Isoquinolones as PI3K inhibitors

Abstract

Novel PI3K inhibitors of general formula (1) are described, along with methods for their preparation and their use in the treatment of diseases associated with upregulation or activation of the PI3K pathway. JPEG2025530234000341.jpg7058 (wherein R1 to R8 are as defined above).

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 525,460, filed July 7, 2023; and U.S. Provisional Application No. 63 / 404,715, filed September 8, 2022, the disclosures of which are incorporated herein in their entireties for all purposes. [Background technology]

[0002] Background of the Invention Phosphatidylinositol lipids (PIs) and their various phosphorylated variants are secondary messengers involved in a wide range of intracellular vesicle transport and signal transduction processes. Phosphoinositide 3' kinases (PI3Ks) are a family of enzymes responsible for phosphorylating the 3' hydroxyl position of the inositol ring of PIs. PI3Ks are subdivided into three classes based on their structure and substrates. Class II PI3Ks (PI3K-C2α, PI3K-C2β, and PI3K-C2γ) and class III PI3Ks (vps34) are monomeric enzymes primarily involved in endocytosis and autophagy (Posor et al., Biochim Biophys Acta 2015, 1851, 794; Backer, Biochem J. 2016, 473, 2251). Class I PI3Ks are heterodimers consisting of a catalytic kinase subunit (p110α, β, γ, δ) and one of several regulatory subunits that determine binding partners and subcellular localization. Class I PI3Ks are activated by interaction with receptor tyrosine kinases (RTKs), Ras-related GTPases, G-protein-coupled receptors, and / or associated adaptor proteins, converting phosphatidylinositol 4,5-bisphosphate (PIP2) to phosphatidyl 3,4,5-triphosphate (PIP3) in its active form (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 pathway has been linked to 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 PI3K as a key upstream regulator of these functions.

[0004] Class I PI3Ks are further subdivided into four isoforms (α, β, γ, and δ) based on the identity of their catalytic (p110α, p110β, p110γ, or p110δ) and regulatory (p85α or its various splice variants, p85β, p55γ, or p101) subunits, which play distinct roles in cell physiology (Vanhaesebroeck et al., J Mol Med (Berl). 2016, 94, 5). PI3Kγ and PI3Kδ are predominantly expressed in leukocytes and play important roles 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). PI3Kα and β are more ubiquitously expressed and share similar, but not identical, roles: for example, PI3Kα has a non-redundant role in angiogenesis (Soler et al., J Exp Med. 2013, 210, 1937), while PI3Kβ is known to exert 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] Elevated or constitutive activation of the PI3K pathway is one of the most frequent events in human cancer. The PI3K pathway becomes hyperactivated through diverse mechanisms, including activating mutations of PI3K isoforms, upregulation 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 encoding PI3Kα or mutations leading to upregulation of PI3Kα have been found to occur in many human cancers, including lung, gastric, 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 p110α subunit of PI3Kα, is frequently mutated or amplified in diverse tumor types. Missense mutations occur throughout the p110α domain but are clustered in two 'hotspots', the most common of which are E542K and E545K in the helical domain and H1047R in the kinase domain. Helical domain mutations reduce the inhibition of p110α by p85 or promote the direct interaction of p110α with insulin receptor substrate 1 (IRS1)37, whereas kinase domain mutations increase the interaction of p110α with lipid membranes, concomitantly upregulating signaling events (Thorpe et al., Nat Rev Cancer 2015, 15, 7).

[0006] Developing inhibitors of the PI3K pathway has been challenging due to the impossibility of achieving a sufficient dose for tumor suppression without adverse events. To date, clinically available PI3K inhibitors (alpelisib, buparlisib, copanlisib, duvelisib, idelalisib, pictilisib, taselisib, and others) have caused dose-dependent adverse events, such as hyperglycemia, rash, fatigue, and diarrhea (Jiang et al., Mol Biol Rep. 2020, 47, 4587), known as on-target toxicity. Hyperglycemia results from the body's insufficient production or abnormal utilization of insulin. The pancreas regulates insulin release in response to changes in blood glucose levels, leading to glucose uptake by muscle and fat cells when insulin levels are high or to gluconeogenesis by the liver when insulin levels are low. Tissue cell responses to insulin require PI3K signaling via the ubiquitously expressed p110α subunit. As a result, targeted pan-PI3K inhibition disrupts tissue glucose metabolism, leading to insulin resistance (Hopkins et al., Nature 2018, 560, 499). To mitigate adverse events, selective PI3K isoform inhibitors have been developed. The severity of adverse events depends on the selected isoform; for example, PI3Kα inhibitors are associated with hyperglycemia and rash due to the role of the p110α subunit in insulin response (Rugo et al., The Breast 2022, 61, 156). Similarly, the use of a selective PI3Kδ inhibitor (idelalisib) causes severe diarrhea and colitis when the p110δ subunit is highly expressed in immune cells. Inhibition with a dual inhibitor (taselisib), a potent PI3Kδ inhibitor with moderate PI3Kα inhibition, leads to gastrointestinal (GI) side effects, whereas no GI-related adverse events have been reported with a highly selective and potent PI3Kδ inhibitor (umbralisib) (Gadkar et al., CPT Pharmacometrics Syst Pharmacol. 2021, 11, 616).The amelioration of these adverse events with highly isoform-selective and potent inhibitors suggests that the development of mutant-selective isoform inhibitors to mitigate toxicity is a promising strategy for reducing the severity of toxicity. Furthermore, selective inhibition of mutant versus wild-type PI3Kα isoforms may suppress cancer signaling with minimal impact on PI3K signaling in healthy cells that only harbor wild-type PI3Kα, potentially reducing the toxicity associated with non-selective PI3K inhibition (Castel et al., Nat Cancer 2021 2, 587). Summary of the Invention [Problem to be solved by the invention]

[0007] There is currently interest in developing PI3K inhibitors for cancer therapy (WO2023 / 081209, WO2023 / 078401, WO2023 / 060262, WO2023 / 056407, WO2021 / 202964, WO2023 / 159155). However, there is a continuing need for novel potent and selective PI3K inhibitors for cancer treatment, either as single agents or in combination therapy. [Means for solving the problem]

[0008] Summary of the Invention One embodiment of the present invention is a compound represented by formula (1) [ka] [During the ceremony, R1 is alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein each of the alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; R2 is H, C1-C4 alkyl, C3-C7 cycloalkyl, CF3, CH2F, or CF2H, where R2 is not H and the carbon atom attached to R2 is a chiral center and exists as an (R)- and (S)-racemic mixture or as an (R)- or (S)-enantiomer; R3 is H or C1-C4 alkyl; R4 is H, F, Cl or CH3; R6 is H, C1-C4 alkyl, C3-C7 cycloalkyl, heteroaryl, CF3, CH2F, or CF2H; R7 is H, C1-C4 alkyl, C3-C7 cycloalkyl, halogen, CN, CF3, OCF3, CH2F or CF2H; each R8 is independently H, C1-C4 alkyl, C3-C7 cycloalkyl, halogen, CN, CF3, OCF3, CH2F, or CF2H; R5 is H; halogen; -O-L1-L2-L3-L4-L5-L6-L7-R9; -S-L1-L2-L3-L4-L5-L6-L7-R9; -S(O)-L1-L2-L3-L5-L6-L7-R9; -S(O)2-L1-L2-L3-L5-L6-L7-R9; -(NR 10 )-L1-L2-L3-L4-L5-L6-L7-R9; or -L8-L9-L 10 -L 11 -L 12 -R 14 where: Each of L1, L2, L3, L6 and L7 independently represents (CHR 11 ), (CHR 11 -O), (CHR 11 -S), (C3-C7 cycloalkyl), (CH2) 1-4 or is a bond; L4 is C=O, C=S or a bond; L5 is NR 10 , S, O or a bond; R9 is H, C(=O)R 12 , C(=O)NR 12 R 13 , N.R. 12 R 13 , C(=O)OR 12, C1-C6 alkyl, C1-C6 fluoroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, where each of the C1-C6 alkyl, C1-C6 fluoroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; or NR 10 When exists, R9 and R 10 may be taken together with the nitrogen atom to which it is attached to form a substituted or unsubstituted ring. or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound thereof. In an exemplary embodiment, the ring is a 4-7 membered substituted or unsubstituted non-aromatic heterocyclic ring containing 0, 1, or 2 heteroatoms, which may be N, O, S, or Si (in addition to the nitrogen atom), with the proviso that when the ring size is 4 or 5, the number of additional heteroatoms is 0 or 1, and when the ring size is 6-7, the number of additional heteroatoms is 0, 1, or 2, and when the ring is substituted, the substituents are CH, F, Cl, CF, CFH, CHF, OCH, cyclopropyl, CHCF, an oxetane ring, or COR. a (where R a is C1-C4 alkyl, O-C1-C4 alkyl or NR b R c and R b and R c are independently H or C1-C4 alkyl; R 10 and R 11 each is independently H or C1-C4 alkyl (e.g., CH3, CH2CH3, or CH(CH3)2), where C1-C4 alkyl is unsubstituted or substituted; R 12 and R 13 each is independently H, C1-C6 alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, where each of the C1-C6 alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; or R 12 and R 13may be taken together with the nitrogen atom to which it is attached to form a substituted or unsubstituted ring. In exemplary embodiments, the ring is a 4-7 membered substituted or unsubstituted non-aromatic heterocyclic ring containing 0, 1, or 2 heteroatoms, which may be N, O, S, or Si (in addition to the nitrogen atom), provided that when the ring size is 4 or 5, the number of additional heteroatoms is 0 or 1, and when the ring size is 6-7, the number of additional heteroatoms is 0, 1, or 2; and when the ring is substituted, the substituents are CH, F, Cl, CF, CFH, CHF, OCH, cyclopropyl, CHCF, oxetane, or COR. a (where R a is C1-C4 alkyl, O-C1-C4 alkyl or NR b R c and R b and R c are independently H or C1-C4 alkyl; L8 is (CHR 15 ), (CHR 15 -O), (CHR 15 -S), (CHR 15 -NR 16 ), C=O, C=S or a bond; L9 is a C3-C7 cycloalkyl, optionally part of a bridged, fused, or spiro ring system, C(R 15 )=C(R 15 ), C≡C or bond; L 10 independently (CHR 15 ), O, S, (NCR 15 ), N(C=O) or a bond; L 11 (CHR 15 ), C=O, C=S or a bond; L 12is H, (C3-C7 cycloalkyl), heterocyclyl, aryl, heteroaryl, or a bond, where each of the (C3-C7 cycloalkyl), heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted, and the (C3-C7 cycloalkyl) and / or heterocyclyl is optionally part of a bridged, fused, or spiro ring system; R 14 is H, CR 15 R 16 R 17 , OR 17 , S.R. 17 , N.R. 16 R 17 , C1-C6 alkyl, C1-C6 fluoroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein each of the C1-C6 alkyl, C1-C6 fluoroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; R 15 and R 16 each is independently H or C1-C3 alkyl; and Each R 17 is independently H, C-C alkyl, C-C fluoroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, where each of the C-C alkyl, C-C fluoroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; or R 16 and R 17 may be taken together with the nitrogen atom to which it is attached to form a substituted or unsubstituted ring. In exemplary embodiments, the ring is a 4-7 membered substituted or unsubstituted non-aromatic heterocyclic ring containing 0, 1, or 2 heteroatoms, which may be N, O, or S (in addition to the nitrogen atom), provided that when the ring size is 4 or 5, the number of additional heteroatoms is 0 or 1, and when the ring size is 6-7, the number of additional heteroatoms is 0, 1, or 2; and when the ring is substituted, the substituents are Me, F, Cl, CF, CFH, CHF, OCH, cyclopropyl, CHCF, an oxetane ring, or COR. a (where R ais C1-C4 alkyl, O-C1-C4 alkyl or NR b R c and R b and R c are independently H or C1-C4 alkyl; However, R5 is -L8-L9-L 10 -L 11 -L 12 -R 14 When L8, L9, L 10 , L 11 , L 12 and R 14 at least one of which is a carbon-containing moiety, and R5 is directly attached to the (isoquinolone) core structure by a carbon atom; or R5 is a non-aromatic N-linked heterocyclic ring [ka] wherein the heterocyclic ring is substituted or unsubstituted, optionally contains one or more additional ring atoms selected from N, O, Si, and S, and is optionally part of a bridged, fused, or spiro ring system. In certain embodiments, the N-linked heterocyclyl ring is 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, or 2-azaadamantane.

[0009] In an exemplary embodiment, R5 is -(NR 10 )-L1-L2-L3-L4-L5-L6-L7-R9, where L1 to L7, R9 and R 10 is as defined.

[0010] In an exemplary embodiment, R5 is -O-L1-L2-L3-L4-L5-L6-L7-R9, where L1 through L7 and R9 are as defined.

[0011] In exemplary embodiments, R5 is -S-L1-L2-L3-L4-L5-L6-L7-R9; -S(O)-L1-L2-L3-L5-L6-L7-R9; or -S(O)2-L1-L2-L3-L5-L6-L7-R9, where L1 through L7 and R9 are as defined.

[0012] In exemplary embodiments, R is a 6-membered aryl ring; or a 5- to 6-membered heteroaryl ring containing 1 to 3 nitrogen atoms; or a non-aromatic 3- to 7-membered carbocyclic ring; or a non-aromatic 3- to 7-membered heterocyclic ring containing 1 to 3 heteroatoms selected from N, O, S, and Si (provided that when the ring size is 4 or 5, the number of heteroatoms is 1 or 2, and when the ring size is 6 or 7, the number of heteroatoms is 1, 2, or 3); or a C-C alkyl group, wherein the aryl ring, heteroaryl ring, carbocyclic ring, heterocyclic ring, and C-C alkyl group are unsubstituted or substituted with CH, F, Cl, CF, CFH, CHF, OCH, -CHCF, cyclopropyl, -CN, N(CH), an oxetane ring, a phenyl or phenoxy group optionally substituted with 1 to 3 halogens (F, Cl, or Br) or CH groups, or COR a (where R a is C1-C4 alkyl, O-C1-C4 alkyl or NR b R c where R b and R c are independently H or C1-C4 alkyl).

[0013] In exemplary embodiments of R5, L8, L 10 and L 11 Each of is a bond and L9 is not a bond.

[0014] In exemplary embodiments of R5, L8, L 10 and L 11 each of which is a bond and L9 is cycloalkyl which is optionally part of a bridged, fused or spiro ring system.

[0015] In exemplary embodiments of R5, L8, L 10 and L 11 Each of is a bond and L9 is a cycloalkyl that is part of a bridged ring system.

[0016] In exemplary embodiments of R5, L8, L 10 and L 11 Each of is a bond and L9 is a cycloalkyl that is part of a fused ring system.

[0017] In exemplary embodiments of R5, L8, L 10 and L 11 Each of is a bond and L9 is a cycloalkyl that is part of a spiro ring system.

[0018] In exemplary embodiments of R5, L8, L 10 and L 11 Each of is a bond, and L9 is C(R 15 )=C(R 15 )

[0019] In exemplary embodiments of R5, L8, L 10 and L 11 Each of is a bond and L9 is C≡C.

[0020] In an exemplary embodiment of R5, L8, L9, L 10 and L 11 Each of the is a bond, and L 12 is cycloalkyl, heterocyclyl, aryl, or heteroaryl, where each of the cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted.

[0021] In an exemplary embodiment of R5, L8, L9, L 10 and L 11 Each of the is a bond, and L 12 is cycloalkyl, heterocyclyl, aryl, or heteroaryl, where each of the cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; R 14 is H.

[0022] In an exemplary embodiment of R5, L8, L9, L 10 and L 11 Each of the is a bond, and L 12 is cycloalkyl, heterocyclyl, aryl, or heteroaryl, where each of the cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; R 14 is cycloalkyl, heterocyclyl, aryl, or heteroaryl, where each of the C1-C6 alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted.

[0023] In an exemplary embodiment of R5, L8, L9, L 10 and L 11 Each of the is a bond, and L 12 is cycloalkyl, heterocyclyl, aryl, or heteroaryl, where each of the cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; R 14 Ha-CR 14 R 15 R 16 is.

[0024] In an exemplary embodiment of R5, L8, L9, L 10 and L 11 Each of the is a bond, and L 12 is cycloalkyl, heterocyclyl, aryl, or heteroaryl, where each of the cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; R 14 HA-OR 16 -OR 17 is.

[0025] In an exemplary embodiment of R5, L8, L9, L 10 and L 11 Each of the is a bond, and L 12is cycloalkyl, heterocyclyl, aryl, or heteroaryl, where each of the cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; R 14 Ha-SR 17 is.

[0026] In an exemplary embodiment of R5, L8, L9, L 10 and L 11 Each of the is a bond, and L 12 is cycloalkyl, heterocyclyl, aryl, or heteroaryl, where each of the cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; R 14 Ha-NR 16 R 17 is.

[0027] In an exemplary embodiment, R5 is [ka] where R d is H or CH3, and R e is CH3 optionally substituted with CH3, F, Cl, CF3, CF2H, CH2F, OCH3, cyclopropyl, CN or N(CH3)2, C3-C6 cycloalkyl, a 6-membered aromatic or heteroaromatic ring containing 0, 1 or 2 nitrogen atoms, or R d and R e may, together with the nitrogen atom to which it is attached, form a 4- to 7-membered non-aromatic heterocycle containing 1 to 2 heteroatoms which may be N or O, provided that when the ring size is 4 or 5, the number of heteroatoms is 1, and when the ring size is 6 to 7, the number of heteroatoms is 1 or 2, and wherein the ring is unsubstituted or is selected from the group consisting of CH, F, Cl, CF, CFH, CHF, OCH, cyclopropyl, CHCF, oxetane ring, and COR. a (where R a is C1-C4 alkyl, O-C1-C4 alkyl or NR b R c and R b and Rc are independently H or C1-C4 alkyl).

[0028] In an exemplary embodiment, R5 is an N-linked non-aromatic heterocyclyl ring. [ka] wherein the heterocyclyl ring is substituted or unsubstituted, optionally contains one or more additional atoms selected from N, O, Si, and S, and is optionally part of a bridged, fused, or spiro ring system. In certain embodiments, the N-linked heterocyclyl ring is azetidine, pyrrolidine, imidazoline, imidazolidine, piperazine, morpholine, thiomorpholine, piperidine, indoline, tetrahydroquinoline, decahydroquinoline, 2-oxa-7-azaspiro[3.5]nonane, 1,3,8-triazaspiro[4.5]-decan-4-one, 1,4-dioxa-7-azaspiro[4.4]nonane.

[0029] In an exemplary embodiment, the N-linked non-aromatic heterocyclyl ring is substituted or unsubstituted, optionally contains one or more additional atoms selected from N, O, Si, and S, and is not part of a bridged, fused, or spiro ring system.

[0030] In an exemplary embodiment, the N-linked non-aromatic heterocyclyl ring is substituted or unsubstituted, optionally contains one or more additional atoms selected from N, O, Si and S, and is part of a bridged, fused or spiro ring system.

[0031] In an exemplary embodiment, the N-linked non-aromatic heterocyclyl ring is substituted or unsubstituted, does not contain an additional atom selected from N, O, Si and S, and is not part of a bridged, fused or spiro ring system.

[0032] In an exemplary embodiment, the N-linked non-aromatic heterocyclyl ring is substituted or unsubstituted, does not contain additional atoms selected from N, O, Si and S, and is part of a bridged, fused, or spiro ring system.

[0033] In an exemplary embodiment, the N-linked non-aromatic heterocyclyl ring is substituted or unsubstituted, contains at least one sulfur ring atom, and is not part of a bridged, fused, or spiro ring system.

[0034] In an exemplary embodiment, the N-linked non-aromatic heterocyclyl ring is substituted or unsubstituted, contains at least one sulfur ring atom, and is part of a bridged, fused, or spiro ring system.

[0035] In an exemplary embodiment, the N-linked non-aromatic heterocyclyl ring is substituted or unsubstituted, contains at least one oxygen ring atom, and is not part of a bridged, fused, or spiro ring system.

[0036] In an exemplary embodiment, the N-linked non-aromatic heterocyclyl ring is substituted or unsubstituted, contains at least one oxygen ring atom, and is part of a bridged, fused, or spiro ring system.

[0037] In an exemplary embodiment, the N-linked non-aromatic heterocyclyl ring is substituted or unsubstituted, contains at least one additional nitrogen ring atom, and is not part of a bridged, fused, or spiro ring system.

[0038] In exemplary embodiments, the N-linked non-aromatic heterocyclyl ring is substituted or unsubstituted, contains at least one additional nitrogen ring atom, and is part of a bridged, fused, or spiro ring system.

[0039] In an exemplary embodiment of the compound of Formula (1), R 1 is heterocyclyl, aryl, or heteroaryl, wherein each of the heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted.

[0040] In an exemplary embodiment of the compound of Formula (1), R 1 is heterocyclyl, wherein the heterocyclyl is unsubstituted or substituted.

[0041] In an exemplary embodiment of the compound of Formula (1), R 1 is aryl, wherein the aryl is unsubstituted or substituted.

[0042] In an exemplary embodiment of the compound of Formula (1), R 1 is heteroaryl, wherein heteroaryl is unsubstituted or substituted.

[0043] In an exemplary embodiment of the compound of Formula (1), R2 is CH3.

[0044] In an exemplary embodiment of the compound of Formula (1), R3 is H.

[0045] In an exemplary embodiment of the compound of Formula (1), R 4 is H.

[0046] In an exemplary embodiment of the compound of Formula (1), R5 is -O-L1-L2-L3-L4-L5-L6-L7-R9.

[0047] In an exemplary embodiment of the compound of Formula (1), R5 is -S-L1-L2-L3-L4-L5-L6-L7-R9.

[0048] In an exemplary embodiment of the compound of Formula (1), R5 is -S(O)-L1-L2-L3-L5-L6-L7-R9.

[0049] In an exemplary embodiment of the compound of Formula (1), R5 is -S(O)2-L1-L2-L3-L5-L6-L7-R9.

[0050] In an exemplary embodiment of the compound of Formula (1), R5 is —(NR 10 )-L1-L2-L3-L4-L5-L6-L7-R9.

[0051] In an exemplary embodiment of the compound of Formula (1), R5 is -L8-L9-L 10 -L 11 -L 12 -R 14 is.

[0052] In an exemplary embodiment of the compound of Formula (1), R6 is CH3.

[0053] In an exemplary embodiment of the compound of Formula (1), R7 is CH3.

[0054] In an exemplary embodiment of the compound of Formula (1), R 8 is H.

[0055] In an exemplary embodiment of the compound of Formula (1), R 1 is heterocyclyl, wherein the heterocyclyl is unsubstituted or substituted, R 2 is CH 3 , and R 3 is H.

[0056] In an exemplary embodiment of the compound of Formula (1), R 1 is aryl, where aryl is unsubstituted or substituted, R 2 is CH 3 , and R 3 is H.

[0057] In an exemplary embodiment of the compound of Formula (1), R 1 is heteroaryl, where heteroaryl is unsubstituted or substituted, R 2 is CH 3 , and R 3 is H.

[0058] In an exemplary embodiment of the compound of Formula (1), R 1 is heterocyclyl, wherein the heterocyclyl is unsubstituted or substituted, R 2 is CH 3 , R 3 is H, and R 8 is H.

[0059] In an exemplary embodiment of the compound of Formula (1), R 1 is aryl, where aryl is unsubstituted or substituted, R 2 is CH 3 , R 3 is H, and R 8 is H.

[0060] In an exemplary embodiment of the compound of Formula (1), R 1 is heteroaryl, wherein heteroaryl is unsubstituted or substituted, R 2 is CH 3 , R 3 is H, and R 8 is H.

[0061] In an exemplary embodiment of the compound of Formula (1), R1 is heterocyclyl, wherein the heterocyclyl is unsubstituted or substituted, R2 is CH3, R3 is H, R8 is H, and R6 is CH3.

[0062] In an exemplary embodiment of the compound of Formula (1), R1 is aryl, where aryl is unsubstituted or substituted, R2 is CH3, R3 is H, R8 is H, and R6 is CH3.

[0063] In an exemplary embodiment of the compound of Formula (1), R1 is heteroaryl, where heteroaryl is unsubstituted or substituted, R2 is CH3, R3 is H, R8 is H, and R6 is CH3.

[0064] In an exemplary embodiment of the compound of Formula (1), R1 is heterocyclyl, where the heterocyclyl is unsubstituted or substituted, R2 is CH3, R3 is H, and R5 is -O-L1-L2-L3-L4-L5-L6-L7-R9.

[0065] In an exemplary embodiment of the compound of Formula (1), R1 is aryl, where aryl is unsubstituted or substituted, R2 is CH3, R3 is H, and R5 is -S-L1-L2-L3-L4-L5-L6-L7-R9.

[0066] In an exemplary embodiment of the compound of Formula (1), R1 is heteroaryl, where heteroaryl is unsubstituted or substituted, R2 is CH3, R3 is H, and R5 is -S(O)-L1-L2-L3-L5-L6-L7-R9.

[0067] In an exemplary embodiment of the compound of Formula (1), R1 is heterocyclyl, where the heterocyclyl is unsubstituted or substituted, R2 is CH3, R3 is H, and R5 is -S(O)2-L1-L2-L3-L5-L6-L7-R9.

[0068] In an exemplary embodiment of the compound of Formula (1), R1 is aryl, where aryl is unsubstituted or substituted, R2 is CH3, R3 is H, and R5 is -(NR 10 )-L1-L2-L3-L4-L5-L6-L7-R9.

[0069] In an exemplary embodiment of the compound of Formula (1), R1 is heteroaryl, where heteroaryl is unsubstituted or substituted, R2 is CH3, R3 is H, and R5 is -L8-L9-L 10 -L 11 -L 12 -R 14 is.

[0070] In an exemplary embodiment of the compound of Formula (1), R1 is heterocyclyl, where the heterocyclyl is unsubstituted or substituted, R2 is CH3, R3 is H, R5 is -O-L1-L2-L3-L4-L5-L6-L7-R9, and R8 is H.

[0071] In an exemplary embodiment of the compound of Formula (1), R1 is aryl, where aryl is unsubstituted or substituted, R2 is CH3, R3 is H, R5 is -S-L1-L2-L3-L4-L5-L6-L7-R9, and R8 is H.

[0072] In an exemplary embodiment of the compound of Formula (1), R is heteroaryl, where heteroaryl is unsubstituted or substituted, R is CH, R is H, and R は -S(O)-L1-L2-L3-L5-L6-L7-R 9であり、R8 is H.

[0073] In an exemplary embodiment of the compound of Formula (1), R1 is heterocyclyl, where the heterocyclyl is unsubstituted or substituted, R2 is CH3, R3 is H, R5 is -S(O)2-L1-L2-L3-L5-L6-L7-R9, and R8 is H.

[0074] In an exemplary embodiment of the compound of Formula (1), R1 is aryl, where aryl is unsubstituted or substituted, R2 is CH3, R3 is H, and R5 is -(NR 10 )-L1-L2-L3-L4-L5-L6-L7-R9, and R8 is H.

[0075] In an exemplary embodiment of the compound of Formula (1), R1 is heteroaryl, where heteroaryl is unsubstituted or substituted, R2 is CH3, R3 is H, and R5 is -L8-L9-L 10 -L 11 -L 12 -R 14 and R8 is H.

[0076] In an exemplary embodiment of the compound of Formula (1), R1 is heterocyclyl, wherein the heterocyclyl is unsubstituted or substituted, R2 is CH3, R3 is H, R5 is -O-L1-L2-L3-L4-L5-L6-L7-R9, R8 is H, and R6 is CH3.

[0077] In an exemplary embodiment of the compound of Formula (1), R1 is aryl, where aryl is unsubstituted or substituted, R2 is CH3, R3 is H, R5 is -S-L1-L2-L3-L4-L5-L6-L7-R9, R8 is H, and R6 is CH3.

[0078] In an exemplary embodiment of the compound of Formula (1), R1 is aryl, where aryl is unsubstituted or substituted, R2 is CH3, R3 is H, R5 is -S(O)-L1-L2-L3-L4-L5-L6-L7-R9, R8 is H, and R6 is CH3.

[0079] In an exemplary embodiment of the compound of Formula (1), R1 is heteroaryl, where heteroaryl is unsubstituted or substituted, R2 is CH3, R3 is H, R5 is -S(O)2-L1-L2-L3-L5-L6-L7-R9, R8 is H, and R6 is CH3.

[0080] In an exemplary embodiment of the compound of Formula (1), R1 is heterocyclyl, where heterocyclyl is unsubstituted or substituted, R2 is CH3, R3 is H, and R5 is -(NR 10 )-L1-L2-L3-L4-L5-L6-L7-R9, where R8 is H and R6 is CH3.

[0081] In an exemplary embodiment of the compound of Formula (1), R1 is aryl, where aryl is unsubstituted or substituted, R2 is CH3, R3 is H, and R5 is -L8-L9-L 10 -L 11 -L 12 -R 14 where R8 is H and R6 is CH3.

[0082] In an exemplary embodiment, the compound of formula (1) is represented by formula (2): [ka] [During the ceremony, Each of X1, X2, and X3 is independently N, CH, or substituted C; R5 and R8 are as defined in the compound of formula (1), and Carbons marked with an * are chiral centers and exist as (R)- and (S)-racemic mixtures or as (R)- or (S)-enantiomers.] or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound or pharmaceutically acceptable salt thereof.

[0083] In an exemplary embodiment, the compound of formula (1) is represented by formula (3): [ka] [During the ceremony, R1 is heterocyclyl, aryl, or heteroaryl, wherein the heterocyclyl, aryl, or heteroaryl ring directly attached to the nitrogen atom attached to the asymmetric center attached to the isoquinoline moiety contains a carboxylic acid substituent ortho to the point of attachment and optionally one or more further substituents; R5 and R8 are as defined in the compound of formula (1), and Carbons marked with an * are chiral centers and exist as (R)- and (S)-racemic mixtures or as (R)- or (S)-enantiomers.] or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound or pharmaceutically acceptable salt thereof.

[0084] In an exemplary embodiment of the compound of Formula (3), R1 is heteroaryl.

[0085] In an exemplary embodiment of the compound of Formula (3), R1 is aryl.

[0086] In an exemplary embodiment of the compound of Formula (3), R1 is heterocyclyl and R5 is -O-L1-L2-L3-L4-L5-L6-L7-R9.

[0087] In an exemplary embodiment of the compound of Formula (3), R1 is heteroaryl and R5 is -O-L1-L2-L3-L4-L5-L6-L7-R9.

[0088] In an exemplary embodiment of the compound of Formula (3), R1 is aryl, where aryl is unsubstituted or substituted, and R5 is -O-L1-L2-L3-L4-L5-L6-L7-R9.

[0089] In an exemplary embodiment of the compound of Formula (3), R1 is heterocyclyl, where the heterocyclyl is unsubstituted or substituted, and R5 is -S-L1-L2-L3-L4-L5-L6-L7-R9.

[0090] In an exemplary embodiment of the compound of Formula (3), R1 is heteroaryl, where heteroaryl is unsubstituted or substituted, and R4 is -S-L1-L2-L3-L4-L5-L6-L7-R9.

[0091] In an exemplary embodiment of the compound of Formula (3), R1 is aryl, where aryl is unsubstituted or substituted, and R5 is -S-L1-L2-L3-L4-L5-L6-L7-R9.

[0092] In an exemplary embodiment of the compound of Formula (3), R1 is heterocyclyl, where the heterocyclyl is unsubstituted or substituted, and R5 is -S(O)-L1-L2-L3-L4-L5-L6-L7-R9.

[0093] In an exemplary embodiment of the compound of Formula (3), R1 is heteroaryl, where heteroaryl is unsubstituted or substituted, and R5 is -S(O)-L1-L2-L3-L4-L5-L6-L7-R9.

[0094] In an exemplary embodiment of the compound of Formula (3), R1 is aryl, where aryl is unsubstituted or substituted, and R5 is -S(O)-L1-L2-L3-L4-L5-L6-L7-R9.

[0095] In an exemplary embodiment of the compound of Formula (3), R1 is heterocyclyl, where the heterocyclyl is unsubstituted or substituted, and R5 is -S(O)2-L1-L2-L3-L4-L5-L6-L7-R9.

[0096] In an exemplary embodiment of the compound of Formula (3), R1 is heteroaryl, where heteroaryl is unsubstituted or substituted, and R5 is -S(O)2-L1-L2-L3-L4-L5-L6-L7-R9.

[0097] In an exemplary embodiment of the compound of Formula (3), R1 is aryl, where aryl is unsubstituted or substituted, and R5 is -S(O)2-L1-L2-L3-L4-L5-L6-L7-R9.

[0098] In an exemplary embodiment of the compound of Formula (3), R1 is heterocyclyl, wherein the heterocyclyl is unsubstituted or substituted, and R5 is -(NR 10 )-L1-L2-L3-L4-L5-L6-L7-R9.

[0099] In an exemplary embodiment of the compound of Formula (3), R1 is heteroaryl, where heteroaryl is unsubstituted or substituted, and R5 is -(NR 10 )-L1-L2-L3-L4-L5-L6-L7-R9.

[0100] In an exemplary embodiment of the compound of Formula (3), R1 is aryl, where aryl is unsubstituted or substituted, and R5 is -(NR 10 )-L1-L2-L3-L4-L5-L6-L7-R9.

[0101] In an exemplary embodiment of the compound of Formula (3), R1 is heterocyclyl, where the heterocyclyl is unsubstituted or substituted, and R5 is -L8-L9-L 10 -L 11 -L 12 -R 14 is.

[0102] In an exemplary embodiment of the compound of Formula (3), R1 is heteroaryl, where heteroaryl is unsubstituted or substituted, and R5 is -L8-L9-L 10 -L 11 -L 12 -R 14 is.

[0103] In an exemplary embodiment of the compound of Formula (3), R1 is aryl, where aryl is unsubstituted or substituted, and R5 is -L8-L9-L 10 -L 11 -L12 -R 14 is.

[0104] One aspect of the present invention is a pharmaceutical composition comprising any of the compounds of the invention described herein (e.g., any of Formulas (1), (2), (3), (4), (5), or (6)) or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0105] In an exemplary embodiment, a pharmaceutical composition comprising any of the compounds of the invention described herein (e.g., any of Formulas (1), (2), (3), (4), (5), or (6)) or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, further comprises one or more anti-cancer agents.

[0106] Another aspect of the invention is a method of treating a disease associated with PI3K activity in a subject in need thereof, comprising administering a therapeutically effective amount of any of the compounds of the invention described herein (e.g., any of Formulas (1), (2), (3), (4), (5), or (6)) or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof.

[0107] In an exemplary embodiment, the disease to be treated is cancer. In a particular embodiment, the disease is a cancer carrying a PI3Kα H1047 mutation (e.g., H1047R). DETAILED DESCRIPTION OF THE INVENTION

[0108] Detailed Description of the Invention As used herein, the term "at risk" refers to a medical condition or set of medical conditions exhibited by a patient that may predispose the patient to a particular disease or affliction. For example, these conditions may result from influences, including, but not limited to, behavioral, emotional, chemical, biochemical, or environmental influences.

[0109] As used herein, the term "effective amount" refers to a specific amount of a pharmaceutical composition containing a therapeutic agent that achieves a clinically beneficial result (i.e., reduction of symptoms). The toxicity and therapeutic efficacy of such a composition can be determined, for example, by the LD 50 (the dose that is lethal to 50% of the population) and ED 50 The dose ratio between toxic and therapeutic effects is the therapeutic index, which can be determined by standard pharmaceutical procedures in cell cultures or experimental animals for determination of the LD (the dose which is therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index, which is the ratio LD (LD ). 50 / ED 50 Compounds with 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 dosage for human use. The dosage of such compounds is preferably administered with little or no toxicity and with an ED 50 The dosage will vary within this range depending on the dosage form used, patient sensitivity, and route of administration.

[0110] As used herein, the term "symptom" refers to any subjective or objective evidence of disease or physical abnormality observed by a patient. For example, subjective evidence is typically based on patient self-report and may include, but is not limited to, pain, headache, visual disturbances, nausea, and / or vomiting. Alternatively, objective evidence is typically the results of medical tests, including, but not limited to, temperature, complete blood count, lipid panel, thyroid panel, blood pressure, heart rate, electrocardiogram, tissue body imaging scans, and other medical test results.

[0111] As used herein, the term "disease" refers to any disturbance in the normal function of a living animal or part thereof that disrupts or modifies the production of vital functions. Typically manifested by noticeable signs and symptoms, diseases are usually a response to: i) environmental factors (e.g., nutritional deficiencies, industrial hazards, or climate); ii) specific infectious agents (e.g., parasites, bacteria, or viruses); iii) an inherent defect of the organism (e.g., genetic abnormalities); and / or iv) a combination of these factors.

[0112] The terms "reduce," "inhibit," "lower," "suppress," "reduce," "prevent," and grammatical equivalents (such as "low," "small"), when used in reference to the occurrence of some symptom in a treated versus untreated subject, indicate that the amount and / or intensity of the symptom in the treated subject is lower than in the untreated subject by some amount that would be recognized as clinically relevant by any medical professional. In certain embodiments, the amount and / or intensity of the symptom in the treated subject is at least 10% lower, at least 25% lower, at least 50% lower, at least 75% lower, and / or at least 90% lower than the amount and / or intensity of the symptom in the untreated subject.

[0113] As used herein, the term "inhibitory compound" refers to any compound that can interact with (i.e., attach, bind, etc.) a binding partner under conditions such that the binding partner is unresponsive to its natural ligand. Inhibitory compounds can include, but are not limited to, small organic molecules, antibodies, and proteins / peptides.

[0114] As used herein, the term "adhesion" refers to any interaction between a vehicle (or carrier) and a drug. Adhesion may be reversible or irreversible. Such attachment includes, but is not limited to, covalent bonds, ionic bonds, van der Waals forces, or friction. A drug is attached to a vehicle (or carrier) if it is impregnated, incorporated, coated, suspended, dissolved, mixed, or the like.

[0115] As used herein, the term "drug" or "compound" refers to any pharmacologically active substance that can be administered to achieve a desired effect. Drugs or compounds can be synthetic or naturally occurring, non-peptides, proteins or peptides, oligonucleotides or nucleotides, polysaccharides or sugars.

[0116] As used herein, the term "administration" or "administering" refers to any method of providing a composition to a patient so that the composition has its intended effect in the patient. Exemplary methods of administration are by direct mechanisms such as local tissue administration (i.e., extravascular administration, e.g., subcutaneous, intramuscular, or intraperitoneal), intravenous, oral ingestion, transdermal patch, topical, inhalation, suppository, etc.

[0117] As used herein, the term "patient" refers to a human or animal, and need not be hospitalized. For example, outpatients and nursing home residents are "patients." Patients may be humans or non-human animals of any age, and thus include adults and juveniles (i.e., children). The term "patient" is not intended to imply the need for medical treatment. Thus, patients may voluntarily be the subject of experimentation, whether clinical or to support basic science testing.

[0118] As used herein, the term "subject" includes, but is not limited to, humans (e.g., humans of any age, male or female, e.g., a pediatric subject (e.g., infant, child, adolescent) or an adult subject (e.g., a young adult, middle-aged adult, or elderly adult)) and / or other primates (e.g., monkeys); non-human mammals such as cows, pigs, horses, sheep, mice, goats, cats, dogs, etc.; and / or birds such as chickens, ducks, and / or geese.

[0119] As used herein, the term "affinity" refers to any force of attraction between substances or particles that allows them to become and remain part of a chemical combination. For example, an inhibitory compound with high affinity for a receptor will provide greater effectiveness in preventing the receptor from interacting with its natural ligand than a low-affinity inhibitor.

[0120] As used herein, the term "derived" refers to the source of a compound or sequence. In some respects, the compound or sequence may be derived from an organism or a particular species. In other respects, the compound or sequence may be derived from a larger complex or sequence.

[0121] As used herein, the term "test compound" refers to any compound or molecule that is considered a candidate inhibitory compound.

[0122] As used herein, the term "combination therapy" refers to a dosing regimen of two or more different therapeutically active agents within a given period of time, where the therapeutically active agents are administered together or separately. In certain embodiments, the combination therapy is a non-fixed combination.

[0123] As used herein, the term "non-fixed combination" refers to two or more different therapeutic agents that are formulated as separate compositions or dosages so that they can be administered simultaneously or sequentially at varying intervals to a subject in need thereof.

[0124] As used herein, the term "synergistic effect" or "synergistic" refers to the phenomenon where the combination of two therapeutic agents in a combination therapy results in a measured effect that is greater than the sum of the effects of each agent when administered alone.

[0125] As used herein, the term "in vivo" refers to events that take place inside the body of a subject.

[0126] As used herein, the term "in vitro" refers to an event that takes place outside a subject's body.

[0127] As used herein, the term "protein" refers to any of a number of naturally occurring, highly complex substances (e.g., enzymes or antibodies) that contain amino acid residues joined by peptide bonds and contain carbon, hydrogen, nitrogen, oxygen, and typically sulfur. Generally, proteins contain on the order of several hundred amino acids.

[0128] As used herein, the term "peptide" refers to any of a variety of amides derived from two or more amino acids by the combination of the amino group of one with the carboxyl group of another, usually obtained by partial hydrolysis of proteins. Generally, peptides contain several dozen amino acids.

[0129] As used herein, the terms "pharmaceutically acceptable" or "pharmacologically acceptable" refer to molecular entities and compositions that do not produce adverse, allergic or other untoward reactions when administered to animals or humans.

[0130] As used herein, the term "pharmaceutically acceptable carrier" includes any and all solvents or dispersion media, including, but not limited to, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, vegetable oils, coatings, isotonic and absorption delaying agents, liposomes, commercially available detergents, etc. Supplementary biologically active ingredients can also be incorporated into such carriers.

[0131] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that is suitable for use in contact with the tissues of a subject without adversely affecting the biological activity and properties of the compound, without undue toxicity, irritation, and / or allergic response, etc. Pharmaceutically acceptable salts include those derived from suitable inorganic acids and bases, including 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, naphthalenesulfonic acid, lactic acid, succinic acid, oxalic acid, stearic acid, and the like. In some cases, pharmaceutically acceptable salts are obtained by reacting a compound having an acidic group, as described herein, with a base to form a salt, such as ammonium salts, alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., calcium or magnesium salts), salts formed with organic bases, and amino acid salts. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, and ammonium and quaternary ammonium compounds. Specific metals include, but are not limited to, sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc. Organic bases from which salts can be prepared include, for example, primary, secondary, and tertiary amines.

[0132] As used herein, the term "prodrug" refers to a compound that is converted 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 to an active compound in vivo. In various cases, the physicochemical properties (e.g., bioavailability) and / or delivery properties of the prodrug are improved over the parent compound. Prodrugs are typically designed to enhance pharmaceutical and / or pharmacokinetic-based properties relative to the parent compound. Prodrug compounds often offer advantages of solubility, tissue compatibility, or delayed release in a subject. Prodrugs include compounds in which a hydroxy, amino, or mercapto group is bonded to any group that cleaves to form a free hydroxy, free amino, or free mercapto group, respectively, when the prodrug is administered to a subject. It is well known to prepare prodrugs from carboxylic acids, for example, in the form of a carboxylic acid ester or thioester.

[0133] As used herein, the terms "purified" or "isolated" can refer to a composition (e.g., a peptide composition) that has been subjected to a process to remove various other components (e.g., fractionation), wherein the composition substantially retains its expressed biological activity.

[0134] The term "sample" as used herein includes, for example, environmental and biological samples. Environmental samples include materials 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., feces), tissues, liquid foods (e.g., milk), and solid foods (e.g., plants). For example, a lung sample may be collected by bronchoalveolar lavage (BAL), which contains fluid and cells from lung tissue. Biological samples may include cells, tissue extracts, body fluids, chromosomal or extrachromosomal elements isolated from cells, 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.

[0135] As used herein, the term "biologically active" refers to any molecule that has a structural, regulatory, or biochemical function. For example, biological activity can be determined by, for example, restoring wild-type growth in cells lacking protein activity. Cells lacking protein activity can be produced by a number of methods (i.e., point mutations and frameshift mutations, for example). Complementation is achieved by transfecting cells lacking protein activity with an expression vector that expresses the protein, its derivative, or a portion thereof.

[0136] As used herein, the term "label" or "detectable label" refers to any composition detectable by spectral, photochemical, biochemical, immunochemical, electrical, optical, or chemical means. Such labels include biotin for staining with labeled streptavidin conjugates, magnetic beads (e.g., Dynabeads (登録商標) ), fluorescent dyes (e.g., fluorescein, Texas Red (登録商標) , rhodamine, green fluorescent protein, etc.), radiolabels (e.g., 3 H, 125 I, 35 S, 14 C or 32P), enzymes (e.g., horseradish peroxidase, alkaline phosphatase, and others commonly used in 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. Patents 3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149; and 4,366,241 (all of which are incorporated herein by reference in their entirety). Labels contemplated by the present invention can be detected by conventional methods. For example, radiolabels can be detected with photographic film or a scintillation counter, and fluorescent markers can be detected using a photodetector to detect emitted light. Enzyme labels typically involve providing an enzyme and a substrate and detecting the reaction product produced by the action of the enzyme on the substrate, while calorimetric labels are detected simply by visualization of the colored label.

[0137] As used herein, the term "conjugate" refers to any compound formed by the joining of two or more moieties.

[0138] As used herein, a "moiety" or "group" refers to any type of molecular configuration 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 moieties is linked and substituted at a position to become part of the molecular configuration of the conjugate. While multiple moieties can be directly covalently linked, it is not intended that the linkage of two or more moieties must be direct to each other. A linking group, bridging group, or bonding group refers to any molecular configuration that connects moieties by a covalent bond, such as, but not limited to, one or more amide groups. Furthermore, while a conjugate can be unsubstituted, a conjugate can have a variety of additional substituents attached to the linking group and / or attached to the moieties.

[0139] As used herein, "polymer" or "polymeric group" refers to a chemical species or group composed of repeatedly linked moieties. Within certain embodiments, the number of repeating moieties is preferably three or more, or more than ten. The linked moieties may be identical in structure or may vary in structure. A "monomeric polymer" or "homopolymer" is a polymer containing the same repeating, asymmetric subunits. A "copolymer" is a polymer derived from two or more types of monomeric species (i.e., two or more different chemically asymmetric subunits). A "block copolymer" is a polymer composed of two or more types of polymeric subunits covalently linked.

[0140] As used herein, the term "substituted" refers to at least one hydrogen atom in a molecular configuration being replaced with a non-hydrogen substituent. The number of substituents present depends on the number of hydrogen atoms available for substitution, including the replacement of more than one hydrogen atom bonded to a single atom (e.g., in the case of a carbon or silicon atom, which may be mono-, di-, or tri-substituted, or in the case of a nitrogen atom, which may be mono-, di-, or tri-substituted, or in the case of an oxygen or sulfur atom, which may be mono-substituted). In the case of an oxo substituent ("=O"), two hydrogen atoms are replaced (this results in, for example, -(CH)-C(=O)-CH when two hydrogen atoms on the central carbon atom of -CH-CH-CH are replaced). When substituted, one or more of the following groups are "substituents": Substituents may be halogen (e.g., F, Cl, Br, I), hydroxy (OH), oxo, cyano (CN), 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., CH2Cl, CH(CH3)Br, etc.), O-alkyl(alkoxy), 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., CH2Cl, CH(CH3)Br, etc.), O-alkyl(alkoxy), cycloalkyl (e.g., cyclopropyl ... O-cycloalkyl (e.g., OCH3, OCH2CH3, OCH(CH3)2, 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 OCH2Cl), O-aryl (e.g., O-phenyl), O-heteroaryl, O-heterocyclyl, thioalkyl (e.g., S-CH3), hydroxyalkyl (e.g., CH2OH), alkyl ether (e.g., CHOCH3), alkynyl (e.g., -C≡CR f ), alkenyl (e.g., -CR f=CR f R 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 -NR f R g , -NR f C(=O)R g , -NR f C(=O)NR f NR g , -NR f -C(=O)OR f SO2R g , -C(=O)R f , -C(=O)OR f , -OR f , -C(=O)NR f R g , -OC(=O)NR f R g , -SR f , -SOR f , -S(=O)2R f , -OS(=O)2R f and -S(=O)OR f (where each R f and R g may be the same or different and are independently hydrogen, alkyl (e.g., CH), substituted alkyl, haloalkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heterocyclyl, substituted heterocyclyl, heterocycloalkyl, substituted heterocycloalkyl, heteroaryl, or substituted heteroaryl. Additionally, the above substituents may be further substituted with one or more substituents as defined above, such that the substituent may comprise, for example, a substituted alkyl, substituted aryl, substituted arylalkyl, substituted heterocyclyl, or substituted heterocycloalkyl.

[0141] As used herein, the term "unsubstituted" refers to any compound that does not contain additional substituents attached to the compound. For example, an unsubstituted compound refers to the chemical structure of a compound that does not have any additional substituents attached (e.g., no non-hydrogen substituents). For example, unsubstituted proline is a proline amino acid, even though the amino group of the proline can be considered to be disubstituted with alkyl groups.

[0142] As used herein, the term "bond" in describing a substituent having atoms on both sides refers to the absence of the substituent. For example, in the four-atom sequence ABCD, when B and C are both listed as bonds, the resulting sequence is AD, which is two atoms. When only B is listed as a bond, the resulting sequence is ACD, which is three atoms.

[0143] As used herein, the term "alkyl" refers to any straight-chain or branched, acyclic or cyclic, unsaturated or saturated aliphatic hydrocarbon containing 1 to 10 carbon atoms, while the term "lower alkyl" has the same meaning as alkyl, except containing 1 to 3 carbon atoms. The term "higher alkyl" has the same meaning as alkyl, except containing 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, tert-butyl, isopentyl, and the like. As used herein, a methyl substituent may be depicted as "CH" or "Me," or as a terminal bond where no specific atom is designated.

[0144] As used herein, the term "cycloalkyl" refers to saturated and unsaturated cyclic alkyls. Representative saturated cyclic alkyls include C-C 14(e.g., C3-C7)cycloalkyls include, but are not limited to, 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 may also be referred to herein as "homocycles" or "homocyclic rings."

[0145] As used herein, the term "bicyclic compound" encompasses the described "bridged," "fused," and "spiro" compounds.

[0146] As used herein, the term "spiro" or "spirocyclic" refers to a chemical structure having at least two rings that share one common atom. The rings may be cycloalkyl, heterocyclyl, or a combination thereof, and may contain one or more aryl or heteroaryl rings. Examples include, but are not limited to, spirocyclic cyclopropane, spirocyclic aziridine, spirocyclic cyclobutane, spirocyclic azetidine, spirocyclic oxetane, spirocyclic cyclopentane, spirocyclic pyrrolidine, spirocyclic 1,3-dioxolane, spirocyclic dioxane, spirocyclic oxathiolane, spirocyclic thiazolidine, spirocyclic cyclohexane, spirocyclic piperidine, and spirocyclic piperidine, 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, 1,4-dioxaspiro[4.5]decane, 1,4-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, 1,3-dihydrospiro[indene-2,3′-pyrrolidine], and 3,4-dihydro-2H-spiro[naphthalene-1,4′-piperidine].

[0147] As used herein, the term "bridged" refers to a compound containing two non-adjacent atoms common to two rings. Exemplary embodiments include, but are not limited to, norbornane, bicyclo[1.1.1]pentane, bicyclo[2.2.1]heptane, 1,4-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, and other bridged piperazines and bridged piperidines.

[0148] As used herein, the term "fused" 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 the rings contain two and only two adjacent atoms common to each of two or more rings in a consecutive series of ortho-fused rings (ortho- and peri-fused). Exemplary embodiments are pentalene and dibenzoxepin (ortho-fused) and pyrene (ortho- and peri-fused). Ortho-fused systems have "n" common sides and "2n" common atoms, while peri-fused systems have "n" common sides and fewer than "2n" common atoms. 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 the same or a different ring, such as a pyrrolidine ring fused to another pyrrolidine ring (e.g., octohydropyrrolo[3,4-c]pyrrole) or a cyclohexane ring (e.g., octohydro-1H-indole or octohydro-1H-isoindole). Other examples include fused aryl or heteroaryl rings, such as a pyridine ring fused to a cycloalkyl ring (e.g., cyclopentane or cyclohexane) or a heterocyclyl ring (e.g., tetrahydrofuran or tetrahydropyran).

[0149] As used herein, the term "aromatic" or "aryl" refers to any aromatic carbocyclic (i.e., all of the ring atoms are carbon) substituent, including, but not limited to, phenyl (from benzene), tolyl (from toluene), xylyl (from xylene), or polycyclic ring systems (e.g., naphthyl (from naphthalene) and anthracenyl (from anthracene)).

[0150] As used herein, the term "arylalkyl" or "aralkyl" refers to any alkyl having at least one alkyl hydrogen atom replaced with an aryl moiety, such as, but not limited to, benzyl, -(CH)phenyl, -(CH)phenyl, -CH(phenyl)phenyl.

[0151] As used herein, the term "halogen" refers to any fluoro, chloro, bromo, or iodo moiety.

[0152] As used herein, the term "haloalkyl" refers to any alkyl in which at least one hydrogen atom (and inclusive of all hydrogen atoms) has been replaced with a halogen atom, such as, for example, trifluoromethyl, dichloromethyl, difluoromethyl, monofluoromethyl, monobromomethyl, 1,1,1-trifluoroethyl, etc.

[0153] As used herein, the terms "heteroaromatic" or "heteroaryl" refer to any aromatic heterocyclic ring, including, but not limited to, both monocyclic and bicyclic ring systems, having 5 to 10 or more ring members, at least one heteroatom selected from nitrogen, oxygen, or sulfur, and containing at least one carbon atom. Heteroaryl rings may be attached as substituents via ring heteroatoms or carbon atoms. Representative heteroaromatics include 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, and 1,2,3-triazole. and the like.

[0154] The term "heteroarylalkyl" as used herein means any alkyl having at least one alkyl hydrogen atom replaced with a heteroaryl moiety, such as -CH2 pyridinyl, -CH2 pyrimidinyl, etc.

[0155] As used herein, the term "heterocycle" or "heterocyclyl" or "heterocyclic ring" refers to a non-aromatic ring that is saturated or unsaturated and contains one or more heteroatoms independently selected from nitrogen, oxygen, sulfur, and silicon, where each of the nitrogen and sulfur heteroatoms may be in an oxidized state, each of the nitrogen and silicon heteroatoms may be substituted or unsubstituted, and the nitrogen heteroatom may be optionally quaternized, including bicyclic rings in which any of the above heterocycles is fused to an aryl or heteroaryl ring. Heterocyclic rings may be attached as substituents via ring heteroatoms or carbon atoms. In various embodiments, the heterocycle can contain 3 to 14 or more ring atoms (e.g., a 3- to 7-membered monocyclic ring or a 7- to 10-membered bicyclic ring) and includes 2H-azirine, azetidine, 2,3-dihydroazeto, 1,3-diazetidine, 2H-oxete, thietane, 2H-thiette, azetidin-2-one, morpholine, thiomorpholine, pyrrolidinone, pyrrolidinine, 2-pyrroline, 3-pyrroline, pyrazolidine, 2-pyridinone ... Imidazolidine, 2-imidazoline, imidazolidine, piperidine, piperazine, pyridin-2-one (e.g., 2-pyridone and 1-methyl-2-pyridone), ethylene oxide (oxirane), ethyleneimine (aziridine), ethylene sulfide (thiirane), oxetane, propylene oxide, 1,3-dioxolane, 1,2-oxathiolane, 1,3-oxathiolane, sulfolane, 2,4-thiazolidinedione ion, succinimide, 2-oxazolidone, dioxane, hydantoin, valerolactam, tetrahydrofuran, tetrahydropyran, 2H-pyran, 4H-pyran, thiane, 2H-thiopyran, 1,3-dithiane, 1,4-dithiane, 1,3,5-trithiane, pyrrolizidine, 1,4,5,6-tetrahydrocyclopenta[b]pyrrole, tetrahydropyridine, tetrahydropyrimidine, tetrahydrothio Phen, tetrahydrothiopyran, indoline, isoindoline, decahydroisoquinoline, decahydroquinoline, 1,2,3,4-tetrahydroquinoline, 1,2-dihydroquinoline, 2H-benzo[e][1,3]oxazine, 2H-benzo[b][1,4]oxazine, quinolin-2(1H)-one, isoquinolin-1(2H)-one, quinuclidine, 1-azaadamantane, 2-azaadamantane, 2,Examples of dihydroazepines include, but are not limited to, 3-dihydroazepines, 2,5-dihydroazepines, oxepanes, azonanes, spiro[cyclobutane-1,3'-indole], 1-oxaspiro[4,5]decane, 1,6-dioxaspiro[3,4]octane, 2-oxa-7-azaspiro[3,5]nonane, 1,4-dioxa-7-azaspiro[4,4]nonane, 1,3-diazaspiro[4,4]non-2-en-4-one, 2,9-diazaspiro[5,5]undecan-1-one, 8-azaspiro[4,5]decane-7,9-dione, 1,4-dithia-7-azaspiro[4,4]nonane, and the like.

[0156] The term "heterocycloalkyl" as used herein refers to any alkyl having at least one alkyl hydrogen atom replaced with a heterocycle, such as -CH2morpholinyl.

[0157] As used herein, the term "alkylamino" refers to at least one alkyl moiety attached through a nitrogen bridge (i.e., -N-(alkyl)amino), such as, but not limited to, methylamino, ethylamino, dimethylamino, diethylamino, and the like. n , where n=1 or 2, e.g., alkylamino or dialkylamino).

[0158] The term "alkyloxy" or "alkoxy" as used herein means any alkyl moiety attached through an oxygen bridge (ie, --O-alkyl), such as, but not limited to, methoxy, ethoxy, and the like.

[0159] The term "thioalkyl," as used herein, means any alkyl moiety attached through a sulfur bridge (ie, --S-alkyl), such as, but not limited to, methylthio, ethylthio, and the like.

[0160] As used herein, the term "alkenyl" refers to an unbranched or branched hydrocarbon chain having one or more carbon-carbon double bonds, and may also be referred to herein 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, and 4-(2-methyl-3-butene)-pentenyl. Alkenyl groups can be unsubstituted or substituted with one or two suitable substituents.

[0161] The term "alkynyl," as used herein, refers to an unbranched or branched hydrocarbon chain having one or more carbon-carbon triple bonds, and may also be referred to herein 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-1-butynyl, 4-propyl-2-pentynyl, and 4-butyl-2-hexynyl. Alkynyl groups can be unsubstituted or substituted with one or two suitable substituents.

[0162] As used herein, "reactive group" refers to a nucleophile, electrophile, or radically active group, i.e., a group that reacts in the presence of radicals. A nucleophile is a moiety that forms a chemical bond with its reaction partner (electrophile) by donating both bonding electrons. An electrophile accepts these electrons. Nucleophiles can participate in nucleophilic substitution, whereby the nucleophile is attracted to a full or partial positive charge on an element and displaces the group to which it is attached. Alternatively, nucleophiles participate in the displacement of carbonyl groups. Carboxylic acids are often rendered electrophilic by forming succinyl esters, and these esters react with aminoalkyls to form amides. Other common nucleophilic groups are thiolalkyls, hydroxylalkyls, primary and secondary amines, and carbon nucleophiles, such as enols and alkylmetal complexes. Other preferred methods for ligating proteins, oligosaccharides, and cells using reactive groups have been disclosed (Lemieux et al., Trends in Biotechnology 1998, 16, 506, incorporated herein by reference in its entirety). In yet another preferred method, reactive groups for Staudinger ligation, i.e., "click chemistry" of an azide-containing moiety with an alkynyl reactive group to form a triazole, are provided. Michael addition of a carbon nucleophile enolate with an electrophilic carbonyl or Schiff base formation of a nucleophilic primary or secondary amine with an aldehyde or ketone can also be utilized. Other methods of bioconjugation are provided (Hang et al., Accounts of Chemical Research 2001, 34, 727 and Kiick et al., Proc Natl Acad Sci USA 2002, 99, 19, both incorporated herein by reference in their entirety).

[0163] As used herein, the term "biocompatible" refers to any material that does not induce a substantial adverse response in the host. When introducing a foreign object into a living organism, there is always the concern that the object will induce an immune response, such as an inflammatory response, that will have a negative impact on the host. In the context of the present invention, biocompatibility is evaluated according to the application for which it is designed: for example, a bandage is considered biocompatible with the skin, and an implanted medical device is considered biocompatible with the body's internal tissues. Preferably, biocompatible materials include, but are not limited to, biodegradable and biostable materials. A substantial adverse response will not occur if an implant containing the material closely integrates with the implant site within the host animal and the response is better than the tissue response recognized and established as suitable for the material as provided by ASTM. The ASTM Subcommittee on Biocompatibility Test Methods, F04.16, develops biocompatibility standards for medical and surgical materials and devices, including E1262-88, F612-20, F719-20e1, F720-17, F748-16, F749-20, F750-20, F756-17; F763-04, F813-20, F895-11, and F98 These include F1-04, 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 used in contact with the bloodstream must be made of materials that meet hemocompatibility standards. One of these tests is for hemolysis, i.e., damage to red blood cells that can result in destruction of the cells, as described in F756-17 Standard Practice for Evaluation of Hemolytic Characterization of Materials.

[0164] As used herein, the term "bioactive agent" refers to any of a variety of chemical moieties that bind to biomolecules, including, but not limited to, peptides, proteins, enzymes, receptors, substrates, lipids, antibodies, antigens, and nucleic acids. In certain preferred embodiments, the bioactive agent is a biomolecule, although it is not intended to limit bioactive agents to biomolecules. In other preferred embodiments, the bioactive agent provides hydrophobic, hydrophilic, or electrostatic interactions, such as polycarboxylic acids that are anionic at physiological pH. In other preferred embodiments, alkaline growth factors (having an isoelectric point greater than 7) are retained by favorable electrostatic interactions with polycarboxylates and subsequently released in a controlled and sustained manner.

[0165] "Cancer" is a term used to describe the 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 specific 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, kidney 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, gastric cancer, urothelial carcinoma (including locally advanced or metastatic urothelial carcinoma), bladder cancer, hepatocellular carcinoma, breast cancer, and head and neck cancer.

[0166] The term "stereoisomers" refers to compounds that have the same atomic connectivity but differ in the arrangement of the atoms 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 is present in an enantiomeric excess of more than 95%, preferably more than 97%, with respect to the absolute configuration of the chiral centers.

[0167] The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomeric isomers, (D)-isomers, (L)-isomers, atropisomers, tautomers, and racemic and other mixtures thereof, such as enantiomeric or diastereomeric adduct mixtures, all of which are within the scope of the present invention. To the extent that the compounds of the present invention as defined herein exist as optically active or racemic due to one or more asymmetric carbon atoms, the present invention includes within its definition any such optically active or racemic form. Synthesis of optically active compounds can be carried out by standard techniques of organic synthesis well known in the art, such as synthesis from optically active starting materials or resolution of racemic compounds. Similarly, the enantiomeric or diastereomeric purity of a compound can be assessed using standard laboratory techniques.

[0168] The pharmaceutical compositions of the present invention may be in a form appropriate for the desired route of administration. When the compositions are administered orally, any suitable orally deliverable dosage form may be used, including oral liquids such as suspensions, syrups, elixirs, emulsions, and solutions, including, but not limited to, water, glycols, oils, alcohols, and the like; or solid carriers such as starches, sugars, kaolin, diluents, lubricants, binders, disintegrants, and the like, for powders, pills, capsules, and tablets. Due to ease of administration, tablets and capsules represent the most advantageous oral dosage unit form. 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 saline solution, glucose solution, or a mixture of saline and 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 with other oils. In compositions suitable for transdermal administration, the carrier optionally comprises a penetration enhancer and / or a suitable wetting agent, optionally in combination with suitable additives, where the additives can facilitate application of the composition to the skin and / or facilitate preparation of the composition to be delivered.These compositions can be administered in various ways, for example, as a transdermal patch or ointment.The acid or base addition salts of the compounds of the present invention have increased water solubility compared to the corresponding neutral forms of the compounds, and therefore are typically more suitable for aqueous compositions.

[0169] The pharmaceutical compositions of the present invention may contain one or more fillers, diluents, adjuvants, vehicles or other additives to facilitate the preservation and / or administration of the active ingredients contained therein.

[0170] In exemplary embodiments, the pharmaceutical compositions of the present invention can contain one or more additional therapeutic agents, for example, to increase efficacy or reduce unwanted side effects.In certain embodiments, the pharmaceutical compositions further contain one or more additional therapeutic agents useful for treating or preventing diseases directly or indirectly mediated by PI3K.Examples of such agents include, but are not limited to, agents for treating or preventing cancer, Huntington's disease, cystic fibrosis, liver fibrosis, kidney fibrosis, pulmonary fibrosis, skin fibrosis, rheumatoid arthritis, diabetes or heart failure.

[0171] In certain embodiments, the additional therapeutic agent that is included is anticancer drug.The example of anticancer drug includes but is not limited to DNA damaging cytotoxic drug, alkylating agent, for example cyclophosphamide, dacarbazine and cisplatin; antimetabolite, for example methotrexate, mercaptopurine, thioguanine, fluorouracil and cytarabine; plant alkaloid, for example vinblastine and paclitaxel; antitumor antibiotic, for example doxorubicin, bleomycin and mitomycin; hormone / antihormone, for example prednisone, tamoxifen and flutamide; other types of anticancer drug, for example asparaginase, rituximab, trastuzumab, imatinib, retinoic acid and derivative, colony stimulating factor, amifostine, camptothecin, topotecan, thalidomide analogue, for example lenalidomide and proteasome inhibitor, for example velcade.

[0172] In another embodiment, the present invention provides a method for preventing or treating a disease caused by abnormal cell proliferation and / or differentiation in a subject in need of treatment, comprising administering to the subject a therapeutically effective amount of one or more compounds of the present invention.In some embodiments, the method for preventing or treating a disease comprises administering to a subject in need thereof a composition comprising an effective amount of one or more compounds of the present invention and a pharmaceutically acceptable carrier.The administered composition may further comprise a therapeutic agent, for example, an anti-cancer agent.

[0173] The compounds of the invention are defined herein by chemical structure and / or chemical name, and are generally listed according to the IUPAC or CAS nomenclature system. Abbreviations well known to those skilled in the art may be used. When a compound is described by both a chemical structure and a chemical name, and the chemical structure and chemical name conflict, it is intended that the chemical structure be determinative of the compound's identity.

[0174] The present invention includes compounds labeled with various radioactive or non-radioactive isotopes. Examples of atomic isotopes include deuterium ( 2 H), tritium ( 3 H), iodine-125( 125 I), carbon-14( 14 C), nitrogen-15( 15 N), sulfur-35( 35 S) and chlorine-36( 36 In exemplary embodiments, one or more hydrogen atoms in the compounds of the invention may be replaced with deuterium. In various embodiments, the compounds of the invention may contain at least one deuterium atom, or two or more deuterium atoms, or three or more deuterium atoms, etc. As described herein, the compounds of the invention may contain tritium ( 3 H), iodine-125( 125 I) and carbon-14( 14 Radiolabeled compounds may also be radiolabeled with radioactive isotopes such as C. Radiolabeled compounds are useful as therapeutic or prophylactic agents, provide research reagents such as in assays, and / or provide diagnostic agents for techniques such as in vivo imaging. Synthetic methods for incorporating isotopes into organic compounds are well known in the art.

[0175] In certain embodiments of the invention, a compound of the invention as defined herein (e.g., a compound of any of Formula (1), (2), (3), (4), (5) or (6)) or a pharmaceutically acceptable salt thereof is present as a single enantiomer in an enantiomeric excess (% ee) of ≥ 95%, such as ≥ 98%, for example ≥ 99%.

[0176] In certain embodiments of the invention, in a pharmaceutical composition comprising a compound of the invention as defined herein (e.g., a compound of Formula I) or a pharmaceutically acceptable salt thereof, the compound is present as a single enantiomer in an enantiomeric excess (% ee) of ≥ 95%, such as ≥ 98%, for example ≥ 99%.

[0177] In exemplary embodiments of the invention, the disease or disorder treated by the compounds of the invention is selected from congenital lipomatous overgrowth, vascular malformations, epidermal nevi, scoliosis / skeletal and spinal cord syndrome (CLOVES), mosaic tissue overgrowth syndrome, venous malformations and brain malformations associated with severe epilepsy, or PIK3CA-associated overgrowth syndrome (PROS) (Keppler-Noreuil et al., Am J Med Genet A. 2015, 167A, 287; Kurek et al. Am. J. Hum. Genet. 2012, 90, 1108).

[0178] In an exemplary embodiment of the invention, the cancer to be treated is a cancer carrying a PI3K H1047 mutation (e.g., H1047R) (Thorpe et al., Nat Rev Cancer 2015, 15, 7).

[0179] The compounds of the present invention (e.g., defined by formulas (1) to (6)) are PI3Kα H1047R mutant-selective inhibitors that typically exhibit greater selectivity for the H1047R mutation than for the wild-type. That is, the compounds can selectively reduce the amount of phosphorylated AKT (pAKT) and reduce proliferation in PI3Kα H1047R mutant cell lines, preferably across several tumor types.

[0180] The PI3K H1047R mutant selective inhibitors of the present invention (e.g., as defined by Formulas (1)-(6)) administered in combination with a selective estrogen receptor degrader (SERD), such as, but not limited to, fulvestrant, elakestrant, camizestrant, or bepdegestrant, may show a combination benefit leading to tumor regression in ER+ / PI3K H1047R mutant tumors, such as, but not limited to, the breast cancer xenograft model T47D, at doses where either agent alone results in little or no regression.

[0181] The PI3K H1047R mutant selective inhibitors of the present invention (e.g., as defined by Formulas (1)-(6)) administered in combination with a HER2 inhibitor, such as, but not limited to, tucatinib or trastuzumab, may show 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 either agent alone results in little or no regression.

[0182] The compounds of formula (1) of the present invention may generally be prepared according to the synthetic routes defined in Schemes 1-11.

[0183] In Scheme 1, the synthesis can begin with an appropriately substituted 2,3-dihydro-1H-inden-1-one. When R7 is methyl, 1 is commercially available. Otherwise, the starting material can be prepared via established methods known to those skilled in the art. Nitrosation of indenone 1 to convert it to oxime derivative 2 can be achieved using established methods (see, e.g., Touster, O.; Org. Reactions, VII, 1953, 327). A Beckmann-type rearrangement mediated by phosphorus pentachloride can convert oxime 2 to chloroisoquinolone 3 (Cushman, M.; Dekow, FW Tetrahedron 1978, 34(10), 1435-9). Alkylation of isoquinolone intermediate 3 with an appropriate electrophile and base can provide nitrogen-substituted isoquinolone 4. When R6 is methyl, this can be achieved with methyl iodide and a suitable base (e.g., sodium hydride). Other electrophiles and alkylating agents known to those skilled in the art can also be used. To convert the bromide of isoquinolone 4 to methyl ketone 5, a Stille coupling reaction can be carried out using an appropriate tin reagent, such as (α-ethoxyvinyl)-tributyltin, followed by acid hydrolysis (Sugiyama, et al., Bull. Chem. Soc. Jpn. 1987, 60(2), 767-768). Alternatively, conversion of 4 to 5 can be achieved by other established methods, such as Heck coupling with an appropriate enol-ether, followed by acid hydrolysis (Mingcui, L. et al., Org. Biomol. Chem., 2010, 8, 2012-2015). Reduction of ketone 5 to secondary alcohol 6 can then be achieved using an appropriate hydride reducing agent, such as sodium borohydride.It is understood that, in addition to the methods described in Scheme 1, there are other reported methods available for the preparation of isoquinolones such as 3 or 4 and their derivatives (see, e.g., Li, B. et al., Tetrahedron Letters 2010, 51(29), 3748-3751 and Wang, R.; et al., Organic & Biomolecular Chemistry 2011, 9(16), 5802-5808). [ka]

[0184] In Scheme 2, alcohol 6 can be converted to anthranilic acid derivative 8 via several different methods. The alcohol functionality is first converted to a leaving group such as bromide or mesylate using commonly known methods. Nucleophilic displacement of 7 with an anthranilic acid ester (methyl, t-butyl, or other commonly used esters can be used) affords compound 8. An alternative method is a direct Mitsunobu-type reaction of anthranilic acid ester 9 with alcohol 6 to directly afford 8. In some cases, the temporary use of an activating group for the anthranilic acid amine functionality (e.g., 2,4-dinitrobenzenesulfonyl group) can facilitate the Mitsunobu reaction. 2,3-Dichloro-5,6-dicyanobenzoquinone (DDQ) and triphenylphosphine can be used for the direct reaction of alcohol 6 with anthranilic esters (Shalit, T.; et al., Tetrahedron Letters 2010, 51, 5988-5991; Iranpoor, N.; et al., Tetrahedron 2009, 65, 3893-3899; Panday, SK, Mini-Reviews in Organic Chemistry 2019, 16(2), 127-140; Fukuyama, Tohru; et al., Tetrahedron Letters 1997, 38(33), 5831-5834). [ka]

[0185] An alternative synthesis of a single-enantiomer intermediate similar to 8 is described in Scheme 3. This reaction sequence utilizes the formation of a chiral sulfinyl imine for stereochemical control. Such methods have been widely reported. Ketone 5 can be converted to chiral sulfinyl-imine 10 by known procedures, which can then be reduced to sulfinyl-amine 11 in a stereocontrolled manner using an appropriate 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). Use of the R isomer of the sulfinyl group generally results in predominantly the R,R isomer of the product 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 for the reduction of imines and can often provide enhanced stereocontrol in similar reductions (Hua et al., Synthesis 1991, (11), 970-4; Zhu et al., Journal of Chemical Research 2015, 39(7), 390-393). The major isomer can be separated from the other minor isomers by standard chromatographic means. As shown in the above cited references, by judicious selection of the antipodes of the sulfinyl-imines and reducing agents, either antipodes of the sulfinyl-amines can be obtained. The sulfinyl-amines can be cleaved into single enantiomers of the chiral amines 12 using standard conditions (e.g., hydrogen chloride in dioxane).Standard coupling reactions (e.g., Ullmann or Buchwald-Hartwig coupling) of amine 1 with aryl iodides 13 can then provide anthranilic acid derivatives 14 (Yang et al., Organic Process Research & Development 2022, 26(6), 1690-1750; Surry and Buchwald, Chemical Science 2011, 2(1), 27-50). [ka]

[0186] Certain final compounds may be prepared as outlined in Scheme 4. Intermediate 8 may be converted to bromide or iodide 15. 8 or 15 may then be reacted with an amine under appropriate coupling conditions, where R I and R II can be alkyl or aryl or R I and R II One of R can be hydrogen. I and R II may be joined together to form a ring. In some cases, R I and / or R II The group can be further elaborated before subsequent steps. Alternatively, halogen-substituted isoquinolones 8 or 15 can be reacted with alkyl or aromatic boronic acids or boronic acids or alkyl carboxylic acids or alkynes under appropriate coupling conditions to give carbon-linked versions of 16. Ester 16 can then be converted to carboxylic acid 17 using standard ester hydrolysis conditions. The resulting products can then be resolved into their individual enantiomers (18 and 19) using chiral chromatography (HPLC or SFC). [ka]

[0187] Single enantiomers of isoquinoline 21 can be purified directly from enantiomerically pure intermediate 20 using chemistry similar to that shown in Scheme 4, as shown in Scheme 5. [ka]

[0188] In some cases, the reaction sequence can be adjusted as shown in Scheme 6. In this case, Buchwald-Hartwig coupling of intermediate 10 with an appropriately substituted amine can provide amine-substituted isoquinolone 21. The sulfinyl group can then be removed from compound 21 to give amine 22, which can then undergo Buchwald-Hartwig coupling with (for example) 2-iodobenzoate (or other appropriately substituted benzoic acid derivative). Ester hydrolysis of the product can then provide a benzoic acid such as 23. Alternatively, intermediate 22 can be coupled to an electrophile-containing heterocycle (e.g., methyl 6-chloro-3-fluoropyridine-2-carboxylate 24 or other appropriately substituted heterocycle) via S-coupling. N A subsequent ester hydrolysis step can then provide carboxylic acid compounds such as 25. [ka]

[0189] Compounds with oxygen or sulfur bond substitution from the 3-position of the isoquinolone ring can be prepared as shown in Scheme 7. Intermediate 14 can undergo Buchwald-Hartwig coupling with an appropriate thiol 26 to give thioether 27. Ester hydrolysis can then provide carboxylic acid-containing compounds such as 28. Alternatively, intermediate 14 can be condensed with an appropriate alcohol 29 to give S-substituted thioethers 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, N The resulting ether 30 can then be subjected to ester hydrolysis conditions to give compounds such as 31. [ka]

[0190] When the substitution from the 3-position of the isoquinolone ring is an alkyl or alkenyl group, these compounds can be prepared as shown in Scheme 8. Intermediate 8 can be converted via Suzuki coupling reaction with the appropriate alkenyl-boronate (or boronic acid) 32 (Stanforth, SP Tetrahedron 1998, 54(3 / 4), 263-303), followed by removal of the ester to give carboxylic acid 33. Racemic 33 can then be resolved by chiral chromatographic methods to give the individual enantiomers 34 and 35. Alternatively, the double bond of 33 can be reduced under standard hydrogenation conditions (e.g., hydrogen and palladium catalyst). After this chiral chromatographic separation, enantiomers 36 and 37 can be obtained. R V and R VI When does not form a symmetric configuration, additional isomers may be obtained, which may also be separated by chromatography. [ka]

[0191] 2-Fluoroisoquinolones can be prepared as shown in Scheme 9. Isoquinolone intermediate 38 can be prepared using Selectfluor TM Reaction with a suitable electrophilic fluorinating agent such as (1-chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane bis(tetrafluoroborate) followed by hydrolysis can provide 2-fluoroisoquinolone 39. [ka]

[0192] An alternative method for synthesizing carboxylic acid-containing isoquinolones is shown in Scheme 10. Suzuki coupling reaction between chloro-containing intermediate 10 and an appropriate aryl or heteroaryl boronate (or boronic acid), followed by removal of the sulfinyl group, can provide aryl-substituted isoquinolone amine 40. Ullmann or Buchwald-Hartwig coupling between amine 40 and a halogen-substituted benzoate (e.g., 2-iodobenzoic acid methyl ester or other appropriately substituted aryl halide), followed by ester hydrolysis, can provide carboxylic acid compounds such as 41. Alternatively, intermediate 40 can be converted to an S-type isoquinolone by S-coupling with a halogen-substituted heteroaryl ester (e.g., methyl 6-chloro-3-fluoropyridine-2-carboxylate or other appropriately substituted heterocycle). N Ar reaction can provide carboxylic acid compounds such as 42 after ester hydrolysis. [ka]

[0193] Further synthesis of carboxylic acid-containing isoquinolones is described in Scheme 11. Chloro intermediate 14 can be converted to boronate 43 under appropriate conditions, such as treatment with PdCl(dppf) and bis(pinacolato)diboron. Boronate 43 can then be reacted with an aryl or heteroaryl halide to afford functionalized isoquinolone 44 after hydrolysis. [ka]

[0194] The chemistry depicted in Schemes 1-11 illustrates various methods for synthesizing the described compounds. It is understood that other variations of these methods can be used, and that the exact protecting groups, reaction order, or particular types of transition metal-catalyzed coupling reactions can be selected as needed and known to those skilled in the art.

[0195] The following compounds of formula (4) represent various embodiments of the present invention, with variations in the structures of R1' and R5 and all other atoms shown. The carbon atom marked with an * is a chiral center and exists as an (R)- and (S)-racemic mixture or as either the (R)- or (S)-enantiomer. Substituents listed in parentheses (for listed embodiments of formula (4) or R5 or R1') indicate individual compounds containing any one of the substituents. When present in an embodiment of R5 or R1', each X is independently N or CH; each X a are independently O or CH2; each X b are independently O, CH2 or N k N m and each R h and each R i are independently selected from H, CH, c-Pr, c-Bu, CF, and OH; each R j are independently selected from CF3, CH2CF3, CH2CF2H, OCH3, OCF3, OCH2CF3, Oct-Pr, aryl, heteroaryl, COCH3, and CO2CH3; each R k and R m is CH3, CH2CH3, CH2CH2CH3, CH2CH2OH, CH2CH2N(CH3)2, COCH3; and each "A" is selected from O, S, S(O), and S(O)2. All chiral centers in the R5 structures below that are not specified exist as (R)- and (S)-racemic mixtures or as (R)- or (S)-enantiomers. [ka] wherein R5 is selected from: [ka] [ka] [ka] [ka] [ka] [ka] And R1' is [ka] is selected from

[0196] The following compounds of formula (5) represent various embodiments of the present invention, with variations in the structure of R5 and all other atoms shown. The carbon atom marked with an * is a chiral center and exists as an (R)- and (S)-racemic mixture or as either the (R)- or (S)-enantiomer. Substituents listed in parentheses (for listed embodiments of formula (5) or R5) indicate individual compounds containing any one of the substituents. When present in an embodiment of R5, each R 18 are independently H, C1-C4 alkyl, C3-C7 cycloalkyl, halogen, CN, CF3, OCF3, CFH2, or CF2H; and each X is independently N or CH. All chiral centers in the R5 structures below that are not specified exist as (R)- and (S)-racemic mixtures or as (R)- or (S)-enantiomers. [ka] wherein R5 is: [ka] is selected from the following.

[0197] The following compounds of formula (6) represent various embodiments of the present invention, with variations in the R5 structure and all other atoms shown. The carbon atoms marked with an * are chiral centers and exist as (R)- and (S)-racemic mixtures or as (R)- or (S)-enantiomers. Substituents listed in parentheses (for listed embodiments of formula (6) or R5) indicate individual compounds containing any one of the substituents. When a dashed line (----) is present in an R5 embodiment, the bond can be saturated or unsaturated. All chiral centers in the R5 structures below that are not specified exist as (R)- and (S)-racemic mixtures or as (R)- or (S)-enantiomers. [ka] wherein R5 is: [ka] [ka] is selected from the following.

[0198] experiment All commercially available solvents and reagents were used as received. 1 H NMR spectra were recorded using a Bruker Avance III HD 300 MHz or a Bruker Avance III HD 400 MHz. MS samples were analyzed on a Shimadzu LCMS-2020 mass spectrometer using electrospray ionization operating in positive and negative ion mode. Samples were introduced into the mass spectrometer using chromatography. The purity of all final products was ≥90% unless otherwise indicated in the experimental details. HPLC purity was determined on a Shimadzu Acquity HPLC system.

[0199] The following abbreviations are used in the experimental section for well-known chemical solvents, reagents, parameters and techniques: 1H NMR: Proton nuclear magnetic resonance spectroscopy ACN: acetonitrile AcOH: acetic acid c-Bu: cyclobutyl c-Pr: cyclopropyl CeCl3: Cerium(III) chloride CH2Cl2: Dichloromethane CHCl3: Chloroform Cs2CO3: Cesium carbonate DBAD: Di-tert-butyl azodicarboxylate DCM: dichloromethane DIBAL: Diisobutylaluminum hydride DIEA: N,N-diisopropylethylamine DMF: N,N-dimethylformamide DMSO: dimethyl sulfoxide DTAD: Di-tert-butyl azodicarboxylate EA: Ethyl acetate ee: enantiomeric excess Et2O: Diethyl ether Et3N: Triethylamine EtOAc: ethyl acetate EtOH: ethanol FA: Formic acid h: time H2O: Water HATU: 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate HCl: Hydrochloric acid Hex: Hexane HPLC: High-performance liquid chromatography IPA: Isopropanol K2CO3: Potassium carbonate KOAc: Potassium acetate LiOH: Lithium hydroxide mCPBA: meta-chloroperoxybenzoic acid Me: Methyl MeCN: acetonitrile MeOH: Methanol mg: milligram min:minutes mL: milliliter MsCl: methanesulfonyl chloride Ms2O: methanesulfonic anhydride NaBH4: sodium borohydride N2: Nitrogen NaCl: Sodium chloride Na2CO3: Sodium carbonate NaH: sodium hydride NaOH: Sodium hydroxide NaHCO3: Sodium bicarbonate NaH2PO4: Monosodium phosphate Na2SO4: Sodium sulfate NH3: Ammonia NH4HCO3: Ammonium bicarbonate NMP: N-methylpyrrolidone oxetane four-membered ring containing three carbon ring atoms and one oxygen ring atom PBr3: Phosphorus tribromide PCl5: Phosphorus pentachloride Pd-PEPPSI-IHeptCl 3-Chloropyridine: Dichloro[1,3-bis(2,6-di-4-heptylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) Pd(dppf)Cl2: (1,1'-bis(diphenylphosphino)ferrocene)palladium(II) dichloride Pd(PPh3)4: Tetrakis(triphenylphosphine)palladium(0) Pd2(dba)3: Tris(dibenzylideneacetone)dipalladium(0) PdCl2(PPh3)2: Bis(triphenylphosphine)palladium(II) dichloride PE: Petroleum ether POCl3: Phosphorus oxychloride PPh3: 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 [Example]

[0200] Example 1: 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0201] Example 2: 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0202] Project 1: Preparation of (2Z)-4-bromo-2-(hydroxyimino)-6-methyl-3H-inden-1-one [ka] To a stirred solution of 4-bromo-6-methyl-2,3-dihydroinden-1-one (2 g, 8.89 mmol) in 12 M aqueous HCl (10 mL) and EtO (10 mL) was slowly added dropwise at 0 °C. The resulting solution was stirred for 4 h at room temperature. The mixture was cooled to 0 °C, and the precipitated solid was collected by filtration and washed with HO (3 × 50 mL) and EtO (2 × 20 mL). The collected solid was concentrated and dried under vacuum. The crude product was used directly in the next step without further purification. This gave (2Z)-4-bromo-2-(hydroxyimino)-6-methyl-3H-inden-1-one (1.5 g, 66%) as an off-white solid. MS: (ES + ) m / z = 253.9 [M+H] + .

[0203] Project 2: Preparation of 5-bromo-3-chloro-7-methyl-2H-isoquinolin-1-one [ka] To a stirred solution of (2Z)-4-bromo-2-(hydroxyimino)-6-methyl-3H-inden-1-one (1.5 g, 5.90 mmol) in CHCl (30 mL) was added PCl (2.46 g, 11.81 mmol) slowly in portions at 0 °C. The resulting mixture was stirred for 3 hours at room temperature and then concentrated under reduced pressure. To the crude product was added a 4 M solution of HCl 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 with 5:1 PE / EtOAc to give 5-bromo-3-chloro-7-methyl-2H-isoquinolin-1-one (1 g, 61%) as a yellow solid. MS: (ES - ) m / z = 269.9 [M-1] - .

[0204] Project 3: Preparation of 5-bromo-3-chloro-2,7-dimethylisoquinolin-1-one [ka] To a stirred solution of 5-bromo-3-chloro-7-methyl-2H-isoquinolin-1-one (1.3 g, 4.77 mmol) in DMF (10 mL) was slowly added NaH (0.17 g, 7.16 mmol) portionwise at 0 °C. The resulting solution was stirred for 20 minutes at 0 °C. Iodomethane (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 × 50 mL). The combined organic layers were washed with brine (3 × 40 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA = 4:1) to give 5-bromo-3-chloro-2,7-dimethylisoquinolin-1-one (900 mg, 65%) as a reddish-brown solid. MS: (ES + ) m / z = 286.0 [M+H] + .

[0205] Project 4: Preparation of 5-bromo-3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethylisoquinolin-1-one [ka] A mixture of 5-bromo-3-chloro-2,7-dimethylisoquinolin-1-one (900 mg, 3.14 mmol), 4,4-dimethylpiperidine (940 mg, 6.28 mmol), and K2CO3 (1.30 g, 9.42 mmol) in NMP (15 mL) was stirred overnight at 140 °C under a nitrogen atmosphere. The mixture was cooled to room temperature, diluted with water (40 mL), and extracted with EtOAc (3 × 80 mL). The combined organic layers were washed with brine (3 × 40 mL), dried over anhydrous Na2SO4, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA = 3:1) to give 5-bromo-3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethylisoquinolin-1-one (700 mg, 61%) as a yellow solid. MS: (ES + ) m / z = 363.1 [M+H] + .

[0206] Project 5: Preparation of 5-acetyl-3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethylisoquinolin-1-one [ka] A solution of 5-bromo-3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethylisoquinolin-1-one (700 mg, 1.93 mmol), tributyl(1-ethoxyethenyl)stannane (835 mg, 2.31 mmol), and Pd(PPh3)4 (222 mg, 0.19 mmol) in anhydrous 1,4-dioxane (12 mL) was stirred overnight at 100 °C under a nitrogen atmosphere. The reaction mixture was cooled to room temperature. 1N aqueous HCl (2 mL) was added, and the mixture was stirred for 20 minutes at 50 °C, and then the mixture was cooled to room temperature. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were washed with water (3 × 20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA=2:1) ​​to give 5-acetyl-3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethylisoquinolin-1-one (550 mg, 87%) as a yellow solid. MS: (ES + ) m / z = 327.1 [M+H] + .

[0207] Project 6: Preparation of 3-(4,4-dimethylpiperidin-1-yl)-5-(1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one [ka] To a stirring solution of 5-acetyl-3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethylisoquinolin-1-one (550 mg, 1.69 mmol) in MeOH (10 mL) was added NaBH (127 mg, 3.37 mmol) portionwise slowly at 0 °C. The resulting solution was stirred for 2 h at room temperature. The resulting mixture was quenched with HO (20 mL) and extracted with EtOAc (3 × 80 mL). The combined organic layers were washed with brine (2 × 50 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA = 2:1) to give 3-(4,4-dimethylpiperidin-1-yl)-5-(1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one (400 mg, 72%) as a light yellow solid. MS: (ES + ) m / z = 329.2 [M+H] + .

[0208] Project 7: Preparation of methyl 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate [ka] To a solution of 3-(4,4-dimethylpiperidin-1-yl)-5-(1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one (400 mg, 1.22 mmol), methyl anthranilate (920 mg, 6.090 mmol), and PPh3 (798 mg, 3.05 mmol) in THF (10 mL) was added a solution of (E)-N-[[(tert-butoxy)carbonyl]imino](tert-butoxy)formamide (560 mg, 2.44 mmol) in tetrahydrofuran (1 mL) at 0° C. under a nitrogen atmosphere. The resulting mixture was stirred at room temperature overnight and then concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA=6:1) to give methyl 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (280 mg, 49%) as a light yellow solid. MS: (ES + ) m / z = 462.2 [M+H] + .

[0209] Project 8: Preparation of 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka] A stirred solution of methyl 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (280 mg, 0.61 mmol) and NaOH (242 mg, 6.07 mmol) in MeOH (5 mL) and HO (5 mL). The resulting mixture was stirred overnight at 50 °C, cooled to room temperature, and then acidified to pH 5-6 with 2 M aqueous HCl. The resulting mixture was extracted with ethyl acetate (3 × 60 mL). The combined organic layers were washed with water (2 × 30 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by preparative TLC (DCM / MeOH=15:1) to give 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (150 mg, 55%) as an off-white solid. MS: (ES + ) m / z = 448.2 [M+H] + .

[0210] Project 9: Preparation of 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka] The racemic mixture of 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (150 mg, 0.34 mmol) was separated by preparative chiral HPLC (column: Chiralpak IC-3, 2 × 25 cm, 5 μm; mobile phase A: Hex (0.1% FA)-HPLC, mobile phase B: IPA-HPLC; flow rate: 20 mL / min; gradient: 20% B to 20% B for 20 min; wavelength: 220 / 254 nm) to give the enantiomers of 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid:

[0211] 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 1, (20.9 mg, 14%, ∼99.9% ee, white solid). MS: (ES + ) m / z = 448.2 [M+H] + . 1 H NMR: (300 MHz, DMSO-d6) δ 12.76 (s, 1H), 8.40 (s, 1H), 7.94 - 7.71 (m, 2H), 7.40 (d, J=2.0 Hz, 1H), 7.23 - 7.12 (m, 1H), 6.57 - 6.48 (m, 1H), 6.39 (d, J=8.9 Hz, 2H), 5.22 - 5.10 (m, 1H), 3.47 (s, 3H), 3.00 - 2.80 (m, 4H), 2.32 (s, 3H), 1.71 - 1.44 (m, 7H), 1.00 (s, 6H).

[0212] 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid, enantiomer 2, 24.1 mg, 16%, 98.6% ee), light yellow solid, MS: (ES + ) m / z = 448.2 [M+H] + . 1 H NMR: (300 MHz, DMSO-d6) δ 12.77 (s, 1H), 8.47 (s, 1H), 8.06 - 7.76 (m, 2H), 7.41 (d, J=1.9 Hz, 1H), 7.23 - 7.12 (m, 1H), 6.57 - 6.48 (m, 1H), 6.38 (d, J=5.6 Hz, 2H), 5.22 - 5.10 (m, 1H), 3.47 (s, 3H), 3.00 - 2.80 (m, 4H), 2.32 (s, 3H), 1.70 - 1.46 (m, 7H), 1.00 (s, 6H).

[0213] Example 3: 6-chloro-3-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid (Enantiomer 1)

[0214] Example 4: 6-chloro-3-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid (Enantiomer 2) [ka]

[0215] Project 1: Preparation of methyl 6-chloro-3-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinate To a stirred solution of 3-(4,4-dimethylpiperidin-1-yl)-5-(1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one (500 mg, 1.52 mmol) and TEA (924 mg, 9.13 mmol) in DCM (10 mL) was added methanesulfonic anhydride (1.06 g, 6.09 mmol) in portions at 0 °C. The resulting mixture was stirred for 1 h at 0 °C. Methyl 3-amino-6-chloropyridine-2-carboxylate (341 mg, 1.83 mmol) was added, and the mixture was stirred for an additional 12 h at 50 °C. The resulting mixture was diluted with HO (50 mL) and extracted with DCM (3 × 50 mL). The combined organic layers were washed with brine (3 × 50 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (Regular C18 column; 330 g, 20-40 μm; gradient: 10% to 100% aqueous ACN over 30 min, flow rate: 100 mL / min) to give methyl 6-chloro-3-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinate (410 mg, 54%) as a yellow solid. MS: (ES + ) m / z = 497.2 [M+H] + .

[0216] Project 2: Preparation of 6-chloro-3-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid (enantiomers 1 and 2) To a stirred solution of methyl 6-chloro-3-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinate (290 mg, 0.58 mmol) in MeOH (6 mL) and HO (3 mL) was added NaOH (233 mg, 5.83 mmol). The resulting mixture was stirred for 12 h at room temperature. The mixture was acidified to pH 5 with 1 N aqueous HCl and diluted with HO (20 mL). The resulting mixture was extracted with EA (3 × 30 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by reverse flash chromatography (column: Regular C18; 330 g, 20–40 μm; gradient: 10%–100% aqueous ACN over 30 min, flow rate: 100 mL / min), followed by chiral HPLC under the following conditions: column: Chiralpak IC-3, 4.6 × 50 mm, 3 μm; mobile phase: hexane (0.1% TFA):EtOH = 80:20; flow rate: 20 mL / min), and then further purified by preparative HPLC (column: XBridge Shield RP18 OBD column; 30 × 150 mm, 5 μm; gradient: mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3·H2O), mobile phase B: ACN; 10% to 100% ACN for 30 min, 25% B to 55% for 9 min, flow rate: 60 mL / min) to give each enantiomer 6-chloro-3-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid as a white solid:

[0217] 6-Chloro-3-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid enantiomer 1 (16.6 mg, 5% yield, >99% ee). 1H-NMR (300 MHz, methanol-d4) δ 7.97 (s, 1H), 7.52 (d, J=1.9 Hz, 1H), 7.09 (d, J=8.8 Hz, 1H), 6.79 (d, J=8.9 Hz, 1H), 6.48 (s, 1H), 5.05-5.15 (m, 1H), 3.61 (s, 3H), 3.10 - 2.81 (m, 4H), 2.37 (s, 3H), 1.65 (d, J=6.7 Hz, 7H), 1.06 (s, 6H); MS: (ES + ) m / z = 483.1 [M+H] + .

[0218] 6-chloro-3-((1-(3-(4,4-dimethylpiperidin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid enantiomer 2 (14.4 mg, 5% yield, >99% ee), 1 H-NMR (300 MHz, methanol-d4) δ 7.98 (s, 1H), 7.52 (d, J=1.8 Hz, 1H), 7.13 (d, J=8.8 Hz, 1H), 6.83 (d, J=8.9 Hz, 1H), 6.48 (s, 1H), 5.05-5.15 (m, 1H), 3.61 (s, 3H), 3.10 - 2.81 (m, 4H), 2.37 (s, 3H), 1.66 (d, J=6.7 Hz, 7H), 1.06 (s, 6H); MS: (ES + ) m / z = 483.1 [M+H] + .

[0219] Example 5: 2-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0220] Example 6: 2-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0221] Project 1: Preparation of 5-acetyl-3-chloro-2,7-dimethylisoquinolin-1-one [ka] A mixture of 5-bromo-3-chloro-2,7-dimethylisoquinolin-1-one (2.6 g, 9.07 mmol), tributyl(1-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 1N aqueous HCl (10 mL), and stirred for 15 minutes. The resulting mixture was diluted with water (60 mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with water (3 × 60 mL) and brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 5-acetyl-3-chloro-2,7-dimethylisoquinolin-1-one (1.9 g, 83%) as a yellow solid. MS: ES + (m / z)=250.1 [M+H] + .

[0222] Project 2: Preparation of 3-chloro-5-(1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one [ka] To a stirred solution of 5-acetyl-3-chloro-2,7-dimethylisoquinolin-1-one (1.9 g, 7.61 mmol) in MeOH (20 mL) was added NaBH (0.58 g, 15.218 mmol) portionwise slowly at 0 °C. The resulting mixture was stirred for 2 h at room temperature. The reaction was quenched with water (30 mL) and extracted with EtOAc (3 × 80 mL). The combined organic layers were washed with brine (2 × 60 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA = 1:1) to give 3-chloro-5-(1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one (1.5 g, 78%) as an off-white solid. MS: (ES + ) m / z = 252.1 [M+H] + .

[0223] Project 3: Preparation of tert-butyl 2-((N-(1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)-2,4-dinitrophenyl)sulfonamido)benzoate [ka] To a solution of 3-chloro-5-(1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one (1.4 g, 5.56 mmol), tert-butyl 2-(2,4-dinitrobenzenesulfonamido)benzoate (2.59 g, 6.12 mmol), and PPh3 (3.65 g, 13.91 mmol) in THF (20 mL) was added a solution of (E)-N-[[(tert-butoxy)carbonyl]imino](tert-butoxy)formamide (3.84 g, 16.69 mmol) in tetrahydrofuran (1.5 mL) at 0° C. under a nitrogen atmosphere. The resulting solution was stirred at room temperature overnight. The residue was concentrated under reduced pressure and purified by silica gel column chromatography (PE / EA=3:1) to give tert-butyl 2-((N-(1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)-2,4-dinitrophenyl)sulfonamido)benzoate (2.4 g, 65%) as a yellow solid. MS: (ES + ) m / z = 657.3 [M+H] + .

[0224] Project 4: Preparation of tert-butyl 2-{[1-(3-chloro-2,7-dimethyl-1-oxoisoquinolin-5-yl)ethyl]amino}benzoate [ka] To a stirred solution of tert-butyl 2-((N-(1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)-2,4-dinitrophenyl)sulfonamido)benzoate (2.4 g, 3.65 mmol) in DCM (12 mL) was added EtN (0.74 g, 7.30 mmol) and 2-sulfanylacetic acid (0.50 g, 5.48 mmol) slowly dropwise at 0 °C. The resulting mixture was stirred for 3 h at rt. The reaction was quenched with water (30 mL) and extracted with DCM (3 × 50 mL). The combined organic layers were washed with brine (2 × 30 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA=6:1) to give tert-butyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (1.2 g, 76%) as a light yellow solid. MS: (ES + ) m / z = 427.1 [M+H] + .

[0225] Project 5: Preparation of tert-butyl 2-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate [ka] A mixture of tert-butyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (200 mg, 0.47 mmol), 2-(4,4-dimethylcyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (221 mg, 0.94 mmol), NaCO (149 mg, 1.40 mmol), and Pd(dppf)Cl (38 mg, 0.047 mmol) in 1,4-dioxane (10 mL) and HO (2 mL) was stirred overnight at 100 °C under a nitrogen atmosphere. The reaction mixture was cooled to room temperature. The residue was concentrated under reduced pressure and purified by preparative TLC (PE / EA=3:1) to give tert-butyl 2-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (140 mg, 59%) as a light yellow solid. MS: (ES + ) m / z = 501.3 [M+H] + .

[0226] Project 6: Preparation of 2-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka] A solution of tert-butyl 2-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (120 mg, 0.24 mmol) in 4 M HCl in 1,4-dioxane (10 mL) was stirred overnight at room temperature and then concentrated under reduced pressure. The residue was purified by preparative TLC (DCM / MeOH=25:1) to give 2-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (80 mg, 75%) as a yellow solid. MS: (ES+ ) m / z = 445.3 [M+H] + .

[0227] Project 7: Preparation of 2-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (enantiomers 1 and 2) The racemic mixture of 2-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (80 mg, 0.18 mmol) was purified by chiral HPLC (Chiralpak column). Separation on AD-H, 2 x 25 cm, 5 μm; mobile phase A: Hex (0.1% FA)-HPLC, mobile phase B: EtOH-HPLC; flow rate: 20 mL / min; gradient: 10% B to 10% B over 12 min; wavelength: 220 / 254 nm; RT1 (min): 6.58; RT2 (min): 10.85; sample solvent: EtOH-HPLC; injection volume: 0.5 mL) gave the enantiomers of 2-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid as light yellow solids:

[0228] 2-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 1 (29.3 mg, 36%, 99.9% ee). MS: (ES + ) m / z = 445.1 [M+H] + . 1H NMR: (300 MHz, methanol-d4) δ 8.01 (s, 1H), 7.92 - 7.88 (m, 1H), 7.55 (d, J=1.9 Hz, 1H), 7.11- 7.10 (m, 1H), 6.75 (s, 1H), 6.51 - 6.50 (m, 1H), 6.36-6.27 (m, 1H), 5.96 - 5.87 (m, 1H), 5.20 - 5.10 (m, 1H), 3.54 (s, 3H), 2.39 (s, 3H), 2.36 - 2.25 (m, 2H), 2.10 - 2.02 (m, 2H), 1.65 - 1.53 (m, 5H), 1.04 (d, J=1.1 Hz, 6H).

[0229] 2-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 2 (29.1 mg, 37%, 99.9% ee). MS: (ES + ) m / z = 445.1 [M+H] + . 1 H NMR: (300 MHz, methanol-d4) δ 8.02 (s, 1H), 7.92 - 7.88 (m, 1H), 7.55 (d, J=1.8 Hz, 1H), 7.12 - 7.11 (m, 1H), 6.75 (s, 1H), 6.57 - 6.45 (m, 1H), 6.36 - 6.27 (m, 1H), 5.96 - 5.87 (m, 1H), 5.20 - 5.10 (m, 1H), 3.53 (s, 3H), 2.39 (s, 3H), 2.36 - 2.25 (m, 2H), 2.10 - 2.02 (m, 2H), 1.65 - 1.52 (m, 5H), 1.03 (d, J=1.3 Hz, 6H).

[0230] Example 7: 2-((1-(3-(5-Fluoro-1H-indol-2-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid. [ka]

[0231] Project 1: Preparation of tert-butyl 2-((1-(3-(5-fluoro-1H-indol-2-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate [ka] A mixture of tert-butyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (450 mg, 1.05 mmol), 5-fluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (330 mg, 1.27 mmol), NaCO (335 mg, 3.16 mmol), and Pd(dppf)Cl (129 mg, 0.16 mmol) in 1,4-dioxane (8 mL) and HO (1 mL) was stirred overnight at 100 °C under a nitrogen atmosphere. The reaction mixture was cooled to room temperature. The solid was filtered and washed with DCM (2 × 50 mL). The filtrate was concentrated under reduced pressure and purified by preparative TLC (PE / EA=3:1) to give tert-butyl 2-((1-(3-(5-fluoro-1H-indol-2-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (300 mg, 54%) as a yellow solid. MS: (ES + ) m / z = 526.2 [M+H] + .

[0232] Project 2: Preparation of 2-((1-(3-(5-fluoro-1H-indol-2-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka] A solution of tert-butyl 2-((1-(3-(5-fluoro-1H-indol-2-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (100 mg, 0.19 mmol) in TFA (6 mL) and DCM (6 mL) was stirred overnight at room temperature. The resulting mixture was concentrated under reduced pressure and purified by preparative TLC (CHCl / MeOH = 15:1). The desired product was further purified by reverse flash chromatography (column, C18 silica gel; mobile phase, ACN in HO, 30% to 50% gradient over 10 min; detector, UV 254 nm). Finally, the desired product was further purified by preparative HPLC (column: YMC-Actus Triart C18 ExRS, 30 × 150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3 HO), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 8% B to 38% B for 9 min, 38% B; wavelength: 254 nm) to give 2-((1-(3-(5-fluoro-1H-indol-2-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (18.1 mg, 20%) as a white solid. MS: (ES) + ) m / z = 470.1 [M+H] + . 1 H NMR: (400 MHz, DMSO-d6) δ 11.87 (s, 1H), 8.76 (s, 1H), 8.00 (s, 1H), 7.83 (d, J=7.9 Hz, 1H), 7.56 - 7.44 (m, 2H), 7.42 - 7.36 (m, 1H), 7.21 - 7.00 (m, 3H), 6.84 (d, J=2.0 Hz, 1H), 6.50 - 6.46 (m, 1H), 6.32 (d, J=8.5 Hz, 1H), 5.31 - 5.25 (m, 1H), 3.56 (s, 3H), 2.38 (s, 3H), 1.53 (d, J=6.6 Hz, 3H).

[0233] Intermediate 1 [ka]

[0234] Project 1: Preparation of 5-(1-bromoethyl)-3-chloro-2,7-dimethylisoquinolin-1(2H)-one To a stirring solution of 3-chloro-5-(1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one (1 g, 3.97 mmol) in DCM (12 mL) was added PBr (2.15 g, 7.95 mmol) at 0 °C. The resulting mixture was stirred for 4 h at room temperature under a nitrogen atmosphere. The reaction was quenched with HO (30 mL) at 0 °C, and the mixture was adjusted to approximately pH 7 with saturated aqueous NaHCO. The resulting mixture was extracted with DCM (3 × 20 mL). The combined organic layers were washed with brine (2 × 20 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure to give 5-(1-bromoethyl)-3-chloro-2,7-dimethylisoquinolin-1(2H)-one, which was used in the next step without further purification. MS: (ES + ) m / z = 314.0 [M+H] + .

[0235] Project 2: Preparation of methyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate To a stirred solution of 5-(1-bromoethyl)-3-chloro-2,7-dimethylisoquinolin-1(2H)-one (1 g, 3.18 mmol) in ACN (30 mL) and THF (10 mL) was added methyl anthranilate (1.20 g, 7.95 mmol). The resulting mixture was stirred for 3 hours at 80°C under a nitrogen atmosphere and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA=4:1) to give methyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (0.8 g, 65%) as a light yellow solid. MS: (ES + ) m / z = 385.2 [M+H] + .

[0236] Example 8: 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0237] Example 9: 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0238] Project 1: Preparation of methyl 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate [ka] To a stirred solution of methyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (200 mg, 0.52 mmol), 1-(2,2,2-trifluoroethyl)piperazine dihydrochloride (187 mg, 0.78 mmol), and CsCO (508 mg, 1.56 mmol) in dioxane (3 mL) was added Pd(dba) (95 mg, 0.10 mmol) and Ruphos (60 mg, 0.10 mmol) in portions at room temperature under a nitrogen atmosphere. The resulting mixture was stirred overnight at 100 °C under a nitrogen atmosphere. The resulting mixture was quenched with 30 mL of H2O. The mixture was extracted with EtOAc (3 × 30 mL). The combined organic layers were washed with brine (3 × 20 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA = 1:1) to give methyl 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (120 mg, 44%) as a yellow oil. MS: (ES + ) m / z = 517.6 [M+H] + .

[0239] Project 2: Preparation of 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka] A mixture of methyl 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (30 mg, 0.058 mmol), MeOH (600 μL), HO (120 μL), and NaOH (4.65 mg, 0.12 mmol) was stirred overnight at 50 °C. The mixture was acidified to pH 5 with aqueous NaHPO. The resulting mixture was diluted with HO (20 mL) and extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine (3 × 20 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column: C18 silica gel; mobile phase: MeCN in water (0.1% FA), 5% to 100% gradient over 40 min; detector: UV 254 nm) to give 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (14.4 mg, 49%) as an off-white solid. MS: (ES + ) m / z = 503.5 [M+H] + .

[0240] Project 3: Preparation of 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (enantiomers 1 and 2) The racemic mixture of 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (60 mg, 0.119 mmol) was separated by preparative chiral HPLC (column: Chiralpak IC-3, 4.6×50 mm, 3 μm; mobile phase: Hex (0.1% FA):IPA=80:20; flow rate: 1 mL / min) to give the enantiomers of 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid as light yellow solids:

[0241] 2-((1-(2,7-Dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 1 (21.2 mg, 35%, >99.9% ee). MS: (ES + ) m / z = 503.5 [M+H] + . 1 H NMR: (300 MHz, DMSO-d6) δ 12.86 (s, 1H), 8.46 (s, 1H), 7.87 (s, 1H), 7.86 - 7.80 (m, 1H), 7.41 (d, J=1.9 Hz, 1H), 7.20 - 7.17 (m, 1H), 6.57 - 6.45 (m, 1H), 6.44 - 6.32 (m, 2H), 5.22 - 5.15 (m, 1H), 3.49 (s, 3H), 3.50 - 3.34 (m, 2H), 3.21 - 2.61 (m, 8H), 2.32 (s, 3H), 1.53 (d, J=6.5 Hz, 3H).

[0242] 2-((1-(2,7-Dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 2 (24.0 mg, 39%, >99.9% ee). MS: (ES +) m / z = 503.5 [M+H] + . 1 HNMR: (300 MHz, DMSO-d6) δ 12.76 (s, 1H), 8.43 (s, 1H), 7.87 (s, 1H), 7.86 - 7.80 (m, 1H), 7.41 (d, J=1.9 Hz, 1H), 7.20 - 7.18 (m, 1H), 6.57 - 6.45 (m, 1H), 6.44 - 6.32 (m, 2H), 5.22 - 5.15 (m, 1H), 3.49 (s, 3H), 3.50 - 3.31 (m, 2H), 3.21 - 2.60 (m, 8H), 2.32 (s, 3H), 1.53 (d, J=6.5 Hz, 3H).

[0243] Example 10: 2-((1-(3-(3-azabicyclo[3.2.1]octan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0244] Example 11: 2-((1-(3-(3-azabicyclo[3.2.1]octan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0245] A solution of methyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (300 mg, 0.78 mmol), 3-azabicyclo[3.2.1]octane hydrochloride (288 mg, 1.95 mmol), potassium 2-methyl-2-propanolate (350 mg, 3.12 mmol), and Pd-PEPPSI-IHeptCl 3-chloropyridine (379 mg, 0.39 mmol) in anhydrous dioxane (20 mL) was stirred overnight at 100° C. under a nitrogen atmosphere. The reaction mixture was cooled to room temperature. The solid was filtered and washed with DCM (2×50 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (DCM / MeOH=10:1) to give 2-((1-(3-(3-azabicyclo[3.2.1]octan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (80 mg, 23%) as a yellow solid. MS: (ES + ) m / z = 446.6 [M+H] + .

[0246] The racemic mixture of 2-((1-(3-(3-azabicyclo[3.2.1]octan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (80 mg, 0.18 mmol) was separated by preparative chiral HPLC (column: Chiralcel OD-3, 4.6 × 50 mm, 3 μm; mobile phase: Hex (0.1% FA):EtOH = 80:20; flow rate: 1 mL / min) to give each enantiomer of 2-((1-(3-(3-azabicyclo[3.2.1]octan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid as a white solid:

[0247] 2-((1-(3-(3-azabicyclo[3.2.1]octan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 1 (4.8 mg, 6%, >99% ee). MS: (ES + ) m / z = 446.6 [M+H]+ . 1 H NMR: (300 MHz, DMSO-d6) δ 12.74 (s, 1H), 8.35 (d, J=5.9 Hz, 1H), 7.87 (d, J=1.8 Hz, 1H), 7.80 (s, 1H), 7.41 (d, J=1.9 Hz, 1H), 7.25 - 7.20 (m, 1H), 6.58 - 6.47 (m, 1H), 6.44 (s, 1H), 6.37 (d, J=8.3 Hz, 1H), 5.20 - 5.12 (m, 1H), 3.56 (s, 3H), 3.02 - 2.95 (m 2H), 2.80 - 2.76 (m, 2H), 2.32 - 2.28 (m, 5H), 1.82 (d, J=7.7 Hz, 2H), 1.77 - 1.62 (m, 2H), 1.53 (d, J=6.7 Hz, 3H).

[0248] 2-((1-(3-(3-アザビシクロ[3.2.1]オクタン-3-イル)-2,7-ジメチル-1-オキソ-1,2 - ジヒドロイソキノリン-5-イル) エチル) アミノ) benzoic acid エナンチオマー2 (4.7mg, 6%, >99%ee). MS: (ES + ) m / z = 446.6 [M+H] + . 1 H NMR: (300 MHz, DMSO-d6) δ 12.74 (s, 1H), 8.35 (d, J=5.9 Hz, 1H), 7.87 (d, J=1.8 Hz, 1H), 7.80 (s, 1H), 7.41 (d, J=1.9 Hz, 1H), 7.25 - 7.20 (m, 1H), 6.58 - 6.47 (m, 1H), 6.44 (s, 1H), 6.37 (d, J=8.3 Hz, 1H), 5.20 - 5.12 (m, 1H), 3.56 (s, 3H), 3.02 - 2.95 (m 2H), 2.80 - 2.76 (m, 2H), 2.32 - 2.28 (m, 5H), 1.82 (d, J=7.7 Hz, 2H), 1.77 - 1.62 (m, 2H), 1.53 (d, J=6.7 Hz, 3H).

[0249] Example 12: 2-((1-(2,7-dimethyl-3-(4-methylpiperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0250] Example 13: 2-((1-(2,7-dimethyl-3-(4-methylpiperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0251] Prepared analogously to Examples 8 and 9, using 1-(2,2,2-trifluoroethyl)piperazine dihydrochloride in place of 1-methylpiperazine in step 1:

[0252] 2-((1-(2,7-dimethyl-3-(4-methylpiperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 1 (>99.9% ee). MS: (ES + ) m / z = 435.5 [M+H] + . 1 H NMR: (300 MHz, DMSO-d6) δ 8.46 (s, 1H), 7.87 (s, 1H), 7.80 (s, 1H), 7.42 (d, J=1.9 Hz, 1H), 7.24 - 7.12 (m, 1H), 6.61 - 6.51 (m, 1H), 6.43 - 6.32 (m, 2H), 5.20 - 5.12 (m, 1H), 4.20 - 3.91 (m, 4H), 3.48 (s, 3H), 3.10 - 2.95 (m, 4H), 2.29 (s, 3H), 2.26 (s, 3H), 1.54 (d, J=6.5 Hz, 3H).

[0253] 2-((1-(2,7-dimethyl-3-(4-methylpiperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 2 (>99.9% ee). MS: (ES + ) m / z = 435.5 [M+H] + . 1 H NMR: (300 MHz, DMSO-d6) δ 8.46 (s, 1H), 7.87 (s, 1H), 7.80 (s, 1H), 7.42 (d, J=1.9 Hz, 1H), 7.24 - 7.12 (m, 1H), 6.61 - 6.51 (m, 1H), 6.43 - 6.32 (m, 2H), 5.20 - 5.12 (m, 1H), 4.20 - 3.91 (m, 4H), 3.48 (s, 3H), 3.10 - 2.95 (m, 4H), 2.29 (s, 3H), 2.26 (s, 3H), 1.54 (d, J=6.5 Hz, 3H).

[0254] Example 14: 2-((1-(3-((1R,5S,6r)-6-cyano-3-azabicyclo[3.1.0]hexan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0255] Example 15: 2-((1-(3-((1R,5S,6r)-6-cyano-3-azabicyclo[3.1.0]hexan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0256] Prepared in a manner analogous to Examples 8 and 9, except that 1-(2,2,2-trifluoroethyl)piperazine dihydrochloride was used in place of (1R,5S,6r)-3-azabicyclo[3.1.0]hexane-6-carbonitrile in Step 1;

[0257] 2-((1-(3-((1R,5S,6r)-6-cyano-3-azabicyclo[3.1.0]hexan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 1 (99.9% ee). MS: (ES - ) m / z)=441.0 [M−H] - . 1 H NMR: (400 MHz, methanol-d4) δ 7.96 (s, 1H), 7.91 (d, J=7.8 Hz, 1H), 7.57 (d, J=2.0 Hz, 1H), 7.10 - 6.95 (m, 1H), 6.61 (s, 1H), 6.55 - 6.50 (m, 1H), 6.33 (d, J=8.4 Hz, 1H), 5.18 - 4.95 (m, 1H), 4.63 (s, 3H), 3.52 - 3.35 (m, 5H), 3.15 - 3.05 (m, 2H), 2.35 - 2.20 (m, 5H), 1.63 (d, J=6.7 Hz, 3H).

[0258] 2-((1-(3-((1R,5S,6r)-6-cyano-3-azabicyclo[3.1.0]hexan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 2 (99.9% ee). MS: (ES - ) m / z)=441.0 [M−H] - . 1 H NMR: (400 MHz, methanol-d4) δ 7.97 (s, 1H), 7.91 (d, J=7.8 Hz, 1H), 7.55 (d, J=2.0 Hz, 1H), 7.10 - 6.95 (m, 1H), 6.61 (s, 1H), 6.55 - 6.50 (m, 1H), 6.33 (d, J=8.4 Hz, 1H), 5.18 - 4.95 (m, 1H), 4.63 (s, 3H), 3.52 - 3.35 (m, 5H), 3.15 - 3.05 (m, 2H), 2.35 - 2.20 (m, 5H), 1.63 (d, J=6.7 Hz, 3H).

[0259] Example 16: 2-((1-(3-((1R,5S,6r)-6-cyano-3-azabicyclo[3.1.0]hexan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0260] Example 17: 2-((1-(3-((1R,5S,6r)-6-cyano-3-azabicyclo[3.1.0]hexan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0261] Prepared analogously to Examples 8 and 9, using 1-(2,2,2-trifluoroethyl)piperazine dihydrochloride in place of 3-(4-methoxyphenyl)azetidine in step 1:

[0262] 2-((1-(3-((1R,5S,6r)-6-cyano-3-azabicyclo[3.1.0]hexan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 1 (99.9% ee). MS: (ES + ) m / z = 498.2 [M+H] + . 1H NMR: (300 MHz, DMSO-d6): δ 8.47 (s, 1H), 7.90 - 7.70 (m, 2H), 7.39 - 7.28 (m, 3H), 7.25 - 7.15 (m, 1H), 7.00 - 6.90 (m, 2H), 6.55 - 4.45 (m, 1H), 6.36 (d, J=8.5 Hz, 1H), 5.94 (s, 1H), 5.15 - 5.05 (m, 1H), 4.40 - 4.30 (m, 2H), 3.85 - 3.75 (m, 3H), 3.73 (s, 3H), 3.44 (s, 3H), 2.28 (s, 3H), 1.52 (d, J=6.5 Hz, 3H).

[0263] 2-((1-(3-((1R,5S,6r)-6-cyano-3-azabicyclo[3.1.0]hexan-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 1 (99.9% ee). MS: (ES + ) m / z = 498.2 [M+H] + . 1 H NMR (300 MHz, DMSO-d6): δ 8.45 (s, 1H), 7.90 - 7.70 (m, 2H), 7.39 - 7.28 (m, 3H), 7.25 - 7.15 (m, 1H), 6.99 - 6.89 (m, 2H), 6.55 - 4.45 (m, 1H), 6.37 (d, J=8.5 Hz, 1H), 5.93 (s, 1H), 5.15 - 5.05 (m, 1H), 4.40 - 4.30 (m, 2H), 3.85 - 3.75 (m, 3H), 3.73 (s, 3H), 3.44 (s, 3H), 2.28 (s, 3H), 1.53 (d, J=6.4 Hz, 3H).

[0264] Example 18: 2-((1-(3-(5,7-dihydro-6H-pyrrolo[3,4-b]pyridin-6-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0265] Example 19: 2-((1-(3-(5,7-dihydro-6H-pyrrolo[3,4-b]pyridin-6-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0266] Prepared analogously to Examples 8 and 9, using 1-(2,2,2-trifluoroethyl)piperazine dihydrochloride in place of 6,7-dihydro-5H-pyrrolo[3,4-b]pyridine in step 1:

[0267] 2-((1-(3-(5,7-dihydro-6H-pyrrolo[3,4-b]pyridin-6-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 1 (99.9% ee). MS: (ES + ) m / z = 455.1 [M+H] + . 1 H NMR: (400 MHz, DMSO-d6) δ 13.35 - 12.00 (m, 1H), 8.56 - 8.44 (m, 2H), 7.88 (d, J=1.7 Hz, 1H), 7.85 - 7.80 (m, 2H), 7.41 (d, J=1.9 Hz, 1H), 7.35 - 7.25 (m, 1H), 7.21 - 7.09 (m, 1H), 6.74 (s, 1H), 6.52 -6.48 (m, 1H), 6.36 (d, J=8.4 Hz, 1H), 5.24 - 5.20 (m, 1H), 4.76 - 4.53 (m, 4H), 3.59 (s, 3H), 2.32 (s, 3H), 1.52 (d, J=6.4 Hz, 3H).

[0268] 2-((1-(3-(5,7-dihydro-6H-pyrrolo[3,4-b]pyridin-6-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 2 (99.9% ee). MS: (ES + ) m / z = 455.1 [M+H] + . 1 H NMR: (400 MHz, DMSO-d6) δ 13.35 - 12.00 (m, 1H), 8.56 - 8.44 (m, 2H), 7.88 (d, J=1.7 Hz, 1H), 7.85 - 7.80 (m, 2H), 7.41 (d, J=1.9 Hz, 1H), 7.35 - 7.25 (m, 1H), 7.21 - 7.09 (m, 1H), 6.74 (s, 1H), 6.52 -6.48 (m, 1H), 6.36 (d, J=8.4 Hz, 1H), 5.24 - 5.20 (m, 1H), 4.76 - 4.53 (m, 4H), 3.59 (s, 3H), 2.32 (s, 3H), 1.52 (d, J=6.4 Hz, 3H).

[0269] Example 20: 2-((1-(2,7-dimethyl-3-(6-methylpyridin-3-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0270] Example 21: 2-((1-(2,7-dimethyl-3-(6-methylpyridin-3-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0271] Project 1: Preparation of methyl 2-((1-(2,7-dimethyl-3-(6-methylpyridin-3-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate A solution of methyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (100 mg, 0.26 mmol) and (6-methylpyridin-3-yl)boronic acid (89 mg, 0.65 mmol) in dioxane (5 mL) and HO (1 mL) was treated with NaCO (83 mg, 0.78 mmol) and Pd(dppf)Cl (29 mg, 0.039 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred overnight at 100 °C under a nitrogen atmosphere, then concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA=1:1) to give methyl 2-((1-(2,7-dimethyl-3-(6-methylpyridin-3-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (58 mg, 50%) as an off-white solid. MS: (ES + ) m / z = 442.2 [M+H] + .

[0272] Project 2: Preparation of 2-((1-(2,7-dimethyl-3-(6-methylpyridin-3-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid A solution of methyl 2-((1-(2,7-dimethyl-3-(6-methylpyridin-3-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (58 mg, 0.13 mmol) in MeOH (10 mL) and HO (2 mL) was treated with NaOH (26 mg, 0.66 mmol) at room temperature, and the resulting mixture was stirred at 50° C. overnight. The mixture was adjusted to pH 7 with 1 M aqueous HCl and extracted with EtOAc (3 × 25 mL). The combined organic layers were washed with brine (3 × 15 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The crude residue was purified by preparative HPLC (Column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3·H2O), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 10% B to 38% B for 9 min, 38% B; Wavelength: 254 nm) to give 2-((1-(2,7-dimethyl-3-(6-methylpyridin-3-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (11.5 mg, 20%) as a white solid. MS: (ES) + ) m / z = 428.1 [M+H] + .

[0273] Project 3: Preparation of 2-((1-(2,7-dimethyl-3-(6-methylpyridin-3-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (enantiomers 1 and 2) A racemic mixture of 2-((1-(2,7-dimethyl-3-(6-methylpyridin-3-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (20 mg, 0.047 mmol) was analyzed by chiral HPLC (CHIRAL ART) Separation on Cellulose-SC, 2 x 25 cm, 5 μm; Mobile phase A: Hex (0.1% FA)-HPLC, Mobile phase B: EtOH-HPLC; Flow rate: 20 mL / min; Gradient: 15% B to 15% B in 20 min; Wavelength: 220 / 254 nm; RT1 (min): 20.317; RT2 (min): 22.801; Sample solvent: EtOH--HPLC; Injection volume: 1 mL) gave the enantiomers of 2-((1-(2,7-dimethyl-3-(6-methylpyridin-3-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid as white solids:

[0274] 2-((1-(2,7-dimethyl-3-(6-methylpyridin-3-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 1 (2.8 mg, 13%, 99.7% ee). MS: (ES + ) m / z = 428.1 [M+H] + . 1 H NMR: (400 MHz, DMSO-d6) δ 8.72 (s, 1H), 8.38 (d, J=5.9 Hz, 1H), 8.06 - 7.98 (s, 2H), 7.84 - 7.78 (m, 1H), 7.49 (d, J=2.0 Hz, 2H), 7.17 - 7.15 (m, 1H), 6.92 (s, 1H), 6.56 - 6.48 (m, 1H), 6.34 (d, J=8.4 Hz, 1H), 5.30 - 5.22 (m, 1H), 3.58 - 3.35 (m, 3H), 2.59 (s, 3H), 2.38 (s, 3H), 1.51 (d, J=6.5 Hz, 3H).

[0275] 2-((1-(2,7-dimethyl-3-(6-methylpyridin-3-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 2 (2 mg, 10%, 99.1% ee). MS: (ES + ) m / z = 428.1 [M+H] + . 1 H NMR: (400 MHz, DMSO-d6) δ 8.72 (s, 1H), 8.38 (d, J=5.9 Hz, 1H), 8.06 - 7.98 (m, 2H), 7.84 - 7.78 (m, 1H), 7.49 (d, J=2.0 Hz, 2H), 7.17-7.15 (m, 1H), 6.93 (s, 1H), 6.56 - 6.48 (m, 1H), 6.34 (d, J=8.5 Hz, 1H), 5.30 - 5.22 (m, 1H), 3.58 - 3.35 (m, 3H), 2.59 (s, 3H), 2.38 (s, 3H), 1.51 (d, J=6.5 Hz, 3H).

[0276] Example 22: 6-chloro-3-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid (Enantiomer 1)

[0277] Example 23: 6-chloro-3-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid (Enantiomer 2) [ka]

[0278] Project 1: Preparation of 3-(4,4-dimethylcyclohex-1-en-1-yl)-5-(1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one [ka] A mixture of 3-chloro-5-(1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one (200 mg, 0.80 mmol), 2-(4,4-dimethylcyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (281 mg, 1.19 mmol), sodium permethanesulfonate (170 mg, 1.59 mmol), and Pd(dppf)Cl (324 mg, 0.40 mmol) in dioxane (2 mL) and water (1 mL) was stirred for 2 hours at 100 °C under a nitrogen atmosphere. The resulting mixture was extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with brine (2 × 30 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA=1:1) to give 3-(4,4-dimethylcyclohex-1-en-1-yl)-5-(1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one (160 mg, 61%) as a yellow oil. MS: (ES + ) m / z = 326.5 [M+H] + .

[0279] Project 2: Preparation of methyl 6-chloro-3-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinate [ka] A solution of 3-(4,4-dimethylcyclohex-1-en-1-yl)-5-(1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one (120 mg, 0.37 mmol) in DCM (2 mL) was treated with PBr (200 mg, 0.74 mmol) at 0 °C. The resulting mixture was stirred for 2 h at room temperature. The resulting mixture was quenched with HO (20 mL). The resulting mixture was extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with brine (2 × 20 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The crude intermediate was taken up in ACN (3 mL), and methyl 3-amino-6-chloropyridine-2-carboxylate (172 mg, 0.92 mmol) was added portionwise at room temperature. The resulting mixture was stirred overnight at 80 °C. The resulting mixture was quenched with HO (20 mL). The resulting mixture was extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with brine (2 × 20 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA = 1:1) to give methyl 6-chloro-3-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinate (120 mg, 65%) as a yellow oil. MS: (ES + ) m / z = 494.0 [M+H] + .

[0280] Project 3: Preparation of 6-chloro-3-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid [ka] A mixture of methyl 6-chloro-3-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinate (80 mg, 0.16 mmol), MeOH (2.5 mL), HO (0.5 mL), and NaOH (13 mg, 0.32 mmol) was stirred overnight at 50 °C. The mixture was adjusted to pH 5 by the addition of aqueous NaHPO. The resulting mixture was extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with brine (2 × 20 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column: C18 silica gel; mobile phase: MeCN in water (0.1% FA), 5% to 100% gradient over 40 min; detector: UV 254 nm) to give 6-chloro-3-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid (40 mg, 51%) as a colorless oil. MS: (ES + ) m / z = 480.2 [M+H] +

[0281] Project 4: Preparation of 6-chloro-3-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid (enantiomers 1 and 2) The racemic mixture of 6-chloro-3-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid (100 mg, 0.21 mmol) was purified by preparative chiral HPLC (column: Chiralpak IC-3, 4.6 × 50 mm, 3 μm; mobile phase: Hex (0.1% TFA): EtOH = 60:40; flow rate: 1 mL / min) to give each enantiomer of 6-chloro-3-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid as an off-white solid:

[0282] 6-chloro-3-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid enantiomer 1 (15.3 mg, 15%, >93.1% ee). MS: (ES + ) m / z = 480.2 [M+H] + . 1 H NMR: (300 MHz, DMSO-d6) δ 13.08 (s, 1H), 8.41 (s, 1H), 7.94 (s, 1H), 7.43 (s, 1H), 7.32 (d, J=8.9 Hz, 1H), 6.89 (d, J=9.0 Hz, 1H), 6.61 (s, 1H), 5.85 (s, 1H), 5.25 - 5.15 (m, 1H), 3.41 (s, 3H), 2.35 (s, 3H), 2.31 - 2.23 (m, 2H), 2.10 - 1.99 (m, 2H), 1.61 - 1.51 (m, 5H), 1.10 - 0.99 (m, 6H).

[0283] 6-chloro-3-((1-(3-(4,4-dimethylcyclohex-1-en-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid enantiomer 2 (5.8 mg, 5%, >97.5% ee). MS: (ES+ ) m / z = 480.2 [M+H] + . 1 H NMR: (300 MHz, DMSO-d6) δ 13.08 (s, 1H), 8.55 (s, 1H), 7.93 (s, 1H), 7.43 (s, 1H), 7.32 (d, J=8.9 Hz, 1H), 6.86 (d, J=8.9 Hz, 1H), 6.61 (s, 1H), 5.86 (s, 1H), 5.25 - 5.15 (s, 1H), 3.41 (s, 3H), 2.35 (s, 3H), 2.31 - 2.23 (m, 2H), 2.10 - 1.99 (m, 2H), 1.57 - 1.45 (m, 5H), 1.10 - 0.99 (m, 6H).

[0284] Intermediate 2 [ka]

[0285] Project 1: Preparation of methyl (R)-2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate To a stirred solution of 5-acetyl-3-chloro-2,7-dimethylisoquinolin-1-one (9.8 g, 39.2 mmol) and (R)-2-methylpropane-2-sulfinamide (23.78 g, 196.2 mmol) in THF (200 mL) was added Ti(Oi-Pr) (55.78 g, 196.2 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 × 300 mL). The filtrate was extracted with ethyl acetate (3 × 300 mL). The combined organic layers were washed with HO (3 × 200 mL), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA=3:2) to give (R)-N-[(1E)-1-(3-chloro-2,7-dimethyl-1-oxoisoquinolin-5-yl)ethylidene]-2-methylpropane-2-sulfinamide (9 g, 64%) as a yellow solid. MS: (ES + ) m / z = 353.1 [M+H] + . Project 2: Preparation of (R)-N-[1-(3-chloro-2,7-dimethyl-1-oxoisoquinolin-5-yl)ethyl]-2-methylpropane-2-sulfinamide

[0286] To a stirred solution of (R)-N-[(1E)-1-(3-chloro-2,7-dimethyl-1-oxoisoquinolin-5-yl)ethylidene]-2-methylpropane-2-sulfinamide (10.5 g, 29.76 mmol) and CeCl3·7H2O (16.63 g, 44.63 mmol) in MeOH (120 mL) was added NaBH4 (2.81 g, 74.39 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 × 200 mL). The combined organic layers were washed with brine (2 × 200 mL), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA = 1:2), followed by further purification by HP-Flash (25%-55% ACN in HO (0.1% FA) for 45 min) to give (R)-N-[1-(3-chloro-2,7-dimethyl-1-oxoisoquinolin-5-yl)ethyl]-2-methylpropane-2-sulfinamide (6.5 g, 61%) as an off-white solid. MS: (ES + ) m / z = 355.0 [M+H] + .

[0287] Project 3: (R)-5-(1-aminoethyl)-3-chloro-2,7-dimethylisoquinolin-1-one hydrochloride A mixture of (R)-N-[(1R)-1-(3-chloro-2,7-dimethyl-1-oxoisoquinolin-5-yl)ethyl]-2-methylpropane-2-sulfinamide (240 mg, 0.68 mmol) in a 1:1 4 M HCl solution in 1,4-dioxane / CHCl (10 mL) was stirred overnight at room temperature under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure to give 5-[(1R)-1-aminoethyl]-3-chloro-2,7-dimethylisoquinolin-1-one hydrochloride (165 mg, 85%) as a white solid. The crude product was used directly in the next step without further purification. MS: (ES + ) m / z = 250.0 [M+H] + .

[0288] Project 4: Preparation of methyl (R)-2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate A solution of 5-[(1R)-1-aminoethyl]-3-chloro-2,7-dimethylisoquinolin-1-one hydrochloride (165 mg, 0.66 mmol), methyl 2-iodobenzoate (344 mg, 1.20 mmol), CsCO (1.27 g, 3.6 mmol), Xantphos (138 mg, 0.24 mmol), and Pd(dba) (109.8 mg, 0.12 mmol) in 1,4-dioxane (15 mL) was stirred at 90 °C for 3 h under a nitrogen atmosphere. The reaction mixture was cooled to room temperature, concentrated under reduced pressure, and purified by preparative TLC (PE / EA=4:1) to give methyl (R)-2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (170 mg, 67%) as a yellow solid. MS: (ES + ) m / z = 385.0 [M+H] + .

[0289] Example 24: (R)-6-chloro-3-((1-(3-(4-(4-fluorophenyl)piperazin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid [ka]

[0290] Project 1: Preparation of (R)-N-((R)-1-(3-(4-(4-fluorophenyl)piperazin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)-2-methylpropane-2-sulfinamide A solution of (R)-N-[(1R)-1-(3-chloro-2,7-dimethyl-1-oxoisoquinolin-5-yl)ethyl]-2-methylpropane-2-sulfinamide (280 mg, 0.79 mmol) in dioxane (10 mL) was treated with CsCO (771 mg, 2.37 mmol), Ruphos (73 mg, 0.16 mmol), p-fluorophenylpiperazine (284 mg, 1.58 mmol), and Pd(dba) (72 mg, 0.079 mmol). The resulting mixture was stirred overnight at 100 °C under a nitrogen atmosphere and then concentrated under reduced pressure. The residue was purified by preparative TLC (CHCl / MeOH=12:1) to give (R)-N-((R)-1-(3-(4-(4-fluorophenyl)piperazin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)-2-methylpropane-2-sulfinamide (200 mg, 50%) as a yellow solid. MS: (ES + ) m / z = 499.2 [M+H] + .

[0291] Project 2: Preparation of (R)-5-(1-aminoethyl)-3-(4-(4-fluorophenyl)piperazin-1-yl)-2,7-dimethylisoquinolin-1(2H)-one A mixture of (R)-N-((R)-1-(3-(4-(4-fluorophenyl)piperazin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)-2-methylpropane-2-sulfinamide (200 mg, 0.40 mmol) in 4 M HCl in dioxane (15 mL) was stirred for 2 hours at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (CHCl / MeOH=10:1) to give (R)-5-(1-aminoethyl)-3-(4-(4-fluorophenyl)piperazin-1-yl)-2,7-dimethylisoquinolin-1(2H)-one (150 mg, 94%) as a yellow solid. MS: (ES + ) m / z = 395.2 [M+H] + .

[0292] Project 3: Preparation of methyl (R)-6-chloro-3-((1-(3-(4-(4-fluorophenyl)piperazin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinate To a stirred solution of (R)-5-(1-aminoethyl)-3-(4-(4-fluorophenyl)piperazin-1-yl)-2,7-dimethylisoquinolin-1(2H)-one (150 mg, 0.38 mmol) and methyl 6-chloro-3-fluoropyridine-2-carboxylate (108 mg, 0.57 mmol) in DMSO (10 mL) was added dropwise at room temperature. The resulting mixture was stirred overnight at 100 °C. The reaction was quenched with HO (20 mL) and extracted with EtOAc (3 × 30 mL). The combined organic layers were washed with brine (3 × 15 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by preparative TLC (CHCl / MeOH=15:1) to give methyl (R)-6-chloro-3-((1-(3-(4-(4-fluorophenyl)piperazin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinate (130 mg, 60%) as a yellow solid. MS: (ES + ) m / z = 564.2 [M+H] + .

[0293] Project 4: Preparation of (R)-6-chloro-3-((1-(3-(4-(4-fluorophenyl)piperazin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid A solution of methyl (R)-6-chloro-3-((1-(3-(4-(4-fluorophenyl)piperazin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinate (120 mg, 0.21 mmol) in MeOH (5 mL) and HO (1 mL) was treated with NaOH (43 mg, 1.07 mmol). The resulting mixture was stirred overnight at 50 °C. The mixture was then acidified to pH 5 with 1 N aqueous HCl. The resulting mixture was diluted with HO (15 mL) and extracted with EtOAc (3 × 30 mL). The combined organic layers were washed with brine (2 × 10 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by preparative TLC (CHCl / MeOH = 5:1). The crude product was purified by preparative HPLC (Column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3 HO), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 21% B to 51% B in 9 min, 51% B; Wavelength: 254 nm; RT1 (min): 7.00) to give (R)-6-chloro-3-((1-(3-(4-(4-fluorophenyl)piperazin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid (34 mg, 28%) as a white solid. MS: (ES) + ) m / z = 550.1 [M+H] + . 1 H NMR: (400 MHz, methanol-d4) δ 8.02 (s, 1H), 7.54 (d, J=1.9 Hz, 1H), 7.17 (d, J=8.9 Hz, 1H), 7.11 - 6.98 (m, 4H), 6.89 (d, J=8.9 Hz, 1H), 6.56 (s, 1H), 5.17 - 5.02 (m, 1H), 3.69 (s, 3H), 3.68 - 3.55 (m, 2H), 3.15 - 2.80 (m, 6H), 2.40 (s, 3H), 1.69 (d, J=6.6 Hz, 3H).

[0294] Example 25: (R)-6-chloro-3-((1-(3-(4-(4-fluorobenzyl)piperazin-1-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid [ka]

[0295] Prepared in a manner similar to Example 24, using para-fluorophenylpiperazine instead of 1-(4-fluorobenzyl)piperazine in Step 1. MS: (ES + ) m / z = 564.1 [M+H] + . 1 H NMR: (300 MHz, methanol-d4): δ 8.01 - 7.94 (m, 1H), 7.59 - 7.49 (m, 3H), 7.19 - 6.92 (m, 3H), 6.71 (d, J=8.8 Hz, 1H), 6.51 (s, 1H), 5.05 - 4.95 (m, 1H), 4.06 (s, 2H), 3.61 (s, 3H), 3.25- 2.90 (m, 8H), 2.37 (s, 3H), 1.63 (d, J=6.6 Hz, 3H).

[0296] Example 26: (R)-6-chloro-3-((1-(3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid [ka]

[0297] Project 1: Preparation of 2-(4-fluorophenyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine To a stirred solution of 5-bromo-2-(4-fluorophenyl)pyridine (300 mg, 1.19 mmol) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (604 mg, 2.38 mmol) in 1,4-dioxane was added Pd(dppf)Cl (87 mg, 0.12 mmol) and KOAc (233 mg, 2.38 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 90 °C for 1 hour and then concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA = 1:2) to give 2-(4-fluorophenyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (290 mg, 81%) as a white solid. MS: (ES + ) m / z = 300.2 [M+H] + .

[0298] Project 2: Preparation of (R)-N-((R)-1-(3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)-2-methylpropane-2-sulfinamide To a stirred solution of (R)-N-[(1R)-1-(3-chloro-2,7-dimethyl-1-oxoisoquinolin-5-yl)ethyl]-2-methylpropane-2-sulfinamide (290 mg, 0.82 mmol) and 2-(4-fluorophenyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (293 mg, 0.98 mmol) in 1,4-dioxane (16 mL) and HO (2 mL) was added Pd(dppf)Cl (59 mg, 0.082 mmol) and NaCO (173 mg, 1.63 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred for 2 h at 100 °C and then concentrated under reduced pressure. The residue was purified by preparative TLC (EA) to give (R)-N-((R)-1-(3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)-2-methylpropane-2-sulfinamide (320 mg, 79%) as a yellow solid. MS: (ES + ) m / z = 492.2 [M+H] + .

[0299] Project 3: Preparation of (R)-5-(1-aminoethyl)-3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethylisoquinolin-1(2H)-one hydrochloride [ka] A solution of (R)-N-((R)-1-(3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)-2-methylpropane-2-sulfinamide (300 mg, 0.61 mmol) in 4N HCl in 1,4-dioxane (5 mL) was stirred for 2 hours at room temperature under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure to give (R)-5-(1-aminoethyl)-3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethylisoquinolin-1(2H)-one hydrochloride (200 mg, 77%). The crude product was used directly in the next step without further purification. MS: (ES+ ) m / z = 388.2 [M+H] + .

[0300] Project 4: Preparation of methyl (R)-6-chloro-3-((1-(3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinate To a stirred solution of (R)-5-(1-aminoethyl)-3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethylisoquinolin-1(2H)-one hydrochloride (200 mg, 0.47 mmol) and methyl 6-chloro-3-fluoropyridine-2-carboxylate (391 mg, 2.06 mmol) in DMSO (5 mL) was added KCO (195 mg, 1.41 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred for 3 hours at 120 °C. The reaction was quenched with HO (10 mL). The aqueous layer was extracted with CHCl (3 × 10 mL). The combined organic layers were washed with brine (2 × 10 mL), dried over anhydrous NaSO, and concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA=1:1) to give methyl (R)-6-chloro-3-((1-(3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinate (140 mg, 48%) as a yellow solid. MS: (ES + ) m / z = 557.2 [M+H] + .

[0301] Project 5: Preparation of (R)-6-chloro-3-((1-(3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid To a stirred solution of methyl (R)-6-chloro-3-((1-(3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinate (130 mg, 0.23 mmol) and NaOH (46 mg, 1.16 mmol) in MeOH (9 mL) was added HO (1.6 mL) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred overnight at 50° C. The mixture was adjusted to pH 6 with 1 N aqueous HCl and concentrated under reduced pressure. The crude product was purified by preparative HPLC (Column: XSelect CSH Prep C18 OBD Column, 19 × 250 mm, 5 μm; Mobile phase A: water (0.1% FA), Mobile phase B: ACN; Flow rate: 20 mL / min; Gradient: 13% B to 40% B in 8 min, 40% B; Wavelength: 254 nm; RT1 (min): 8.00) to give (R)-6-chloro-3-((1-(3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)picolinic acid (28 mg, 21%) as a white solid. MS: (ES + ) m / z = 543.0 [M+H] + . 1 H NMR: (300 MHz, DMSO-d6): δ 8.94 - 8.88 (m, 1H), 8.44 (d, J=6.1 Hz, 1H), 8.32 - 8.20 (m, 2H), 8.25 - 8.10 (m, 2H), 8.04 (d, J=1.8 Hz, 1H), 7.49 (d, J=1.8 Hz, 1H), 7.45 - 7.28 (m, 3H), 7.00 (s, 1H), 6.92 (d, J=9.0 Hz, 1H), 5.40 - 5.30 (m, 1H), 3.42 (s, 3H), 2.40 (s, 3H), 1.54 (d, J=6.5 Hz, 3H).

[0302] Example 27: (R)-5-chloro-2-((1-(3-(6-(4-fluorophenyl)pyridin-3-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka]

[0303] Prepared analogously to Example 26, using 2-(4-fluorophenyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine in place of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl)-1,2,3,6-tetrahydropyridine in Step 2. MS: (ES + ) m / z = 535.0 [M+H] + . 1 H NMR: (300MHz, CD3OD): δ 8.02 (s, 1 H), 7.54 (s, 1H), 7.08 (d, J=8.7 Hz, 1H), 6.84 (s, 1H), 6.77 (d, J=9.0 Hz, 1H), 5.98 (s, 1H), 5.18-5.11 (m, 1H), 3.54 (s, 3H), 3.43 (s, 3H), 3.31-3.20 (m, 2H), 2.98-2.96 (m, 2H), 2.48-2.37 (m, 2H), 2.37 (s, 3H), 1.63 (d, J=6.6 Hz, 3H).

[0304] Example 28: 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0305] Example 29: 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0306] Project 1:Preparation of tert-butyl 2-((N-(1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)-2,4-dinitrophenyl)sulfonamido)benzoate To a solution of 3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-5-(1-hydroxyethyl)-7-methylisoquinolin-1-one (prepared in a manner similar to Example 1, using iodomethane instead of ethyl iodide in Step 3, 400 mg, 1.17 mmol), tert-butyl 2-(2,4-dinitrobenzenesulfonamido)benzoate (494 mg, 1.17 mmol), and triphenylphosphine (612 mg, 2.34 mmol) in tetrahydrofuran (12 mL) was added a solution of DTAD (672 mg, 2.92 mmol) in tetrahydrofuran (3 mL) at 0° C. under a nitrogen atmosphere. The resulting solution was stirred overnight at room temperature and then concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA=3:1) to give tert-butyl 2-((N-(1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)-2,4-dinitrophenyl)sulfonamido)benzoate (864 mg, 98%) as a yellow solid. MS: (ES + ) m / z = 748.3 [M+H] + .

[0307] Project 2: Preparation of tert-butyl 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate A mixture of tert-butyl 2-((N-(1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)-2,4-dinitrophenyl)sulfonamido)benzoate (850 mg, 1.14 mmol), thioglycolic acid (0.16 mL, 2.27 mmol), and triethylamine (0.47 mL, 3.41 mmol) in methylene chloride (20 mL) was stirred at room temperature for 3 hours under a nitrogen atmosphere. The mixture was adjusted to pH 7-8 with saturated aqueous NaHCO3. The reaction mixture was extracted with methylene chloride (3 × 100 mL). The combined organic layers were washed with brine (2 × 100 mL), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA=4:1) to give tert-butyl 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (466 mg, 79%) as a yellow solid. MS: (ES + ) m / z = 518.3 [M+H] + .

[0308] Project 3: Preparation of 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid A mixture of tert-butyl 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (466 mg, 0.90 mmol) in 4 M HCl in 1,4-dioxane (20 mL) was stirred overnight at room temperature under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was diluted with 5 mL DCM, neutralized with 7 M NH in MeOH, and purified by preparative TLC (DCM / MeOH=15:1) to give 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (250 mg, 60%) as a yellow solid. MS: (ES + ) m / z = 462.3 [M+H] + .

[0309] Project 4: Preparation of 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (enantiomers 1 and 2) The racemic mixture of 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (160 mg, 0.35 mmol) was separated by preparative chiral HPLC (column: CHIRAL ART Purification by Cellulose-SC, 2 x 25 cm, 5 μm; Mobile phase A: Hex (0.1% FA)-HPLC, Mobile phase B: IPA-HPLC; Flow rate: 20 mL / min; Gradient: 5% B to 5% B in 33 min; Wavelength: 220 / 254 nm; RT1 (min): 22.187; RT2 (min): 29.177; Sample solvent: EtOH-HPLC; Injection volume: 0.5 mL) gave the enantiomers of 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid as light yellow solids:

[0310] 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid Enantiomer 1 (29 mg, 18%, 99.9% ee). MS: (ES + ) m / z = 462.2 [M+H] + . 1 H NMR: (300 MHz, methanol-d4): δ 7.98 (s, 1H), 7.93 - 7.76 (m, 1H), 7.52 (d, J=1.9 Hz, 1H), 7.22 - 7.10 (m, 1H), 6.61 (s, 1H), 6.58 - 6.47 (m, 1H), 6.41 (d, J=8.4 Hz, 1H), 5.15 - 5.05 (m, 1H), 4.33 - 4.17 (m, 2H), 3.00 - 2.80 (m, 4H), 2.38 (s, 3H), 1.74 - 1.37 (m, 7H), 1.28 (t, J=6.9 Hz, 3H), 1.04 (s, 6H).

[0311] 2-((1-(3-(4,4-dimethylpiperidin-1-yl)-2-ethyl-7-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 2 (29 mg, 17%, 99.3% ee). MS: (ES + ) m / z = 462.2 [M+H] + . 1 H NMR: (300 MHz, methanol-d4): δ 7.98 (s, 1H), 7.93 - 7.76 (m, 1H), 7.52 (d, J=1.9 Hz, 1H), 7.20 - 7.10 (m, 1H), 6.61 (s, 1H), 6.58 - 6.47 (m, 1H), 6.41 (d, J=8.4 Hz, 1H), 5.15 - 5.05 (m, 1H), 4.34 - 4.18 (m, 2H), 3.00 - 2.80 (m, 4H), 2.38 (s, 3H), 1.78 - 1.36 (m, 7H), 1.28 (t, J=6.9 Hz, 3H), 1.03 (s, 6H).

[0312] Example 30: 2-((1-(2,7-dimethyl-1-oxo-3-((3aR,6aS)-5-(2,2,2-trifluoroethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0313] Example 31: 2-((1-(2,7-dimethyl-1-oxo-3-((3aR,6aS)-5-(2,2,2-trifluoroethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0314] Project 1: Preparation of tert-butyl (3aR,6aS)-5-(5-(1-((2-(methoxycarbonyl)phenyl)amino)ethyl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-3-yl)hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate [ka] A solution of methyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (200 mg, 0.52 mmol), tert-butyl (3aR,6aS)-hexahydro-1H-pyrrolo[3,4-c]pyrrole-2-carboxylate (275 mg, 1.30 mmol), CsCO (509 mg, 1.56 mmol), Xantphos (60 mg, 0.10 mmol), and Pd(dba) (47 mg, 0.052 mmol) in 1,4-dioxane (10 mL) was stirred overnight at 100 °C under a nitrogen atmosphere. The reaction mixture was cooled to room temperature, concentrated under reduced pressure, and purified by preparative TLC (DCM / MeOH=25:1) to give tert-butyl (3aR,6aS)-5-(5-(1-((2-(methoxycarbonyl)phenyl)amino)ethyl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-3-yl)hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate (200 mg, 68%) as a yellow solid. MS: (ES + ) m / z = 561.3 [M+H] + .

[0315] Project 2: Preparation of methyl 2-((1-(3-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate [ka] A mixture of tert-butyl (3aR,6aS)-5-(5-(1-((2-(methoxycarbonyl)phenyl)amino)ethyl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-3-yl)hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate (200 mg, 0.36 mmol) in a 1:1 mixture of 4 M HCl in 1,4-dioxane and DCM (10 mL) was stirred at room temperature for 1 hour under a nitrogen atmosphere. The mixture was concentrated under reduced pressure. The residue was diluted with 5 mL of DCM, neutralized with 7 M NH in MeOH, and purified by preparative TLC (DCM / MeOH=15:1) to give methyl 2-((1-(3-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (156 mg, 94%) as a yellow solid. MS: (ES + ) m / z = 461.1 [M+H] + .

[0316] Project 3: Preparation of methyl 2-((1-(2,7-dimethyl-1-oxo-3-((3aR,6aS)-5-(2,2,2-trifluoroethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate [ka] A solution of methyl 2-((1-(3-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (156 mg, 0.34 mmol), KCO (141 mg, 1.02 mmol), and 2,2,2-trifluoroethyl trifluoromethanesulfonate (0.059 mL, 0.41 mmol) in acetonitrile (20 mL) was stirred overnight at room temperature under a nitrogen atmosphere. The mixture was concentrated under reduced pressure and purified by preparative TLC (PE / EA=1:2) to give methyl 2-((1-(2,7-dimethyl-1-oxo-3-((3aR,6aS)-5-(2,2,2-trifluoroethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (110 mg, 59%) as a yellow solid. MS: (ES + ) m / z = 543.2 [M+H] + .

[0317] Project 4: Preparation of 2-((1-(2,7-dimethyl-1-oxo-3-((3aR,6aS)-5-(2,2,2-trifluoroethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka] A mixture of methyl 2-((1-(2,7-dimethyl-1-oxo-3-((3aR,6aS)-5-(2,2,2-trifluoroethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (110 mg, 0.20 mmol), NaOH (121 mg, 3.05 mmol), methanol (10 mL), and water (1 mL) was stirred overnight at 50 °C under a nitrogen atmosphere. The mixture was adjusted to pH 5-6 with 1 M aqueous HCl. The reaction mixture was diluted with HO (30 mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were washed with brine (2 × 30 mL), dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The residue was purified by preparative TLC (DCM / MeOH = 15:1) and then further purified by preparative HPLC (Column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3·H2O), Mobile phase B: ACN (gradient: 24% to 54% in 9 min); Flow rate: 60 mL / min; UV detector at 254 nm) to give 2-((1-(2,7-dimethyl-1-oxo-3-((3aR,6aS)-5-(2,2,2-trifluoroethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (10 mg, 9%) as a white solid. MS: (ES + ) m / z = 529.2 [M+H] + .

[0318] Project 5: Preparation of 2-((1-(2,7-dimethyl-1-oxo-3-((3aR,6aS)-5-(2,2,2-trifluoroethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (enantiomers 1 and 2) The racemic mixture of 2-((1-(2,7-dimethyl-1-oxo-3-((3aR,6aS)-5-(2,2,2-trifluoroethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (30 mg, 0.057 mmol) was separated by preparative chiral HPLC (column: CHIRAL ART) under the following conditions: Separation on Cellulose-SB, 2 × 25 cm, 5 μm; mobile phase A: Hex (0.1% FA)-HPLC, mobile phase B: EtOH-HPLC; flow rate: 20 mL / min; gradient: 20% B to 20% B over 13.5 min; wavelength: 220 / 254 nm; RT1 (min): 7.385; RT2 (min): 12.31; sample solvent: EtOH-HPLC; injection volume: 0.5 mL gave the enantiomers of 2-((1-(2,7-dimethyl-1-oxo-3-((3aR,6aS)-5-(2,2,2-trifluoroethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid as off-white solids.

[0319] 2-((1-(2,7-dimethyl-1-oxo-3-((3aR,6aS)-5-(2,2,2-trifluoroethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 1 (14.4 mg, 46%, 99.9% ee). MS: (ES + ) m / z = 529.2 [M+H] + . 1 H NMR: (400 MHz, methanol-d4): δ 7.96 - 7.80 (m, 2H), 7.51 (d, J=1.9 Hz, 1H), 7.16 - 7.10 (m, 1H), 6.56 - 6.44 (m, 2H), 6.36 (d, J=8.5 Hz, 1H), 5.11 - 5.03 (m, 1H), 3.64 (s, 3H), 3.23 - 3.07 (m, 4H), 3.07 - 2.86 (m, 6H), 2.70-2.55 (m, 2H), 2.36 (s, 3H), 1.62 (d, J=6.6 Hz, 3H).

[0320] 2-((1-(2,7-dimethyl-1-oxo-3-((3aR,6aS)-5-(2,2,2-trifluoroethyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 2 (14.0 mg, 45%, 99.9% ee). MS: (ES + ) m / z = 529.2 [M+H] + . 1 H NMR: 1 H NMR (400 MHz, methanol-d4): δ 7.96 - 7.80 (m, 2H), 7.51 (d, J=1.9 Hz, 1H), 7.16 - 7.05 (m, 1H), 6.54 - 6.45 (m, 2H), 6.37 (d, J=8.5 Hz, 1H), 5.12 - 5.01 (m, 1H), 3.64 (s, 3H), 3.23 - 3.09 (m, 4H), 3.07 - 2.86 (m, 6H), 2.68 - 2.63 (m, 2H), 2.36 (s, 3H), 1.63 (d, J=6.6 Hz, 3H).

[0321] Example 32: 2-((1-(2,7-dimethyl-3-(4-(3-methylbutanoyl)piperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0322] Example 33: 2-((1-(2,7-dimethyl-3-(4-(3-methylbutanoyl)piperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0323] Project 1:Preparation of methyl 2-((1-(2,7-dimethyl-3-(4-(3-methylbutanoyl)piperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate [ka] 3-Methylbutanoyl chloride (42.5 μL, 0.023 mmol) was added to a mixture of methyl 2-((1-(2,7-dimethyl-1-oxo-3-(piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (150 mg, 0.35 mmol, prepared in a manner analogous to Examples 30 and 31, using tert-butyl piperazine-1-carboxylate instead of tert-butyl (3aR,6aS)-hexahydro-1H-pyrrolo[3,4-c]pyrrole-2-carboxylate in 1), CHCl (3 mL), and TEA (240 μL, 1.73 mmol) at 0 °C. The resulting mixture was stirred for 2 h at room temperature. The resulting mixture was quenched with HO (30 mL) and extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with brine (2 × 20 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by preparative TLC (EA) to give methyl 2-((1-(2,7-dimethyl-3-(4-(3-methylbutanoyl)piperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (120 mg, 67%) as a yellow oil. MS: (ES + ) m / z = 519.3 [M+H] + .

[0324] Project 2: Preparation of 2-((1-(2,7-dimethyl-3-(4-(3-methylbutanoyl)piperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (enantiomers 1 and 2) [ka] A solution of methyl 2-((1-(2,7-dimethyl-3-(4-(3-methylbutanoyl)piperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (120 mg, 0.23 mmol) and NaOH (28 mg, 0.69 mmol) in 5:1 MeOH / HO (6 mL) was stirred overnight at 50 °C. The mixture was adjusted to pH 5 with aqueous NaHPO. The resulting mixture was extracted with EtOAc (2 × 20 mL). The combined organic layers were washed with brine (2 × 20 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography (column: C18 silica gel; mobile phase: MeCN in water (0.1% FA), 5% to 100% gradient over 30 min; detector: UV 254 nm) to give 2-((1-(2,7-dimethyl-3-(4-(3-methylbutanoyl)piperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (100 mg, 85%) as a yellow solid. MS: (ES + ) m / z = 505.3 [M+H] + .

[0325] The racemic mixture of 2-((1-(2,7-dimethyl-3-(4-(3-methylbutanoyl)piperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (100 mg, 0.198 mmol) was purified by preparative chiral HPLC (column: chiral cellulose-SB, 4.6 × 100 mm, 3 μm; mobile phase: Hex (0.1% FA): EtOH = 70:30; flow rate: 1.0 mL / min) to give each enantiomer of 2-((1-(2,7-dimethyl-3-(4-(3-methylbutanoyl)piperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid as a white solid:

[0326] 2-((1-(2,7-dimethyl-3-(4-(3-methylbutanoyl)piperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 1 (24.2 mg, 24%, >99.9% ee) as a white solid. MS: (ES + ) m / z = 505.3 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 12.76 (s, 1H), 8.43 (s, 1H), 7.90 - 7.88 (m, 1H), 7.85 - 7.80 (m, 1H), 7.42 (d, J=1.8 Hz, 1H), 7.25 - 7.18 (m, 1H), 6.55 - 6.52 (m, 1H), 6.42 - 6.33 (m, 2H), 5.22 - 5.14 (m, 1H), 4.46 - 3.92 (m, 1H), 3.54 (s, 3H), 3.20 - 2.84 (m, 3H), 2.34 (s, 3H), 2.29 - 2.26 (m, 5H), 2.05 - 2.02 (m, 1H), 1.53 (d, J=6.5 Hz, 3H), 0.93 (d, J=6.6 Hz, 6H).

[0327] 2-((1-(2,7-dimethyl-3-(4-(3-methylbutanoyl)piperazin-1-yl)-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid enantiomer 2 (24.0 mg, 24%, >99.9% ee) as a white solid. MS: (ES+) m / z = 505.3 [M+H] + . 1H NMR: (400 MHz, DMSO-d6) δ 12.72 (s, 1H), 8.48 (s, 1H), 7.90 - 7.88 (m, 1H), 7.85 - 7.80 (m, 1H), 7.42 (d, J=1.9 Hz, 1H), 7.25 - 7.18 (m, 1H), 6.55 - 6.52 (m, 1H), 6.42 - 6.33 (m, 2H), 5.22 - 5.14 (m, 1H), 4.46 - 3.92 (m, 1H), 3.54 (s, 3H), 3.20 - 2.84 (m, 3H), 2.34 (s, 3H), 2.29 - 2.26 (m, 5H), 2.05 - 2.02 (m, 1H), 1.53 (d, J=6.5 Hz, 3H), 0.93 (d, J=6.6 Hz, 6H).

[0328] Example 34: (R)-2-((1-(3-(benzylthio)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka]

[0329] Project 1: Preparation of methyl (R)-2-((1-(3-(benzylthio)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate To a stirred solution of methyl (R)-2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (100 mg, 0.26 mmol) and benzyl mercaptan (32 mg, 0.26 mmol) in dioxane (2 mL) was added DIEA (101 mg, 0.78 mmol), Xantphos (30 mg, 0.052 mmol), and Pd(dba) (24 mg, 0.026 mmol). The resulting mixture was stirred for 12 h at 100 °C under a N atmosphere. The resulting mixture was diluted with HO (20 mL) and extracted with EA (3 × 30 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The residue was purified by preparative TLC (PE / EA=3:1) to give methyl (R)-2-((1-(3-(benzylthio)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (110 mg, 89%) as a yellow solid. MS (ES + ) m / z = 473.1 [M+H] + .

[0330] Project 2: Preparation of (R)-2-((1-(3-(benzylthio)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid To a stirred solution of methyl (R)-2-((1-(3-(benzylthio)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (110 mg, 0.23 mmol) in MeOH (4 mL) was added a solution of NaOH (93 mg, 2.33 mmol) in HO (2 mL). The resulting mixture was stirred for 12 h at 50 °C. The mixture was adjusted to pH 5 with 2 M aqueous HCl. The resulting mixture was diluted with HO (30 mL) and extracted with DCM (3 × 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, and then concentrated under reduced pressure. The crude product was purified by preparative HPLC using the following conditions: XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3·HO); mobile phase B: ACN; flow rate: 60 mL / min; gradient: 26% B to 46% B over 8.5 min, 46% B; wavelength: 254°C to give (R)-2-((1-(3-(benzylthio)-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (59.4 mg, 55%) as an off-white solid. MS (ES) + ) m / z = 481.1 [M+Na] + . 1 H NMR: (400 MHz, methanol-d4) δ 8.0-7.92 (m, 1H), 7.92-7.85 (m, 1H), 7.53 (d, J=1.9 Hz, 1H), 7.32 - 7.14 (m, 5H), 7.10-7.0 (m, 1H), 6.90 (s, 1H), 6.55-6.45 (m, 1H), 6.15-6.1 (m, 1H), 4.95-4.85 (m, 1H), 4.18 (s, 2H), 3.75 (s, 3H), 2.34 (s, 3H), 1.43 (d, J=6.7 Hz, 3H).

[0331] Example 35: 2-((1-(2,7-dimethyl-1-oxo-3-(phenylethynyl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 1)

[0332] Example 36: 2-((1-(2,7-dimethyl-1-oxo-3-(phenylethynyl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (Enantiomer 2) [ka]

[0333] Project 1: Preparation of methyl 2-((1-(2,7-dimethyl-1-oxo-3-(phenylethynyl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate To a stirred solution of methyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (450 mg, 1.17 mmol) and phenylacetylene (239 mg, 2.34 mmol) in 1-butyl-3-methyl-1H-imidazol-3-ium tetrafluoroborate (3 mL) was added pyrrolidine (166 mg, 2.34 mmol) at room temperature under a nitrogen atmosphere. To the above mixture were added PPh (31 mg, 0.12 mmol) and allylpalladium chloride dimer (43 mg, 0.12 mmol) at room temperature. The resulting mixture was stirred for 2 h at 100 °C under a nitrogen atmosphere. The resulting mixture was quenched with HO (20 mL) and extracted with EA (3 × 10 mL). The combined organic layers were washed with brine (1 × 10 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE / EA (8:2)) to give methyl 2-((1-(2,7-dimethyl-1-oxo-3-(phenylethynyl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (230 mg, 44% yield) as a light yellow solid. LCMS m / z 451.3 [M+H] + .

[0334] Project 2:Preparation of 2-((1-(2,7-dimethyl-1-oxo-3-(phenylethynyl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (enantiomers 1 and 2) To a stirring mixture of methyl 2-((1-(2,7-dimethyl-1-oxo-3-(phenylethynyl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (230 mg, 0.51 mmol) in MeOH (5 mL) and HO (1 mL) was added NaOH (408 mg, 10.2 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred for 1 h at 70 °C under a nitrogen atmosphere. The residue was adjusted to pH 6 with 1 N aqueous HCl. The residue was purified by reverse-phase flash chromatography to give 2-((1-(2,7-dimethyl-1-oxo-3-(phenylethynyl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (110 mg, 49% yield) as an off-white solid. LCMS m / z 437.4 [M+H] + .

[0335] The racemic product 2-((1-(2,7-dimethyl-1-oxo-3-(phenylethynyl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (108 mg) was separated into its individual enantiomers by chiral SFC to give 2-((1-(2,7-dimethyl-1-oxo-3-(phenylethynyl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (enantiomer 1, 29 mg, 27% yield) as a white solid, LCMS m / z 437.1 [M+H]. + , 1H NMR (300 MHz, chloroform-d) δ 8.28 - 8.17 (m, 2H), 8.01 (dd, J=8.1, 1.7 Hz, 1H), 7.65 - 7.59 (m, 2H), 7.54 (d, J=1.8 Hz, 1H), 7.48 - 7.36 (m, 3H), 7.25 - 7.13 (m, 2H), 6.60 (ddd, J=8.1, 7.1, 1.0 Hz, 1H), 6.27 (d, J=8.5 Hz, 1H), 5.15-5.0 (m, 1H), 3.86 (s, 3H), 2.42 (s, 3H), 1.68 (d, J=6.7 Hz, 3H); and 2-((1-(2,7-dimethyl-1-oxo-3-(phenylethynyl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (enantiomer 2, 28 mg, 26% yield) as a white solid, LCMS m / z 437.0 [M+H] + , 1 H NMR (300 MHz, chloroform-d) δ 8.25–8.11 (m, 2H), 8.01 (dd, J=8.1, 1.7 Hz, 1H), 7.69–7.58 (m, 2H), 7.57–7.53 (m, 1H), 7.48–7.35 (m, 3H), 7.25–7.13 (m, 2H), 6.66–6.55 (m, 1H), 6.27 (d, J=8.5 Hz, 1H), 5.15–5.0 (m, 1H), 3.86 (s, 3H), 2.42 (s, 3H), 1.68 (d, J=6.7 Hz, 3H).

[0336] Example 37: 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoic acid (Enantiomer 1)

[0337] Example 38: 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoic acid (Enantiomer 1) [ka]

[0338] Project 1: Preparation of 3-chloro-5-(1-ethoxyvinyl)-2,7-dimethylisoquinolin-1(2H)-one A solution of 5-bromo-3-chloro-2,7-dimethylisoquinolin-1-one (5 g, 17.4 mmol) in dioxane (80 mL) was treated portionwise with tributyl(1-ethoxyethenyl)stannane (5.67 g, 15.7 mmol) and Pd(PPh3)4 (2.02 g, 1.7 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 (1:1)) to give 3-chloro-5-(1-ethoxyethenyl)-2,7-dimethylisoquinolin-1-one (4.2 g, 87% yield) as a yellow solid. MS: (ES+) m / z = 278.1 [M+H] + .

[0339] Project 2: Preparation of 3-chloro-5-(2-fluoroacetyl)-2,7-dimethylisoquinolin-1(2H)-one Selectfluor during stirring TMTo a solution of 3-chloro-5-(1-ethoxyethenyl)-2,7-dimethylisoquinolin-1-one (1.42 g, 4 mmol) in ACN (10 mL) and HO (5 mL) was added dropwise a solution of 3-chloro-5-(1-ethoxyethenyl)-2,7-dimethylisoquinolin-1-one (740 mg, 2.7 mmol) in CHCN (10 mL) at 0 °C. The resulting mixture was stirred at room temperature for 1 h. The reaction was then quenched at room temperature with saturated aqueous NaHCO (10 mL). The resulting mixture was extracted with EA (3 × 50 mL). The combined organic layers were washed with brine (2 × 200 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE / EA (1:1)) to give 3-chloro-5-(2-fluoroacetyl)-2,7-dimethylisoquinolin-1-one (500 mg, 70% yield) as a light yellow solid. MS: (ES+) m / z = 268.1 [M+H] + .

[0340] Project 3: Preparation of 3-chloro-5-(2-fluoro-1-hydroxyethyl)-2,7-dimethylisoquinolin-1(2H)-one To a stirred solution of 3-chloro-5-(2-fluoroacetyl)-2,7-dimethylisoquinolin-1-one (500 mg, 1.9 mmol) in MeOH (6 mL) was added NaBH (141 mg, 3.7 mmol) slowly in portions at 0 °C. The resulting solution was stirred for 1 h at room temperature. The reaction was quenched by the addition of saturated aqueous ammonium chloride (20 mL) at room temperature. The resulting mixture was extracted with EA (3 × 30 mL). The combined organic layers were washed with brine (3 × 20 mL), dried over anhydrous NaSO, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE / EA (1:1)) to give 3-chloro-5-(2-fluoro-1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one (198 mg, 39% yield) as a white solid. MS: (ES + ) m / z = 270.1 [M+H] + .

[0341] Project 4:Preparation of methyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoate To a stirred solution / mixture of 3-chloro-5-(2-fluoro-1-hydroxyethyl)-2,7-dimethylisoquinolin-1-one (180 mg, 0.67 mmol) in DCM, TEA (405 mg, 4 mmol) and methanesulfonic anhydride (486 mg, 2.8 mmol) were added sequentially at 0°C. The resulting solution was stirred for 1 hour at 0°C. To the solution, methyl anthranilate (486 mg, 3.2 mmol) was added. The resulting solution was stirred overnight at room temperature. The residue was purified by silica gel column chromatography (eluted with PE / EA (1:1)) to give methyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoate (170 mg, 63% yield) as a light yellow solid. MS: (ES + ) m / z = 403.2 [M+H] + .

[0342] Project 5: Preparation of methyl 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoate To a stirred solution of methyl 2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoate (170 mg, 0.42 mmol) in 1,4-dioxane (10 mL) was added 1-(2,2,2-trifluoroethyl)piperazine (177 mg, 1.1 mmol), CsCO (550 mg, 1.7 mmol), BINAP (26 mg, 0.04 mmol), and Pd(OAc) (9 mg, 0.042 mmol) in portions at room temperature under a nitrogen atmosphere. The resulting solution 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 aqueous layer was extracted with EA (3 × 20 mL). The combined organic layers were dried over anhydrous NaSO and then filtered. The filtrate was then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE / EA (1:1)) to give methyl 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoate (173 mg, 77% yield) as a light yellow oil. MS: (ES + ) m / z = 535.3 [M+H] + .

[0343] Project 6: Preparation of 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoic acid (enantiomers 1 and 2) To a stirred solution of methyl 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoate (163 mg, 0.31 mmol) in MeOH (2 mL) and water (2 mL) was added NaOH (125 mg, 3.1 mmol) portionwise at room temperature. The resulting mixture was stirred for 1 h at 50° C. under a nitrogen atmosphere. The mixture was neutralized to pH 6 with aqueous HCl (1 N), and the mixture was extracted with EA (3 × 30 mL). The organic phase was then washed with brine (2 × 30 mL). The combined organic phases were dried over Na2SO4, filtered, and concentrated. The residue was purified by reverse-phase flash chromatography to give 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoic acid (110 mg, 67% yield) as a white solid. MS: (ES + ) m / z = 521.3 [M+H] + .

[0344] The racemic product 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoic acid (110 mg) was purified by chiral HPLC (column: Chiralcel OD-3 4.6 × 50 mm, 3 μm; mobile phase A: hexane (0.1% FA):EtOH = 90:10; gradient: isocratic) to give 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoic acid (enantiomer 1, 38 mg, 32% yield, 99% ee) as a light yellow solid. 1H NMR (300 MHz, methanol-d4) δ 8.05 (s, 1H), 7.96 - 7.92 (m, 1H), 7.62 - 7.59 (m, 1H), 7.23 - 7.18 (m, 1H), 6.68 - 6.55 (m, 1H), 6.49 (m, 2H), 5.42 - 5.38 (m, 1H), 4.97 - 4.80 (m, 2H), 3.65 (s, 3H), 3.19 - 3.05 (m, 2H), 2.89 - 2.85 (m, 8H), 2.41 (s, 3H), MS: (ES + ) m / z = 521.1 [M+H] + and 2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)-2-fluoroethyl)amino)benzoic acid (enantiomer 2, 34 mg, 31% yield, >99% ee) as a white solid, 1 H NMR (300 MHz, methanol-d4) δ 8.05 (s, 1H), 7.96 - 7.92 (m, 1H), 7.62 - 7.59 (m, 1H), 7.23 - 7.18 (m, 1H), 6.68 - 6.55 (m, 1H), 6.49 (m, MS: (ES) + ) m / z = 521.1 [M+H] + was obtained as.

[0345] Example 39: (R)-2-fluoro-6-((1-(7-fluoro-2-methyl-1-oxo-3-(3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka]

[0346] Project 1: Preparation of 1,3-dioxoisoindolin-2-yl 3-(trifluoromethyl)bicyclo[1.1.1]pentane-1-carboxylate To a stirred solution of 3-(trifluoromethyl)bicyclo[1.1.1]pentane-1-carboxylic acid (1.0 g, 5.6 mmol), DMAP (102 mg, 0.83 mmol), and N-hydroxyphthalimide (1.0 g, 6.0 mmol) in DCM (20 mL) was added N,N'-diisopropylcarbodiimide (0.95 mL, 6.1 mmol) dropwise at 0 °C under an argon atmosphere. The resulting mixture was stirred overnight. The resulting mixture was filtered, and the filter cake was washed with DCM (3 × 30 mL). The filtrate was concentrated under reduced pressure. The resulting mixture was dissolved in EtOAc (50 mL). The combined organic layers were washed with aqueous Na2CO3 (3 × 50 mL) and 1 N aqueous HCl (3 × 50 mL), then dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give 1,3-dioxoisoindol-2-yl 3-(trifluoromethyl)bicyclo[1.1.1]pentane-1-carboxylate (1.6 g, 89% yield) as an off-white solid. 1 H NMR (400 MHz, chloroform-d) δ 7.93-7.87 (m, 2H), 7.83-7.78 (m, 2H), 2.50 (s, 6H).

[0347] Project 2: Preparation of methyl (R)-2-((1-(3-chloro-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-fluorobenzoate To a 100 mL round-bottom flask was added 5-[(1R)-1-aminoethyl]-3-chloro-7-fluoro-2-methylisoquinolin-1-one (1.0 g, 3.9 mmol), dioxane (30 mL), methyl 2-fluoro-6-iodobenzoate (0.6 mL, 3.9 mmol), CsCO (3.84 g, 11.8 mmol), Xantphos (454 mg, 0.78 mmol), and Pd(dba) (360 mg, 0.39 mmol) at room temperature. The resulting mixture was stirred for 3 hours at 90 °C under an argon atmosphere. The resulting mixture was poured into water (60 mL). The resulting mixture was extracted with EtOAc (3 × 60 mL). The combined organic layers were washed with brine and then dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give methyl (R)-2-((1-(3-chloro-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-fluorobenzoate (922 mg, 58% yield) as a light yellow solid. LCMS (ES + ) m / z = 407.0 [M+H] + .

[0348] Project 3: Preparation of methyl (R)-2-fluoro-6-((1-(7-fluoro-3-iodo-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate To a 40 mL vial, methyl (R)-2-((1-(3-chloro-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-fluorobenzoate (729 mg, 1.8 mmol), HI (20 mL), and NaI (1.34 g, 9 mmol) were added at room temperature. The resulting mixture was stirred overnight at room temperature under an argon atmosphere. The mixture was adjusted to pH 7 by the addition of saturated aqueous NaHCO3 (50 mL). The resulting mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine and then dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give methyl (R)-2-fluoro-6-((1-(7-fluoro-3-iodo-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (526 mg, 63% yield) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.79 - 7.71 (m, 2H), 7.66 (s, 1H), 7.50 - 7.43 (m, 1H), 7.22 - 7.14 (m, 1H), 6.43-6.36 (m, 1H), 6.11 (d, J=8.6 Hz, 1H), 5.30 - 5.22 (m, 1H), 3.90 (s, 3H), 3.77 (s, 3H), 1.52 (d, J=6.4 Hz, 3H).

[0349] Project 4: Preparation of methyl (R)-2-fluoro-6-((1-(7-fluoro-2-methyl-1-oxo-3-(3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate To a 20 mL vial was added methyl (R)-2-fluoro-6-((1-(7-fluoro-3-iodo-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (200 mg, 0.4 mmol), DMA (5 mL), 1,3-dioxoisoindol-2-yl 3-(trifluoromethyl)bicyclo[1.1.1]pentane-1-carboxylate (261 mg, 0.8 mmol), (Z)-4,4'-di-tert-butyl-N'-cyano-[2,2'-bipyridine]-6-carboximidamide (27 mg, 0.08 mmol), Zn (105 mg, 1.6 mmol), and NiBr₂·DME (14 mg, 0.04 mmol) at room temperature. The resulting mixture was stirred overnight at 50 °C under an argon atmosphere. The reaction contents were then poured into water (60 mL), and the resulting mixture was extracted with EtOAc (3 x 60 mL). The combined organic layers were washed with brine and then dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford methyl (R)-2-fluoro-6-((1-(7-fluoro-2-methyl-1-oxo-3-(3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (52 mg, 26% yield) as an off-white solid. LCMS (ES) + ) m / z = 507.3 [M+H] + .

[0350] Project 5: Preparation of (R)-2-fluoro-6-((1-(7-fluoro-2-methyl-1-oxo-3-(3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid To an 8 mL vial, methyl (R)-2-fluoro-6-((1-(7-fluoro-2-methyl-1-oxo-3-(3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (52 mg, 0.1 mmol), THF (2 mL), and trimethyl(potassiooxy)silane (40 mg, 0.3 mmol) were added at room temperature. The resulting mixture was stirred for 3 hours at 70 °C under an argon atmosphere. The resulting mixture was then poured into water (10 mL) and extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC to give (R)-2-fluoro-6-((1-(7-fluoro-2-methyl-1-oxo-3-(3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (5.5 mg, 11% yield) as a white solid. LCMS (ES + ) m / z = 493.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.08 (d, J=5.9 Hz, 1H), 7.80-7.74 (m, 1H), 7.50-7.47 (m, 1H), 7.20-7.14 (m, 1H), 6.68 (s, 1H), 6.40-6.33 (m, 1H), 6.16 (d, J=8.6 Hz, 1H), 5.29-5.21 (m, 1H), 3.62 (s, 3H), 2.52 (s, 6H), 1.56 (d, J=6.6 Hz, 3H).

[0351] Example 40: (R)-2-((1-(3-isobutoxy-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka]

[0352] Project 1:Preparation of methyl (R)-2-((1-(3-isobutoxy-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate A stirring solution of methyl (R)-2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (200 mg, 0.52 mmol) in DMF (10 mL) was treated with NaH (37 mg, 1.6 mmol) and stirred for 15 minutes at 0° C. under a nitrogen atmosphere. Isobutanol (193 mg, 2.6 mmol) was then added portionwise at 0° C. The resulting mixture was stirred for 2 hours at 80° 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 CHCl (3×15 mL), and the combined organic layers were dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give methyl (R)-2-((1-(3-isobutoxy-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (100 mg, 47% yield) as a white solid. MS: (ES + ) m / z = 423.2 [M+H] + .

[0353] Project 2: Preparation of (R)-2-((1-(3-isobutoxy-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid To a stirred solution of methyl (R)-2-((1-(3-isobutoxy-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (100 mg, 0.24 mmol) in MeOH (5 mL) and HO (1 mL) was added NaOH (95 mg, 2.4 mmol) at room temperature. The reaction mixture was stirred overnight at 50 °C. The mixture was acidified to pH 3 with 1 N HCl (aq). The aqueous layer was extracted with EtOAc (3 × 10 mL), and the combined organic layers were dried over anhydrous NaSO and then concentrated under reduced pressure. The crude product was purified by preparative HPLC to afford (R)-2-((1-(3-isobutoxy-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (9.4 mg, 10% yield, >99% ee) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.86 - 7.77 (m, 2H), 7.40 - 7.35 (m, 1H), 7.13 (s, 1H), 6.50 - 6.45 (m, 1H), 6.32 - 6.28 (m, 1H), 6.17 (s, 1H), 5.14 (s, 1H), 3.95 - 3.88 (m, 2H), 3.45 (s, 3H), 2.30 (s, 3H), 2.15 - 2.05 (m, 1H), 1.51 - 1.45 (m, 3H), 1.05 - 1.00 (m, 6H). MS: (ES - ) m / z = 407.2 [M−H] - .

[0354] Example 41: (R)-5-fluoro-2-((1-(4-fluoro-2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka]

[0355] Project 1:Preparation of (R)-5-(1-aminoethyl)-3-chloro-2,7-dimethylisoquinolin-1(2H)-one To a stirred solution of (R)-N-[(1R)-1-(3-chloro-2,7-dimethyl-1-oxoisoquinolin-5-yl)ethyl]-2-methylpropane-2-sulfinamide (1 g, 2.8 mmol) in MeOH (10 mL) was added a solution of HCl in 1,4-dioxane (4 M, 10 mL) at room temperature. The resulting mixture was stirred for 1 hour at room temperature. The resulting mixture was concentrated under reduced pressure. The crude product was used directly in the next step without further purification. MS: (ES + ) m / z = 250.9 [M+H] + .

[0356] Project 2: Preparation of methyl (R)-2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-5-fluorobenzoate A solution of (R)-5-(1-aminoethyl)-3-chloro-2,7-dimethylisoquinolin-1(2H)-one (1.32 g, 4 mmol) in 1,4-dioxane (20 mL) was treated with methyl 5-fluoro-2-iodobenzoate (2.95 g, 10.5 mmol), Xantphos (0.61 g, 1.1 mmol), CsCO (6.86 g, 21 mmol), and Pd(dba) (0.48 g, 0.52 mmol) at room temperature. The mixture was stirred overnight at 90 °C under a nitrogen atmosphere. The reaction was quenched with water (30 mL) at room temperature. The aqueous layer was extracted with EtOAc (3 × 150 mL), and the combined organic layers were dried over anhydrous NaSO and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give methyl (R)-2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-5-fluorobenzoate (530 mg, 25% yield) as a brownish yellow solid. MS: (ES + ) m / z = 403.0 [M+H] + .

[0357] Project 3:Preparation of methyl (R)-2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-5-fluorobenzoate To a stirred solution of methyl (R)-2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-5-fluorobenzoate (100 mg, 0.25 mmol) and 1-(2,2,2-trifluoroethyl)piperazine (104 mg, 0.62 mmol) in 1,4-dioxane (1 mL) was added CsCO (162 mg, 0.5 mmol), Pd(OAc) (5.6 mg, 0.025 mmol), and BINAP (15.5 mg, 0.025 mmol) at room temperature. The resulting mixture was stirred overnight at 100 °C under a nitrogen atmosphere. The resulting mixture was concentrated in vacuo. The residue was purified by preparative TLC (PE:EA=1:1) to give methyl (R)-2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-5-fluorobenzoate (90 mg, 68% yield) as a yellow solid. MS: (ES + ) m / z = 535.3 [M+H] + .

[0358] Project 4: Preparation of methyl (R)-5-fluoro-2-((1-(4-fluoro-2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate To a stirring solution of methyl (R)-2-((1-(2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-5-fluorobenzoate (180 mg, 0.34 mmol) in acetonitrile (1 mL) was added Selectfluor TM(107 mg, 0.3 mmol) and acetic acid (4 mg, 0.07 mmol) were added at room temperature. The resulting mixture was stirred at 60° C. overnight. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (PE:EA=1:1) to give methyl (R)-5-fluoro-2-((1-(4-fluoro-2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (69 mg, 37% yield) as a yellow solid. MS: (ES + ) m / z = 553.2 [M+H] + .

[0359] Project 5: Preparation of (R)-5-fluoro-2-((1-(4-fluoro-2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid To a stirred solution of methyl (R)-5-fluoro-2-((1-(4-fluoro-2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (69 mg, 0.13 mmol) in MeOH (5 mL) was added HO (2 mL) and NaOH (100 mg, 0.25 mmol) dropwise at room temperature. The resulting mixture was stirred for 30 minutes at 70 °C. The reaction was diluted with water (10 mL) at room temperature. The aqueous layer was extracted with CHCl (3 × 20 mL). The organic layers were combined, dried over NaSO, filtered, and concentrated under reduced pressure. The crude product was purified by preparative HPLC to give (R)-5-fluoro-2-((1-(4-fluoro-2,7-dimethyl-1-oxo-3-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (20 mg, 30% yield, >99% ee) as a yellow solid. 1H-NMR (400 MHz, methanol-d4) δ 8.07 (m, 1H), 7.67 (s, 1H), 7.59 (m, 1H), 6.94 (m, 1H), 6.28 (m, 1H), 5.37 (m, 1H), 3.65 (s, 3H), 3.54 - 3.40 (m, 2H), 3.23 - 3.03 (m, 4H), 3.05 - 2.95 (m, 2H), 2.76 - 2.55 (m, 2H), 2.36 (s, 3H), 1.65 - 1.55 (m, 3H). MS: (ES + ) m / z = 539.1 [M+H] + .

[0360] Example 42: (R)-2-((1-(3-(3,5'-difluoro-1'-methyl-2'-oxo-1',2'-dihydro-[2,4'-bipyridin]-5-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka]

[0361] Project 1: Preparation of 5-fluoro-1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one In a 40 mL vial, a solution of 4-bromo-5-fluoro-1-methylpyridin-2-one (500 mg, 2.4 mmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (924 mg, 3.6 mmol), KOAc (715 mg, 7.3 mmol), and Pd(dppf)Cl (53 mg, 0.073 mmol) in dioxane (10 mL) was stirred for 1 h at 100° C. under a nitrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with EtOAc. The filtrate was concentrated under reduced pressure to give 5-fluoro-1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one (500 mg, 81% yield) as a black oil, which was used in the next step without further purification. MS: (ES + ) m / z = 254.1 [M+H] + .

[0362] Project 2: Preparation of 5-bromo-3,5'-difluoro-1'-methyl-[2,4'-bipyridin]-2'(1'H)-one In a 40 mL vial, a solution of 5-fluoro-1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one (500 mg, 2 mmol), 2,5-dibromo-3-fluoropyridine (2.52 g, 9.9 mmol), Pd(dppf)Cl (289 mg, 0.4 mmol), and KPO (1.26 g, 5.9 mmol) in dioxane (5 mL) and HO (1 mL) was stirred for 1 h at 100 °C under a nitrogen atmosphere. The resulting mixture was quenched with 20 mL HO and extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine (3 × 10 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (CH2Cl2:MeOH=15:1) to give 5-bromo-3,5'-difluoro-1'-methyl-[2,4'-bipyridin]-2'(1'H)-one (300 mg, 50% yield) as a yellow oil. MS: (ES +) m / z = 301.0 [M+H] + .

[0363] Project 3: Preparation of methyl (R)-2-((1-(7-fluoro-2-methyl-1-oxo-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate In a 40 mL vial, a solution of methyl (R)-2-((1-(3-chloro-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (500 mg, 1.3 mmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (653 mg, 2.6 mmol), KOAc (379 mg, 3.9 mmol), and Pd(dppf)Cl (94 mg, 0.13 mmol) in dioxane (10 mL) was stirred overnight at 100 °C under a nitrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with EtOAc (3 × 5 mL). The filtrate was concentrated under reduced pressure to give methyl (R)-2-((1-(7-fluoro-2-methyl-1-oxo-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (500 mg, 81% yield) as a black oil. The crude product was used directly in the next step without further purification. MS: (ES + ) m / z = 481.2 [M+H] + .

[0364] Project 4: Preparation of methyl (R)-2-((1-(3-(3,5'-difluoro-1'-methyl-2'-oxo-1',2'-dihydro-[2,4'-bipyridin]-5-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate In a 40 mL vial, a solution of methyl (R)-2-((1-(7-fluoro-2-methyl-1-oxo-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (500 mg, 1 mmol), 5-bromo-3,5'-difluoro-1'-methyl-[2,4'-bipyridin]-2'-one (376 mg, 1.25 mmol), Pd(PPh) (241 mg, 0.21 mmol), and KCO (288 mg, 2.1 mmol) in dioxane (6 mL) and HO (1 mL) was stirred for 3 h at 100 °C under a nitrogen atmosphere. The resulting mixture was quenched with 20 mL HO, and the mixture was extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine (3 × 20 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (CHCl:MeOH = 20:1) to give methyl (R)-2-((1-(3-(3,5'-difluoro-1'-methyl-2'-oxo-1',2'-dihydro-[2,4'-bipyridin]-5-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (300 mg, 50% yield) as a yellow oil. MS: (ES + ) m / z = 575.2 [M+H] + .

[0365] Project 5: Preparation of (R)-2-((1-(3-(3,5'-difluoro-1'-methyl-2'-oxo-1',2'-dihydro-[2,4'-bipyridin]-5-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid To a 20 mL vial, methyl (R)-2-((1-(3-(3,5'-difluoro-1'-methyl-2'-oxo-1',2'-dihydro-[2,4'-bipyridin]-5-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (300 mg, 0.52 mmol), potassium trimethylsilanolate (670 mg, 5.2 mmol), and THF (10 mL) were added at room temperature. The resulting mixture was stirred overnight at room temperature. The mixture was acidified to pH 5 by the addition of saturated aqueous NaHPO solution. The resulting mixture was extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine (3 × 10 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by preparative TLC (DCM:MeOH=15:1) followed by preparative achiral SFC to give (R)-2-((1-(3-(3,5′-difluoro-1′-methyl-2′-oxo-1′,2′-dihydro-[2,4′-bipyridin]-5-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (74 mg, 25% yield, 97.9% ee) as an off-white solid. 1 H NMR (400 MHz, DMSO-d6) δ 12.82 (s, 1H), 8.90 - 8.85 (m, 1H), 8.40 (d, J=5.9 Hz, 1H), 8.35 - 8.30 (m, 1H), 8.20 (d, J=6.5 Hz, 1H), 7.90 - 7.85 (m, 1H), 7.85 - 7.82 (m, 1H), 7.55 - 7.48 (m, 1H), 7.33 (s, 1H), 7.19 - 7.08 (m, 1H), 6.80 (d, J=7.5 Hz, 1H), 6.58 - 6.50 (m, 1H), 6.29 - 6.20 (m, 1H), 5.34 - 5.27 (m, 1H), 3.48 (s, 3H), 3.40 (s, 3H), 1.54 (d, J=6.5 Hz, 3H); MS: (ES + ) m / z = 561.2 [M+H] + .

[0366] Example 43: (R)-2-((1-(3-cyclopropyl-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka]

[0367] Project 1: Preparation of methyl (R)-2-((1-(3-cyclopropyl-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate To a 250 mL round-bottom flask was added methyl (R)-2-((1-(3-chloro-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (2.3 g, 1.3 mmol), cyclopropylboronic acid (1.0 g, 0.1 mmol), KPO (3.7 g, 3.9 mmol), dioxane (25 mL), HO (25 mL), and Pd(dppf)Cl (428 mg, 0.13 mmol) at room temperature. The resulting mixture was stirred overnight at 100 °C under a nitrogen atmosphere. The resulting mixture was quenched with 20 mL HO and extracted with CHCl (3 × 50 mL). The combined organic layers were washed with brine (2 × 30 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give methyl (R)-2-((1-(3-cyclopropyl-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (1.1 g, 47% yield) as a yellow solid. MS: (ES + ) m / z = 391.3 [M+H] + .

[0368] Project 2: Preparation of (R)-2-((1-(3-cyclopropyl-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid To a 20 mL vial, methyl (R)-2-((1-(3-cyclopropyl-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (150 mg, 0.38 mmol), MeOH (3 mL), HO (2 mL), THF (3 mL), and NaOH (154 mg, 3.8 mmol) were added at room temperature. The resulting mixture was stirred at 50 °C for 4 h. The mixture was acidified to pH 4 with aqueous HCl (1 N), and the resulting mixture was extracted with CHCl (3 × 50 mL). The combined organic layers were washed with brine (2 × 50 mL) and then dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC to afford (R)-2-((1-(3-cyclopropyl-2,7-dimethyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (55 mg, 35% yield, >99% ee) as a white solid. 1 H-NMR (400 MHz, DMSO-d6) δ 8.54 (s, 1H), 7.92 (s, 1H), 7.80 (d, J=6.8 Hz, 1H), 7.45 (d, J=1.9 Hz, 1H), 7.15 - 7.13 (m, 1H), 6.65 (s, 1H), 6.52 - 6.50 (m, 1H), 6.33 (d, J=8.4 Hz, 1H), 5.18 - 5.10 (m, 1H), 3.69 (s, 3H), 2.34 (s, 3H), 2.06 - 2.04 (m, 1H), 1.51 (d, J=6.5 Hz, 3H), 1.06 - 0.94 (m, 2H), 0.84 - 0.83 (m, 2H). MS (ES-) m / z = 375.1 [MH] - .

[0369] Example 44: 2-(((1R)-1-(3-(3-ethoxy-3,8-diazabicyclo[3.2.1]octan-8-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka]

[0370] Project 1: Preparation of 1-benzylpyrrolidine-2,5-dicarbaldehyde To a stirring solution of oxalyl chloride (3.44 g, 27.1 mmol) in DCM (50 mL) was added dropwise DMSO (4.24 g, 54.2 mmol) at −78° C. under a nitrogen atmosphere. The resulting mixture was stirred for 15 minutes at −78° C. To the above mixture was added dropwise a solution of [1-benzyl-5-(hydroxymethyl)pyrrolidin-2-yl]methanol (1 g, 4.5 mmol) in DCM (5 mL) at −78° C. The resulting mixture was stirred for 30 minutes. To the above mixture was added dropwise a solution of EtN (5.49 g, 54.2 mmol) in DCM (5 mL) at −78° C. The resulting mixture was stirred for 1 hour at room temperature. The resulting mixture was quenched with saturated aqueous sodium bicarbonate (100 mL) and extracted with EtOAc (2 × 100 mL). The combined organic layers were washed with brine (2 × 100 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure to give the crude product (1-benzylpyrrolidine-2,5-dicarbaldehyde (980 mg) which was used directly in the next step without further purification. MS (ES+) m / z = 218.1 [M+H] + .

[0371] Project 2: Preparation of 8-benzyl-3-ethoxy-3,8-diazabicyclo[3.2.1]octane To a stirred solution of 1-benzylpyrrolidine-2,5-dicarbaldehyde (1.46 g, 6.7 mmol) and O-ethylhydroxylamine hydrochloride (1.31 g, 13.4 mmol) in MeOH (40 mL) was added HOAc (2.42 g, 40.3 mmol) and NaBHCN (2.53 g, 40.3 mmol) at 0 °C. The resulting mixture was stirred for 12 h at room temperature, and then the reaction was quenched with HO (100 mL). The resulting mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (2 × 50 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reverse-phase flash chromatography to give 8-benzyl-3-ethoxy-3,8-diazabicyclo[3.2.1]octane (750 mg, 45% yield) as a yellow oil. MS: (ES + ) m / z = 247.2 [M+H] + .

[0372] Project 3: Preparation of 3-ethoxy-3,8-diazabicyclo[3.2.1]octane To a stirring solution of 8-benzyl-3-ethoxy-3,8-diazabicyclo[3.2.1]octane (1 g, 4.1 mmol) in MeOH (100 mL) was added Pd / C (2.0 g, 18.8 mmol) in a pressure tank reactor. The mixture was hydrogenated at room temperature under 50 atmospheres of hydrogen pressure for 12 hours. The reaction mixture was filtered through a pad of Celite and concentrated under reduced pressure. The crude product was purified by preparative HPLC to give 3-ethoxy-3,8-diazabicyclo[3.2.1]octane (216 mg, 34% yield) as a yellow oil. MS: (ES + ) m / z = 157.1 [M+H] + .

[0373] Project 4: Preparation of methyl 2-(((1R)-1-(3-(3-ethoxy-3,8-diazabicyclo[3.2.1]octan-8-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate To a stirred solution of methyl (R)-2-((1-(3-chloro-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (180 mg, 0.46 mmol) and 3-ethoxy-3,8-diazabicyclo[3.2.1]octane (216 mg, 1.4 mmol) in dioxane (5 mL) was added Pd(dba) (42 mg, 0.046 mmol), Ruphos (43 mg, 0.092 mmol), and CsCO (451 mg, 1.4 mmol). The resulting mixture was stirred for 12 h at 100 °C under a nitrogen atmosphere. The resulting mixture was diluted with HO (20 mL). The resulting mixture was extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine (1 × 50 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give methyl 2-(((1R)-1-(3-(3-ethoxy-3,8-diazabicyclo[3.2.1]octan-8-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (170 mg, 73% yield) as a yellow solid. MS: (ES + ) m / z = 509.3 [M+H] + .

[0374] Project 5: Preparation of 2-(((1R)-1-(3-(3-ethoxy-3,8-diazabicyclo[3.2.1]octan-8-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid To a stirred solution of methyl 2-(((1R)-1-(3-(3-ethoxy-3,8-diazabicyclo[3.2.1]octan-8-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (170 mg, 0.33 mmol) in THF (10 mL) was added trimethyl(potassiooxy)silane (429 mg, 3.3 mmol). The resulting mixture was stirred for 2 h at room temperature. The mixture was acidified to pH 6 with aqueous HCl (1 N). The resulting mixture was diluted with HO (20 mL) and extracted with EtOAc (3 × 30 mL). The combined organic layers were washed with brine (2 × 50 mL) and then dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC to give 2-(((1R)-1-(3-(3-ethoxy-3,8-diazabicyclo[3.2.1]octan-8-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (95 mg, 57% yield, >99% ee) as an off-white solid. 1 H NMR (400 MHz, methanol-d4) δ 7.97 - 7.90 (m, 1H), 7.80 - 7.73 (m, 1H), 7.48 - 7.40 (m, 1H), 7.24 - 7.16 (m, 1H), 6.63 - 6.54 (m, 1H), 6.41 (d, J=8.4 Hz, 1H), 6.33 (s, 1H), 5.14 - 5.04 (m, 1H), 3.88 (m, 2H), 3.80 - 3.73 (m, 2H), 3.71 (s, 3H), 3.32 - 3.25 (m, 2H), 2.92 - 2.83 (m, 2H), 2.11 - 1.84 (m, 4H), 1.65 (d, J=6.7 Hz, 3H), 1.23 - 1.16 (m, 3H). MS (ES-) m / z = 493.2 [MH] - .

[0375] Example 45: 3-(((R)-1-(3-(trans-[1,1'-bi(cyclopropane)]-2-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinic acid, diastereomer 1

[0376] Example 46: -(((R)-1-(3-(trans-[1,1'-bi(cyclopropane)]-2-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinic acid, diastereomer 2 [ka]

[0377] Project 1: Preparation of 2-{[1,1'-bi(cyclopropane)]-2-yl}-4,4,5,5-tetramethyl-1,3,2-dioxaborolane To a stirred solution of 1-methyl-1-nitrosourea (2.55 g, 24.7 mmol) in diethyl ether (20 mL), 20% aqueous potassium hydroxide (20 mL) was added at 0 °C, and the mixture was stirred at this temperature for 1 hour. Under ice bath conditions, the organic phase of the above mixture was added to a solution of 2-[(E)-2-cyclopropylethenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1 g, 5.2 mmol) in diethyl ether (20 mL), and then Pd(OAc) (12 mg, 0.052 mmol) was added to the mixture. The mixture was then stirred for 30 minutes at 0 °C. The resulting mixture was then filtered and washed with DCM (3 × 20 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give 2-{[1,1'-bi(cyclopropane)]-2-yl}-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (500 mg, 47% yield) as a colorless oil. GCMS: 208.1.

[0378] Project 2:Preparation of methyl (R)-3-((1-(3-chloro-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinate To a stirred solution of 5-isopropyl-2,3,7-trimethyl-1-methylidene-2H-naphthalene (400 mg, 1.8 mmol), methyl 3-bromo-6-methylpyridine-2-carboxylate (1.22 g, 5.3 mmol), Xantphos (205 mg, 0.35 mmol), and CsCO (1.73 g, 5.3 mmol) in toluene (20 mL) was added Pd(dba) (162 mg, 0.18 mmol) at room temperature. The resulting mixture was stirred overnight at 110 °C under a nitrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with EtOAc (3 × 30 mL). The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC to give methyl (R)-3-((1-(3-chloro-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinate (270 mg, 38% yield) as a yellow solid. MS: (ES+) m / z = 404.2 [M+H] + .

[0379] Project 3: Preparation of methyl 3-(((R)-1-(3-(trans-[1,1'-bi(cyclopropane)]-2-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinate To a stirred mixture of methyl (R)-3-((1-(3-chloro-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinate (240 mg, 0.59 mmol), 2-{[1,1'-bi(cyclopropane)]-2-yl}-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (619 mg, 2.97 mmol), and KCO (246 mg, 1.78 mmol) was added a solution of Pd(PPh) (69 mg, 0.059 mmol) in dioxane (10 mL) and HO (2 mL) at room temperature. The resulting mixture was stirred overnight at 100 °C. The resulting mixture was quenched with HO (20 mL) and extracted with EtOAc (3 × 30 mL). The combined organic layers were washed with brine (1 × 30 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC to give methyl 3-(((R)-1-(3-(trans-[1,1'-bi(cyclopropane)]-2-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinate (170 mg, 64% yield) as a yellow solid. MS: (ES + ) m / z = 450.2 [M+H] + .

[0380] Project 4: Preparation of 3-(((R)-1-(3-(trans-[1,1'-bi(cyclopropane)]-2-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinic acid To a stirred solution of methyl 3-(((R)-1-(3-(trans-[1,1'-bi(cyclopropane)]-2-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinate (170 mg, 0.38 mmol) in MeOH (5 mL) and HO (1 mL) was added LiOH·HO (79 mg, 1.9 mmol) at room temperature. The resulting mixture was stirred for 2 h at room temperature. The mixture was neutralized to pH 6 with 1 M aqueous HCl. The resulting mixture was extracted with EtOAc (3 × 30 mL). The combined organic layers were washed with brine (1 × 30 mL) and dried over anhydrous NaSO. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC to give 3-(((R)-1-(3-(trans-[1,1'-bi(cyclopropane)]-2-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinic acid (88 mg, 53% yield) as a yellow oil. MS: (ES + ) m / z = 436.4 [M+H] + .

[0381] The diastereomers of 3-(((R)-1-(3-(trans-[1,1'-bi(cyclopropane)]-2-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinic acid (80 mg) were separated by chiral HPLC to give 3-(((R)-1-(3-(trans-[1,1'-bi(cyclopropane)]-2-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinic acid, diastereomer 1 (23 mg, 29% yield, >99% ee) as a white solid, 1H NMR (400 MHz, methanol-d4) δ 7.82 - 7.81 (m, 1H), 7.45 (d, J=9.2 Hz, 1H), 7.25 - 6.86 (m, 2H), 6.74 (s, 1H), 5.23 - 5.22 (m, 1H), 3.82 (s, 3H), 2.48 - 2.27 (m, 3H), 1.88 - 1.87 (m, 1H), 1.64 (d, J=6.5 Hz, 3H), 1.35 - 1.27 (m, 1H), 1.07 - 0.89 (m, 3H), 0.57 - 0.45 (m, 2H), 0.29 - 0.20 (m, 2H), MS: (ES + ) m / z = 436.4 [M+H] + and 3-(((R)-1-(3-(trans-[1,1'-bi(cyclopropane)]-2-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)-6-methylpicolinic acid, diastereomer 2 (22 mg, 27% yield, >98% ee) as a white solid, 1 H NMR (400 MHz, methanol-d4) δ 7.83 - 7.82 (m, 1H), 7.45 (d, J=8.7 Hz, 1H), 7.30 - 6.92 (m, 2H), 6.73 (s, 1H), 5.25 - 5.24 (m, 1H), 3.83 (s, 3H), 2.55 - 2.22 (m, 3H), 1.89 - 1.87 (m, 1H), 1.66 (d, J=6.5 Hz, 3H), 1.33 - 1.30 (m, 1H), 1.11 - 0.91 (m, 3H), 0.57 - 0.47 (m, 2H), 0.29 - 0.21 (m, 2H). MS: (ES + ) m / z = 436.4 [M+H] + was obtained as.

[0382] Example 47: (R)-2-((1-(3-(2-([1,1'-bi(cyclopropane)]-1-yl)pyrimidin-5-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka]

[0383] Project 1: Preparation of methyl (R)-2-((1-(3-chloro-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate A solution of (R)-5-(1-aminoethyl)-3-chloro-7-fluoro-2-methylisoquinolin-1(2H)-one (2.00 g, 7.85 mmol), methyl 2-iodobenzoate (2.47 g, 9.4 mmol), Pd(dba) (1.44 g, 1.6 mmol), Xantphos (909 mg, 1.6 mmol), and CsCO (6.4 g, 19.6 mmol) in dioxane (50 mL) was degassed three times and purged with nitrogen. The mixture was then stirred at 90 °C for 12 h under a nitrogen atmosphere. The solution was filtered through a Celite pad, and the filtrate was concentrated. The residue was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 100 / 1 to 30 / 1) to give methyl (R)-2-((1-(3-chloro-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (1.2 g, 39% yield) as a white solid. LCMS (ESI) m / z = 389.1 (M+H).

[0384] Project 2: Preparation of 5-bromo-2-(1-cyclopropylvinyl)pyrimidine A solution of 5-bromo-2-iodo-pyrimidine (2.10 g, 7.4 mmol), 2-(1-cyclopropylvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.3 g, 6.7 mmol), Pd(dppf)Cl CHCl (547 mg, 0.67 mmol), and NaCO (2 g, 18.9 mmol) in dioxane (15 mL) and HO (1.5 mL) was degassed three times and purged with nitrogen. The mixture was then stirred at 90 °C for 24 h under a nitrogen atmosphere. The reaction mixture was partitioned between HO (100 mL) and EtOAc (100 mL). The organic phase was separated, washed with brine (30 mL), dried over NaSO, filtered, and then concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (SiO2, PE / EA=20 / 1) to give 5-bromo-2-(1-cyclopropylvinyl)pyrimidine (440 mg, 17% yield) as a colorless liquid. LCMS (ESI) m / z = 225.1 (M+H); 1 H NMR (400 MHz, CDCl3): δ 8.77 (s, 2H), 6.35 (s, 1H), 5.31 (s, 1H), 2.19-2.08 (m, 1H), 0.94-0.86 (m, 2H), 0.60-0.58 (m, 2H).

[0385] Project 3: Preparation of 2-([1,1'-bi(cyclopropane)]-1-yl)-5-bromopyrimidine To a stirring solution of 5-bromo-2-(1-cyclopropylvinyl)pyrimidine (440 mg, 1.95 mmol) and t-BuOK (329 mg, 2.9 mmol) in THF (10 mL) was added trimethylsulfoxonium iodide (645 mg, 2.93 mmol) in portions at 25 °C under a nitrogen atmosphere. The resulting mixture was stirred for 12 h at 25 °C under a nitrogen atmosphere. The reaction mixture was filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was purified by preparative TLC (SiO, PE / EA = 20 / 1) to give 2-([1,1'-bi(cyclopropane)]-1-yl)-5-bromopyrimidine (150 mg, 19% yield) as a colorless liquid. LCMS (ESI) m / z = 238.9 (M+H).

[0386] Project 4: Preparation of 2-([1,1'-bi(cyclopropane)]-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine A solution of 2-([1,1'-bi(cyclopropane)]-1-yl)-5-bromopyrimidine (150 mg, 627 μmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (319 mg, 1.25 mmol), Pd(dppf)Cl·CHCl (51 mg, 63 μmol), and KOAc (185 mg, 1.88 mmol) in dioxane (4 mL) was degassed three times and purged with nitrogen. The mixture was then stirred at 90 °C for 12 h under a nitrogen atmosphere. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (petroleum ether / ethyl acetate=20 / 1) to give 2-([1,1′-bi(cyclopropane)]-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (179 mg, 99% yield) as a gray solid. 1 H NMR (400 MHz, CDCl3): δ 8.90 (s, 2H), 1.92-1.84 (m, 1H), 1.35 (s, 12H), 1.25 (s, 2H), 0.83-0.75 (m, 2H), 0.60-0.53 (m, 2H), 0.09-0.05 (m, 2H).

[0387] Project 5: Preparation of methyl (R)-2-((1-(3-(2-([1,1'-bi(cyclopropane)]-1-yl)pyrimidin-5-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate A solution of 2-([1,1'-bi(cyclopropane)]-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (170 mg, 594 μmol), methyl (R)-2-((1-(3-chloro-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (150 mg, 386 μmol), Pd(dppf)Cl2·CHCl2 (32 mg, 39 μmol), and Na2CO3 (123 mg, 1.2 mmol) in dioxane (3 mL) and HO (0.3 mL) was degassed three times and purged with nitrogen. The mixture was then stirred at 90 °C for 4 h under a nitrogen atmosphere. The reaction mixture was then partitioned between HO (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 (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1) to give methyl (R)-2-((1-(3-(2-([1,1'-bi(cyclopropane)]-1-yl)pyrimidin-5-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (210 mg, 87% yield) as a yellow oil. LCMS (ESI) m / z = 513.3 (M+H); 1 H NMR (400 MHz, CDCl3): δ 8.77 (s, 2H), 8.30 (d, J=5.2 Hz, 1H), 8.03-7.98 (m, 1H), 7.94 (d, J=8.4 Hz, 1H), 7.50-7.45 (m, 1H), 7.19-7.11 (m, 1H), 6.69 (s, 1H), 6.60-6.55 (m, 1H), 6.16 (d, J=8.4 Hz, 1H), 4.98-4.95 (m, 1H), 3.93 (s, 3H), 3.52 (s, 3H), 1.96-1.92 (m, 1H), 1.64 (d, J=6.8 Hz, 3H), 1.36-1.32 (m, 2H), 0.93-0.86 (m, 2H), 0.66-0.58 (m, 2H), 0.19-0.11 (m, 2H).

[0388] Project 6: Preparation of (R)-2-((1-(3-(2-([1,1'-bi(cyclopropane)]-1-yl)pyrimidin-5-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid To a solution of methyl (R)-2-((1-(3-(2-([1,1'-bi(cyclopropane)]-1-yl)pyrimidin-5-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (200 mg, 390 μmol) in THF (1 mL) and MeOH (1 mL) was added aqueous LiOH·HO (2 M, 1 mL). The mixture was then stirred at 60 °C for 1 h. The reaction mixture was adjusted to pH 5-6 by the addition of aqueous HCl (1 N) at 25 °C and then diluted with HO (5 mL). The mixture was then extracted with EtOAc (3 × 5 mL). The combined organic layers were dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC to give (R)-2-((1-(3-(2-([1,1'-bi(cyclopropane)]-1-yl)pyrimidin-5-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (99 mg, 51% yield) as a yellow solid. LCMS (ESI) m / z = 499.1 (M+H); 1 H NMR (400 MHz, MeOD-d4): δ 8.87 (s, 2H), 8.01-7.83 (m, 2H), 7.50-7.45 (m, 1H), 7.26-7.09 (m, 1H), 7.06 (s, 1H), 6.56 (t, J=8.0 Hz, 1H), 6.30 (d, J=8.0 Hz, 1H), 5.26-5.20 (m, 1H), 3.50 (s, 3H), 1.92-1.87 (m, 1H), 1.63 (d, J=6.8 Hz, 3H), 1.40-1.25 (m, 2H), 0.94-0.78 (m, 2H), 0.65-0.54 (m, 2H), 0.23-0.09 (m, 2H).

[0389] Example 48: (R)-2-((1-(3-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)oxy)-5-fluoropyridin-3-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid [ka]

[0390] Project 1: Preparation of 1-(difluoromethyl)-1H-pyrazol-4-ol To a solution of 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole (2 g, 8.20 mmol) in THF (30 mL) was added HO (30% pure, 1.86 g, 16.4 mmol) and NaOH (2 M, 8.2 mL). The mixture was stirred at 0 °C for 30 min. The reaction was quenched with water (50 mL), and the pH of the mixture was adjusted to 5–6 by adding aqueous hydrochloric acid (1 N). The mixture was extracted with EA (3 × 60 mL). The combined organic layers were washed successively with saturated sodium sulfite solution (200 mL) and brine (200 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to give 1-(difluoromethyl)pyrazol-4-ol (900 mg, 82% yield) as a yellow solid. 1 H NMR (400 MHz, CDCl3): δ 7.43 (s, 1H), 7.40 (s, 1H), 7.04 (t, J=60.4 Hz, 1H).

[0391] Project 2: Preparation of 5-bromo-2-((1-(difluoromethyl)-1H-pyrazol-4-yl)oxy)-3-fluoropyridine To a solution of 5-bromo-2,3-difluoro-pyridine (1.29 g, 6.67 mmol) and 1-(difluoromethyl)pyrazol-4-ol (895 mg, 6.67 mmol) in DMF (15 mL) was added K2CO3 (2.77 g, 20.0 mmol). The mixture was stirred at 60 °C for 1 h. The reaction was quenched with water (20 mL) and extracted with EA (3 × 15 mL). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to give 5-bromo-2-((1-(difluoromethyl)-1H-pyrazol-4-yl)oxy)-3-fluoropyridine (1.6 g, 77% yield) as a white solid. LCMS (ESI) m / z = 308.0 (M+H).

[0392] Project 3: Preparation of 2-((1-(difluoromethyl)-1H-pyrazol-4-yl)oxy)-3-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine A solution of 5-bromo-2-((1-(difluoromethyl)-1H-pyrazol-4-yl)oxy)-3-fluoropyridine (500 mg, 1.62 mmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (618 mg, 2.43 mmol), Pd(dppf)Cl.CHCl (133 mg, 162 μmol), and potassium acetate (319 mg, 3.25 mmol) in dioxane (10 mL) was degassed three times and purged with nitrogen. The mixture was then stirred at 90 °C for 12 h under a nitrogen atmosphere. The reaction mixture was filtered, and the filtrate was concentrated to give 2-((1-(difluoromethyl)-1H-pyrazol-4-yl)oxy)-3-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (620 mg, 45% yield) as a yellow solid. LCMS (ESI) m / z = 356.2 (M+H).

[0393] Project 4:Preparation of methyl (R)-2-((1-(3-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)oxy)-5-fluoropyridin-3-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate A solution of 2-((1-(difluoromethyl)-1H-pyrazol-4-yl)oxy)-3-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (365 mg, 1.03 mmol), methyl (R)-2-((1-(3-chloro-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (200 mg, 514 μmol), Pd(dppf)Cl·CHCl (42 mg, 51.4 μmol), and KCO (142 mg, 1.03 mmol) in dioxane (8 mL) and HO (1 mL) was degassed three times and purged with nitrogen. The mixture was then stirred at 90 °C for 12 h under a nitrogen atmosphere. The reaction was quenched with water (10 mL) and extracted with DCM (3 × 15 mL). The combined organic layers were washed with brine (20 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography to give methyl (R)-2-((1-(3-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)oxy)-5-fluoropyridin-3-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (220 mg, 73% yield) as a yellow solid. LCMS (ESI) m / z = 582.2 (M+H).

[0394] Project 5: Preparation of (R)-2-((1-(3-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)oxy)-5-fluoropyridin-3-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid To a solution of methyl (R)-2-((1-(3-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)oxy)-5-fluoropyridin-3-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoate (210 mg, 361 μmol) in MeOH (1 mL), THF (1 mL), and HO (0.4 mL) was added LiOH·HO (45 mg, 1.08 mmol) at 60° C. for 1 h. The reaction was quenched with water (2 mL), and the pH of the mixture was adjusted to 7 with aqueous citric acid. The mixture was then extracted with EA (3 × 2 mL). The combined organic layers were washed with brine (5 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC and lyophilized to give (R)-2-((1-(3-(6-((1-(difluoromethyl)-1H-pyrazol-4-yl)oxy)-5-fluoropyridin-3-yl)-7-fluoro-2-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)ethyl)amino)benzoic acid (29 mg, 14% yield) as a white solid. LCMS (ESI) m / z = 590.2 (M+Na); 1 H NMR (400 MHz, CDCl3): δ 8.23 ​​(s, 2H), 8.15 (d, J=2.0 Hz, 1H), 8.08-7.98 (m, 2H), 7.83 (s, 1H), 7.61-5.56 (m, 1H), 7.52-7.45 (m, 1H), 7.34-7.03 (m, 2H), 6.71 (s, 1H), 6.63 (t, J=7.6 Hz, 1H), 6.20 (d, J=8.4 Hz, 1H), 5.01-4.95 (m, 1H), 3.51 (s, 3H), 1.67 (d, J=6.8 Hz, 3H).

[0395] Examples 49 to 595 The compounds of Examples 49-595 were prepared analogously to the methods described in Examples 1-48 and are identified and characterized below in Table 1. Unless otherwise noted, all depicted chiral centers exist as (R)- and (S)-racemic mixtures or as either the (R)- or (S)-enantiomers.

Table 1

Table 2

Table 3

Table 4

Table 5

Table 6

Table 7

Table 8

Table 9

Table 10

Table 11

Table 12

Table 13

Table 14

Table 15

Table 18

Table 19

Table 33

Table 91

Table 93

Table 95

Table 99

Table 100

Table 199

[0396] Assays and Compound Testing In vitro cell proliferation: Determination of the growth inhibition values of T47D cells expressing the mutant PI3Ka (H1047R) mutation and SKBR3 cells expressing WT PI3Ka. EC 50 Value determination.

[0397] T47D or SKBR3 cells were trypsinized, resuspended in culture medium, and seeded into assay-ready plates. T47D culture medium consisted of RPMI, 10% FBS, and insulin (0.2 units / mL). SKBR3 culture medium consisted of McCoys 5a and 10% FBS. Cells were seeded at a density of 1,500 cells / well and dispensed in 50 μL into 384-well assay plates (Corning, 89089-790). Assay plates were pre-stamped with 10-point dilutions of the compound of interest and controls. Echo655 was used to stamp 40 nL of compound or DMSO. Cells were grown for 72 hours at 37°C and 5% CO2. After 72 hours, cells were equilibrated at room temperature for 15 minutes. 30 μL of CellTiter-Glo reagent was added to the plate, followed by shaking at 300-500 rpm for 30 minutes. The cells were then read on an Envision plate reader. The percentage of growth inhibition was calculated using the following formula: % inhibition = 100 × (Lum D -Lum サンプル ) / (Lum D -Lum Inh ) (where D is from cells treated with 0.1% DMSO only; Inh is from cells treated with 10 μM alpelisib). The effective concentration (EC 50 ) with Xlfit(v5.3.1.3), formula 201: Y = base + (top - base) / (1 + 10^((LogEC 50 Calculated by curve fitting using (x) × Hillslope). [Table 210]

[0398] In vitro cell pAKT: IC determination of AKT phosphorylation (pAKT) inhibition in T47D cells expressing the mutant PI3Ka (H1047R) mutation and SKBR3 cells expressing WT PI3Ka 50 Value determination .

[0399] T47D or SKBR3 cells were trypsinized, resuspended in culture medium, and seeded into assay plates. T47D culture medium consisted of RPMI, 10% FBS, and insulin (0.2 units / mL). SKBR3 culture medium consisted of McCoy's 5a and 10% FBS. Cells were seeded at a density of 5000 cells / well and dispensed into 384-well assay plates (Perkin Elmer, 6008238) at 12.5 μL. Assay plates were pre-stamped with 10-point dilutions of the compound of interest and controls. Echo655 was used to stamp 12.5 nL of compound or DMSO. Cells were grown for 6 hours at 37°C and 5% CO2. After 6 hours, 4 μL of lysis buffer reagent was added to the plate, which was then centrifuged at 1000 rpm for 1 minute. The plate was then incubated at room temperature for 30 minutes. After 30 minutes, 4 μL of antibody mixture containing Eu cryptate, d2 cryptate, and detection buffer was added to the plate. The plate was centrifuged at 1000 rpm for 1 minute and then incubated overnight at room temperature. The plate was read on an Envision plate reader using the HTRF protocol. The percentage inhibition of AKT phosphorylation was calculated using the following formula: % inhibition = 100 × (pAKTHC - pAKT sample) / (pAKTHC - pAKTLC) (where pAKTHC is from cells treated with 0.1% DMSO only; pAKTLC is from cells treated with 10 μM alpelisib). IC 50 The concentration (concentration achieving 50% inhibition of pAKT) was calculated using Xlfit (v5.3.1.3), using the formula: Y = nadir + (top-nadir) / (1 + 10^((LogIC 50 Calculated by curve fitting using (x) × Hillslope). [Table 211]

[0400] Table EC50 EC values ​​shown 50 For values, "A" is 1nM <EC 50 <500nM; "B" is 500nM <EC 50 <2 μM; "C" indicates 2 μM <EC 50 <15 μM; and "D" refers to the EC 50 >15μM. [Table 212] [Table 213] [Table 214] [Table 215] [Table 216] [Table 217] [Table 218] [Table 219] [Table 220] [Table 221] [Table 222] [Table 223] [Table 224] [Table 225] [Table 226]

[0401] Table IC 50 IC values ​​shown 50 For values, "A" is 1nM <IC 50 <500nM; "B" is 500nM <IC 50 <2 μM; "C" indicates 2 μM <IC 50 <15 μM; and "D" refers to the IC 50 >15μM. [Table 227] [Table 233] [Table 233] [Table 233] [Table 233] [Table 233] [Table 233] Table 233 Table 233 Table 233 Table 233 Table 233 Table 233 Table 233 Table 233

[0402] CD1 mice were administered a single IV or PO dose, followed by serial plasma sampling at 0.0833 h (IV only), 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, and 24 h post-dose. Desired serial concentrations of working solution were achieved by diluting the analyte stock solution with 50% aqueous acetonitrile. 10 µL of working solution (0.5, 1, 2, 5, 10, 50, 100, 500, and 1000 ng / mL) was added to 10 µL of blank female CD1 mouse plasma to achieve calibration standards ranging from 0.5 to 1000 ng / mL (0.5, 1, 2, 5, 10, 50, 100, 500, and 1000 ng / mL) in a total volume of 20 µL. Five quality control samples of plasma at concentrations of 1 ng / mL, 2 ng / mL, 5 ng / mL, 50 ng / mL, and 800 ng / mL were prepared independently from those used in the calibration curve. These QC samples were prepared on the day of analysis in the same manner as the calibration standards. 20 μL of the standard, 20 μL of the QC sample, and 20 μL of the unknown sample (10 μL of plasma and 10 μL of blank solution) were each added to 200 μL of acetonitrile containing the IS mixture to precipitate proteins. After centrifugation at 4000 rpm for 15 minutes at 4°C, the samples were vortexed for 30 seconds. The supernatant was diluted 1:2 (V / V, 1:2) with water. Five μL of the diluted supernatant was then injected into the LC / MS / MS system for quantitative analysis. The results are shown in Table 4. [Table 240]

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Claims

1. Formula (1) [Chemical Formula 1] [During the ceremony, R １ is alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein each of the alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; R ２ is H, C １ -C ４ Alkyl, C ３ -C ７ Cycloalkyl, CF ３ , C.H. ２ F or CF ２ H, where R ２ is not H, but R ２ is a chiral center and exists as an (R)- and (S)-racemic mixture or as an (R)- or (S)-enantiomer; R ３ is H or C １ -C ４ is alkyl; R ４ is H, F, Cl or CH ３ and R ６ is H, C １ -C ４ Alkyl, C ３ -C ７ Cycloalkyl, heteroaryl, CF ３ , C.H. ２ F or CF ２ H; R ７ is H, C １ -C ４ Alkyl, C ３ -C ７ Cycloalkyl, halogen, CN, CF ３ , OCF ３ , C.H. ２ F or CF ２ H; Each R ８ are independently H, C １ -C ４ Alkyl, C ３ -C ７ Cycloalkyl, halogen, CN, CF ３ , OCF ３ , C.H. ２ F or CF ２ H; R ５ teeth H; halogen; -O-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ ; -S-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ ; -S(O)-L １ -L ２ -L ３ -L ５ -L ６ -L ７ -R ９ ; -S(O) ２ -L １ -L ２ -L ３ -L ５ -L ６ -L ７ -R ９ ; -(NR １０ )-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ ;または -L ８ -L ９ -L １０ -L １１ -L １２ -R １４ and where: L １ , L ２ , L ３ , L ６ and L ７ Each of the is independently (CHR １１ ), (CHR １１ -O), (CHR １１ -S), (C ３ -C ７ cycloalkyl), (CH ２ ) １－４ or a bond; L ４ is C=O, C=S or a bond; L ５ is NR １０ , S, O or a bond; R ９ is H, C(=O)R １２ , C(═O)NR １２ R １３ , N.R. １２ R １３ , C(=O)OR １２ , C １ -C ６ Alkyl, C １ -C ６ fluoroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, where C １ -C ６ Alkyl, C １ -C ６ Each of the fluoroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; or NR １０ When exists, R ９ and R １０ may be taken together with the nitrogen atom to which it is attached to form a substituted or unsubstituted ring; R １０ and R １１ each independently represents H or C １ -C ４ Alkyl (e.g. CH ３ , C.H. ２ CH ３ or CH(CH ３ ) ２ ) where C １ -C ４ The alkyl is unsubstituted or substituted; R １２ and R １３ each independently represents H, C １ -C ６ alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein C １ -C ６ Each of alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; or R １２ and R １３ may be taken together with the nitrogen atom to which it is attached to form a substituted or unsubstituted ring; L ８ (CHR １５ ), (CHR １５ -O), (CHR １５ -S), (CHR １５ -NR １６ ), C═O, C═S or a bond; L ９ is optionally part of a bridged, fused or spiro ring system; ３ -C ７ Cycloalkyl, C(R １５ ) = C(R １５ ), C≡C or a bond; L １０ is independently (CHR １５ ), O, S, (NCR １５ ), N(C═O) or a bond; L １１ (CHR １５ ), C═O, C═S or a bond; L １２ is H, (C ３ -C ７ cycloalkyl), heterocyclyl, aryl, heteroaryl, or a bond, ３ -C ７ Each of (cycloalkyl), heterocyclyl, aryl or heteroaryl is unsubstituted or substituted; ３ -C ７ cycloalkyl) and / or heterocyclyl are optionally part of a bridged, fused, or spiro ring system; R １４ H, CR １５ R １６ R １７ , OR １７ , S.R. １７ , N.R. １６ R １７ , C １ -C ６ Alkyl, C １ -C ６ fluoroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, where C １ -C ６ Alkyl, C １ -C ６ each of the fluoroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; R １５ and R １６ each independently represents H or C １ -C ３ is alkyl; and Each R １７ are independently H, C １ -C ６ Alkyl, C １ -C ６ fluoroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, where C １ -C ６ Alkyl, C １ -C ６ Each of the fluoroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl is unsubstituted or substituted; or R １６ and R １７ may be taken together with the nitrogen atom to which it is attached to form a substituted or unsubstituted ring; However, R ５ Ga-L ８ -L ９ -L １０ -L １１ -L １２ -R １４ When L ８ , L ９ , L １０ , L １１ , L １２ and R １４ at least one of R is a carbon-containing moiety; ５ is directly attached to the (isoquinolone) core structure by a carbon atom; or R ５ is a non-aromatic N-linked heterocyclic ring 【Chemistry 2】 wherein the heterocyclic ring is substituted or unsubstituted, optionally contains one or more additional ring atoms selected from N, O, Si and S, and is optionally part of a bridged, fused or spiro ring system. or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof.

2. R ５ -(NR １０ )-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ where L １ ~L ７ , R ９ and R １０ 2. The compound of claim 1, wherein:

3. R ５ Ga-O-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ where L １ ~L ７ and R ９ 2. The compound of claim 1, wherein:

4. R ５ Ga-S-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ ;-S(O)-L １ -L ２ -L ３ -L ５ -L ６ -L ７ -R ９ or -S(O) ２ -L １ -L ２ -L ３ -L ５ -L ６ -L ７ -R ９ where L １ ~L ７ and R ９ 2. The compound of claim 1, wherein:

5. R ５ Ga-L ８ -L ９ -L １０ -L １１ -L １２ -R １４ where L ８ ~L １２ and R １４ 2. The compound of claim 1, wherein:

6. R ５ is a non-aromatic N-linked heterocyclyl ring 【Chemistry 3】 2. The compound of claim 1, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound or pharmaceutically acceptable salt thereof, wherein the heterocyclyl ring is substituted or unsubstituted, optionally contains one or more additional ring atoms selected from N, O, Si and S, and optionally is part of a bridged, fused or spiro ring system.

7. R ６ is CH ３ 2. The compound of claim 1, wherein:

8. R ７ is CH ３ 2. The compound of claim 1, wherein:

9. R ８ 2. The compound of claim 1, wherein is H, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound, or pharmaceutically acceptable salt thereof.

10. R １ is heterocyclyl, where heterocyclyl is unsubstituted or substituted, and R ２ is CH ３ and R ３ 2. The compound of claim 1, wherein is H, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound, or pharmaceutically acceptable salt thereof.

11. R １ is aryl, where aryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ 2. The compound of claim 1, wherein is H, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound, or pharmaceutically acceptable salt thereof.

12. R １ is heteroaryl, where heteroaryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ 2. The compound of claim 1, wherein is H, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound, or pharmaceutically acceptable salt thereof.

13. R １ is heterocyclyl, where heterocyclyl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ８ 2. The compound of claim 1, wherein is H, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound, or pharmaceutically acceptable salt thereof.

14. R １ is aryl, where aryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ８ 2. The compound of claim 1, wherein is H, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound, or pharmaceutically acceptable salt thereof.

15. R １ is heteroaryl, where heteroaryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ８ 2. The compound of claim 1, wherein is H, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound, or pharmaceutically acceptable salt thereof.

16. R １ is heterocyclyl, where heterocyclyl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ８ is H and R ６ is CH ３ 2. The compound of claim 1, wherein:

17. R １ is aryl, where aryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ８ is H and R ６ is CH ３ 2. The compound of claim 1, wherein:

18. R １ is heteroaryl, where heteroaryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ８ is H and R ６ is CH ３ 2. The compound of claim 1, wherein:

19. R １ is heterocyclyl, where heterocyclyl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ５ Ga-O-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ 2. The compound of claim 1, wherein:

20. R １ is aryl, where aryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ５ Ga-S-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ８ 2. The compound of claim 1, wherein:

21. R １ is aryl, where aryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ５ -(NR １０ )-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ 2. The compound of claim 1, wherein:

22. R １ is heteroaryl, where heteroaryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ５ Ga-L ８ -L ９ -L １０ -L １１ -L １２ -R １４ 2. The compound of claim 1, wherein:

23. R １ is heterocyclyl, where heterocyclyl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ５ Ga-O-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ and R ８ 2. The compound of claim 1, wherein is H, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound, or pharmaceutically acceptable salt thereof.

24. R １ is aryl, where aryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ５ Ga-S-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ and R ８ 2. The compound of claim 1, wherein is H, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound, or pharmaceutically acceptable salt thereof.

25. R １ is aryl, where aryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ５ -(NR １０ )-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ and R ８ 2. The compound of claim 1, wherein is H, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound, or pharmaceutically acceptable salt thereof.

26. R １ is heteroaryl, where heteroaryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ５ Ga-L ８ -L ９ -L １０ -L １１ -L １２ -R １４ and R ８ 2. The compound of claim 1, wherein is H, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound, or pharmaceutically acceptable salt thereof.

27. R １ is heterocyclyl, where heterocyclyl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ５ Ga-O-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ and R ８ is H and R ６ is CH ３ 2. The compound of claim 1, wherein:

28. R １ is aryl, where aryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ５ Ga-S-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ and R ８ is H and R ６ is CH ３ 2. The compound of claim 1, wherein:

29. R １ is heterocyclyl, where heterocyclyl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ５ -(NR １０ )-L １ -L ２ -L ３ -L ４ -L ５ -L ６ -L ７ -R ９ and R ８ is H and R ６ is CH ３ 2. The compound of claim 1, wherein:

30. R １ is aryl, where aryl is unsubstituted or substituted, and R ２ is CH ３ and R ３ is H and R ５ Ga-L ８ -L ９ -L １０ -L １１ -L １２ -R １４ and R ８ is H and R ６ is CH ３ 2. The compound of claim 1, wherein:

31. The compound of formula (1) is represented by formula (2) 【Chemistry 4】 [During the ceremony, X １ , X ２ and X ３ each independently is N, CH, or substituted C; R ５ and R ８ is as defined in the compound of formula (1), and The carbons marked with an * are chiral centers and exist as (R)- and (S)-racemic mixtures or as (R)- or (S)-enantiomers.] or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound or pharmaceutically acceptable salt thereof.

32. The compound of formula (1) is represented by formula (3) 【Chemistry 5】 [During the ceremony, R １ is heterocyclyl, aryl, or heteroaryl, wherein the heterocyclyl, aryl, or heteroaryl ring directly attached to the nitrogen atom attached to the asymmetric center to which the isoquinoline moiety is attached contains a carboxylic acid substituent ortho to the point of attachment, and optionally one or more further substituents; R ５ and R ８ is as defined in the compound of formula (1), and The carbons marked with an * are chiral centers and exist as (R)- and (S)-racemic mixtures or as (R)- or (S)-enantiomers.] or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound or pharmaceutically acceptable salt thereof.

33. A pharmaceutical composition comprising the compound of any one of claims 1 to 32, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

34. 34. The pharmaceutical composition of claim 33, further comprising one or more anti-cancer agents.

35. 35. The pharmaceutical composition of claim 34, 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, CDK inhibitors and proteasome inhibitors.

36. 33. A method of treating a disease associated with PI3K activity in a subject in need thereof, comprising administering a therapeutically effective amount of a compound of any of claims 1 to 32, or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof.

37. 37. The method of claim 36, wherein the disease is cancer.

38. 37. The method of claim 36, wherein the disease is congenital lipomatous overgrowth, vascular malformation, epidermal nevus, scoliosis / skeletal and spinal cord syndrome (CLOVES), mosaic tissue overgrowth syndrome, venous malformations and brain malformations associated with severe epilepsy, or PIK3CA-related overgrowth syndrome (PROS).

39. 38. The method of claim 37, wherein the disease is a cancer carrying the PI3Kα H1047R mutation.

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