro-2,3-dihydroinden-l-one (1 g, 4.07 mmol) in THF (2.5 mL) and CH2CI2 (10 mL) was added aqueous HCI (12 M, 0.12 mL, 1.42 mmol) and 3- methylbutyl nitrite (7.16 g, 61.1 mmol) dropwise at 0 °C. The resulting mixture was stirred for 4 h at ambient temperature and the resulting mixture was then concentrated under reduced pressure. The resulting solids were collected by filtration, washed with Et₂O (3 x 10 mL), and dried under reduced pressure to afford 4-bromo-6-chloro-2-(hydroxyimino)-2,3-dihydro-lH- inden-l-one (1 g, 89% yield) as an off-white solid. LCMS: (ESP) m/z = 273.8 (M+H).

Step 2: Preparation of 5-bromo-3,7-dichloroisoquinolin-l(2H)-one

[000224] To a stirred mixture 4-bromo-6-chloro-2-(hydroxyimino)-2,3-dihydro-lH-inden-l- one (1 g, 3.44 mmol) in CH2CI2 (10 mL) under a nitrogen atmosphere was added thionyl chloride (2.05 g, 17.2 mmol) at 0 °C. The resulting mixture was stirred overnight at ambient temperature and then concentrated under reduced pressure. The resulting solids were collected by filtration and washed with PE:EA = 1:1 (3 x 10 mL) and dried under reduced pressure to afford 5-bromo- 3,7-dichloroisoquinolin-l(2H)-one (0.65 g, 61% yield) as a red solid. LCMS: (ESP) m/z = 291.9 (M+H).

Step 3: Preparation of 5-bromo-3,7-dichloro-2-methylisoquinolin-l(2H)-one

[000225] To a stirred mixture of 5-bromo-3,7-dichloroisoquinolin-l(2H)-one (0.65 g, 2.22 mmol) in THF (6 mL) and NMP (2 mL) were added sodium tert-pentoxide (0.50 g, 4.44 mmol) and iodomethane (0.17 mL, 3.33 mmol) at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred for 4 h at ambient temperature and diluted with water (10 mL). The resulting mixture was extracted with EtOAc (3 x 15 mL) and the combined organics were washed with brine (3 x 15 mL), dried over anhydrous Na2SCU, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA = 4:1 to afford 5- bromo-3,7-dichloro-2-methylisoquinolin-l(2H)-one (0.42 g, 62% yield) as a white solid. LCMS: (ES ) m/z = 307.9 (M+H).

Step 4: Preparation of 5-acetyl-3,7-dichloro-2-methylisoquinolin-l(2H)-one

[000226] To a stirred mixture of 5-bromo-3,7-dichloro-2-methylisoquinolin-l(2H)-one (0.42 g, 1.37 mmol) and tributyl(l-ethoxyethenyl)stannane (0.59 g, 1.64 mmol) in NMP (10 mL) were added Pd₂(dba)3»CHCl3 (35 mg, 0.034 mmol) and 3-(tert-nutyl)-4-(2,6-dimethoxyphenyl)-2,3- dihydrobenzo[d][l,3]oxaphosphole (110 mg, 0.342 mmol) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 4 h at 60 °C, cooled to ambient temperature, and then acidified to pH 4 with aqueous 1 N HCI. The resulting mixture was extracted with EtOAc (3 x 20 mL) and the combined organics were washed with brine (3 x 20 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA = 4:1, to afford 5- acetyl-3,7-dichloro-2-methylisoquinolin-l(2H)-one (0.2 g, 54% yield) as a light yellow solid. LCMS: (ESP) m/z = 270.0 (M+H).

[000227] Intermediate 20: (R)-5-(l-aminoethyl)-3,7-dichloro-2-methylisoquinolin-l(2H)-one

[000228] This intermediate was prepared using methods similar to those described in

Intermediate 11, Steps 3-8 and Intermediate 12 using 4-bromo-6-chloro-2,3-dihydro-lH-inden-l- one in place of 4-bromo-6-methyl-2,3-dihydro-lH-inden-l-one. LCMS: (ESP) m/z = 270.9 (M+H). [000229] Intermediate 21: 4-(5-bromopyrimidin-2-yl)-3-chloro-l-methylpyridin-2(lH)-one

Step 1: Preparation of 5-bromo-2-(3-chloro-2-methoxypyridin-4-yl)pyrimidine

[000230] To mixture of 3-chloro-2-methoxypyridin-4-ylboronic acid (1.00 g, 5.34 mmol) and 5- bromo-2-iodopyrimidine (1.52 g, 5.34 mmol) in dioxane (10 mL) and H2O (1 mL) were added Pd(dppf)Ck (780 mg, 1.07 mmol) and K2CO3 (2.21 g, 16.0 mmol) under a nitrogen atmosphere.

The resulting mixture was stirred at 100 °C overnight and then cooled to room temperature. The mixture was extracted with CH2CI2 (3 x 20 mL) and the combined organics were washed with brine (1 x 30 mL), dried over anhydrous Na2SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA = 19:1, to afford 5-bromo-2-(3-chloro-2-methoxypyridin-4-yl)pyrimidine (590 mg, 37% yield) as a white solid. LCMS: (ES ) m/z = 300.0 (M+H).

Step 2: Preparation of 4-(5-bromopyrimidin-2-yl)-3-chloropyridin-2-ol

[000231] A mixture of 5-bromo-2-(3-chloro-2-methoxypyridin-4-yl)pyrimidine (750 mg, 2.50 mmol) and hydrobromic acid (40% in H2O, 10 mL) was stirred at 100 °C for 1 h under a nitrogen atmosphere. The mixture was cooled to room temperature and then diluted with water (15 mL). The precipitated solids were collected by filtration and washed with water (5 x 5 mL). The solids were dried under reduced pressure to afford 4-(5-bromopyrimidin-2-yl)-3-chloropyridin-2-ol (680 mg, 95% yield). LCMS: (ES ) m/z = 285.9 (M+H).

Step 3: Preparation of 4-(5-brornopyrimidin-2-yl)-3-chloro-l-rnethylpyridin-2(lH)-one

[000232] A mixture of 4-(5-bromopyrimidin-2-yl)-3-chloropyridin-2-ol (1.68 g, 5.86 mmol), CS2CO3 (4.78 g, 14.7 mmol), and Mel (0.91 mL, 14.7 mmol) in DMF (17 mL) was stirred at room temperature for 30 min under a nitrogen atmosphere. The resulting mixture was diluted with EtOAc (3 x 20 mL). The combined organics were washed with brine (5 x 20 mL), dried over anhydrous Na2SCU, filtered, and concentrated under reduced pressure. The crude product was slurried with methyl tert-butyl ether (20 mL) for 30 minutes at room temperature and the resulting solids were collected via filtration. The filter cake was washed with cold methyl tertbutyl ether (2 x 5 mL) and then dried to afford 4-(5-bromopyrimidin-2-yl)-3-chloro-l- methylpyridin-2(lH)-one (1.59 g, 90% yield) as a light-yellow solid. LCMS: (ESP) m/z = 299.9 (M+H).

[000233] Example 1: (R)-5-(l-((2-(4H-l,2,4-triazol-4-yl)phenyl)amino)ethyl)-2,7-dimethyl-3-(4-

(2-methylpyrimidin-5-yl)piperazin-l-yl)isoquinolin-l(2H)-one

Step 1: Preparation of 4-(2-iodophenyl)-l,2,4-triazole

[000234] To a stirred solution of 2-iodoaniline (1 g, 4.5 mmol) and diformylhydrazine (1.2 g, 13.6 mmol) in pyridine (10.0 mL) were added chlorotrimethylsilane (7.4 g, 68.4 mmol) and TEA (3.1 g, 31 mmol) in portions at room temperature under an argon atmosphere. The resulting mixture was stirred for 4 h at 100 °C. The reaction was then quenched with ice-water (50 mL) at 0 °C and the resulting mixture was extracted with EtOAc (3 x 50 mL) and the combined organics were dried over anhydrous Na2SC After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA (3:1), to afford 4-(2-iodophenyl)-l,2,4-triazole (620 mg, 50% yield) as an off-white solid. LCMS (ESI) m/z = 271.9 (M+H).

Step 2: Preparation of (R)-5-(l-((2-(4H-l,2,4-triazol-4-yl)phenyl)amino)ethyl)-2,7-dimethyl-3-(4-(2- methylpyrimidin-5-yl)piperazin-l-yl)isoquinolin-l(2H)-one (Example 1)

[000235] To a stirred solution of 4-(2-iodophenyl)-l,2,4-triazole (40 mg, 0.1 mmol) and 5- [(lR)-l-aminoethyl]-2,7-dimethyl-3-[4-(2-methylpyrimidin-5-yl)piperazin-l-yl]isoquinolin-l-one (87 mg, 0.2 mmol) in dioxane (2.0 mL) were added CS2CO3 (144 mg, 0.4 mmol) and {l,3-bis[2,6- bis(pentan-3-yl)phenyl]-4,5-dichloro-2,3-dihydro-lH-imidazol-2-yl}dichlorO (2-methyl-llambda4- pyridin-l-yl)palladium (12 mg, 0.01 mmol) in portions at room temperature under an argon atmosphere. The resulting mixture was stirred overnight at 100 °C. The reaction was quenched with ice-water (20 mL) at 0 °C. The resulting mixture was extracted with EtOAc (3 x 20 mL) and dried over anhydrous Na2SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA (1:1), to afford 30 mg of a crude product. The crude product (30 mg) was purified by prep-HPLC to afford (R)-5-(l-((2-(4H-l,2,4-triazol-4-yl)phenyl)amino)ethyl)-2,7-dimethyl-3-(4-(2- methylpyrimidin-5-yl)piperazin-l-yl)isoquinolin-l(2H)-one (6.5 mg, 8% yield) as a white solid. LCMS (ESI) m/z = 536.2 (M+H); ^TH NMR (400 MHz, DMSO-d₆) 68.76 (s, 2H), 8.47 (s, 2H), 7.87 (s, 1H), 7.52 (d, J = 1.9 Hz, 1H), 7.15- 7.10 (m, 1H), 7.09- 7.06 (m 1H), 6.68- 6.63 (m, 1H), 6.48 - 6.44 (m, 1H), 6.40 (s, 1H), 5.57 (d, J = 6.3 Hz, 1H), 5.03- 4.97 (m, 1H), 3.55 (s, 3H), 3.26-2.99 (m, 8H),2.52 (s, 3H), 2.35 (s, 3H), 1.43 (d, J = 6.6 Hz, 3H).

[000236] Example 2: (R)-2,7-dimethyl-5-(l-((2-(oxazol-2-yl)phenyl)amino)ethyl)-3-(4-(2,2,2- trifluoroethyl)piperazin-l-yl)isoquinolin-l(2H)-one

Step 1: Preparation of (R)-3-chloro-2,7-dimethyl-5-(l-((2-(oxazol-2- yl)phenyl)amino)ethyl)isoquinolin-l(2H)-one [000237] Into a 20 mL vial were added 5-[(lR)-l-aminoethyl]-3-chloro-2,7- dimethylisoquinolin-l-one (200 mg, 0.8 mmol), 2-(2-bromophenyl)-l,3-oxazole (357 mg, 1.6 mmol), CS2CO3 (182 mg, 2.4 mmol), toluene (10 mL), Pd2(dba)s (73 mg, 0.08 mmol), and RuPhos (74 mg, 0.16 mmol) at room temperature. The resulting mixture was stirred overnight at 110 °C under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by prep-TLC (PE:EA = 1:1) to afford (R)-3-chloro-2,7-dimethyl- 5-(l-((2-(oxazol-2-yl)phenyl)amino)ethyl)isoquinolin-l(2H)-one (180 mg, 57% yield) as a yellow solid. MS (ES⁺) m/z = 394.1 [M+H]⁺.

Step 2: Preparation of (R)-2,7-dimethyl-5-(l-((2-(oxazol-2-yl)phenyl)amino)ethyl)-3-(4-(2,2,2- trifluoroethyl)piperazin-l-yl)isoquinolin-l(2H)-one (Example 2)

[000238] Into a 20 mL sealed tube were added (R)-3-chloro-2,7-dimethyl-5-(l-((2-(oxazol-2- yl)phenyl)amino)ethyl)isoquinolin-l(2H)-one (130 mg, 0.33 mmol), l-(2,2,2- trifluoroethyl)piperazine (111 mg, 0.66 mmol), CS2CO3 (323 mg, 0.99 mmol), dioxane (10 mL), Xantphos (38 mg, 0.066 mmol), and Pd2(dba)s (30 mg, 0.033 mmol) at room temperature. The resulting mixture was stirred overnight at 100 °C under a nitrogen atmosphere. The reaction was quenched by the addition of water (20 mL) at room temperature. The resulting mixture was extracted with CH2CI2 (3 x 20 mL). The combined organic layers were washed with brine (2 x 20 mL) and dried over anhydrous Na2SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by prep-TLC (PE:EA = 1:1) to afford crude product. The crude product was purified by prep-HPLC to afford (R)-2,7-dimethyl-5-(l-((2- (oxazol-2-yl)phenyl)amino)ethyl)-3-(4-(2,2,2-trifluoroethyl)piperazin-l-yl)isoquinolin-l(2H)-one (14 mg, 8% yield) as a white solid. ^TH NMR (400 MHz, DMSO-d₆) 6 8.58 (d, J = 6.0 Hz, 1H), 8.22 (d, J = 0.9 Hz, 1H), 7.90 - 7.85 (m, 1H), 7.82 - 7.81 (m, 1H), 7.51 (d, J = 0.9 Hz, 1H), 7.43 (d, J = 1.9 Hz, 1H), 7.13 - 7.11 (m, 1H), 6.68 - 6.60 (m, 1H), 6.46 (d, J = 8.2 Hz, 2H), 5.24 (m, 1H), 3.49 (s, 3H), 3.31 - 3.27 (m, 2H), 3.24 - 2.67 (m, 8H), 2.30 (s, 3H), 1.60 (d, J = 6.6 Hz, 3H). MS (ES⁺) m/z = 526.4 [M+H]⁺.

[000239] Example 3: (R)-5-(l-((6-chloro-2-(l-methyl-lH-l,2,4-triazol-3-yl)pyridin-3- yl)amino)ethyl)-2,7-dimethyl-3-(4-(2,2,2-trifluoroethyl)piperazin-l-yl)isoquinolin-l(2H)-one Step 1: Preparation of (R)-6-chloro-3-((l-(2,7-dimethyl-l-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-l- yl)-l,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinonitrile nitrile

[000240] Into a 20 mL vial were added 6-chloro-3-fluoropyridine-2-carbonitrile (123 mg, 0.78 mmol), (R)-5-(l-aminoethyl)-2,7-dimethyl-3-(4-(2,2,2-trifluoroethyl)piperazin-l-yl)isoquinolin- l(2H)-one (300 mg, 0.78 mmol), DMF (10 mL) and DIEA (203 mg, 1.6 mmol) at room temperature. The resulting mixture was stirred for 2 h at 90 °C. The reaction was quenched by the addition of water (20 mL) at room temperature. The resulting mixture was extracted with EtOAc (2 x 20 mL). The combined organic layers were washed with brine (3 x 50 mL) and dried over anhydrous Na₂SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA (1:1), to afford (R)-6-chloro-3-((l-(2,7-dimethyl-l-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-l-yl)-l,2- dihydroisoquinolin-5-yl)ethyl)amino)picolinonitrile nitrile (330 mg, 81% yield) as a yellow solid. MS (ES⁺) m/z = 519.3 [M+H]⁺.

Step 2: Preparation of (R)-6-chloro-3-((l-(2,7-dimethyl-l-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-l- yl)-l,2-dihydroisoquinolin-5-yl)ethyl)amino)-N'-methylpicolinimidohydrazide

[000241] Into a 20 mL vial were added (R)-6-chloro-3-((l-(2,7-dimethyl-l-oxo-3-(4-(2,2,2- trifluoroethyl)piperazin-l-yl)-l,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinonitrile nitrile (210 mg, 0.4 mmol), methylhydrazine sulfuric acid salt (58 mg, 0.4 mmol), EtOH (10 mL), and DIEA (105 mg, 0.8 mmol) at room temperature. The resulting mixture was stirred overnight at 90 °C under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA (4:1) to afford (R)-6-chloro-3-((l-(2,7-dimethyl-l-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-l-yl)-l,2- dihydroisoquinolin-5-yl)ethyl)amino)-N'-methylpicolinimidohydrazide (130 mg, 57% yield) as a yellow solid. MS: (ES⁺) m/z = 565.2 [M+H]⁺.

Step 3: Preparation of (R)-5-(l-((6-chloro-2-(l-methyl-lH-l,2,4-triazol-3-yl)pyridin-3-yl)amino)ethyl)- 2,7-dimethyl-3-(4-(2,2,2-trifluoroethyl)piperazin-l-yl)isoquinolin-l(2H)-one (Example 3)

[000242] Into a 20 mL vial were added (R)-6-chloro-3-((l-(2,7-dimethyl-l-oxo-3-(4-(2,2,2- trifluoroethyl)piperazin-l-yl)-l,2-dihydroisoquinolin-5-yl)ethyl)amino)-N'- methylpicolinimidohydrazide (130 mg, 0.23 mmol) and HCOOH (5 mL) at room temperature. The resulting mixture was stirred for 2 h at room temperature under a nitrogen atmosphere. The mixture was adjusted to pH 8 with saturated NaHCO₃ aqueous solution. The aqueous layer was extracted with CH2CI2 (3 x 20 mL) and dried over anhydrous Na2SO₄. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA (1:1), to afford the intermediate as a yellow solid. Into a 20 mL vial were added MeOH (5 mL) and the intermediate at room temperature. The resulting mixture was stirred for 2 days at 60 °C under a nitrogen atmosphere. The crude product was purified by prep-achiral-SFC to afford crude product as a yellow solid. The crude product was purified by prep-HPLC to afford (R)-5-(l-((6-chloro-2-(l-methyl-lH-l,2,4-triazol-3-yl)pyridin-3- yl)amino)ethyl)-2,7-dimethyl-3-(4-(2,2,2-trifluoroethyl)piperazin-l-yl)isoquinolin-l(2H)-one (19 mg, 14% yield) as a white solid. ^TH NMR (400 MHz, DMSO-d₆) 6 8.76 (s, 1H), 8.48 (d, J = 5.9 Hz, 1H), 7.88 (s, 1H), 7.42 (s, 1H), 7.15 (d, J = 8.7 Hz, 1H), 6.84 (d, J = 8.8 Hz, 1H), 6.44 (s, 1H), 5.35 - 5.10 (m, 1H), 4.03 (s, 3H), 3.49 (s, 3H), 3.35 - 3.25 (m, 2H), 3.20 - 2.55 (m, 8H), 2.30 (s, 3H), 1.59 (d, J = 6.5 Hz, 3H). MS (ES⁺) m/z = 575.4 [M+H]⁺.

[000243] Example 4: (R)-2,7-dimethyl-5-(l-((2-(5-methyl-l,2,4-oxadiazol-3- yl)phenyl)amino)ethyl)-3-(4-(2,2,2-trifluoroethyl)piperazin-l-yl)isoquinolin-l(2H)-one

Step 1: Preparation of (R)-2-((l-(2,7-dimethyl-l-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-l-yl)-l,2- dihydroisoquinolin-5-yl)ethyl)amino)-N'-hydroxybenzimidamide:

[000244] To a solution of (R)-2-((l-(2,7-dimethyl-l-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-l- yl)-l,2-dihydroisoquinolin-5-yl)ethyl)amino)benzonitrile (190 mg, 0.4 mmol) in EtOH (3 mL) was added Na2CO₃ (83 mg, 0.8 mmol) and hydroxylamine hydrochloride (41 mg, 0.6 mmol). The mixture was stirred at 60 °C for 12 h. The reaction mixture was partitioned between H2O (10 mL) and EtOAc (10 mL). The organic phase was separated, dried over Na2SO₄, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography to give (R)-2-((l-(2,7-dimethyl-l-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-l-yl)- l,2-dihydroisoquinolin-5-yl)ethyl)amino)-N'-hydroxybenzimidamide (150 mg, 71% yield) as a yellow solid. LCMS (ESI) m/z = 539.3 [M+Na],

Step 2: Preparation of (R)-2,7-dimethyl-5-(l-((2-(5-methyl-l,2,4-oxadiazol-3-yl)phenyl)amino)ethyl)- 3-(4-(2,2,2-trifluoroethyl)piperazin-l-yl)isoquinolin-l(2H)-one (Example 4) [000245] To a solution of (R)-2-((l-(2,7-dimethyl-l-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-l- yl)-l,2-dihydroisoquinolin-5-yl)ethyl)amino)-N'-hydroxybenzimidamide (140 mg, 0.27 mmol) in toluene (4 mL) was added AcjO (33 mg, 0.33 mmol) and TEA (73 mg, 0.72 mmol). The mixture was stirred at 120 °C for 2 h. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC to afford (R)-2,7-dimethyl-5-(l-((2-(5- methyl-l,2,4-oxadiazol-3-yl)phenyl)amino)ethyl)-3-(4-(2,2,2-trifluoroethyl)piperazin-l- yl)isoquinolin-l(2H)-one (55 mg, 37% yield) as a yellow solid. LCMS (ESI) m/z = 563.3 [M+Na], ¹H NMR (400 MHz, DMSO-d₆): 5 7.98-7.96 (m, 1H), 7.88 (s, 1H), 7.43 (d, J = 1.6 Hz, 1H), 7.25 - 7.16 (m, 2H), 6.69 (t, J = 7.2 Hz, 1H), 6.54 - 6.43 (m, 2H), 5.33 - 5.20 (m, 1H), 3.49 (s, 3H), 3.30 - 2.92 (m, 10H), 2.71 (s, 3H), 2.33 - 2.28 (m, 3H), 1.60 (d, J = 6.6 Hz, 3H).

[000246] Example 5: (R)-5-(l-((6-chloro-2-(l-methyl-lH-l,2,4-triazol-3-yl)pyridin-3- yl)amino)ethyl)-2,7-dimethyl-3-(4-(2-methylpyrimidin-5-yl)piperazin-l-yl)isoquinolin-l(2H)-one

Step 1: Preparation of (R)-6-chloro-3-((l-(2,7-dimethyl-3-(4-(2-methylpyrimidin-5-yl)piperazin-l-yl)- l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinonitrile

[000247] Into a 10 mL vial were added 6-chloro-3-fluoropyridine-2-carbonitrile (359 mg, 2.3 mmol), (R)-5-(l-aminoethyl)-2,7-dimethyl-3-(4-(2-methylpyrimidin-5-yl)piperazin-l- yl)isoquinolin-l(2H)-one (300 mg, 0.76 mmol), DMF (2 mL) and DIEA (198 mg, 1.53 mmol) at room temperature. The resulting mixture was stirred for 16 hours at 100 °C under a nitrogen atmosphere. The reaction was quenched with water (20 mL) at room temperature. The resulting mixture was extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine (3 x 20 mL) and dried over anhydrous Na2SC The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with CH2CI2 / MeOH (9:1) to afford (R)-6-chloro-3-((l-(2,7-dimethyl-3-(4-(2-methylpyrimidin-5-yl)piperazin-l- yl)-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinonitrile (249 mg, 62% yield) as a yellow solid. MS (ES⁺) m/z = 529.2 [M+H]⁺.

Step 2: Preparation of (R)-6-chloro-3-((l-(2,7-dimethyl-3-(4-(2-methylpyrimidin-5-yl)piperazin-l-yl)- l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)amino)-N'-methylpicolinimidohydrazide [000248] Into a 10 mL vial were added EtOH (2 mL), methylhydrazine sulfuric acid salt (136 mg, 0.95 mmol), DIPEA (183 mg, 1.4 mmol) and (R)-6-chloro-3-((l-(2,7-dimethyl-3-(4-(2- methylpyrimidin-5-yl)piperazin-l-yl)-l-oxo-l,2-dihydroisoquinolin-5- yl)ethyl)amino)picolinonitrile (250 mg, 0.47 mmol) at room temperature. The resulting mixture was stirred for 2 h at 85 °C under a nitrogen atmosphere. The resulting mixture was concentrated under vacuum. The residue was purified by reversed-phase flash chromatography to give (R)-6-chloro-3-((l-(2,7-dimethyl-3-(4-(2-methylpyrimidin-5-yl)piperazin-l-yl)-l-oxo-l,2- dihydroisoquinolin-5-yl)ethyl)amino)-N'-methylpicolinimidohydrazide (114 mg, 42% yield) as a yellow solid. MS (ES⁺) m/z = 575.4 [M+H]⁺.

Step 3: Preparation of (R)-5-(l-((6-chloro-2-(l-methyl-lH-l,2,4-triazol-3-yl)pyridin-3-yl)amino)ethyl)- 2,7-dimethyl-3-(4-(2-methylpyrimidin-5-yl)piperazin-l-yl)isoquinolin-l(2H)-one (Example 5)

[000249] Into a 10 mL vial were added (R)-6-chloro-3-((l-(2,7-dimethyl-3-(4-(2- methylpyrimidin-5-yl)piperazin-l-yl)-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)amino)-N'- methylpicolinimidohydrazide (114 mg, 0.2 mmol), MeOH (1 mL) and FA (1 mL) at room temperature. The resulting mixture was stirred for 24 h at room temperature under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EA (1:1), to afford the intermediate as a yellow solid, to which were added MeOH (3 mL) at room temperature. The resulting mixture was stirred at 70 °C for 3 days under a nitrogen atmosphere. The reaction mixture was concentration, and the residual was purified by prep-achiral SFC to afford (R)-5-(l-((6-chloro-2- (l-methyl-lH-l,2,4-triazol-3-yl)pyridin-3-yl)amino)ethyl)-2,7-dimethyl-3-(4-(2-methylpyrimidin- 5-yl)piperazin-l-yl)isoquinolin-l(2H)-one (36 mg, 45% yield) as a white solid. ¹H NMR (400 MHz, DMSO-dg) 6 8.56 (s, 1H), 8.48 (d, J = 1.0 Hz, 2H), 8.06 - 8.01 (m, 1H), 7.56 (d, J = 1.8 Hz, 1H), 7.08 - 7.05 (m, 1H), 6.86-6.60 (m, 1H), 6.60 (s, 1H), 5.23 - 5.18 (m, 1H), 4.08 (s, 3H), 3.85-3.80 (m, 2H), 3.69 (s, 3H), 3.33 - 3.01 (m, 6H), 2.62 (s, 3H), 2.39 (s, 3H), 1.74 (d, J = 6.8 Hz, 3H). MS (ES⁺) m/z = 585.2 [M+H]⁺.

[000250] Example 20: (R)-4-((l-(2,7-dimethyl-3-(2-(l-methyl-6-oxo-l,6-dihydropyridin-2- yl)pyrimidin-5-yl)-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)amino)pyrazolo[l,5-a]pyridine-3- carbonitrile

Step 1: Preparation of tert-Butyl (R)-(l-(3-chloro-2,7-dimethyl-l-oxo-l,2-dihydroisoquinolin-5- yl)ethyl)carbamate

[000251] A solution of (R)-5-(l-aminoethyl)-3-chloro-2,7-dimethylisoquinolin-l(2H)-one (Intermediate 12, 3 g, 12.0 mmol), DIEA (1.55 g, 12.0 mmol) and (Boc O (3.92 g, 18.0 mmol) in THF (30 mL) was stirred at 25 °C for 0.5 h. The reaction mixture was then concentrated under reduced pressure to give the crude product. The crude product was triturated with PE (50 mL) at 25 °C for 30 min to give tert-butyl (R)-(l-(3-chloro-2,7-dimethyl-l-oxo-l,2-dihydroisoquinolin-5- yl)ethyl)carbamate (3.4 g, 81% yield) as a light-yellow solid. LCMS (ES ) m/z = 351.2 (M+H).

Step 2: Preparation of tert-butyl (R)-(l-(2,7-dimethyl-3-(2-(l-methyl-6-oxo-l,6-dihydropyridin-2- yl)pyrimidin-5-yl)-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)carbamate

[000252] A mixture of (2-(l-methyl-6-oxo-l,6-dihydropyridin-2-yl)pyrimidin-5-yl)boronic acid (Intermediate 14, 590 mg, 1.88 mmol), tert-butyl (R)-(l-(3-chloro-2,7-dimethyl-l-oxo-l,2- dihydroisoquinolin-5-yl)ethyl)carbamate (661 mg, 1.88 mmol), mesylate[(di(l-adamantyl)-n- butylphosphine)-2-(2'-amino-l,l'-biphenyl)]palladium(ll) (137 mg, 0.19 mmol), and K2CO3 (781 mg, 5.65 mmol) in H2O (1 mL) in dioxane (10 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 100 °C for 1 h, then cooled to ambient temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EA = 10:1 to 0:1) to afford tert-butyl (R)-(l-(2,7-dimethyl-3-(2-(l-methyl-6- oxo-1, 6-dihydropyridin-2-yl)pyrimidin-5-yl)-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)carbamate (600 mg, 61% yield) as a yellow solid. LCMS ( ES ) m/z = 502.1 (M+H).

Step 3: Preparation of (R)-5-(l-aminoethyl)-2,7-dimethyl-3-(2-(l-methyl-6-oxo-l,6-dihydropyridin-2- yl)pyrimidin-5-yl)isoquinolin-l(2H)-one

[000253] To a mixture of tert-butyl (R)-(l-(2,7-dimethyl-3-(2-(l-methyl-6-oxo-l,6- dihydropyridin-2-yl)pyrimidin-5-yl)-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)carbamate (800 mg, 1.59 mmol) in DCM (6 mL) was added TFA (3.07 g, 26.9 mmol). The mixture was stirred at 25 °C for 1 h and then concentrated under reduced pressure to give a residue. The residue was adjusted to pH 8 via the addition of a saturated aqueous NaHCO₃ solution (5 mL) and then concentrated under reduced pressure to give a residue. Methanol (1 mL) and DCM (5 mL) was added to the residue, which was then dried over Na2SO₄, filtered, and concentrated under reduced pressure to afford (R)-5-(l-aminoethyl)-2,7-dimethyl-3-(2-(l-methyl-6-oxo-l,6- dihydropyridin-2-yl)pyrimidin-5-yl)isoquinolin-l(2H)-one (700 mg, crude yield) as a brown solid. LCMS (ESP) m/z = 402.2 (M+H). Step 4: Preparation of (R)-4-((l-(2,7-dimethyl-3-(2-(l-methyl-6-oxo-l,6-dihydropyridin-2- yl)pyrimidin-5-yl)-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)amino)pyrazolo[l,5-a]pyridine-3- carbonitrile (Example 20)

[000254] A mixture of 4-bromopyrazolo[l,5-a]pyridine-3-carbonitrile (100 mg, 0.45 mmol), (R)-5-(l-aminoethyl)-2,7-dimethyl-3-(2-(l-methyl-6-oxo-l,6-dihydropyridin-2-yl)pyrimidin-5- yl)isoquinolin-l(2H)-one (150 mg, 0.37 mmol), [l,3-bis[2,6-bis(l-ethylpropyl)phenyl]-4,5- dichloro-imidazol-2-ylidene]-dichloro-(2-methylpyridin-l-ium-l-yl)palladium (31 mg, 0.037 mmol) and CS2CO3 (365 mg, 1.12 mmol) in dioxane (3 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 100 °C for 1 h, then cooled to ambient temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EA =100:1 to 0:1) and then purified further via prep-HPLC (column: Welch Ultimate XB-SiOH 250 x 50 mm x 10 pm; mobile phase: hexanes/EtOH; gradient: 15%-55% B over 15.0 min) to afford (R)-4-((l-(2,7-dimethyl-3-(2-(l-methyl-6-oxo-l,6-dihydropyridin-2- yl)pyrimidin-5-yl)-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl)amino)pyrazolo[l,5-a]pyridine-3- carbonitrile (15 mg, 7% yield) as a yellow solid. LCMS (ES ) m/z = 543.3 (M+H); ¹H NMR (400 MHz, CDCI3): 6 9.05 (s, 2H), 8.27 (s, 1H), 8.18 (s, 1H), 7.98 (d, J = 6.8 Hz, 1H), 7.65 (s, 1H), 7.50 - 7.43 (m, 1H), 6.84 - 6.74 (m, 3H), 6.70 - 6.64 (m, 1H), 5.93 (d, J = 7.6 Hz, 1H), 5.39 (d, J = 4.0 Hz, 1H), 5.09 - 4.94 (m, 1H), 3.73 (s, 3H), 3.55 (s, 3H), 2.49 (s, 3H), 1.75 (d, J = 6.8 Hz, 3H).

[000255] Example 21: (R)-7-chloro-3-(2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4- yl)pyrimidin-5-yl)-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)phenyl)amino)ethyl)-2- methylisoquinolin-l(2H)-one

Step 1: Preparation of 3,7-dichloro-5-(l-hydroxyethyl)-2-methylisoquinolin-l(2H)-one

[000256] To a mixture of 5-acetyl-3,7-dichloro-2-methylisoquinolin-l(2H)-one (Intermediate 19) (16 g, 59.2 mmol) in THF (200 mL) was added NaBH₄ (6.81 g, 180 mmol) at 0 °C. The mixture was stirred at 0 °C for 1 h and then slowly poured into a saturated aqueous NH₄CI solution (100 mL) chilled via an ice bath. The mixture was then diluted further with H₂O (100 mL) and extracted with EA (200 mL x 2). The combined organics were washed with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give a residue. The crude product was triturated with PE:EA = 3:1 (50 mL) at 25 °C for 30 min to afford 3,7-dichloro-5-(l- hydroxyethyl)-2-methylisoquinolin-l(2H)-one (14.8 g, 82% yield) as an off-white solid. LCMS (ES ) m/z = 271.9 (M+H).

Step 2: Preparation of l-(3,7-dichloro-2-methyl-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl methanesulfonate [000257] To a mixture of 3,7-dichloro-5-(l-hydroxyethyl)-2-methylisoquinolin-l(2H)-one (1.2 g, 4.41 mmol) and TEA (3.57 g, 35.3 mmol) in DCM (15 mL) was added methanesulfonic anhydride (3.84 g, 22.1 mmol) at 0 °C. The mixture was stirred at 25 °C for 1 h. The reaction mixture containing l-(3,7-dichloro-2-methyl-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl methanesulfonate (1.54 g, expected crude yield) was used as is in the next step without further purification. LCMS (ES ) m/z = 350.2 (M+H).

Step 3: Preparation of 3,7-dichloro-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4- yl)phenyl)amino)ethyl)-2-methylisoquinolin-l(2H)-one

[000258] A mixture of 4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)aniline (845 mg, 4.39 mmol), l-(3,7-dichloro-2-methyl-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl methanesulfonate (1.54 g, 4.40 mmol), DIEA (2.84 g, 22.0 mmol) in DCM (20 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 40 °C for 12 h and then concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE:EA = 75:25) to afford 3,7-dichloro-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)phenyl)amino)ethyl)-2- methylisoquinolin-l(2H)-one (1.3 g, 59% yield) as a yellow solid. LCMS (ESP) m/z = 446.1 (M+H).

Step 4: Preparation of 7-chloro-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4- yl)phenyl)amino)ethyl)-2-methyl-3-(tributylstannyl)isoquinolin-l(2H)-one [000259] A mixture of 3,7-dichloro-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4- yl)phenyl)amino)ethyl)-2-methylisoquinolin-l(2H)-one (400 mg, 0.90 mmol), tributyl(tributylstannyl)stannane (2.60 g, 4.48 mmol), Pd₂(dba)s (82 mg, 0.090 mmol), tricyclohexylphosphane (50 mg, 0.18 mmol) in dioxane (5 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 120 °C for 6 h and then was cooled to ambient temperature, to which a saturated aqueous potassium fluoride solution (5 mL) was added. The resulting mixture was filtered, and the filtrate was diluted further with water (5 mL). The aqueous mixture was extracted with ethyl acetate (10 mL x 3). The combined organics were washed with brine (10 mL), dried with anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (PE:EA = 80:20) to afford 7-chloro-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)phenyl)amino)ethyl)-2- methyl-3-(tributylstannyl)isoquinolin-l(2H)-one (190 mg, 25% yield) as a yellow oil. LCMS (ES ) m/z = 701.9 (M+H).

Step 5: Preparation of 7-chloro-3-(2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)pyrimidin-5- yl)-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)phenyl)amino)ethyl)-2-methylisoquinolin- l(2H)-one

[000260] A mixture of 7-chloro-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4- yl)phenyl)amino)ethyl)-2-methyl-3-(tributylstannyl)isoquinolin-l(2H)-one (230 mg, 0.328 mmol), 4-(5-bromopyrimidin-2-yl)-3-chloro-l-methylpyridin-2(lH)-one (Intermediate 21) (118 mg, 0.393 mmol), Cui (125 mg, 0.654 mmol), and Pd(PPha)₄ (189 mg, 0.164 mmol) in ACN (5 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 80 °C for 0.5 h and then was cooled to ambient temperature, to which a saturated aqueous potassium fluoride solution (1 mL) was added. The resulting mixture was filtered, and the filtrate was diluted further with water (2 mL). The aqueous mixture was extracted with DCM (2 mL x 3). The combined organics were washed with brine (2 mL), dried with anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was purified by prep-TLC (silica gel, PE:EA = 0:1) and purified further by prep-TLC (silica gel, EA:MEOH = 50:1) to afford 7-chloro-3-(2- (3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)pyrimidin-5-yl)-5-(l-((4-fluoro-2-(l-methyl-lH- l,2,3-triazol-4-yl)phenyl)amino)ethyl)-2-methylisoquinolin-l(2H)-one (40 mg, 18% yield) as a yellow solid. LCMS (ES ) m/z = 631.1 (M+H).

Step 6: Preparation of (R)-7-chloro-3-(2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)pyrimidin- 5-yl)-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)phenyl)amino)ethyl)-2-methylisoquinolin- l(2H)-one (Example 21) and (S)-7-chloro-3-(2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4- yl)pyrimidin-5-yl)-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)phenyl)amino)ethyl)-2- methylisoquinolin-l(2H)-one

[000261] A racemic mixture of 7-chloro-3-(2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4- yl)pyrimidin-5-yl)-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)phenyl)amino)ethyl)-2- methylisoquinolin-l(2H)-one was separated by chiral SFC (Daicel CHIRALPAK AD (250 mm x 30 mm x 10 pm); mobile phase: CO2-EtOH:ACN = 4:1 (0.1% NH₄OH); B%: 45%, isocratic elution mode) to afford (R)-7-chloro-3-(2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)pyrimidin-5- yl)-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)phenyl)amino)ethyl)-2-methylisoquinolin- l(2H)-one (Example 21, 14.3 mg, 35% yield) and (S)-7-chloro-3-(2-(3-chloro-l-methyl-2-oxo-l,2- dihydropyridin-4-yl)pyrimidin-5-yl)-5-(l-((4-fluoro-2-(l-methyl-lH-l,2,3-triazol-4- yl)phenyl)amino)ethyl)-2-methylisoquinolin-l(2H)-one (13.4 mg, 32% yield) each as a yellow solid.

(R)-7-chloro-3-(2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)pyrimidin-5-yl)-5-(l-((4- fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)phenyl)amino)ethyl)-2-methylisoquinolin-l(2H)-one (Example 21): ^TH NMR (400 MHz, CDCI₃): 6 8.97 (s, 2H), 8.38 (d, J = 2.0 Hz, 1H), 7.83 - 7.75 (m, 2H), 7.41 (d, J = 7.2 Hz, 1H), 7.25 - 7.22 (m, 1H), 7.12-7.06 (m, 1H), 6.85 (s, 1H), 6.75 - 6.70 (m, 1H), 6.67 (d, J = 7.2 Hz, 1H), 6.41 - 6.31 (m, 1H), 5.03 - 4.98 (m, 1H), 4.21 (s, 3H), 3.72 (s, 3H), 3.53 (s, 3H), 1.75 (br d, J = 6.8 Hz, 3H); LCMS (ESP) m/z = 631.1 (M+H). (S)-7-chloro-3-(2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)pyrimidin-5-yl)-5-(l-((4- fluoro-2-(l-methyl-lH-l,2,3-triazol-4-yl)phenyl)amino)ethyl)-2-methylisoquinolin-l(2H)-one: ¹H NMR (400 MHz, CDCI₃): 6 8.99 (s, 2H), 8.41 (d, J = 2.0 Hz, 1H), 7.85 - 7.77 (m, 2H), 7.44 (d, J = 7.2 Hz, 1H), 7.27 (br s, 1H), 7.14-7.07 (m, 1H), 6.89 - 6.77 (m, 2H), 6.70 (d, J = 7.2 Hz, 1H), 6.58 - 6.39 (m, 1H), 5.12 - 4.99 (m, 1H), 4.24 (s, 3H), 3.75 (s, 3H), 3.55 (s, 3H), 1.80 (d, J = 6.8 Hz, 3H); LCMS (ES ) m/z = 631.1 (M+H).

[000262] Example 22: (R)-2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-(7-chloro-5-

(l-(cinnolin-8-ylamino)ethyl)-2-methyl-l-oxo-l,2-dihydroisoquinolin-3-yl)pyrimidine 1-oxide

Step 1: Preparation of (R)-3,7-dichloro-5-(l-(cinnolin-8-ylamino)ethyl)-2-methylisoquinolin-l(2H)- one

[000263] A mixture of (R)-5-(l-aminoethyl)-3,7-dichloro-2-methylisoquinolin-l(2H)-one (Intermediate 20) (519 mg, 1.91 mmol), 8-bromocinnoline (1 g, 4.78 mmol), Pd?(dba)3 (175 mg, 0.191 mmol), Xantphos (111 mg, 0.191 mmol) and CS2CO3 (1.87 g, 5.74 mmol) in dioxane (10 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 100 °C for 12 h and then cooled to ambient temperature and concentrated under reduced pressure to afford a residue. The crude product was purified by silica gel chromatography (PE:EA = 64:36) and then purified further by prep-HPLC (Phenomenex luna C18 250 x 50 mm xlO pm; mobile phase: H2O(0.225% FA)-ACN; gradient: 35%-65% B over 22.0 min) to afford (R)-3,7-dichloro-5-(l- (cinnolin-8-ylamino)ethyl)-2-methylisoquinolin-l(2H)-one (1.34 g, 87% yield) as a yellow solid. LCMS (ESP) m/z = 399.0 (M+H).

Step 2: Preparation of (R)-7-chloro-5-(l-(cinnolin-8-ylamino)ethyl)-2-methyl-3- (tributylstannyl)isoquinolin-l(2H)-one

[000264] A mixture of (R)-3,7-dichloro-5-(l-(cinnolin-8-ylamino)ethyl)-2-methylisoquinolin- l(2H)-one (1.2 g, 3.01 mmol), tributyl(tributylstannyl)stannane (8.72 g, 15.0 mmol), Pd2(dba)s (275 mg, 0.300 mmol), LiCI (764 mg, 18.0 mmol), and tricyclohexylphosphane (169 mg, 0.601 mmol) in dioxane (10 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 120 °C for 5 h, cooled to ambient temperature, and treated with a saturated aqueous potassium fluoride solution (10 mL). The resulting mixture was filtered and the filtrate was further diluted with water (10 mL). The aqueous mixture was extracted with ethyl acetate (10 mL x 3). The combined organics were washed with brine (10 mL x 3), dried with anhydrous Na2SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (PE:EA = 82:18) to afford (R)-7-chloro-5-(l-(cinnolin-8-ylamino)ethyl)-2-methyl- 3-(tributylstannyl)isoquinolin-l(2H)-one (760 mg, 38% yield) as a yellow oil. LCMS (ES ) m/z = 655.1 (M+H).

Step 3: Preparation of (R)-2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-(7-chloro-5-(l-

(cinnolin-8-ylamino)ethyl)-2-methyl-l-oxo-l,2-dihydroisoquinolin-3-yl)pyrimidine 1-oxide

(Example 22)

[000265] A mixture of 5-bromo-2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4- yl)pyrimidine 1-oxide (Intermediate 16) (116 mg, 0.367 mmol), (R)-7-chloro-5-(l-(cinnolin-8- ylamino)ethyl)-2-methyl-3-(tributylstannyl)isoquinolin-l(2H)-one (200 mg, 0.305 mmol), Cui (117 mg, 0.611 mmol), Pd(PPh3)₄ (177 mg, 0.152 mmol) in ACN (3 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 80 °C for 0.5 h, then cooled to room temperature and treated with a saturated aqueous potassium fluoride solution (2 mL). The reaction mixture was filtered and the filtrate was further diluted with water (2 mL). The aqueous mixture was extracted with DCM (2 mL x 3). The combined organics were washed with brine (2 mL x 3), dried with anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was purified by reversed-phase HPLC (Waters Xbridge C18 150 x 25 mm x 5 pm; mobile phase: H₂0(0.05% NH₄OH)-ACN; gradient: 26%-56% B over 11.0 min ) to afford (R)-2-(3- chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-(7-chloro-5-(l-(cinnolin-8-ylamino)ethyl)-2- methyl-l-oxo-l,2-dihydroisoquinolin-3-yl)pyrimidine 1-oxide (19.9 mg, 11% yield) as a yellow solid. ^TH NMR (400 MHz, CDCI₃): 6 9.28 (br d, J = 5.6 Hz, 1H), 8.64 (d, J = 1.6 Hz, 1H), 8.54 - 8.40 (m, 2H), 8.03 - 7.80 (m, 2H), 7.52 - 7.44 (m, 2H), 7.36 - 7.32 (m, 1H), 7.10 (br d, J = 8.4 Hz, 1H), 6.89 (s, 1H), 6.42 (d, J = 7.2 Hz, 1H), 6.36 - 6.26 (m, 1H), 5.24 - 5.11 (m, 1H), 3.74 (s, 3H), 3.63 (s, 3H), 1.85 (d, J = 6.4 Hz, 3H); LCMS (ESP) m/z = 600.1 (M+H).

[000266] Example 23: (R)-2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-(7-chloro-5-

(l-((4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4-yl)phenyl)amino)ethyl)-2-methyl-l-oxo-l,2- dihydroisoquinolin-3-yl)pyrimidine 1-oxide

Step 1: Preparation of 3,7-dichloro-5-(l-((4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4- yl)phenyl)amino)ethyl)-2-methylisoquinolin-l(2H)-one

[000267] To a mixture of l-(3,7-dichloro-2-methyl-l-oxo-l,2-dihydroisoquinolin-5-yl)ethyl methanesulfonate (Example 21, Step 2) (1.56 g, 4.45 mmol) and 4-fluoro-2-(3-fluoro-l-methyl- lH-pyrazol-4-yl)aniline (Intermediate 17) (932 mg, 4.45 mmol) in DCM (20 mL) was added DIEA (2.88 g, 22.3 mmol) at 25 °C. The mixture was then stirred for 12 h at 40 °C and then concentrated to give a residue. The residue was purified by Prep-HPLC (column: Welch Ultimate XB-CN 250 x 50 mm x 10 pm; mobile phase: hexanes-EtOH; gradient: l%-40% B over 15.0 min) to afford 3,7-dichloro-5-(l-((4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4-yl)phenyl)amino)ethyl)-2- methylisoquinolin-l(2H)-one (800 mg, 36% yield) as a white solid. LCMS (ES ) m/z = 485.0 (M+Na).

Step 2: Preparation of 7-chloro-5-(l-((4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4- yl)phenyl)amino)ethyl)-2-methyl-3-(tributylstannyl)isoquinolin-l(2H)-one

[000268] To a mixture of 3,7-dichloro-5-(l-((4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4- yl)phenyl)amino)ethyl)-2-methylisoquinolin-l(2H)-one (550 mg, 1.19 mmol) and tributyl(tributylstannyl)stannane (5.51 g, 9.50 mmol) in dioxane (6 mL) was added Pdjdbas (109 mg, 0.119 mmol) and tricyclohexylphosphane (67 mg, 0.237 mmol) at 25 °C under a nitrogen atmosphere. The mixture was heated to 120 °C for 5 h and then cooled to room temperature and treated with a saturated aqueous potassium fluoride solution (20 mL). The resulting mixture was filtered and the filtrate was further diluted with H2O (20 mL) and EA (20 mL). The mixture was then stirred at 20 °C for 1 h, then filtered and the filtrate mixture was extracted with EA (20 mL x 3). The combined organics were washed with brine (20 mL), dried over NaSO₄, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by silica gel chromatography (PE:EA = 5:1 to 1:1) to afford 7-chloro-5-(l-((4-fluoro-2-(3-fluoro-l-methyl-lH- pyrazol-4-yl)phenyl)amino)ethyl)-2-methyl-3-(tributylstannyl)isoquinolin-l(2H)-one (260 mg, 28% yield) as a yellow oil. LCMS (ESP) m/z = 719.0 (M+H).

Step 3: Preparation of 2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-(7-chloro-5-(l-((4- fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4-yl)phenyl)amino)ethyl)-2-methyl-l-oxo-l,2- dihydroisoquinolin-3-yl)pyrimidine 1-oxide

[000269] To a mixture of 7-chloro-5-(l-((4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4- yl)phenyl)amino)ethyl)-2-methyl-3-(tributylstannyl)isoquinolin-l(2H)-one (250 mg, 0.348 mmol) and Pd(PPh3)4 (121 mg, 0.104 mmol), Cui (133 mg, 0.696 mmol) in MeCN (3 mL) was added 5- bromo-2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)pyrimidine 1-oxide (Intermediate 16) (132 mg, 0.418 mmol) at 25 °C under a nitrogen atmosphere. The mixture was then heated to 80 °C for 10 min and then cooled to room temperature and treated with a saturated aqueous potassium fluoride solution (10 mL). The resulting mixture was extracted with DCM (20 mL x 3). The combined organics were washed with brine (10 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by Prep-HPLC (column: Phenomenex luna C18 150 x 25 mm x 10 pm; mobile phase: H₂O(0.225% FA)-ACN; gradient: 40%-70% B over 10.0 min) to afford 2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-(7- chloro-5-(l-((4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4-yl)phenyl)amino)ethyl)-2-methyl-l-oxo- l,2-dihydroisoquinolin-3-yl)pyrimidine 1-oxide (105 mg, 45% yield) as a white solid. LCMS (ES ) m/z = 664.1 (M+H).

Step 4: Preparation of (R)-2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-(7-chloro-5-(l-((4- fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4-yl)phenyl)amino)ethyl)-2-methyl-l-oxo-l,2- dihydroisoquinolin-3-yl)pyrimidine 1-oxide (Example 23) and (S)-2-(3-chloro-l-methyl-2-oxo-l,2- dihydropyridin-4-yl)-5-(7-chloro-5-(l-((4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4- yl)phenyl)amino)ethyl)-2-rnethyl-l-oxo-l,2-dihydroisoquinolin-3-yl)pyrimidine 1-oxide [000270] A racemic mixture of 2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-(7- chloro-5-(l-((4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4-yl)phenyl)amino)ethyl)-2-methyl-l-oxo- l,2-dihydroisoquinolin-3-yl)pyrimidine 1-oxide (105 mg) was separated by chiral SFC (column: Daicel CHIRALPAK AD (250 mm x 30 mm x 10 pm); mobile phase: CO₂-EtOH:ACN = 4:1 (0.1% NH₄OH); B%: 45%, isocratic elution mode) to afford (R)-2-(3-chloro-l-methyl-2-oxo-l,2- dihydropyridin-4-yl)-5-(7-chloro-5-(l-((4-fluoro-2-(3-fluoro-l-methyl-lH-pyrazol-4- yl)phenyl)amino)ethyl)-2-methyl-l-oxo-l,2-dihydroisoquinolin-3-yl)pyrimidine 1-oxide (Example 23) and (S)-2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-(7-chloro-5-(l-((4-fluoro-2-(3- fluoro-l-methyl-lH-pyrazol-4-yl)phenyl)amino)ethyl)-2-methyl-l-oxo-l,2-dihydroisoquinolin-3- yl)pyrimidine 1-oxide.

(R)-2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-(7-chloro-5-(l-((4-fluoro-2-(3-fluoro-l- methyl-lH-pyrazol-4-yl)phenyl)amino)ethyl)-2-methyl-l-oxo-l,2-dihydroisoquinolin-3- yl)pyrimidine 1-oxide (Example 23): ^TH NMR (400 MHz, CDCI3): 68.62 (d, J = 1.6 Hz, 1H), 8.45 (d, J = 2.0 Hz, 1H), 8.40 (d, J = 2.0 Hz, 1H), 7.79 (d, J = 2.4 Hz, 1H), 7.45-7.4 (m, 2H), 6.92 - 6.87 (m, 1H), 6.81 (s, 1H), 6.75 - 6.70 (m, 1H), 6.38 (d, J = 7.2 Hz, 1H), 6.17 - 6.04 (m, 1H), 4.87 - 4.76 (m, 1H), 3.89 (s, 3H), 3.70 (s, 3H), 3.58 (s, 3H), 1.56 - 1.55 (s, 3H); LCMS (ES ) m/z = 664.1 (M+H).

(S)-2-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-(7-chloro-5-(l-((4-fluoro-2-(3-fluoro-l- methyl-lH-pyrazol-4-yl)phenyl)amino)ethyl)-2-methyl-l-oxo-l,2-dihydroisoquinolin-3- yl)pyrimidine 1-oxide: ^XH NMR (400 MHz, CDCI3): 68.63 (d, J = 1.6 Hz, 1H), 8.46 (d, J = 2.0 Hz, 1H), 8.41 (d, J = 2.0 Hz, 1H), 7.80 (d, J = 2.0 Hz, 1H), 7.48 - 7.42 (m, 2H), 6.95 - 6.88 (m, 1H), 6.83 (s, 1H), 6.79 - 6.69 (m, 1H), 6.39 (d, J = 7.2 Hz, 1H), 6.18 - 6.01 (m, 1H), 4.91 - 4.79 (m, 1H), 3.91 (s, 3H), 3.72 (s, 3H), 3.60 (s, 3H), 1.56 (s, 3H); LCMS (ESP) m/z = 664.1 (M+H).

[000271] Example 24: (R)-7-chloro-3-(l-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4-yl)-6- oxo-l,6-dihydropyridazin-4-yl)-5-(l-(cinnolin-8-ylamino)ethyl)-2-methylisoquinolin-l(2H)-one [000272] A mixture of (R)-7-chloro-5-(l-(cinnolin-8-ylamino)ethyl)-2-methyl-3- (tributylstannyl)isoquinolin-l(2H)-one (Example 22, Step 2) (67 mg, 0.102 mmol), 2-(3-chloro-l- methyl-2-oxo-l,2-dihydropyridin-4-yl)-5-iodopyridazin-3(2H)-one (Intermediate 18) (44.70 mg, 0.123 mmol), Cui (39 mg, 0.205 mmol), Pd(PPh3)₄ (59 mg, 0.051 mmol) in ACN (2 mL) was degassed and purged with nitrogen three times. The mixture was stirred at 80 °C for 0.5 h, cooled to room temperature, and then treated with a saturated aqueous potassium fluoride solution (2 mL). The resulting mixture was filtered, the filtrate was diluted further with water (2 mL) and the aqueous phase was extracted with DCM (2 mL x 3). The combined organics were washed with brine (2 mL x 3), dried with anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by prep-TLC (silica gel, EA), followed by reversed- phase HPLC(Waters Xbridge C18 150 x 25mm x 5 pm; mobile phase: H₂0(0.05% NH₄OH)-ACN; gradient: 23%-53% ACN over 12.0 min), and then further purified by prep-TLC (silica gel, DCM:MeOH = 20:1) to afford (R)-7-chloro-3-(l-(3-chloro-l-methyl-2-oxo-l,2-dihydropyridin-4- yl)-6-oxo-l,6-dihydropyridazin-4-yl)-5-(l-(cinnolin-8-ylamino)ethyl)-2-methylisoquinolin-l(2H)- one (Example 24) (10 mg, 16% yield) as a yellow solid. ¹H NMR (400 MHz, CDCh): 69.25 (d, J = 6.0 Hz, 1H), 8.41 (d, J = 2.0 Hz, 1H), 8.00 (d, J = 2.0 Hz, 1H), 7.88 (d, J = 4.0 Hz, 1H), 7.84 (d, J = 2.0 Hz, 1H), 7.48 - 7.42 (m, 2H), 7.32 - 7.29 (m, 1H), 7.15 (d, J = 2.0 Hz, 1H), 7.06 (d, J = 8.4 Hz, 1H), 6.86 (s, 1H), 6.37 (d, J = 7.2 Hz, 1H), 6.27 (d, J = 7.2 Hz, 1H), 5.17 - 5.15 (m, 1H), 3.70 (s, 3H), 3.61 (s, 3H), 1.82 (d, J = 6.8 Hz, 3H); LCMS (ESP) m/z = 600.1 (M+H).

Examples 6 to 19 and 25 to 57

[000273] Examples 6 to 19 and 25 to 57 were prepared in a manner similar to that described for Examples 1 to 5 and 20 to 24 and are identified and characterized in Table 1 below. If not specified otherwise, all depicted chiral centers exist as a (R)- and (S)-racemic mixture or as either (R)- or (S)- enantiomer.

Table 1. Examples 6 to 19 and 25 to 57

Table 2. NMR and LCMS data for Examples 6 to 19 and 25 to 57

Assays and Compound Testing

[000274] In vitro cell proliferation: determination of EC50 values for inhibition of proliferation in T-47D cells expressing mutant PI3Ka (H1047R) mutation and SK-BR-3 cells express WT PI3Ka.

[000275] T-47D or SK-BR-3 cells were trypsinized, resuspended in culture media and seeded onto assay ready plates. T47D culture media consisted of RPMI, 10% FBS and Insulin (0.2 units/mL). SK-BR-3 culture media consisted of McCoys 5a and 10% FBS. Cells were seeded at a density of 1,500 cells/well and dispensed in 50 pL onto 384 well assay ready plates (Corning, 89089-790). Assay ready plates had previously been stamped with 10-point dilutions of compounds of interest, as well as controls. The Echo655 is used to stamp plates at 40 nL of compound or DMSO. Cells were grown for 72 hours at 37 ° Celsius and 5% CO2. After 72 hours, cells were equilibrated at room temperature for 15 minutes. 30 uL of CellTiter-Glo reagent is added to the plate, which is then shaken for 30 minutes at temperature at 300-500 rpm. Cells are then read on an Envision plate reader. The percentage of inhibition of proliferation was calculated using the following formula: %l nhibition = 100 x (Lum_D - Lumsampie) / (Lum_D -Lummh), where D is obtained from cells treated with 0.1% DMSO only; Inh is obtained from cells treated with 10 uM Alpelisib. The effective concentration achieving 50% inhibition of proliferation (EC50) is calculated by fitting the Curve using Xlfit (v5.3.1.3), equation 201: Y = Bottom + (Top - Bottom)/(1 + 10^A((LogEC50 - X)*HillSlope)).

Reagent table: [000276] In vitro cell pAKT: determination of IC50 values for inhibition of phosphorylation of AKT (pAKT) in the following cell lines: SK-BR-3 cells expressing WT PI3Ka, T-47D cells expressing mutant PI3Ka (H1047R), MCF-7 cells expressing mutant PI3Ka (E545K), BT-483 cells expressing mutant PI3Ka (E542K), EFM-19 cells expressing mutant PI3Ka (H1047L), and MFE-280 cells expressing mutant PI3Ka (H1047Y).

[000277] Cells, grown in culture media (specified below), were harvested, resuspended in assay media (specified below), and seeded onto assay ready plates. Cells were seeded at a density as outlined below and dispensed in 12.5 pL onto 384-well assay ready plates (Perkin Elmer, 6008238). Assay ready plates had previously been stamped with 10-point dilutions of compounds of interest, as well as controls. The Echo655 was used to stamp plates at 12.5 nL of compound or DMSO. Cells, in assay ready plates, were grown for 6 hours at 37°C and 5% CO₂. After 6 hours, 4 pL of lysis buffer reagent were added to the plate, which was then centrifuged for 1 minute at 1000 rpm. Then the plate was incubated at room temperature for 30 minutes. After 30 minutes, 4 pL of antibody mix containing Eu cryptate, d2 cryptate, and detection buffer were added to the plate. The plate was centrifuged for 1 minute at 1000 rpm and then incubated overnight at room temperature with lid, protected from light. The plate was read on an BMG-PHERAstar FSX plate reader using the HTRF protocol. The percentage of inhibition of AKT phosphorylation was calculated using the following formula: % Inhibition = 100 x (pAKTHC - pAKTSample) / (pAKTHC -pAKTLC), where pAKTHC is obtained from cells treated with 0.2% DMSO only, pAKTLC is obtained from cells treated with 10 pM alpelisib, and pAKTSample referring to the well for which % inhibition is being calculated. The IC50 (concentration achieving 50% inhibition of pAKT) is calculated by fitting the curve using Xlfit (v5.3.1.3), equation 201: Y = Bottom + (Top - Bottom)/(1 + 10^A((LoglC50 - X)*HillSlope)).

[000278] SK-BR-3 culture media consisted of McCoy's 5a and 10% FBS, and assay media consisted of DMEM (no phenol red) + 10% FBS. T-47D culture media consisted of RPMI, 10% FBS and insulin (0.2 U/mL), and assay media consisted of RPMI 1640 (no phenol red) + 10% FBS + 0.2 U/ml insulin. MCF-7 culture media consisted of EMEM + 10% FBS, and assay media consisted of DMEM (no phenol red) + 10% FBS. BT-483 culture media consisted of RPMI 1640 + 20% FBS + 10 pg/ml insulin, and assay media consisted of DMEM (no phenol red) + 10% FBS. EFM-19 culture media consisted of RPMI 1640 + 10% FBS, and assay media consisted of DMEM (no phenol red) + 10% FBS. MFE-280 culture media consisted of MEM + 10% FBS, and assay media consisted of DMEM (no phenol red) + 10% FBS. [000279] Cell density for dispensing into assay ready plates as follows: SK-BR-3 at 5,000 cells/well, T-47D at 5,000 cells/well, MCF-7 at 5,000 cells/well, BT-483 at 5,000 cells/well, EFM- 19 at 5,000 cells/well, MFE-280 at 10,000 cells/well.

Reagent table:

[000280] For EC50 values shown in Table 3, "A" refers to EC50 < 10 nM; "B" refers to 10 nM <

EC50 < 100 nM; "C" refers to 100 nM < EC50 < 1 pM; "D" refers to 1 pM < EC50 < 10 pM; "E" refers to EC50 > 10 pM.

Table 3. Cellular proliferation data

[000281] For IC50 values shown in Table 4, "A" refers to IC50 < 10 nM; "B" refers to 10 nM < IC50 < 100 nM; "C" refers to 100 nM < IC50 < 1 pM; "D" refers to 1 ptM < IC50 < 10 pM; "E" refers to IC50 > 10 pM.

Table 4. Cellular pAKT data References

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Claims

IN THE CLAIMS

1. A compound of Formula (1) or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein:

Ri is heteroaryl, aryl, heterocyclyl or cycloalkyl, where each of the heteroaryl, aryl, heterocyclyl and cycloalkyl is substituted or unsubstituted; each of Xi, X2, X3 and X₄ is independently N, CH or substituted C (where C may be substituted with, but not limited to, C1-C4 alkyl, C3-C7 cycloalkyl, halogen, CN, CF3, OCF3, CFH? or CF?H, where the C1-C4 alkyl and C3-C7 cycloalkyl are unsubstituted or substituted), with the proviso that X₄ cannot be a carbon atom substituted with a carboxylic acid or ester thereof;

R2 is H, C1-C4 alkyl, C3-C7 cycloalkyl, CF3, CFH2 or CF2H, where the C1-C4 alkyl and C3-C7 cycloalkyl is unsubstituted or substituted, and where R2 is not H, the carbon atom attached to R2 is a chiral center and exists as a (R)- and (S)-racemic mixture or as either the (R)- or (S)- enantiomer;

R3 is H or C1-C4 alkyl, where the C1-C4 alkyl is unsubstituted or substituted;

R₅ is H, C1-C4 alkyl, C3-C7 cycloalkyl, heteroaryl, CF3, CH2F or CF2H, where each of the C1-C4 alkyl, C3-C7 cycloalkyl and heteroaryl is unsubstituted or substituted; Rs is H, halogen, C1-C4 alkyl, C3-C7 cycloalkyl, CN, CF3, OCF3, CH?F or CF?H, where the C1-C4 alkyl and C3-C7 cycloalkyl is unsubstituted or substituted; each R₇ is independently H, halogen, C1-C4 alkyl, C3-C7 cycloalkyl, CN, CF3, OCF3, CH?F or CF?H, where the C1-C4 alkyl and C3-C7 cycloalkyl is unsubstituted or substituted;

Rs is H, C1-C4 alkyl, C3-C7 cycloalkyl, halogen, CN, CF3, OCF3, CFH? or CF?H, where the C1-C4 alkyl and C3-C7 cycloalkyl is unsubstituted or substituted; and

R₄ is heteroaryl, C3-C7 cycloalkyl or heterocyclyl, where the heterocyclic is optionally part of a bridged, fused or spiro ring system, and where each of the heteroaryl, C3-C7 cycloalkyl, and heterocyclyl is unsubstituted or substituted.

2. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R₄ is unsubstituted or unsubstituted heteroaryl.

3. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R₄ is unsubstituted or unsubstituted C3-C7 cycloalkyl.

4. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R₄ is unsubstituted or unsubstituted heterocyclyl.

5. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R₄ is selected from wherein the above rings may be further substituted and Rg is selected from H, halogen, C1-C4 alkyl, C3-C7 cycloalkyl, aryl, heteroaryl, heterocyclyl, CN, CF3, OCF3, CFH? or CF?H, wherein each of the C1-C4 alkyl, C3-C7 cycloalkyl, aryl, heteroaryl and heterocyclyl is unsubstituted or substituted.

6. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2- azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane.

7. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7-azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4-thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4-b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or triazolo-pyridine.

8. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R₂ is CH₃ or CH₂F.

9. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R₃ is H.

10. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R₅ is CH₃.

11. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R₆ is CH₃.

12. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein each R₇ is independently H or F.

13. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Rg is H or F.

14. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Rj, Rs and Rg are CH₃.

15. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Rj is CHg or CHjF; and R₃ is H.

16. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R2 is CH₃ or CH2F; R₃ is H; and each R₇ is independently H or F.

17. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R2 is CH₃ or CH2F; R₃ is H; each R₇ is independently H or F; and R₅ is CH₃.

18. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R2 is CH₃ or CH2F; R₃ is H; and R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7- azaspiro[4.4]nonane, 2-azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane.

19. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7-azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4-thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4-b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or triazolo-pyridine; R₃ is H; and R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa- 7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2-azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane.

20. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7-azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4-thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4-b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or triazolo-pyridine; R₂ is CH3 or CH₂F; R3 is H; and R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2- azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane.

21. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7-azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4-thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4-b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or triazolo-pyridine; R₂ is CH3 or CH₂F; R3 is H; R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2- azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane; and each R₇ is independently H or F.

22. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7-azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4-thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4-b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or triazolo-pyridine; R2 is CH3 or CH?F; R3 is H; R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2- azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane; each R₇ is independently H or F; and R₅ is CH₃.

23. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7-azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4-thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4-b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or triazolo-pyridine; R2 is CH3 or CH2F; R3 is H; R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2- azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane; R₇ is H; R₅ is CH3; and Rg is CH3.

24. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Ri is heteroaryl, where the heteroaryl is unsubstituted or substituted; R₂ is CH3 or CH2F; R3 is H; R₄ is heterocyclyl, where the heterocyclyl is unsubstituted or substituted; each R₇ is independently H or F; R₅ is CH3; and Rg is CH3.

25. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Ri is heteroaryl, where the heteroaryl is unsubstituted or substituted; R2 is CH3 or CH2F; R3 is H; R₄ is C3- C₇ cycloalkyl, where the Cs-C₇ cycloalkyl is unsubstituted or substituted; each R₇ is independently H or F; R₅ is CH3; and Rg is CH3.

26. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein at least one of Xi, X2, X3 and X₄ is N.

27. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein at least two of Xi, X2, X3 and X₄ is N.

28. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein at least three of Xi, X2, X3 and X₄ is N.

29. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Xi is N.

30. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein X2 is N.

31. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein X3 is N.

32. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein X₄ is N.

33. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein none of Xi, X2, X3 and X₄ is N.

34. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein the compound of Formula (1) is a compound of Formula (2) or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein:

Ri, R₄, XI, X2 X3 and X₄ are defined as described in the compound of Formula (1), the carbon marked with * is a chiral center and exists as a (R)- and (S)-racemic mixture or as either the (R)- or (S)- enantiomer, and the listing of substituents within brackets for a given compound indicates individual compounds containing one of each of the substituents.

35. The compound according to claim 34 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7-azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4-thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4-b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or a triazolo-pyridine.

36. The compound according to claim 34 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2- azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane.

37. The compound according to claim 34 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein Ri is a substituted or unsubstituted furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7-azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazole, 1,2,5-oxadiazole, 1,2,3-oxadiazole, 1,3,4-thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrido[3,4-b]pyrazine, pyrido[2,3-b]pyrazine, pteridine or a triazolo-pyridine; and R₄ is a substituted or unsubstituted aziridine, azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7- azaspiro[3.5]nonane, 1, 4-dioxa-7-azaspiro[4.4]nonane, 2-azaadamantane, bicyclo[l.l.l]pentane or 3-azabicylo[3.1.0]hexane.

38. A pharmaceutical composition comprising the compound of any one of claims 1 to 37 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

39. The pharmaceutical composition according to claim 38, further comprising one or more anticancer agents.

40. The pharmaceutical composition according to claim 39, wherein the one or more anti-cancer agents are selected from the group consisting of cyclophosphamide, dacarbazine, cisplatin, methotrexate, mercaptopurine, thioguanine, fluorouracil, cytarabine, vinblastine, paclitaxel, doxorubicin, bleomycin, mitomycin, prednisone, tamoxifen, flutamide, asparaginase, rituximab, trastuzumab, imatinib, retinoic acid, amifostine, camptothecin, topotecan, thalidomide, lenalidomide, a CDK inhibitor and a proteasome inhibitor.

41. A method of treating a disease in which PI3K activity is implicated in a subject in need of such treatment, the method comprising administering to the subject a therapeutically effective amount of the compound of any one of claims 1 to 37 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof.

42. The method of claim 41, wherein the disease is cancer.

43. The method of claim 41, wherein the disease is congenital lipomatous overgrowth, vascular malformations, epidermal naevi, scoliosis/skeletal and spinal syndrome (CLOVES), mosaic tissue overgrowth syndromes, venous malformations and brain malformations associated with severe epilepsy or PIK3CA-related overgrowth syndrome (PROS).

44. The method of claim 41, wherein the disease is a cancer bearing a PI3Ka H1047R mutation.