Benzopyrimidin-4 (3h)-ones as PI3K inhibitors

Abstract

Novel PI3K inhibitors of general formula (1) wherein R1 to R8 are as defined herein, and methods for their preparation and their use in the treatment of diseases associated with elevated or activated PI3K pathways are described. # imgabs0 #

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 63 / 350,234, filed on June 8, 2022, the contents of which are incorporated herein by reference in their entirety for all purposes. Background Art

[0003] Phosphatidylinositol lipids (PI) and their various phosphorylated subspecies are second messengers involved in various cell vesicle transport and signal transduction processes. Phosphoinositide 3' kinase (PI3K) is an enzyme family responsible for phosphorylating the 3' hydroxyl position of the inositol ring of PI. PI3K is subdivided into 3 categories according to its structure and substrate. Class II PI3K (PI3K-C2α, PI3K-C1β, PI3K-C2γ) and class III PI3K (vps34) are monomeric enzymes mainly associated with endocytosis and autophagy (Posor et al., Biochim Biophys Acta 2015, 1851, 794; Backer, Biochem J. 2016, 473, 2251). Class I PI3K is a heterodimer composed 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 upon interaction with receptor tyrosine kinases (RTKs), Ras-related GTPases, G protein-coupled receptors and / or related adaptor proteins and, in their active form, convert phosphatidylinositol 4,5-bisphosphate (PIP2) to phosphatidyl 3,4,5-triphosphate (PIP3) (Fruman et al., Cell 2017, 170, 605).

[0004] High local concentrations of PIP3 promote the recruitment and activation of downstream signaling partners, including AKT and Mtor. Activation of the AKT / mTOR pathway is implicated in a variety of growth-related effects and pathologies, including glucose regulation, cell survival, angiogenesis, and proliferation (Porta et al., Front Oncol. 2014, 4, 1), suggesting that class I PI3Ks act as key upstream regulators of these functions.

[0005] Class I PI3K is further subdivided into 4 isoforms (α, β, γ and δ) based on the identity of its catalytic subunit (p110α, p110β, p110γ or p110δ) and regulatory subunit (p85α or its various splice variants, p85β, p55γ or p101), which play different roles in cell physiology (Vanhaesebroeck et al., J Mol Med (Berl). 2016, 94, 5). PI3Kγ and PI3Kδ are mainly expressed in leukocytes and play an important role in proinflammatory 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), whereas PI3Kβ is known to play a specific role in platelet aggregation (Liu et al., Nat Rev Drug Discov. 2009, 8, 627; Jackson et al., Nat Med. 2005, 11, 507).

[0006] Elevated or constitutive activation of the PI3K pathway is one of the most common events in human cancer. The PI3K pathway is overactivated through a variety of 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 genes encoding PI3Kα or mutations that lead to upregulation of PI3Kα have been found to occur in many human cancers, such as lung cancer, gastric cancer, endometrial cancer, ovarian cancer, bladder cancer, breast cancer, colon cancer, brain cancer, prostate cancer, and skin cancer (Goncalves et al., N Eng J Med. 2018, 379, 2052). In particular, the gene PIK3CA, which encodes the p110α subunit of PI3Kα, is frequently mutated or amplified in many tumor types. Missense mutations occur in all domains of p110α but are clustered in two “hotspots,” the most common being E542K and E545K in the helical domain and H1047R in the kinase domain. Helical domain mutations reduce p85 inhibition of p110α or promote direct interaction of p110α with insulin receptor substrate 1 (IRS1) 37, whereas kinase domain mutations increase p110α interactions with lipid membranes and upregulate signaling events (Thorpe et al., Nat Rev Cancer 2015, 15, 7).

[0007] The development of PI3K pathway inhibitors has been challenging because it is impossible to achieve a dose sufficient to suppress tumors without adverse events. So far, clinical PI3K inhibitors (Apelis, Bupanisi, Kupanisi, Duvilisi, Idelarisi, Pitilisi, Taselixibu, etc.) have caused dose-dependent adverse events such as hyperglycemia, rash, fatigue, diarrhea, etc. (Jiang et al., Mol Biol Rep. 2020, 47, 4587), which is a known on-target toxicity. Hyperglycemia is the result of the body not producing enough insulin or abnormal utilization. The pancreas regulates the release of insulin in response to changes in blood glucose levels, resulting in muscle and fat cells taking up glucose when insulin levels are high, or the liver performing gluconeogenesis when insulin levels are low. The response of tissue cells to insulin requires PI3K signaling through the ubiquitously expressed p110α subunit. Therefore, pan-PI3K inhibition of the target disrupts glucose metabolism in tissues, leading to insulin resistance (Hopkins et al., Nature 2018, 560, 499). In order to mitigate adverse events, selective PI3K isoform inhibitors have been developed. The severity of adverse events depends on the selected isoform, for example, due to the role of the p110α subunit in insulin response, PI3Kα inhibitors are associated with hyperglycemia and rash (Rugo et al., The Breast 2022, 61, 156). Similarly, the use of a selective PI3Kδ inhibitor (Idelaris), in which the p110δ subunit is highly expressed in immune cells, causes severe diarrhea and colitis. The inhibitory effects of dual inhibitors (taselixib), which are potent PI3Kδ inhibitors with moderate PI3Kα inhibition, lead to gastrointestinal (GI) side effects, but no GI-related adverse events were reported for the highly selective and potent PI3Kδ inhibitor (erbulisib) (Gadkar et al., CPT Pharmacometrics Syst Pharmacol. 2021, 11, 616). This improvement in adverse events using highly isoform-selective and potent inhibitors suggests that strategies to mitigate toxicity by developing mutation-selective isoform inhibitors are expected to reduce the severity of toxicity. In addition, selective inhibition of mutant PI3Kα isoforms relative to wild-type may inhibit cancer signaling while having minimal effects on PI3K signaling in healthy cells carrying only wild-type PI3Kα, resulting in reduced toxicity associated with non-selective PI3K inhibition (Castel et al., Nat Cancer 2021 2, 587).

[0008] There is currently interest in developing PI3K inhibitors for cancer treatment (WO 2023 / 081209, WO 2023 / 078401, WO 2023 / 060262, WO 2023 / 2056407, WO 2021 / 202964). However, there is a continuing need for new potent and selective PI3K inhibitors in the treatment of cancer, either as single agents or as combination therapies. Summary of the invention

[0009] One aspect of the present invention is a compound of formula (1)

[0010]

[0011] or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof,

[0012] in:

[0013] R1 is H, C1-C4 alkyl or C3-C7 cycloalkyl;

[0014] Each R2 is independently H, C1-C4 alkyl, C3-C7 cycloalkyl, R 14 -C≡C-, halogen, CN, CF3, OCF3, CFH2 or CF2H;

[0015] R3 is H, C1-C4 alkyl, C3-C7 cycloalkyl, CF3, CFH2, or CF2H, and when R3 is not H, the carbon atom attached to R3 is a chiral center and exists as a (R)- and (S)-racemic mixture or as an (R)- or (S)-enantiomer;

[0016] R4 is H or C1-C4 alkyl;

[0017] R6 is H, C1-C4 alkyl, C3-C7 cycloalkyl, heteroaryl, CF3, CFH2 or CF2H;

[0018] R7 is H, C1-C4 alkyl, C3-C7 cycloalkyl, halogen, CN, CF3, OCF3, OCH3, CFH2 or CF2H;

[0019] R8 is H, C1-C4 alkyl, C3-C7 cycloalkyl, halogen, CN, CF3, OCF3, OCH3, CFH2 or CF2H;

[0020] R5 is

[0021] -O-L1-L2-L3-L4-L5-L6-L7-R9;

[0022] -S-L1-L2-L3-L4-L5-L6-L7-R9;

[0023] -S(O)-L1-L2-L3-L5-L6-L7-R9;

[0024] -S(O)2-L1-L2-L3-L5-L6-L7-R9; or

[0025] -(NR 10 )-L1-L2-L3-L4-L5-L6-L7-R9,

[0026] in:

[0027] Each of L1, L2, L3, L6 and L7 is independently (CHR 11 )、(CHR 11 -O)、(CHR 11 -S), (C3-C7 cycloalkyl) or a bond;

[0028] L4 is C=O, C=S or a bond;

[0029] L5 is NR 10 , S, O or key;

[0030] R9 is H, C(=O)R 12 、C(=O)NR 12 R 13 、C(=O)OR 12 , C1-C6 alkyl, C1-C6 fluoroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each of C1-C6 alkyl, C1-C6 fluoroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is unsubstituted or substituted; or when NR 10 When present, R9 and R 10 Together with the attached nitrogen atom, a substituted or unsubstituted ring can be formed. In an exemplary embodiment, the ring is a 4-7 membered substituted or unsubstituted non-aromatic heterocyclic ring containing (in addition to the nitrogen atom) 0, 1 or 2 heteroatoms which may be N, O, S or Si, provided that if the ring size is 4 or 5, the number of additional heteroatoms will be 0 or 1, and if the ring size is 6 to 7, the number of additional heteroatoms will be 0, 1 or 2, wherein if the ring is substituted, the substituents include, but are not limited to, CH3, F, Cl, CF3, CF2H, CH2F, OCH3, cyclopropyl, CH2CF3, oxetane ring or COR a One or more of which R a is C1-C4 alkyl, O-C1-C4 alkyl or NR b R c , where R b and Rc are independently H or C1-C4 alkyl;

[0031] R 10 and R 11 Each of is H or C1-C4 alkyl, wherein C1-C4 alkyl is unsubstituted or substituted; and

[0032] R 12 and R 13 Each of R is independently H or C1-C6 alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each of C1-C6 alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is unsubstituted or substituted; or R 12 and R 13 Together with the attached nitrogen atom, a substituted or unsubstituted ring can be formed. In an exemplary embodiment, the ring is a 4-7 membered substituted or unsubstituted non-aromatic heterocyclic ring containing (in addition to the nitrogen atom) 0, 1 or 2 heteroatoms which may be N, O, S or Si, provided that if the ring size is 4 or 5, the number of additional heteroatoms will be 0 or 1, and if the ring size is 6 to 7, the number of additional heteroatoms will be 0, 1 or 2, wherein if the ring is substituted, the substituents include, but are not limited to, CH3, F, Cl, CF3, CF2H, CH2F, OCH3, cyclopropyl, CH2CF3, oxetane ring or COR a One or more of which 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; and

[0033] Each R 14 are independently H, C1-C3 alkyl or C3-C7 cycloalkyl;

[0034] Or R5 is

[0035] Nonaromatic N-linked heterocycles wherein the heterocycle is substituted or unsubstituted, optionally contains one or more additional atoms selected from N (wherein N is substituted or unsubstituted), O, Si (wherein Si is substituted or unsubstituted) and S (wherein S is oxidized or unoxidized), and is optionally part of a bridged, fused or spiro ring system.

[0036] In an exemplary embodiment, R5 is -(NR 10 )-L1-L2-L3-L4-L5-L6-L7-R9, where L1 to L7, R9 and R 10 As defined.

[0037] In an exemplary embodiment, R5 is -O-L1-L2-L3-L4-L5-L6-L7-R9, wherein L1 to L7 and R9 are as defined.

[0038] 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; wherein L1 to L7 and R9 are as defined.

[0039] In an exemplary embodiment, R9 is a 6-membered aromatic ring, or a 5- to 6-membered heteroaromatic ring containing 1-3 nitrogen atoms; or a non-aromatic 3- to 7-membered carbocyclic ring; or a non-aromatic 4- to 7-membered heterocyclic ring containing 1 to 3 heteroatoms selected from N, O, S and Si, provided that if the ring size is 4 or 5, the number of heteroatoms will be 1 or 2, and if the ring size is 6 or 7, the number of heteroatoms will be 1, 2 or 3; or is a C1-C6 alkyl group, wherein the aromatic ring, heteroaromatic ring, carbocyclic ring, heterocyclic ring and alkyl ring are unsubstituted or substituted with one or more of the following: CH3, F, Cl, CF3, CF2H, CH2F, OCH3, -CH2CF3, cyclopropyl, -CN, N(CH3)2, an oxetane ring, phenyl or phenoxy optionally substituted with 1 to 3 halogen (F, Cl or Br) or CH3 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.

[0040] In an exemplary embodiment, R5 is

[0041] Where R d is H or CH3, R e is CH3, C3-C6 cycloalkyl, a 6-membered aromatic ring, or a 5-membered or 6-membered heteroaromatic ring containing 0, 1 or 2 nitrogen atoms, or R d and R eTogether with the attached nitrogen atom, a 4- to 7-membered non-aromatic heterocyclic ring may be formed containing (in addition to the nitrogen atom) 0, 1 or 2 heteroatoms which may be N, O, S or Si, provided that if the ring size is 4 or 5, the number of additional heteroatoms will be 0 or 1, and if the ring size is 6 to 7, the number of additional heteroatoms will be 0, 1 or 2, wherein the aromatic, heteroaromatic and heterocyclic rings are unsubstituted or substituted by one or more of the following, including but not limited to: CH3, F, Cl, CF3, CF2H, CH2F, OCH3, cyclopropyl, CH2CF3, -CN, N(CH3)2, an oxetane ring, phenyl or phenoxy optionally substituted by 1 to 3 halogen (F, Cl or Br) or CH3 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.

[0042] In an exemplary embodiment, R5 is an N-linked non-aromatic heterocyclic ring wherein the heterocycle is substituted or unsubstituted, optionally contains one or more additional atoms selected from N (wherein N is substituted or unsubstituted), O, Si (wherein Si is substituted or unsubstituted) and S (wherein S is oxidized or unoxidized), and is optionally part of a bridged, fused or spiro ring system.

[0043] In exemplary embodiments, the N-linked non-aromatic heterocyclic ring is substituted or unsubstituted, optionally contains one or more additional heteroatoms selected from N, O, Si and S, and is not part of a bridged, fused or spiro ring system.

[0044] In exemplary embodiments, the N-linked non-aromatic heterocyclic ring is substituted or unsubstituted, optionally contains one or more additional heteroatoms selected from N, O, Si and S, and is part of a bridged, fused or spiro ring system.

[0045] In exemplary embodiments, the N-linked non-aromatic heterocyclic ring is substituted or unsubstituted, contains no additional atoms selected from N, O, Si, and S, and is not part of a bridged, fused, or spiro ring system.

[0046] In exemplary embodiments, the N-linked non-aromatic heterocyclic ring is substituted or unsubstituted, contains no additional atoms selected from N, O, Si, and S, and is part of a bridged, fused, or spiro ring system.

[0047] In exemplary embodiments, the N-linked non-aromatic heterocyclic ring is substituted or unsubstituted, contains at least one sulfur ring atom, and is not part of a bridged, fused, or spiro ring system.

[0048] In exemplary embodiments, the N-linked non-aromatic heterocyclic ring is substituted or unsubstituted, contains at least one sulfur ring atom, and is part of a bridged, fused, or spiro ring system.

[0049] In exemplary embodiments, the N-linked non-aromatic heterocyclic ring is substituted or unsubstituted, contains at least one oxygen ring atom, and is not part of a bridged, fused, or spiro ring system.

[0050] In exemplary embodiments, the N-linked non-aromatic heterocyclic ring is substituted or unsubstituted, contains at least one oxygen ring atom, and is part of a bridged, fused, or spiro ring system.

[0051] In exemplary embodiments, the N-linked non-aromatic heterocyclic 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.

[0052] In exemplary embodiments, the N-linked non-aromatic heterocycle is substituted or unsubstituted, contains at least one additional nitrogen ring atom, and is part of a bridged, fused, or spiro ring system.

[0053] In an exemplary embodiment of the compound of Formula (I), R1 is H.

[0054] In an exemplary embodiment of the compound of Formula (I), each R2 is H.

[0055] In an exemplary embodiment of the compound of Formula (I), R3 is CH3.

[0056] In an exemplary embodiment of the compound of Formula (I), R4 is H.

[0057] In an exemplary embodiment of the compound of Formula (I), R6 is CH3.

[0058] In exemplary embodiments of the compound of Formula (I), R7 is CH3 or F.

[0059] In an exemplary embodiment of the compound of Formula (I), R8 is H.

[0060] In an exemplary embodiment of the compound of Formula (I), each of R2 and R4 is H.

[0061] In an exemplary embodiment, the compound of formula (1) is a compound of formula (2)

[0062]

[0063] Or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R3 is CH3, CF3, CFH2 or CF2H; R5 is as defined in the compound of formula (1); R7 is CH3 or F; and the carbon marked with * is a chiral center and exists as a (R)- and (S)-racemic mixture or a (R)- or (S)-enantiomer.

[0064] In an exemplary embodiment, the compound of formula (1) is a compound of formula (3)

[0065]

[0066] or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R2 is as defined in the compound of formula (1); R3 is CH3, CF3, CFH2 or CF2H; R7 is CH3 or F; R 15 is OCH3, OCH2CH3, OCH2CF3, O-cyclopropyl, CH2CF3, CH2CF2H, aryl (wherein the aryl group can be, for example, a 6-membered aromatic ring) or heteroaryl (wherein the heteroaryl group can be, for example, a 5-membered or 6-membered heteroaromatic ring or a fused 6,5-heteroaromatic ring system, such as benzimidazole, indazole, imidazopyridine or triazolopyridine); each R 16 are independently H or C1-C3 alkyl; and the carbon marked with an * is a chiral center and exists as a (R)- and (S)-racemic mixture or as a (R)- or (S)-enantiomer.

[0067] In an exemplary embodiment, the compound of formula (1) is a compound of formula (4)

[0068]

[0069] or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R2 is as defined in the compound of formula (1); R3 is CH3, CF3, CFH2 or CF2H; R7 is CH3 or F; R 15 is OCH3, OCH2CH3, OCH2CF3, O-cyclopropyl, CH2CF3, CH2CF2H, aryl (wherein the aryl group can be, for example, a 6-membered aromatic ring) or heteroaryl (wherein the heteroaryl group can be, for example, a 5-membered or 6-membered heteroaromatic ring or a fused 6,5-heteroaromatic ring system, such as benzimidazole, indazole, imidazopyridine or triazolopyridine); and the carbon marked with * is a chiral center and exists as a (R)- and (S)-racemic mixture or as a (R)- or (S)-enantiomer.

[0070] In an exemplary embodiment, the compound of formula (1) is a compound of formula (5)

[0071]

[0072] or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R2 is as defined in the compound of formula (1); R3 is CH3, CF3, CFH2 or CF2H; R7 is CH3 or F; R 15 is OCH3, OCH2CH3, OCH2CF3, O-cyclopropyl, CH2CF3, CH2CF2H, aryl (wherein the aryl group can be, for example, a 6-membered aromatic ring) or heteroaryl (wherein the heteroaryl group can be, for example, a 5-membered or 6-membered heteroaromatic ring or a fused 6,5-heteroaromatic ring system, such as benzimidazole, indazole, imidazopyridine or triazolopyridine); and the carbon marked with * is a chiral center and exists as a (R)- and (S)-racemic mixture or as a (R)- or (S)-enantiomer.

[0073] One aspect of the present invention is a pharmaceutical composition comprising any compound of the present invention as described herein (e.g., any of formula (1), (2), (3), (4) or (5)) or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0074] In an exemplary embodiment, a pharmaceutical composition comprising any compound of the invention as described herein (e.g., any of Formula (1), (2), (3), (4), or (5)), or a solvate, enantiomer, diastereomer, tautomer, polymorph, or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, further comprises one or more anticancer agents.

[0075] Another aspect of the invention is a method for treating a disease in which PI3K activity is associated with a subject in need of such treatment, the method comprising administering to the subject a therapeutically effective amount of any compound of the invention as described herein (e.g., any of Formula (1), (2), (3), (4) or (5)) or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof.

[0076] In an exemplary embodiment, the disease to be treated is cancer. In a specific embodiment, the disease is a cancer carrying a PI3Kα H1047 mutation (eg, H1047R). DETAILED DESCRIPTION

[0077] As used herein, the term "at risk" refers to a patient exhibiting one or a group of medical conditions that may predispose the patient to a particular disease or affliction. For example, these conditions may be caused by influences including, but not limited to, behavioral, emotional, chemical, biochemical, or environmental influences.

[0078] As used herein, the term "effective amount" refers to a specific amount of a pharmaceutical composition that contains a therapeutic agent that achieves a clinically beneficial result (i.e., for example, symptom relief). The toxicity and therapeutic efficacy of such compositions can be determined by cell culture or experimental animals, such as for determining LD 50 (a dose lethal to 50% of the population) and ED 50 The dose ratio between toxic and therapeutic effects is the therapeutic index, which can be expressed as LD 50 / ED 50 Compounds that exhibit large therapeutic indices are preferred. The data obtained from these cell culture assays and additional animal studies can be used to formulate a range of doses for human use. The dose of such compounds is preferably within the range that includes the ED with little or no toxicity. 50 The dosage varies within this range depending on the dosage form employed, sensitivity of the patient, and the route of administration.

[0079] The term "symptom" as used herein refers to any subjective or objective evidence of a disease or physical disorder observed by a patient. For example, subjective evidence is often based on self-reporting by the patient and may include, but is not limited to, pain, headache, visual disturbances, nausea, and / or vomiting. Alternatively, objective evidence is often the result of a medical test, including, but not limited to, temperature, complete blood count, lipid panel, thyroid panel, blood pressure, heart rate, electrocardiogram, tissue imaging scan, and other medical test results.

[0080] As used herein, the term "disease" refers to any impairment of the normal state of a living animal or a part thereof that interrupts or alters the manifestation of vital functions. Diseases usually manifest themselves as obvious signs and symptoms, and diseases are usually a response to: i) environmental factors (such as malnutrition, industrial hazards, or climate); ii) specific infectious agents (such as insects, bacteria, or viruses); iii) inherent defects of the organism (such as genetic abnormalities); and / or iv) a combination of these factors.

[0081] When the terms "reduce", "inhibit", "reduce", "suppress", "lower", and "prevent" and their grammatical equivalents (including "lower", "lesser", etc.) are used in reference to the expression of any symptom in an untreated subject relative to a treated subject, the terms indicate that the amount and / or extent of the symptom in the treated subject is lower than in the untreated subject by any amount that a medically trained person would consider to be clinically relevant. In one embodiment, the amount and / or extent 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 extent of the symptom in the untreated subject.

[0082] As used herein, the term "inhibitory compound" refers to any compound that is capable of interacting with (i.e., e.g., attaching to, binding to, etc.) a binding partner under conditions that render the binding partner unresponsive to its natural ligand. Inhibitory compounds may include, but are not limited to, small organic molecules, antibodies, and proteins / peptides.

[0083] The term "attachment" as used herein refers to any interaction between a medium (or carrier) and a drug. Attachment may be reversible or irreversible. Such attachment includes, but is not limited to, covalent bonds, ionic bonds, van der Waals forces, or frictional forces, etc. If the drug is impregnated, incorporated, coated, suspended, dissolved, mixed, etc. with a medium (or carrier), the drug is attached to the medium (or carrier).

[0084] As used herein, the term "drug" or "compound" refers to any pharmacologically active substance that can be administered to achieve a desired effect. The drug or compound can be a synthetic or naturally occurring non-peptide, protein or peptide, oligonucleotide or nucleotide, polysaccharide or sugar.

[0085] As used herein, the term "administered" or "administering" refers to any method of providing a composition to a patient so that the composition has its intended effect on the patient. Exemplary methods of administration are by direct mechanisms, such as local tissue administration (i.e., for example, extravascular administration, such as subcutaneous, intramuscular or intraperitoneal), intravenous, oral, transdermal patch, topical, inhalation, suppository, etc.

[0086] The term "patient" as used herein refers to a human or animal, and does not require hospitalization. For example, outpatients and people in nursing homes are "patients". Patients can be humans or non-human animals of any age, thus including adults and adolescents (i.e., children). The term "patient" is not intended to mean that medical treatment is required. Therefore, patients can voluntarily undergo experiments, whether clinical experiments or experiments to support basic scientific research.

[0087] The term "subject" as used herein refers to, but is not limited to, humans (e.g., males or females of any age group, such as pediatric subjects (e.g., infants, children, teenagers) or adult subjects, e.g., young adults, middle-aged adults, or elderly people) and / or other primates (e.g., monkeys); non-human mammals, such as cows, pigs, horses, sheep, mice, goats, cats, dogs; and / or birds, such as chickens, ducks, and / or geese.

[0088] As used herein, the term "affinity" refers to any attractive force between substances or particles that causes them to enter and remain in a chemical combination. For example, an inhibitor compound with a high affinity for a receptor will provide greater efficacy in preventing the receptor from interacting with its natural ligand than an inhibitor with a low affinity.

[0089] As used herein, the term "derived from" refers to the source of a compound or sequence. In one aspect, a compound or sequence can be derived from an organism or a specific species. In another aspect, a compound or sequence can be derived from a larger complex or sequence.

[0090] As used herein, the term "test compound" refers to any compound or molecule that is considered a candidate for an inhibitory compound.

[0091] The term "combination therapy" as used herein refers to a dosing regimen of two or more different therapeutically active agents over a period of time, wherein the therapeutically active agents are administered together or separately. In one embodiment, the combination therapy is a non-fixed combination.

[0092] The term "non-fixed combination" as used herein refers to two or more different therapeutic agents that are formulated into separate compositions or dosages so that they can be administered separately to a subject in need thereof, either simultaneously or sequentially with variable intervals of time.

[0093] As used herein, the term "synergy" or "synergistic" refers to the phenomenon whereby the combination of two therapeutic agents in a combination therapy is better in a measured outcome than the sum of the effects of each agent when administered alone.

[0094] The term "in vivo" as used herein refers to events that occur within the body of a subject.

[0095] The term "in vitro" as used herein refers to events that occur outside the body of a subject.

[0096] As used herein, the term "protein" refers to any of a number of naturally occurring extremely complex substances (such as enzymes or antibodies) that contain amino acid residues linked by peptide bonds and that include carbon, hydrogen, nitrogen, oxygen, and usually sulfur. In general, proteins contain on the order of hundreds of amino acids.

[0097] The term "peptide" as used herein refers to any of various amides derived from two or more amino acids by combining the amino group of one amino acid with the carboxyl group of another amino acid, and is generally obtained by partial hydrolysis of a protein. Generally speaking, a peptide contains amino acids on the order of tens.

[0098] As used herein, the term "pharmaceutically acceptable" or "pharmacologically acceptable" refers to molecular entities and compositions that do not produce adverse, allergic or other untoward reactions when administered to animals or humans.

[0099] As used herein, the term "pharmaceutically acceptable carrier" includes any and all solvents or dispersion media, including, but not limited to, water, ethanol, polyols (such as glycerol, propylene glycol and liquid polyethylene glycol, etc.), suitable mixtures thereof, vegetable oils, coatings, isotonic and absorption delaying agents, liposomes, commercially available detergents, etc. Supplementary biologically active ingredients may also be incorporated into such carriers.

[0100] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that does not adversely affect the biological activity and properties of the compound and is suitable for use in contact with the tissues of a subject without producing excessive toxicity, irritation and / or allergic reactions, etc. Pharmaceutically acceptable salts include salts derived from suitable inorganic acids, organic acids and bases, and include salts derived from 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, etc. In some cases, a pharmaceutically acceptable salt is obtained by reacting a compound having an acidic group as described herein with a base to form a salt, such as an ammonium salt, an alkali metal salt (such as a sodium salt or a potassium salt), an alkaline earth metal salt (such as a calcium salt or a magnesium salt), a salt formed by an organic base, and an amino acid salt. Pharmaceutically acceptable salts derived from appropriate bases include alkali metals, alkaline earth metals, and ammonium and quaternary ammonium compounds. Specific metals include, but are not limited to, sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Organic bases from which salts can be prepared include, for example, primary amines, secondary amines, and tertiary amines.

[0101] The term "prodrug" as used herein refers to a compound that is converted in vivo to produce a disclosed compound or a pharmaceutically acceptable form of the compound. Prodrugs may be inactive when applied to a subject, but are converted into active compounds in vivo. In various cases, prodrugs have improved physicochemical properties (such as bioavailability) and / or delivery properties relative to the parent compound. Prodrugs are generally designed to enhance the pharmacological and / or pharmacokinetic properties associated with the parent compound. Prodrug compounds generally provide advantages of solubility, tissue compatibility, or delayed release in the subject. Prodrugs include compounds in which hydroxyl, amino, or sulfhydryl groups are bonded to any group, and the group is cleaved to form free hydroxyl, free amino, or free sulfhydryl groups when the prodrug is applied to a subject. Prodrugs are well known to be prepared by carboxylic acids in the form of, for example, carboxylic acid esters or thioesters.

[0102] As used herein, the terms "purified" or "isolated" may refer to a composition (eg, a peptide composition) that has been processed (eg, fractionated) to remove various other components, and which composition substantially retains its expressed biological activity.

[0103] As used herein, the term "sample" includes, for example, environment and biological samples. Environmental samples include materials from the environment, such as soil and water. Biological samples include animals (such as humans), fluids (such as blood, plasma and serum), solids (such as feces), tissues, liquid foods (such as milk) and solid foods (such as vegetables). For example, lung samples can be collected by bronchoalveolar lavage (BAL), which includes fluids and cells derived from lung tissue. Biological samples can include cells, tissue extracts, body fluids, chromosomes or extrachromosomal elements separated from cells, genomic DNA (in solution or attached to a solid support, such as for Southern blot analysis), RNA (in solution or attached to a solid support, such as for Northern blot analysis), cDNA (in solution or attached to a solid support) etc.

[0104] 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 restoring wild-type growth in cells that lack protein activity. Cells that lack protein activity can be generated by a variety of methods (e.g., point mutations and frameshift mutations). Complementation is achieved by transfecting cells that lack protein activity with an expression vector that expresses the protein, its derivatives, or a portion thereof.

[0105] As used herein, the term "label" or "detectable label" refers to any composition detectable by spectroscopic, photochemical, biochemical, immunochemical, electrical, optical or chemical means. Such labels include biotin, magnetic beads (such as ), fluorescent dyes (such as fluorescein, Texas rhodamine, green fluorescent protein, etc.), radioactive labels (such as 3 H. 125 I. 35 S. 14 C or 32 P), enzymes (such as horseradish peroxidase, alkaline phosphatase and other substances commonly used in ELISA) and calorimetric labels, such as colloidal gold or colored glass or plastic (such as polystyrene, polypropylene, latex, etc.) beads. Patents that teach the use of such labels include, but are not limited to, U.S. Patents No. 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). The labels contemplated in the present invention can be detected by conventional methods. For example, radioactive labels can be detected using photographic film or scintillation counters, and fluorescent markers can use photodetectors to detect emitted light. Enzyme labels are usually detected by providing a substrate to the enzyme and detecting the reaction product produced by the enzyme acting on the substrate, while calorimetric labels are detected by simply visualizing the colored label.

[0106] The term "conjugate" as used herein refers to any compound formed by the linkage of two or more moieties.

[0107] As used herein, "part" or "group" refers to any type of molecular arrangement specified by a formula, chemical name or structure. In the context of certain embodiments, a conjugate includes one or more parts or chemical groups. This means that the chemical formula of the part is substituted at certain positions so as to connect and become part of the molecular arrangement of the conjugate. Although parts can be directly covalently linked, it is not expected that the connection of two or more parts must be directly connected to each other. A linking group, a cross-linking group or a joining group refers to any molecular arrangement that will connect parts by covalent bonds, such as but not limited to one or more amide groups. In addition, although the conjugate can be unsubstituted, the conjugate can have various additional substituents connected to the linking group and / or to the part.

[0108] As used herein, "polymer" or "polymer group" refers to a chemical species or group consisting of repeatedly connected parts. In certain embodiments, the number of preferably repeated parts is 3 or more or greater than 10. The connected parts can be structurally identical or different in the structure of their parts. "Monopolymer" or "homopolymer" is a polymer containing identical repeated asymmetric subunits. "Copolymer" is a polymer derived from two or more types of monomer species (i.e., two or more different chemical asymmetric subunits). "Block copolymer" is a polymer consisting of two or more polymer subunits connected by covalent bonds.

[0109] As used herein, the term "substituted" refers to at least one hydrogen atom in the molecular arrangement being replaced by a substituent. The number of substituents present depends on the number of hydrogen atoms available for substitution, and includes replacing more than one hydrogen atom bound to a single atom (e.g., in the case of carbon atoms or silicon atoms available for mono-, di- or tri-substitution, or in the case of nitrogen atoms available for mono-, di- or tri-substitution, or in the case of oxygen atoms or sulfur atoms available for mono-substitution). In the case of an oxo substituent ("=O"), two hydrogen atoms are replaced (e.g., when two hydrogen atoms of the middle carbon atom of -CH2-CH2-CH3 are replaced, -(CH2)-C(=O)-CH3 is provided as a substituent). When substituted, one or more of the following groups are "substituents". Substituents include, but are not limited to, 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) ( For example, OCH3, OCH2CH3, OCH(CH3)2, etc.), -O-cycloalkyl (for example, O-cyclopropyl), O-haloalkyl (for example, OCF2H, OCFH2, OCF3, OCH2CF3, OCH2CF2H, OCHFCHF2, OCF2CH2F, OCF2CF3, OCF2CH3, OCF(CH3)2, OCH2CH2CF3, OCF2CH2CF3, OCF2CF2CF3 or OCH2Cl), O-aryl (for example, O-phenyl), O-heteroaryl, O-heterocyclyl, thioalkyl (for example, S-CH3), hydroxyalkyl (for example, CH2OH), alkyl ether (for example, CH2OCH3), alkynyl (for example, -C≡CRf ), alkenyl (e.g. -CR f =CR f R g ), aryl (e.g., phenyl), aralkyl (e.g., CH2Ph), heteroaryl (e.g., pyridyl or any 5- or 6-membered heteroaromatic 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 S02R 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 Can be the same or different, and be hydrogen, alkyl (such as CH ), substituted alkyl, haloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl, heterocyclic radical, substituted heterocyclic radical, heterocycloalkyl, substituted heterocycloalkyl, heteroaryl or substituted heteroaryl independently. In addition, the above-mentioned substituents can be further substituted by one or more of the above-mentioned substituents, so that the substituents can constitute, for example, substituted alkyl, substituted aryl, substituted aralkyl, substituted heterocyclic radical or substituted heterocycloalkyl.

[0110] As used herein, the term "unsubstituted" refers to any compound that does not contain additional substituents attached to the compound. An unsubstituted compound refers to the chemical composition of the compound without additional substituents (e.g., without 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 an alkyl group.

[0111] As used herein, the term "bond" when describing a substituent with atoms on both sides refers to the absence of the substituent. For example, in the 4-atom sequence ABCD, when both B and C are listed as bonds, the result is a 2-atom sequence AD. If only B is listed as a bond, the result is a 3-atom sequence ACD.

[0112] As used herein, the term "alkyl" refers to any straight or branched, non-cyclic or cyclic, unsaturated or saturated aliphatic hydrocarbon containing 1 to 10 carbon atoms, and the term "lower alkyl" has the same meaning as alkyl, but contains 1 to 3 carbon atoms. The term "higher alkyl" has the same meaning as alkyl, but contains 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, etc., and saturated branched alkyls include, but are not limited to, isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, etc. As used herein, methyl substituents may be described as "CH3" or "Me", or as terminal bonds without specific atom indications.

[0113] As used herein, the term "cycloalkyl" refers to saturated and unsaturated cycloalkyl groups. Representative saturated cycloalkyl groups include, but are not limited to, C3-C 14 (e.g. C3-C7) cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclododecyl, etc.; and unsaturated cycloalkyl, but not limited to, cyclobutenyl, cyclopentenyl and cyclohexenyl, cyclohexadiene, etc. Cycloalkyl is also referred to herein as "homocyclic" or "homocyclic".

[0114] As used herein, the term "spiro" or "spirocycle" refers to a chemical structure having at least two rings sharing a common atom. The ring can be a cycloalkyl, a heterocyclic radical, or a combination thereof, and can include one or more aryl or heteroaryl rings. Exemplary embodiments include 1,4-dioxaspiro[4.5]decane, spirocyclic azetidine and spirocyclic pyrrolidine and spirocyclic piperidine, wherein the other ring is a cycloalkyl (e.g., cyclobutane, cyclopentane or cyclohexane) or a heterocyclic radical (e.g., piperidine, tetrahydropyran, tetrahydrofuran, azetidine or pyrrolidine).

[0115] The term "bicyclic compound" as used herein encompasses "bridged," "fused," and "spiro" compounds as described.

[0116] As used herein, the term "bridged" refers to compounds 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, other bridged piperazines, and bridged piperidines.

[0117] 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), as well as polycyclic ring systems in which a ring contains two and only two adjacent atoms in common with each of two or more rings in an adjacent series of ortho-fused rings (ortho-fused and peri-fused). Exemplary embodiments are pentalene and dibenzoxepin. (ortho-fused) and pyrene (ortho- and peri-fused). Ortho-fused systems have "n" common edges and "2n" common atoms, while peri-fused systems have "n" common edges and less than "2n" common atoms. Other exemplary fused systems include fused cyclopropyl rings, fused aziridines, and fused azetidines, such as when these rings are fused to a pyrrolidine ring. Other examples include two fused pyrrolidine rings (octahydropyrrolo[3,4-c]pyrrole), fused pyridine rings, such as pyridine rings fused to cycloalkyl (such as cyclopentane) or to heterocyclic groups (such as tetrahydrofuran or tetrahydropyran).

[0118] As used herein, the term "aromatic" or "aryl" refers to any aromatic carbocyclic (i.e., all ring atoms are carbon) substituent such as, but not limited to, phenyl (from benzene), tolyl (from toluene), xylyl (from xylenes), or polycyclic systems such as naphthyl (from naphthalene) and anthracenyl (from anthracene).

[0119] The term "arylalkyl" or "aralkyl" as used herein refers to any alkyl group having at least one alkyl hydrogen atom replaced by an aryl moiety, such as, but not limited to, benzyl, -(CH2)2phenyl, -(CH2)3phenyl, -CH(phenyl)2, and the like.

[0120] The term "halogen" as used herein refers to any fluoro, chloro, bromo, or iodo moiety.

[0121] As used herein, the term "haloalkyl" refers to any alkyl group in which at least one hydrogen atom (and including all hydrogen atoms) has been replaced by a halogen atom, for example, trifluoromethyl, dichloromethyl, difluoromethyl, monofluoromethyl, monobromomethyl, 1,1,1-trifluoroethyl, and the like.

[0122] As used herein, the term "heteroaromatic" or "heteroaryl" refers to any aromatic heterocycle of 5 to 10 or more members, and it has at least one heteroatom selected from nitrogen, oxygen or sulfur, and contains at least 1 carbon atom, including but not limited to monocyclic and bicyclic systems. The heteroaromatic ring can be attached via a ring heteroatom or a carbon atom as a substituent. Representative heteroaromatics include but are not limited to furan, benzofuran, thiophene, benzothiophene, pyrrole, indole, isoindole, 7-azaindole, 4-azaindole, 5-azaindole, 6-azaindole, 7-azaindazole, pyridine, quinoline, isoquinoline, oxazole, isoxazole, benzoxazole, pyrazole, imidazole, benzimidazole, thiazole, benzothiazole, isothiazole, 1,2,4-triazole, 1,2,3-triazole, tetrazolium ... oxadiazole, 1,2,5-oxadiazole, 1,2-3-oxadiazole, 1,3,4-thiadiazole, pyridazine, pyrimidine, pyrazine, 1,2,4-triazine, 1,3,5-triazine, cinnoline, phthalazine, quinazoline, 1,8-naphthylpyridine, pyrido[3,2-d]pyrimidine, pyrido[4,3-d]pyrimidine, pyrimido[3,4-b]pyrazine, pyrido[2,3-b]pyrazine, pteridine, triazolopyridine, etc.

[0123] As used herein, the term "heteroarylalkyl" refers to any alkyl group having at least one alkyl hydrogen atom replaced by a heteroaryl moiety, such as -CH2pyridyl, -CH2pyrimidinyl, and the like.

[0124] As used herein, the term "heterocycle" or "heterocyclyl" or "heterocyclic ring" refers to a saturated or unsaturated non-aromatic ring containing one or more heteroatoms independently selected from nitrogen, oxygen, sulfur and silicon, wherein each of the nitrogen and sulfur heteroatoms may be in an oxidized state, and each of the nitrogen and silicon heteroatoms may be substituted or unsubstituted, the nitrogen heteroatom may be optionally quaternized, and includes bicyclic rings in which any of the above heterocycles is fused to an aryl or heteroaryl ring. The heterocycle may be attached via a ring heteroatom or a carbon atom as a substituent. In various embodiments, the heterocycle may contain 3 to 14 or more ring atoms (e.g., a 3 to 7 membered monocycle or a 7 to 10 membered bicycle), and includes, but is not limited to, 2H-oxadiene, azetidine, 2,3-dihydroazetadiene, 1,3-diazetidine, 2H-oxetene, thietane, 2H-thietene, azetidin-2-one, morpholine, thiomorpholine, pyrrolidone, p-oxadiene, thiophene ... yrrolidinine, 2-pyrroline, 3-pyrroline, pyrazolidine, 2-pyrazoline, 2-imidazoline, imidazolidine, piperidine, piperazine, oxirane (oxirane), ethyleneimine (aziridine), thioethane (thiirane), oxetane, propylene oxide, 1,3-dioxolane, 1,2-oxathiolane, 1,3-oxathiolane, sulfolane, 2,4-thiazole oxadione, succinimide, 2-oxazolidinone, dioxane, hydantoin, valerolactam, tetrahydrofuran, tetrahydropyran, 2H-pyran, 4H-pyran, cyclopentane sulfide, 2H-thiopyran, 1,3-cyclopentane disulfide, 1,4-cyclopentane disulfide, 1,3,5-cyclopentane trisulfide, pyrrolizidine, 1,4,5,6-tetrahydrocyclopenta[b]pyrrole, tetrahydropyridine, tetrahydropyrimidine, Tetrahydrothiophene, 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, 2,3-dihydroaza 2,5-Dihydroazepine Oxahedrine, azacyclononane, 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, etc.

[0125] As used herein, the term "heterocycloalkyl" refers to any alkyl group having at least one alkyl hydrogen atom replaced by a heterocycle, such as -CH2morpholinyl and the like.

[0126] As used herein, the term "alkylamino" refers to at least one alkyl moiety attached through a nitrogen bridge (ie, -N-(alkyl) n , wherein n=1 or 2, such as alkylamino or dialkylamino), including but not limited to methylamino, ethylamino, dimethylamino, diethylamino and the like.

[0127] The term "alkyloxy" or "alkoxy" as used herein refers to any alkyl moiety attached through an oxygen bridge (ie, -O-alkyl), such as, but not limited to, methoxy, ethoxy, and the like.

[0128] The term "thioalkyl" as used herein refers to any alkyl moiety attached through a sulfur bridge (ie, -S-alkyl), such as, but not limited to, methylthio, ethylthio, and the like.

[0129] As used herein, the term "alkenyl" refers to a straight or branched hydrocarbon chain having one or more carbon-carbon double bonds therein, also referred to as an "unsaturated alkyl". The double bond of an alkenyl group may be non-conjugated or conjugated with another unsaturated group. Suitable alkenyl groups include, but are not limited to, vinyl, allyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, 2-ethylhexenyl, 2-propyl-2-butenyl, 4-(2-methyl-3-butene)-pentenyl. An alkenyl group may be unsubstituted or substituted with one or two suitable substituents.

[0130] As used herein, the term "alkynyl" refers to a straight or branched hydrocarbon chain having one or more carbon-carbon triple bonds therein, also referred to as an "unsaturated alkyl". The triple bond of an alkynyl group may be non-conjugated or conjugated with 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. An alkynyl group may be unsubstituted or substituted with one or two suitable substituents.

[0131] As used herein, "reactive group" refers to a nucleophile, an electrophile, or a radical-active group, i.e., a group that reacts in the presence of a free radical. A nucleophile is a part that forms a chemical bond with its reaction partner (electrophile) by providing two bonding electrons. Electrophiles accept these electrons. Nucleophiles can participate in nucleophilic substitutions, whereby nucleophiles become attracted to all or part of the positive charge on the element and replace the group to which they are bonded. Alternatively, nucleophiles can also participate in the substitution of carbonyl groups. Carboxylic acids usually become electrophilic by producing succinates and reacting these esters with aminoalkyl groups to form amides. Other common nucleophilic groups include thiol alkyls, hydroxyalkyls, primary and secondary amines, and carbon nucleophiles, such as enols and alkyl metal complexes. Other preferred methods for connecting proteins, oligosaccharides, and cells using reactive groups are disclosed (Lemieux et al., Trends in Biotechnology 1998, 16, 506, incorporated herein by reference in their entirety). In yet another preferred method, a reactive group for Staudinger connection is provided, i.e., "click chemistry" of triazoles formed with azide-containing moieties and alkynyl reactive groups. Michael addition of carbon nucleophiles enolates to electrophilic carbonyls, or Schiff base formation of nucleophilic primary or secondary amines to aldehydes or ketones can also be utilized. Other bioconjugation methods are provided (Hang et al., Accounts of Chemical Research 2001, 34, 727, and Kiick et al., Proc Natl Acad Sci US.A. 2002, 99, 19, both of which are incorporated herein by reference in their entirety).

[0132] The term "biocompatibility" as used herein refers to any material that does not cause a substantial adverse reaction in the host. When a foreign object is introduced into a living body, there is always concern that the object will cause an immune response, such as an inflammatory response that negatively affects the host. In the context of the present invention, biocompatibility is assessed based on the application for which it is designed: for example, a bandage is considered biocompatible with the skin, while an implanted medical device is considered biocompatible with internal body tissues. Preferably, biocompatible materials include, but are not limited to, biodegradable and biostable materials. If the implant comprising the material is closely related to its implantation site in the host animal, and the reaction is better than the tissue reactions that have been established and determined to be appropriate from the materials provided in ASTM, then no substantial adverse reaction occurs. ASTM Subcommittee F04.16 on Biocompatibility Test Methods has developed standards for biocompatibility of 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, F981- 04, F1027-86, F1408-20a, F1439-03, F1877-16, F1903-18, F1904-14, F1983-14, F1984-99, F2147-01, F2148-18, F2382-18, F2808-17, F1288-19, and F2909-19, each of which is incorporated herein by reference. For example, materials intended for contact with the bloodstream must be composed of materials that meet hemocompatibility standards. One of these tests is for damage to red blood cells, which may result in hemolysis, i.e., cell rupture, as described in F756-17 Standard Practice for Evaluating Hemolytic Properties of Materials.

[0133] As used herein, "biologically active substances" refers to any of a variety of chemical moieties that bind to biological molecules, such as, but not limited to, peptides, proteins, enzymes, receptors, substrates, lipids, antibodies, antigens, and nucleic acids. In certain preferred embodiments, the biologically active substances are biological molecules, but it is not intended that biologically active substances are limited to biological molecules. In other preferred embodiments, the biologically active substances provide hydrophobicity, hydrophilicity, or electrostatic interactions, such as polycarboxylic acids that are anionic at physiological pH. In other preferred embodiments, basic growth factors (isoelectric points above 7) are retained via favorable electrostatic interactions through polycarboxylates and subsequently released in a controlled and sustained manner.

[0134] "Cancer" is a term for a physiological condition of a mammal, 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, renal cancer (renal cancer) (e.g., advanced renal cell carcinoma), ovarian cancer, liver cancer, lymphoblastic leukemia, lymphocytic leukemia, colorectal cancer, endometrial cancer, kidney 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 cancer), bladder cancer, hepatoma, breast cancer, and head and neck cancer.

[0135] The term "stereoisomers" refers to compounds with the same atomic connectivity but different arrangements 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 center.

[0136] The present disclosure encompasses all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, atropisomers, tautomers and racemates and other mixtures thereof, such as enantiomers or diastereoisomerically enriched mixtures, all of which are within the scope of the present disclosure. As long as the compounds of the present invention as defined herein may exist in optically active or racemic form due to one or more asymmetric carbon atoms, the present invention includes any such optically active or racemic form in its definition. The synthesis of optically active compounds can be carried out by standard techniques of organic chemistry well known in the art, for example by synthesis from optically active starting materials or by resolution of racemic compounds. Similarly, the enantiomeric or diastereomeric purity of the compounds can be assessed using standard laboratory techniques.

[0137] The pharmaceutical composition of the present invention can take any suitable form for the desired route of administration. When the composition is administered orally, any suitable oral delivery dosage form can be used, including but not limited to water, glycols, oils, alcohols, etc. in the case of oral liquid preparations such as suspensions, syrups, elixirs, emulsions and solutions; or in the case of powders, pills, capsules and tablets, including but not limited to solid carriers, such as starch, sugar, kaolin, diluents, lubricants, adhesives, disintegrants, etc. Because they are easy to use, tablets and capsules represent the most favorable 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 generally includes sterile water and possible other ingredients to help dissolve. Injectable solutions can be prepared, wherein the carrier includes a mixture of saline solution, glucose solution or saline and glucose solution. Suitable oils include, for example, peanut oil, sesame oil, cottonseed oil, corn oil, soybean oil, synthetic glycerides of long-chain fatty acids and mixtures of these oils and 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 a suitable additive as required, wherein the additive can facilitate the application of the composition to the skin and / or can facilitate the preparation of the composition to be delivered. These compositions can be applied in various ways, for example as a transdermal patch or ointment. Acid or base addition salts of the compounds of the present invention are generally more suitable for preparing aqueous compositions because they have increased water solubility compared to the corresponding neutral form of the compound.

[0138] The pharmaceutical composition of the present invention may contain one or more of fillers, diluents, adjuvants, vehicles or other excipients to facilitate the storage and / or administration of the active ingredients contained therein.

[0139] In an exemplary embodiment, the pharmaceutical composition according to the present invention may contain one or more additional therapeutic agents, for example, to improve efficacy or reduce undesirable side effects. In a specific embodiment, the pharmaceutical composition also contains one or more additional therapeutic agents that can be used to treat or inhibit diseases mediated directly or indirectly by PI3K. Examples of such agents include, but are not limited to, agents for treating or inhibiting cancer, Huntington's disease, cystic fibrosis, liver fibrosis, renal fibrosis, pulmonary fibrosis, skin fibrosis, rheumatoid arthritis, diabetes or heart failure.

[0140] In a specific embodiment, the additional therapeutic agent to be included is an anticancer agent. Examples of anticancer agents include, but are not limited to, cytotoxic drugs that damage DNA, alkylating agents such as cyclophosphamide, dacarbazine, and cisplatin; antimetabolites such as methotrexate, mercaptopurine, thioguanine, fluorouracil, and cytarabine; plant alkaloids such as vinblastine and paclitaxel; antitumor antibiotics such as doxorubicin, bleomycin, and mitomycin; hormones / antihormones such as prednisone, tamoxifen, and flutamide; other types of anticancer agents such as asparaginase, rituximab, trastuzumab, imatinib, retinoic acid and its derivatives, colony stimulating factors, amifostine, camptothecin, topotecan, thalidomide analogs such as lenalidomide, and proteasome inhibitors such as Velcade.

[0141] In another embodiment, the invention provides a method for inhibiting or treating a disease caused by abnormal cell proliferation and / or differentiation in a subject in need, comprising administering to the subject a therapeutically effective amount of one or more compounds according to the present invention. In one embodiment, the method for inhibiting or treating a disease comprises administering to a subject in need a composition comprising an effective amount of one or more compounds of the present invention and a pharmaceutically acceptable carrier. The composition to be administered may further comprise a therapeutic agent, such as an anticancer agent.

[0142] The compounds of the present invention are defined herein by their chemical structures and / or chemical names, and are generally listed according to the IUPAC or CAS nomenclature system. Abbreviations well known to those of ordinary skill in the art can be used. When a compound is referred to by both a chemical structure and a chemical name, and the chemical structure and the chemical name conflict, the chemical structure is expected to determine the identity of the compound.

[0143] The present invention includes compounds labeled with various radioactive or non-radioactive isotopes. Examples of atomic isotopes may include, but are not limited to, 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 by deuterium. In various embodiments, the compounds of the invention include 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 also be substituted with radioactive isotopes such as tritium ( 3 H), iodine-125 ( 125 I) and carbon-14 ( 14C) Radiolabeling. Radiolabeled compounds can be used as therapeutic or prophylactic agents, to provide reagents for research (e.g., assays), and / or to provide diagnostic agents for technology (e.g., in vivo imaging). Synthetic methods for incorporating isotopes into organic compounds are well known in the art.

[0144] In an embodiment of the present invention, the compound of the present invention as defined herein (e.g., a compound of any one of formula (1), (2), (3), (4) or (5)) or a pharmaceutically acceptable salt thereof exists as a single enantiomer with an enantiomeric excess (%ee) ≥95%, such as ≥98%, such as ≥99%.

[0145] In an embodiment of the present invention, the pharmaceutical composition comprises a compound of the present invention as defined herein (e.g., a compound of formula I) or a pharmaceutically acceptable salt thereof, wherein the compound exists as a single enantiomer and its enantiomeric excess (%ee) is ≥95%, for example ≥98%, for example ≥99%.

[0146] In an exemplary embodiment of the invention, the disease or disorder to be treated by the compounds of the invention is selected from congenital lipomatous overgrowth, vascular malformations, epidermal nevi, scoliosis / skeletal and spine syndrome (CLOVES), mosaic tissue overgrowth syndrome, venous malformations associated with severe epilepsy and brain malformations, 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).

[0147] In an exemplary embodiment of the present invention, the cancer to be treated is a cancer carrying a PI3K H1047 mutation (eg, H1047R) (Thorpe et al., Nat Rev Cancer 2015, 15, 7).

[0148] The compounds of formula I of the present invention are generally prepared according to the synthetic routes identified in the following schemes:

[0149]

[0150] Starting from the generally known or commercially available aminobenzoic acid 1 (e.g., from BLD Pharmatech Ltd.), the intermediate quinazoline-2,4-dione 2 can be produced under cyclization conditions. In an exemplary embodiment, when R6=H, the cyclization occurs by treatment with urea at elevated temperature. In an exemplary embodiment, when R6=alkyl, the cyclization occurs via a two-step process consisting of first coupling the amide with the corresponding alkylamine and HATU, followed by treatment with triphosgene. Chlorination of 2 provides an intermediate of general structure 3. In an exemplary embodiment, the chlorination occurs by refluxing 2 in POCl3, followed by neutralization with an alkaline aqueous solution (NaOH when R6=H, or NaHCO3 when R6=alkyl). Intermediates of general structure 4 are prepared by S NAr substitution is produced. In exemplary embodiments, 3 is treated with an amine, amine hydrochloride, alcohol or thiol (depending on whether R5 is N-attached, O-attached or S-attached, respectively), and in some embodiments, an appropriate base (such as but not limited to DIEA, K2CO3 or NaH) is added in an appropriate solvent (such as but not limited to ACN, NMP or DMF) at room temperature, or in some embodiments at elevated temperatures (up to 140°C) to provide an intermediate of general structure 4. Intermediates of general structure 5 are generated by carbonylation. In some exemplary embodiments, the bromo substituent of 4 is replaced with an acetyl group by treatment with tributyl(1-ethoxyvinyl)tin and a catalytic palladium species (such as but not limited to Pd(PPh3)4 or PdCl2(PPh3)2) at elevated temperatures, followed by hydrolysis with aqueous HCl to provide ketone 5. When R3=H, intermediate 5 can also be produced by formylation of 4 by various known methods (such as but not limited to palladium-catalyzed carbonylation in the presence of H2 (Klaus et al., Angew. Chem. Int. Ed. 2006, 45, 154) or cyanation followed by DIBAL reduction). Intermediate 5 can serve as a platform for further expansion to different R3 substitutions (such as trifluoromethyl, difluoromethyl, fluoromethyl, alkyl, etc.) by various known techniques (including but not limited to Prakash et al., J. Am. Chem. Soc. 1989, 111, 393; Zhao et al., Org. Lett. 2011, 13, 5342; Reichel et al., Angew. Chem. Int. Ed. 2020, 59, 12268, etc.). Alcohol intermediates of general structure 6 can be prepared by reducing 5. In an exemplary embodiment, the reduction is carried out by treating 5 with NaBH4 in MeOH. The conversion of 6 to aromatic amines of general structures 8 and 9 can be achieved via substitution reactions. In some exemplary embodiments, when R4 = H, 6 is converted to 9 via Mitsunobu conditions involving PPh3, DBAD, and intermediate 7. In other exemplary embodiments, when R4 = 2-nitrobenzenesulfonyl or 2,4-dinitrobenzenesulfonyl, 6 is substituted with intermediate 7 under Mitsunobu conditions to give sulfonamide intermediate 8. In other exemplary embodiments, 6 is first converted to a mesylate salt via treatment with Ms2O or MsCl and an amine base such as, but not limited to, Et3N or DIEA, and then treated with amine 7 to provide benzylamine 9. Sulfonamides of general structure 8 can be converted to secondary amines 9 via known desulfonylation protocols. In an exemplary embodiment, 8 is treated with potassium thiophenolate and K2CO3 to give intermediate 9. Intermediates 9 with R5 = thioalkyl or thio(hetero)aryl can be further expanded to the corresponding sulfinyl or sulfonyl compounds via known oxidation methods such as, but not limited to, treatment with mCPBA or potassium peroxymonosulfate.The ester intermediate 9 can be converted to the corresponding carboxylic acid of general structure 10 via known deesterification schemes. In some exemplary embodiments, when R1 = CH3, deesterification is achieved via treatment with a metal hydroxide (such as, but not limited to, LiOH or NaOH) to afford a product of general structure 10. In other exemplary embodiments, when R1 = tert-butyl, intermediate 9 is deesterified by treatment with an appropriate acid (such as, but not limited to, 1,4-dioxane or HCl in water, or TFA) to afford 10. Separation of racemate 10 via known chiral HPLC chromatography techniques (such as, but not limited to, using a DAICEL Chiralpak column) affords enantiomerically enriched compounds of general structures 11 and 12.

[0151] The following compounds (in their racemic or enantiomerically resolved form) represent various embodiments of the present invention. A listing of substituents within parentheses for a given compound indicates that the individual compound contains one of each substituent.

[0152]

[0153] The following compounds represent various embodiments of the present invention in which only R5 has been varied in a structure where all other atoms have been set. Carbon atoms marked with an * in the following general structures are chiral centers and exist as (R)- and (S)-racemic mixtures or (R)- or (S)-enantiomers. The listing of substituents within parentheses for a given compound indicates that the individual compound contains one of each substituent. Where present, each R h and R i is independently selected from H, CH3, c-Pr, c-Bu, CF3 and OH; and each R j Independently selected from CF3, CH2CF3, CH2CF2H, OCH3, OCF3, OCH2CF3, Oc-Pr, aryl, heteroaryl, COCH3 and CO2CH3.

[0154]

[0155] Wherein R5 is selected from

[0156]

[0157]

[0158]

[0159] The following compounds represent various embodiments of the present invention, wherein only R5 has been varied in a structure where all other atoms have been set. Carbon atoms marked with an * in the following general structures are chiral centers and exist as (R)- and (S)-racemic mixtures or (R)- or (S)-enantiomers. The listing of substituents within parentheses for a given compound indicates that the individual compound contains one of each substituent.

[0160]

[0161] Where R5 is Among them, L1 to L7, R 10 and R9 are as defined herein, and, where present, each R k and R m In an exemplary embodiment, R5 is selected from

[0162]

[0163]

[0164] The following compounds represent various embodiments of the present invention in which only R5 has been varied in a structure where all other atoms have been set. Carbon atoms marked with an * in the following general structures are chiral centers and exist as (R)- and (S)-racemic mixtures or (R)- or (S)-enantiomers. The listing of substituents in parentheses for a given compound indicates that a single compound contains one of the various substituents. The variable "A" is selected from O, S, S(O) and S(O)2.

[0165]

[0166] Where R5 is

[0167]

[0168]

[0169] experiment

[0170] All commercially available solvents and reagents were used as received. All data were recorded using a Bruker Avance III HD 300 MHz or a Bruker Avanse III HD 400 MHz 1H NMR spectroscopy. MS samples were analyzed on a SHIM-ADZU LCMS-2020 mass spectrometer with electrospray ionization in both positive and negative ion modes. Samples were introduced into the mass spectrometer using chromatography. All final products had a purity of ≥90% unless otherwise specified in the experimental details. HPLC purity was measured on a SHIM-ADZU Acquity HPLC system.

[0171] The following are abbreviations representing well-known chemical solvents, reagents, parameters and techniques used in the Experimental section:

[0172] 1 H-NMR: Proton Nuclear Magnetic Resonance Spectroscopy

[0173] ACN: Acetonitrile

[0174] AcOH: acetic acid

[0175] c-Bu: cyclobutyl

[0176] c-Pr: Cyclopropyl

[0177] DBAD: di-tert-butyl azodicarboxylate

[0178] DCM: dichloromethane

[0179] DIBAL: diisobutylaluminum hydride

[0180] DIEA: N,N-diisopropylethylamine

[0181] DMF: N, N-dimethylformamide

[0182] DMSO: dimethyl sulfoxide

[0183] ee: enantiomeric excess

[0184] Et3N: triethylamine

[0185] EtOAc: ethyl acetate

[0186] EtOH: ethanol

[0187] FA: Formic acid

[0188] h: hours

[0189] HATU: 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

[0190] HCl: hydrochloric acid

[0191] HPLC: High Performance Liquid Chromatography

[0192] IPA: Isopropyl alcohol

[0193] K2CO3: Potassium carbonate

[0194] LiOH: lithium hydroxide

[0195] mCPBA: meta-chloroperbenzoic acid

[0196] Me:Methyl

[0197] MeOH: Methanol

[0198] mg: milligram

[0199] min: minutes

[0200] mL: milliliters

[0201] MsCl: Methanesulfonyl chloride

[0202] Ms2O: Methanesulfonic anhydride

[0203] NaBH4: Sodium borohydride

[0204] NaH: Sodium hydride

[0205] NaOH: Sodium hydroxide

[0206] NaHCO3: Sodium bicarbonate

[0207] Na2SO4: Sodium sulfate

[0208] NMP: N-methylpyrrolidone

[0209] Oxetane: A 4-membered ring containing 3 carbon ring atoms and 1 oxygen ring atom.

[0210] Pd(PPh3)4:Tetrakis(triphenylphosphine)palladium(0)

[0211] PdCl2(PPh3)2: Bis(triphenylphosphine)palladium(II) dichloride

[0212] PE: Petroleum ether

[0213] POCl3: Phosphorus oxychloride

[0214] PPh3: triphenylphosphine

[0215] TFA: trifluoroacetic acid

[0216] THF: Tetrahydrofuran

[0217] TLC: Thin layer chromatography

[0218] Example

[0219] Intermediate 1: 8-bromo-2-chloro-3,6-dimethylquinazolin-4-one.

[0220]

[0221] Step 1 : Preparation of 2-amino-3-bromo-N,5-dimethylbenzamide.

[0222] To a stirred solution of commercially available (e.g., BLD Pharmatech Ltd.) 2-amino-3-bromo-5-methylbenzoic acid (50 g, 0.22 mmol) and DIEA (227 mL, 1.30 mol) in DCM (100 mL) and DMF (3.36 mL) was added HATU (165 g, 435 mmol). After stirring at room temperature for 5 min, methylamine hydrochloride (66 g, 978 mmol) was added. The resulting solution was stirred at room temperature for 2 h. The mixture was concentrated under reduced pressure and purified by silica gel chromatography (0-50% EtOAc in PE) to give the desired product 2-amino-3-bromo-N,5-dimethylbenzamide (50 g, 95% yield) as a solid. MS: (ES + )m / z=243.0[M+H] + .

[0223] Step 2 : Preparation of 8-bromo-3,6-dimethyl-1H-quinazoline-2,4-dione.

[0224] A mixture of 2-amino-3-bromo-N,5-dimethylbenzamide (19.2 g, 79 mmol), triphosgene (23.4 g, 79 mmol) and DIEA (13.8 mL, 79 mmol) in DCM (250 mL) was stirred at 50 °C overnight under nitrogen atmosphere. The mixture was allowed to cool to room temperature and then concentrated under reduced pressure. The crude product was purified by silica gel chromatography eluting with 0-50% EtOAc in PE to give the title compound 8-bromo-3,6-dimethyl-1H-quinazoline-2,4-dione (18 g, 85% yield) as a solid. MS: (ES + )m / z=269.0[M+H] + .

[0225] Step 3 : Preparation of 8-bromo-2-chloro-3,6-dimethylquinazolin-4-one.

[0226] Under nitrogen atmosphere, 8- bromo-3,6- dimethyl -1H- quinazoline -2,4- dione (2.9g, 10.78mmol) and DIEA (7.51mL, 43.11mmol) in POCl (13.2mL, 141.4mmol) mixture was stirred at 100 ° C overnight. The mixture was concentrated under reduced pressure, and quenched with water (100mL), then neutralized to pH = 7 with NaHCO. The resulting solution was extracted with DCM (3x30mL), and the combined organic layer was washed with brine (20mL), dried over NaSO, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography, eluted with 0-50% EtOAc in PE, to obtain 8- bromo-2- chloro-3,6- dimethylquinazoline -4- ketone (800mg, 26% yield) as a solid. 1 H NMR (chloroform-d, 300 MHz): δ7.90 (dd, J=2.0, 0.9 Hz, 1H), 7.85–7.79 (m, 1H), 3.73 (s, 3H), 2.43 (s, 3H), MS: (ES + )m / z=287.0[M+H] + .

[0227] Intermediate 2: tert-butyl 2-(2-nitrobenzenesulfonamido)benzoate.

[0228]

[0229] The stirring solution of tert-butyl 2-aminobenzoate (5.00g, 25.87mmol) in DCM (100mL) and pyridine (10mL) is cooled to 0 ℃ in an ice bath, then dropwise treated with 2-nitrobenzenesulfonyl chloride (11.47g, 51.74mmol) in DCM (20mL). The gained mixture is stirred at room temperature overnight, then diluted with water (50mL), and extracted with DCM (3x50mL). The organic layer merged is washed with salt water (2x50mL), through anhydrous Na2SO4 drying, filtered and concentrated under reduced pressure. The crude product is purified by silica gel chromatography, eluted with 0-33% EtOAc in PE for 30min, isolates tert-butyl 2-(2-nitrobenzenesulfonamido)benzoate (5.20g, 53% yield) as solid. 1 H NMR (DMSO-d6, 400MHz): δ11.01(s,1H),8.19(dd,J=7.7,1.5Hz,1H),8.07(dd,J=7.8,1.4Hz,1H ),8.00-7.80(m,3H),7.64–7.53(m,2H),7.21(ddd,J=8.2,6.7,1.9Hz,1H),1.55(s,9H),MS(ES -)m / z=376.8[MH] - .

[0230] Intermediate 3: methyl 2-(2,4-dinitrobenzenesulfonamido)benzoate.

[0231]

[0232] The solution of methyl anthranilate (5g, 33.08mmol) in DCM (50mL) and pyridine (5.23g, 66.15mmol) in a 250mL round-bottom flask was cooled to 0 ° C in an ice bath, and then 2,4-dinitrobenzenesulfonyl chloride (10.58g, 39.70mmol) was added. The resulting mixture was stirred overnight under a nitrogen atmosphere, and was warmed to room temperature at the same time, then diluted with water (50mL) and extracted with DCM (3x50mL). The combined organic phase was washed with brine (50mL), dried with Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EtOAc=1:1, and 2-(2,4-dinitrobenzenesulfonyl)benzoic acid methyl ester (8g, 63% yield) was obtained as a solid. 1 H NMR (chloroform-d, 300MHz): δ11.42(s,1H),8.63(d,J=2.2Hz,1H),8.49(dd,J=8.6,2.2Hz,1H),8.38(d,J=8.6Hz,1H),8.02(d d,J=8.0,1.7Hz,1H),7.81(d,J=8.4Hz,1H),7.54(td,J=8.3,7.9,1.7Hz,1H),7.21–7.09(m,1H),3.96(s,3H),MS:(ES - )m / z=380.0[MH] - .

[0233] Intermediates 4, 5, 6 and 7

[0234]

[0235] Intermediates 4, 5, 6 and 7 were prepared in a similar manner to Intermediate 1.

[0236] Example 1: 2-((1-(2-(4,4-dimethyl-1,4-azasilan-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0237] Example 2: 2-((1-(2-(4,4-dimethyl-1,4-azasilan-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0238]

[0239] Step 1 : Preparation of 8-bromo-2-(4,4-dimethyl-1,4-azasilane-1-yl)-3,6-dimethylquinazolin-4-one.

[0240] A solution of 8-bromo-2-chloro-3,6-dimethylquinazolin-4-one (Intermediate 1) (1.20 g, 4.18 mmol) and 4,4-dimethyl-1,4-azasilane (450 mg, 3.48 mmol) in ACN (10 mL) was stirred at 80°C overnight. The reaction mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography, eluting with PE:EtOAc=5:1 to give 8-bromo-2-(4,4-dimethyl-1,4-azasilane-1-yl)-3,6-dimethylquinazolin-4-one (800 mg, 60% yield) as a solid. MS: (ES + )m / z=380.1[M+H] + .

[0241] Step 2 : Preparation of 8-acetyl-2-(4,4-dimethyl-1,4-azasilane-1-yl)-3,6-dimethylquinazolin-4-one.

[0242] Under nitrogen atmosphere, a solution of 8-bromo-2-(4,4-dimethyl-1,4-azasilanyl-1-yl)-3,6-dimethylquinazolin-4-one (800 mg, 2.1 mmol) and tributyl(1-ethoxyvinyl)tin (836 mg, 2.3 mmol) was added at room temperature, followed by addition of Pd(PPh3)4 (266 mg, 0.23 mmol) in dioxane (10 mL) and stirred at 100 ° C overnight. The mixture was cooled to room temperature. 1M HCl (2 mL) was added to the above mixture. The resulting mixture was stirred at 50 ° C for 1 h. The resulting mixture was concentrated under vacuum. The residue was purified by preparative TLC (PE / EA=1:1) to give 8-acetyl-2-(4,4-dimethyl-1,4-azasilan-1-yl)-3,6-dimethylquinazolin-4-one (600 mg, 83% yield) as a pale yellow solid. MS: (ES + )m / z=344.2[M+H] + .

[0243] Step 3 : Preparation of 2-(4,4-dimethyl-1,4-azasilan-1-yl)-8-(1-hydroxyethyl)-3,6-dimethylquinazolin-4-one.

[0244] A solution of 8-acetyl-2-(4,4-dimethyl-1,4-azasilanyl)-3,6-dimethylquinazolin-4-one (600 mg, 1.74 mmol) and NaBH4 (264 mg, 6.98 mmol) in MeOH (5 mL) was stirred at 0 ° C for 2 h. The resulting mixture was concentrated under vacuum. The residue was purified by preparative TLC (PE / EA=1:1) to give 2-(4,4-dimethyl-1,4-azasilanyl)-8-(1-hydroxyethyl)-3,6-dimethylquinazolin-4-one (400 mg, 66% yield) as a white solid. MS: (ES + )m / z=346.6[M+H] + .

[0245] Step 4 : Preparation of tert-butyl 2-((N-(1-(2-(4,4-dimethyl-1,4-azasilane-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)-2-nitrophenyl)sulfonamido)benzoate.

[0246] The stirred solution of 2-(4,4-dimethyl-1,4-azasilylcyclohexane-1-yl)-8-(1-hydroxyethyl)-3,6-dimethylquinazolin-4-one (400mg, 1.15mmol), PPh3(759mg, 2.89mmol) and tert-butyl 2-(2-nitrobenzenesulfonamido)benzoate (intermediate 2) (657mg, 1.73mmol) in THF (5mL) was cooled to 0 ° C, and then DBAD (800mg, 3.47mmol) in THF (5mL) was added dropwise. Under a nitrogen atmosphere, the resulting mixture was stirred at room temperature overnight, then quenched with water (5mL), and extracted with EtOAc (3x10mL). The combined organic layer was washed with brine (5mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The obtained residue was purified by preparative TLC eluting with DCM:MeOH=20:1 to give tert-butyl 2-((N-(1-(2-(4,4-dimethyl-1,4-azasilan-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)-2-nitrophenyl)sulfonamido)benzoate (500 mg, 61% yield) as a solid. MS: (ES + )m / z=706.3[M+H] +.

[0247] Step 5 : Preparation of tert-butyl 2-((1-(2-(4,4-dimethyl-1,4-azasilane-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0248] A mixture of tert-butyl 2-((N-(1-(2-(4,4-dimethyl-1,4-azasilanyl-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)-2-nitrophenyl)sulfonamido)benzoate (300 mg, 0.42 mmol), K2CO3 (174 mg, 1.3 mmol) and potassium thiophenolate (252 mg, 1.7 mmol) in DMF (10 mL) was stirred at 60 ° C for 3 h. The solution was cooled to room temperature, diluted with water (20 mL), and extracted with EtOAc (3×10 mL). The organic layers were combined, washed with water (2×10 mL) and dried over anhydrous MgSO4. After filtration, the filtrate was concentrated under reduced pressure and purified by preparative TLC, eluting with PE:EtOAc=2:1 to give tert-butyl 2-((1-(2-(4,4-dimethyl-1,4-azasilane-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (180 mg, 81% yield) as a solid. MS: (ES + )m / z=521.2[M+H] + .

[0249] Step 6 : Preparation of 2-((1-(2-(4,4-dimethyl-1,4-azasilane-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid.

[0250] A solution of tert-butyl 2-((1-(2-(4,4-dimethyl-1,4-azasilan-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (180 mg, 0.35 mmol) and HCl (4M in 1,4-dioxane, 5 mL) was stirred at room temperature overnight. The resulting mixture was concentrated under reduced pressure and purified by preparative TLC, eluting with DCM:MeOH=10:1 to give 2-((1-(2-(4,4-dimethyl-1,4-azasilan-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (100 mg, 62% yield) as a solid. MS:(ES + )m / z=465.3[M+H]+ .

[0251] Step 7 : Example 1 2-((1-(2-(4,4-dimethyl-1,4-azasilan-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomer 1; and Example 2 2-((1-(2-(4,4-dimethyl-1,4-azasilan-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, preparation of enantiomer 2.

[0252] A racemic mixture of 2-((1-(2-(4,4-dimethyl-1,4-azasilan-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (100 mg, 0.22 mmol) was separated by chiral preparative HPLC (column: CHIRALPAK IF, 2x25 cm, 5 μm; mobile phase: hexane (0.1% FA):ethanol=95:5; flow rate: 20 mL / min) to provide the title compound Example 1: 2-((1-(2-(4,4-dimethyl-1,4-azasilan-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomer 1 (34.6 mg, 39% yield, >99% ee), 1 H NMR (DMSO-d6, 300MHz): δ12.63(s,1H),8.43(s,1H),7.76(d,J=7.9Hz,1H),7.68(s,1H),7.43(s,1H),7.21–7.09(m,1H),6.52– 6.36(m,2H),5.46–5.31(m,1H),3.54–3.39(m,7H),2.30(s,3H),1.53(d,J=6.6Hz,3H),1.02–0.79(m,4H),0.12(s,6H),MS:(ES - )m / z=463.2[MH] - ;and

[0253] Example 2: 2-((1-(2-(4,4-dimethyl-1,4-azasilan-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (21 mg, 23% yield, >99% ee), 1HNMR(DMSO-d6,300MHz): δ12.60(s,1H),8.43(s,1H),7.76(d,J=7.9Hz,1H),7.68(s,1H),7.43(s,1H),7.20–7.09(m,1H),6.51 –6.37(m,2H),5.46–5.31(m,1H),3.53–3.40(m,7H),2.30(s,3H),1.53(d,J=6.6Hz,3H),1.00–0.81(m,4H),0.12(s,6H),MS:(ES - )m / z=463.2[MH] - .

[0254] Example 3: 2-((1-(2-(3,3-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0255] Example 4: 2-((1-(2-(3,3-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0256]

[0257] Step 1 : Preparation of 8-bromo-2-(3,3-dimethylpiperidin-1-yl)-3,6-dimethylquinazolin-4-one.

[0258] Under nitrogen atmosphere, a mixture of 8-bromo-2-chloro-3,6-dimethylquinazolin-4-one (intermediate 1) (700 mg, 2.43 mmol), 3,3-dimethylpiperidine hydrochloride (437 mg, 2.92 mmol) and DIEA (629 mg, 4.87 mmol) in ACN (7 mL) was stirred at 100 ° C overnight. The resulting mixture was diluted with water (20 mL) and extracted with DCM (3x20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE: EtOAc = 2: 1 to give 8-bromo-2- (3,3-dimethylpiperidin-1-yl) -3,6-dimethylquinazolin-4-one (700 mg, 79% yield). MS: (ES + )m / z=364.1[M+H] + .

[0259] Step 2: Preparation of 8-acetyl-2-(3,3-dimethylpiperidin-1-yl)-3,6-dimethylquinazolin-4-one.

[0260] This step was carried out in a manner similar to step 2 of Example 1, using PdCl2(PPh3)2 instead of Pd(PPh3)4 as a catalyst to give 8-acetyl-2-(3,3-dimethylpiperidin-1-yl)-3,6-dimethylquinazolin-4-one (600 mg, 95% yield). MS: (ES + )m / z=328.1[M+H] + .

[0261] Step 3 :Preparation of 2-(3,3-dimethylpiperidin-1-yl)-8-(1-hydroxyethyl)-3,6-dimethylquinazolin-4-one.

[0262] This step was carried out in a similar manner to step 3 of Example 1 to provide 2-(3,3-dimethylpiperidin-1-yl)-8-(1-hydroxyethyl)-3,6-dimethylquinazolin-4-one (350 mg, 86% yield). MS: (ES + )m / z=330.1[M+H] + .

[0263] Step 4 : Preparation of methyl 2-((1-(2-(3,3-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0264] A mixture of 2-(3,3-dimethylpiperidin-1-yl)-8-(1-hydroxyethyl)-3,6-dimethylquinazolin-4-one (600 mg, 1.82 mmol), 2-(2,4-dinitrobenzenesulfonyl)benzoic acid methyl ester (intermediate 3) (1.04 g, 2.73 mmol) and PPh3 (717 mg, 2.73 mmol / L) in THF (6 mL) was cooled to 0 ° C, and DBAD (629 mg, 2.73 mmol) was then added. Under a nitrogen atmosphere, the resulting mixture was stirred at room temperature for two days. During the reaction process, the 2,4-dinitrobenzenesulfonyl group was removed simultaneously. The reaction was diluted with water (20 mL) and extracted with DCM (3x30 mL). The combined organic layer was washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by preparative TLC, eluting with PE:EtOAc=5:2 to give methyl 2-((1-(2-(3,3-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (600 mg, 71% yield). MS: (ES+ )m / z=463.3[M+H] + .

[0265] Step 5 : Example 3 2-((1-(2-(3,3-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomer 1; and Example 4 2-((1-(2-(3,3-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, preparation of enantiomer 2.

[0266] A solution of methyl 2-((1-(2-(3,3-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (540 mg, 1.16 mmol) in THF (6 mL) was treated with a solution of LiOH (1.40 g, 58.35 mmol) in water (6 mL) at room temperature. The resulting mixture was stirred at 90 ° C for two days under a nitrogen atmosphere. The mixture was acidified to pH=3 with 6M HCl aqueous solution and extracted with DCM (5x50 mL). The combined organic layers were washed with water (50 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (column: C18 silica gel; mobile phase: 60-100% ACN in water over 20 min). The racemic product was separated by chiral preparative HPLC (column: CHIRALPAKIF, 2x25 cm, 5 μm; mobile phase: hexane (0.1% FA):ethanol=95:5; flow rate: 20 mL / min) to provide the title compound Example 3: 2-((1-(2-(3,3-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomer 1 (57 mg, 11% yield, >99% ee), 1 H NMR (methanol-d4, 300MHz): δ7.87(d,J=8.0Hz,1H),7.78(s,1H),7.50(s,1H),7.16–7.02(m,1H),6.55–6.39(m,2H),5.61–5.43(m,1H),3.61(s,3H) ,3.27–3.10(m,2H),2.96(s,2H),2.34(s,3H),1.94–1.76(m,2H),1.60(d,J=6.7Hz,3H),1.54–1.43(m,2H),1.08(s,3H),1.07(s,3H),MS:(ES +)m / z=449.2[M+H] + ; and Example 4: 2-((1-(2-(3,3-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (65 mg, 12% yield, 98% ee), 1 H NMR (methanol-d4, 300MHz): δ7.87(d,J=8.0Hz,1H),7.78(s,1H),7.50(s,1H),7.18–7.00(m,1H),6.55–6.39(m,2H),5.57–5.44(m,1H),3.61(s,3H) ,3.27–3.11(m,2H),2.96(s,2H),2.34(s,3H),1.94–1.76(m,2H),1.60(d,J=6.8Hz,3H),1.54–1.41(m,2H),1.08(s,3H),1.07(s,3H),MS:(ES + )m / z=449.2[M+H] + .

[0267] Example 5: 2-((1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1; and

[0268] Example 6: 2-((1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2.

[0269]

[0270] Step 1 : Preparation of 8-bromo-3,6-dimethyl-2-(2-azaspiro[3.5]nonan-2-yl)quinazolin-4(3H)-one.

[0271] Proceeding in a manner similar to Example 1, Step 1 using 2-azaspiro[3.5]nonane as the amine afforded 8-bromo-3,6-dimethyl-2-(2-azaspiro[3.5]-nonan-2-yl)quinazolin-4(3H)-one (1.35 g, 52% yield) as a solid. MS: (ES + )m / z=376.1[M+H] + .

[0272] Step 2: Preparation of 8-acetyl-3,6-dimethyl-2-(2-azaspiro[3.5]nonan-2-yl)quinazolin-4(3H)-one.

[0273] This step was carried out in a similar manner to step 2 of Example 1 to give 8-acetyl-3,6-dimethyl-2-(2-azaspiro[3.5]nonan-2-yl)quinazolin-4(3H)-one (1 g, 82% yield) as a solid. MS: (ES + )m / z=340.2[M+H] + .

[0274] Step 3 : Preparation of 8-(1-hydroxyethyl)-3,6-dimethyl-2-(2-azaspiro[3.5]nonan-2-yl)quinazolin-4(3H)-one.

[0275] This step was carried out in a similar manner to step 3 of Example 1 to give 8-(1-hydroxyethyl)-3,6-dimethyl-2-(2-azaspiro[3.5]nonan-2-yl)quinazolin-4(3H)-one (450 mg, 44% yield) as a solid. MS: (ES + )m / z=342.1[M+H] + .

[0276] Step 4 : Preparation of tert-butyl 2-((N-(1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)-2-nitrophenyl)sulfonamido)benzoate.

[0277] In a manner similar to Example 1, step 4, tert-butyl 2-((N-(1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)-2-nitrophenyl)sulfonamido)benzoate (625 mg, 76% yield) was obtained as a solid. MS: (ES + )m / z=702.3[M+H] + .

[0278] Step 5 : Preparation of tert-butyl 2-((1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0279] In a manner similar to Example 1, step 5, tert-butyl 2-((1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (200 mg, 53% yield) was obtained as a solid. MS: (ES + )m / z=517.3

[0280] [M+H] + .

[0281] Step 6 : Preparation of 2-((1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid.

[0282] In a manner similar to Example 1, step 6, 2-((1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (110 mg, 62% yield) was obtained as a solid. MS: (ES + )m / z=461.2[M+H] + .

[0283] Step 7 : Example 5 2-((1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomer 1; and Example 6 2-((1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, preparation of enantiomer 2.

[0284] The racemic mixture of 2-((1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (110 mg, 0.24 mmol) was purified by chiral preparative HPLC (column: CHIRAL ART Cellulose-SB, 2x25 cm, 5 μm; mobile phase: hexane (0.1% FA):EtOH=93:7; flow rate: 20 mL / min). The relevant fractions were concentrated to provide the title compound Example 5: 2-((1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomer 1 (8.0 mg, 8% yield, >99% ee), 1H NMR(DMSO-d6,300MHz): δ8.44(s,1H),7.85–7.74(m,1H),7.65(s,1H),7.40(s,1H),7.23–7.10(m,1H),6.56–6.41(m,2H),5. 46–5.28(m,1H),3.94(s,4H),3.40(s,3H),2.28(s,3H),1.79–1.60(m,4H),1.54(d,J=6.6Hz,3H),1.50–1.19(m,6H),MS:(ES + )m / z=461.2[M+H] + ;and

[0285] Example 6: (2-((1-(3,6-dimethyl-4-oxo-2-(2-azaspiro[3.5]nonan-2-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (10.2 mg, 10% yield, >99% ee) 1 H NMR(DMSO-d6,300MHz): δ8.43(s,1H),8.00–7.73(m,1H),7.65(s,1H),7.40(s,1H),7.32–7.05(m,1H),6.66–6.34(m,2H),5. 55–5.23(m,1H),3.95(s,4H),3.40(s,3H),2.29(s,3H),1.81–1.60(m,4H),1.55(d,J=6.5Hz,3H),1.50–1.13(m,6H),MS:(ES + )m / z=461.1[M+H] + .

[0286] Example 7: 2-((1-(2-(5-methoxyisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1; and

[0287] Example 8: 2-((1-(2-(5-methoxyisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2.

[0288]

[0289] Step 1 : Preparation of 8-bromo-2-(5-methoxy-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one.

[0290] A mixture of 8-bromo-2-chloro-3,6-dimethylquinazolin-4-one (intermediate 1) (900 mg, 3.13 mmol), 5-methoxy-2,3-dihydro-1H-isoindole (514 mg, 3.44 mmol) and K2CO3 (865 mg, 6.26 mmol) in NMP (20 mL) was stirred overnight at 120 ° C. The resulting mixture was cooled to room temperature, diluted with water (10 mL), and extracted with DCM (3x10 mL). The combined organic layers were washed with brine (3x10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE:EtOAc=9:1 to give 8-bromo-2-(5-methoxy-1,3-dihydroisoindole-2-yl)-3,6-dimethylquinazolin-4-one (900 mg, 72% yield) as a solid. MS:(ES + )m / z=400.0[M+H] + .

[0291] Step 2 : Preparation of 8-acetyl-2-(5-methoxy-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one.

[0292] This step was carried out in a similar manner to step 2 of Example 1, using PdCl2(PPh3)2 instead of Pd(PPh3)4 as a catalyst to give 8-acetyl-2-(5-methoxy-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one (560 mg, 62% yield) as a solid. MS: (ES + )m / z=364.2[M+H] + .

[0293] Step 3 : Preparation of 8-(1-hydroxyethyl)-2-(5-methoxy-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one.

[0294] A mixture of 8-acetyl-2-(5-methoxy-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one (500 mg, 1.38 mmol) and MeOH (5 mL) was cooled to 0 ° C and treated with NaBH4 (260 mg, 6.88 mmol). The mixture was heated to 80 ° C and stirred overnight. The resulting mixture was cooled to room temperature and diluted with water (30 mL), then extracted with DCM (3x10 mL). The combined organic layer was washed with brine (3x10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by reverse phase flash chromatography (C18 silica gel; 10%-80% MeOH in water over 40 min) to give 8-(1-hydroxyethyl)-2-(5-methoxy-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one (200 mg, 40% yield) as a solid. MS: (ES + )m / z=366.2[M+H] + .

[0295] Step 4 : Preparation of methyl 2-((1-(2-(5-methoxyisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0296] 8-(1-hydroxyethyl)-2-(5-methoxy-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one (200 mg, 0.547 mmol), THF (5 mL), methyl anthranilate (124 mg, 0.82 mmol) and PPh3 (359 mg, 1.37 mmol) were added to a 40 mL vial. DBAD (315 mg, 1.37 mmol) in THF (5 mL) was added to the mixture at 0 ° C, and the mixture was stirred overnight at room temperature under N2. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with CH2Cl2 (3x10 mL). The combined organic layer was washed with brine (3x10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (PE:EA=3:1) to give a crude product. The crude product was purified by reverse phase flash under the following conditions to give methyl 2-((1-(2-(5-methoxyisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (40 mg, 15% yield) as a white solid. MS: (ES + )m / z=499.2[M+H] + .

[0297] Step 5: Example 11 2-((1-(2-(5-methoxyisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1; and Example 12 Preparation of 2-((1-(2-(5-methoxyisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2.

[0298] A mixture of methyl 2-((1-(2-(5-methoxyisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (60 mg, 0.12 mmol) and LiOH hydrate (101 mg, 2.40 mmol) in THF (1 mL) and water (1 ml) was stirred at room temperature overnight. The mixture was acidified to pH=6 with 1M HCl aqueous solution and extracted with DCM (3x10 mL). The combined organic layers were washed with brine (3x10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (column: C18 silica gel; mobile phase: 5-100% ACN in water, over 40 min) to give a racemic product, which was further purified by chiral preparative HPLC (column: CHIRALPAK IF, 2x25 cm, 5 μm; mobile phase; hexane (0.1% FA): EtOH = 9: 1; flow rate: 20 mL / min) to give the title compound Example 7: 2- ((1- (2- (5-methoxyisoindolin-2-yl) -3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl) ethyl) amino) benzoic acid, enantiomer 1 (5.8 mg, 10% yield, 98% ee), 1 H NMR (methanol-d4, 300MHz): δ7.87 (dd, J=8.0, 1.7Hz, 1H), 7.76 (dd, J=2.1, 1.0Hz, 1H), 7.48 (d, J =2.1Hz,1H),7.24(d,J=8.3Hz,1H),7.06(ddd,J=8.7,7.1,1.7Hz,1H),6.93(d,J=2.3Hz,1 H),6.87(dd,J=8.3,2.4Hz,1H),6.52–6.39(m,2H),5.50(q,J=6.6Hz,1H),5.03(td,J=19. 0,18.6,14.7Hz,4H),3.80(s,3H),3.70(s,3H),2.32(s,3H),1.61(d,J=6.7Hz,3H),MS:(ES + )m / z=485.2[M+H] +; and Example 8: 2-((1-(2-(5-methoxyisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (6.8 mg, 11% yield, 99% ee), 1 H NMR (methanol-d4, 300MHz): δ7.87 (dd, J=8.0, 1.7Hz, 1H), 7.76 (dd, J=2.1, 1.0Hz, 1H), 7.48 (d, J =2.1Hz,1H),7.24(d,J=8.3Hz,1H),7.05(ddd,J=8.7,7.1,1.7Hz,1H),6.92(d,J=2.3Hz,1 H),6.87(dd,J=8.3,2.4Hz,1H),6.52–6.39(m,2H),5.50(q,J=6.6Hz,1H),5.04(td,J=19. 0,18.6,14.7Hz,4H),3.79(s,3H),3.69(s,3H),2.32(s,3H),1.61(d,J=6.7Hz,3H),MS:(ES + )m / z=485.2[M+H] + .

[0299] Example 9: 2-((1-(2-(5-chloroisoindol-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1; and

[0300] Example 10: 2-((1-(2-(5-Chloroisoindol-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2.

[0301]

[0302] Step 1 : Preparation of 8-bromo-2-(5-chloro-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one.

[0303] Proceeding in a manner similar to Example 1, Step 1 using 5-chloro-2,3-dihydro-1H-isoindole hydrochloride as the amine gave 8-bromo-2-(5-chloro-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one (2.8 g, 99% yield) as a solid. MS: (ES + )m / z=406.0[M+H] + .

[0304] Step 2: Preparation of 8-acetyl-2-(5-chloro-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one.

[0305] This step was carried out in a similar manner to Example 1, Step 2 to give 8-acetyl-2-(5-chloro-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one (2 g, 78% yield) as a solid. MS: (ES + )m / z=368.1[M+H] + .

[0306] Step 3 : Preparation of 2-(5-chloro-1,3-dihydroisoindol-2-yl)-8-(1-hydroxyethyl)-3,6-dimethylquinazolin-4-one.

[0307] In a manner similar to Example 1, step 3, 2-(5-chloro-1,3-dihydroisoindole-2-yl)-8-(1-hydroxyethyl)-3,6-dimethylquinazolin-4-one (483 mg, 24% yield) was obtained as a solid. MS: (ES + )m / z=370.1[M+H] + .

[0308] Step 4 : Preparation of methyl 2-((1-(2-(5-chloroisoindoline-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0309] Proceed in a manner similar to Example 1, Step 4 to provide methyl 2-((1-(2-(5-chloroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (181 mg, 44% yield) as an oil. MS: (ES + )m / z=503.1[M+H] + .

[0310] Step 5 : Example 9 2-((1-(2-(5-chloroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomer 1; and Example 10 2-((1-(2-(5-chloroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, preparation of enantiomer 2.

[0311] The preparation of racemic 2-((1-(2-(5-chloroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid was carried out in a manner similar to that of Step 5 of Example 1, which was then separated by chiral preparative HPLC (column: CHIRALPAK IF, 2x25 cm, 5 μm; mobile phase: hexane (0.1% FA):IPA=9:1; flow rate: 20 mL / min) to give the title compound Example 9: 2-((1-(2-(5-chloroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomer 1 (17 mg, 11% yield, >99% ee), 1 HNMR (methanol-d4, 300MHz): δ7.86 (dd, J=7.9, 1.7Hz, 1H), 7.76 (dd, J=2.0, 1.0Hz, 1H), 7.48 (d, J=2.1Hz, 1H), 7.39–7.23 (m, 3H), 7.03 (ddd, J=8 .5,7.1,1.6Hz,1H),6.50–6.35(m,2H),5.47(q,J=6.7Hz,1H),5.14–4.91(m,4H),3.67(s,3H),2.32(s,3H),1.59(d,J=6.6Hz,3H),MS:(ES + )m / z=489.2[M+H] + ;and

[0312] Example 10: 2-((1-(2-(5-chloroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (14 mg, 9% yield, 99% ee), 1 H NMR (methanol-d4, 300MHz): δ7.86 (dd, J=7.9, 1.7Hz, 1H), 7.76 (dd, J=2.0, 1.0Hz, 1H), 7.48 (d, J=2.1Hz, 1H), 7.40–7.21 (m, 3H), 7.04 (ddd, J=8 .6,7.1,1.6Hz,1H),6.51–6.34(m,2H),5.48(q,J=6.6Hz,1H),5.11–4.91(m,4H),3.68(s,3H),2.32(s,3H),1.59(d,J=6.7Hz,3H),MS:(ES + )m / z=489.2[M+H] + .

[0313] Example 11: 2-((1-(2-isobutoxy-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1; and

[0314] Example 12: 2-((1-(2-isobutoxy-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2.

[0315]

[0316] Step 1 : Preparation of 8-bromo-2-isobutoxy-3,6-dimethylquinazolin-4(3H)-one.

[0317] A solution of 8-bromo-2-chloro-3,6-dimethylquinazolin-4-one (intermediate 1) (170 mg, 0.59 mmol) in DMF (6 mL) was treated with NaH (36 mg, 0.89 mmol) and stirred at room temperature for 30 min, then isobutanol (53 mg, 0.71 mmol) was added. The reaction was stirred at room temperature for 1 h, then quenched with water (20 mL) and extracted with EtOAc (3x20 mL). The combined organic phase was washed with brine (20 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography, eluting with PE:EtOAc=10:1 to give the title product 8-bromo-2-isobutoxy-3,6-dimethylquinazolin-4(3H)-one (160 mg, 83% yield) as a solid. MS: (ES+) m / z=325.0[m+H]+.

[0318] Step 2 : Preparation of 8-acetyl-3,6-dimethyl-2-(2-methylpropoxy)quinazolin-4-one.

[0319] Prepared in a manner similar to Example 1, Step 2, the title product 8-acetyl-3,6-dimethyl-2-(2-methylpropoxy)quinazolin-4-one (120 mg, 71% yield) was obtained as a solid. MS: (ES + )m / z=289.1[M+H] + .

[0320] Step 3 : Preparation of 8-(1-hydroxyethyl)-3,6-dimethyl-2-(2-methylpropoxy)quinazolin-4-one.

[0321] Prepared in a manner similar to Example 1, Step 3, the title product 8-(1-hydroxyethyl)-3,6-dimethyl-2-(2-methylpropoxy)quinazolin-4-one (120 mg, 99% yield) was obtained as a solid. MS: (ES + )m / z=291.1[M+H] + .

[0322] Step 4 : Preparation of methyl 2-((1-(2-isobutoxy-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0323] The mixture of 8-(1-hydroxyethyl)-3,6-dimethyl-2-(2-methylpropoxy)quinazoline-4-one (110mg, 0.38mmol) and EtN (230mg, 2.27mmol) in DCM (6mL) was cooled to 0°C, then MsO (427mg, 1.52mmol) was added and stirred for 1h, and then heated to room temperature. Methyl o-aminobenzoate (54mg, 0.36mmol) was added, and the reactant was heated to 50°C and stirred overnight. The reaction was quenched by adding water (20mL) and extracted with DCM (3x30mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and purified by silica gel chromatography eluting with DCM:MeOH=10:1 to give the title product methyl 2-((1-(2-isobutoxy-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (128 mg, 80% yield) as a solid. MS: (ES + )m / z=424.2[M+H] + .

[0324] Step 5 : Example 11: 2-((1-(2-isobutoxy-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1; and Example 12: Preparation of 2-((1-(2-isobutoxy-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2.

[0325] The racemic product 2-((1-(2-isobutoxy-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid was prepared in a manner similar to that in Step 5 of Example 1 and then separated via chiral preparative HPLC (column: CHIRALPAK IF, 2x25 cm, 5 μm; mobile phase: hexane (0.1% FA):IPA=98:2; flow rate: 20 mL / min) to give Example 11: 2-((1-(2-isobutoxy-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomer 1 (3.5 mg, 19% yield, >99% ee), 1 HNMR (methanol-d4, 300MHz): δ7.88(dd,J=7.9Hz,1H),7.79(s,1H),7.52(s,1H),7.10(t,J=7.7Hz,1H),6.48(t,J=7.5Hz,1H),6.43(d,J=8.4Hz,1H),5 .45(q,J=13.5Hz,1H),4.37(dd,J=6.6Hz,2H),3.54(s,3H),2.34(s,3H),2.30–2.15(m,1H),1.60(d,J=6.6Hz,3H),1.08(d,J=1.2Hz,6H),MS(ES + )m / z=410.2[M+H] + ; and Example 12: 2-((1-(2-isobutoxy-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (4.5 mg, 25% yield, 95% ee), 1 H NMR (methanol-d4, 300MHz): δ7.88(dd,J=7.9Hz,1H),7.79(s,1H),7.52(s,1H),7.10(t,J=7.7Hz,1H),6.48(t,J=7.5Hz,1H),6.43(d,J=8.4Hz,1H),5 .45(q,J=13.5Hz,1H),4.37(dd,J=6.6Hz,2H),3.54(s,3H),2.34(s,3H),2.30–2.15(m,1H),1.60(d,J=6.6Hz,3H),1.08(d,J=1.2Hz,6H),MS(ES + )m / z=410.2[M+H] + .

[0326] Example 13: 5-chloro-2-((1-(2-(5-fluoroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1; and

[0327] Example 14: 5-Chloro-2-((1-(2-(5-fluoroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2.

[0328]

[0329] Step 1 : Preparation of 8-bromo-2-(5-fluoro-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one.

[0330] In a manner similar to Example 1, Step 1, with 5-fluoroisoindoline hydrochloride as the amine hydrochloride, the title compound 8-bromo-2-(5-fluoro-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one (800 mg, 90% yield) was obtained as a solid. MS: (ES + )m / z=388.0[M+H] + .

[0331] Step 2 : Preparation of 8-acetyl-2-(5-fluoro-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one.

[0332] In a manner similar to Example 1, step 2, the title compound 8-acetyl-2-(5-fluoro-1,3-dihydroisoindol-2-yl)-3,6-dimethylquinazolin-4-one (300 mg, 40% yield) was obtained as a solid. MS: (ES + )m / z=352.1[M+H] + .

[0333] Step 3 :Preparation of 2-(5-fluoro-1,3-dihydroisoindol-2-yl)-8-(1-hydroxyethyl)-3,6-dimethylquinazolin-4-one.

[0334] In a manner similar to Example 1, step 3, 2-(5-fluoro-1,3-dihydroisoindole-2-yl)-8-(1-hydroxyethyl)-3,6-dimethylquinazolin-4-one (284 mg, 91% yield) was obtained as an oil. MS: (ES + )m / z=354.2[M+H] + .

[0335] Step 4 : Preparation of methyl 5-chloro-2-((1-(2-(5-fluoroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0336] Proceeding in a manner similar to Example 7, Step 4, methyl 5-chloro-2-((1-(2-(5-fluoroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (334 mg, 65% yield) was obtained as an oil. MS: (ES + )m / z=521.1[M+H] + .

[0337] Step 5 : Example 13: 5-chloro-2-((1-(2-(5-fluoroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomer 1; and Example 14: Preparation of 5-chloro-2-((1-(2-(5-fluoroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomer 2.

[0338] The racemic product 5-chloro-2-((1-(2-(5-fluoroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid was prepared in a manner similar to step 5 of Example 1 and then separated by chiral preparative HPLC (column: Lux 5μm Cellulose-4, 2.12×25 cm, 5μm; mobile phase: hexane (0.1% FA):ethanol=80:20; flow rate: 20 mL / min) to give the title compound Example 13 as a solid: 5-chloro-2-((1-(2-(5-fluoroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomer 1 (32.6 mg, 10% yield, >99% ee), 1 H NMR (methanol-d4, 300MHz): δ7.84–7.65(m,2H),7.44(d,J=2.1Hz,1H),7.31(dd,J=8.3,4.9Hz,1H),7.13–6.82(m,3H),6.4 0(d,J=9.1Hz,1H),5.43(q,J=6.6Hz,1H),5.07–4.87(m,4H),3.65(s,3H),2.31(s,3H),1.58(d,J=6.6Hz,3H),MS:(ES +)m / z=507.2[M+H] + ; and Example 14: 5-chloro-2-((1-(2-(5-fluoroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (51 mg, 15% yield, 99% ee), 1 H NMR (methanol-d4, 300MHz): δ7.84–7.65(m,2H),7.44(d,J=2.1Hz,1H),7.31(dd,J=8.4,4.9Hz,1H),7.13–6.92(m,3H),6.4 0(d,J=9.1Hz,1H),5.43(q,J=6.6Hz,1H),5.14–4.89(m,4H),3.65(s,3H),2.31(s,3H),1.58(d,J=6.7Hz,3H),MS:(ES + )m / z=507.2[M+H] + .

[0339] Example 15: 5-cyano-2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1; and

[0340] Example 16: 5-Cyano-2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2.

[0341]

[0342] Step 1 : Preparation of methyl 5-cyano-2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0343] Prepared in a manner similar to Example 7, Step 4, from 2-(4,4-dimethylpiperidin-1-yl)-8-(1-hydroxyethyl)-3,6-dimethylquinazolin-4-one (prepared in Example 19, Step 3) and methyl 2-amino-5-cyanobenzoate to give methyl 5-cyano-2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (130 mg, 29% yield) as a solid. MS: (ES + )m / z=488.3[M+H] + .

[0344] Step 2 : Preparation of 5-cyano-2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomers 1 and 2.

[0345] To a stirred solution of methyl 5-cyano-2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (130 mg, 0.27 mmol) in THF (5 mL) and water (5 mL) was added LiOH hydrate (447 mg, 10.7 mmol). The reaction was warmed to 50 °C and stirred for 12 h, then diluted with water (20 mL) and extracted with EtOAc (3x30 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography (column: C18 silica gel; mobile phase: 10-100% ACN in water over 30 min) to isolate the racemic product 5-cyano-2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)amino)benzoic acid (90 mg, 71% yield) as a solid, which was then purified by chiral preparative HPLC (column: CHIRALPAK IE, 2x25 cm, 5 μm; mobile phase: hexanes (0.1% FA): EtOH = 80:20; flow rate: 20 mL / min) to give Example 15 as a solid: 5-cyano-2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (36.4 mg, 29% yield, >99% ee), 1 H NMR (methanol-d4, 300MHz): δ8.18 (d, J = 2.1Hz, 1H), 7.81 (dd, J = 2.0, 0.9Hz, 1H), 7.49 (d, J = 2.1Hz, 1H), 7.40 (dd, J = 8.9, 2.1Hz, 1H), 6.63 (d, J = 9. 0Hz,1H),5.53(q,J=6.7Hz,1H),3.58(s,3H),3.32–3.23(m,4H),2.37(s,3H),1.66(d,J=6.7Hz,3H),1.63–1.53(m,4H),1.06(s,6H),MS: (ES + )m / z=474.2[M+H] +; and Example 16: 5-cyano-2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (34.3 mg, 26% yield, 99% ee), 1 H NMR (methanol-d4, 300MHz): δ8.18 (d, J = 2.1 Hz, 1H), 7.81 (dd, J = 2.1, 0.9 Hz, 1H), 7.49 (d, J = 2.1 Hz, 1H), 7.40 (dd, J = 8.9, 2.1 Hz, 1H), 6.62 (d, J = 9. 0Hz,1H),5.53(q,J=6.7Hz,1H),3.58(s,3H),3.33–3.23(m,4H),2.37(s,3H),1.66(d,J=6.7Hz,3H),1.63–1.54(m,4H),1.06(s,6H)MS:(ES + )m / z=474.2[M+H] + .

[0346] Example 17: 2-((1-(2-(5-cyanoisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1; and

[0347] Example 18: 2-((1-(2-(5-Cyanoisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2.

[0348]

[0349] Step 1 : Preparation of methyl 2-((1-(2-(5-cyanoisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0350] To a stirred solution of methyl 2-((1-(2-(5-chloroisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (550 mg, 1.09 mmol) and tBuXphos Pd G3 (174 mg, 0.22 mmol) in H2O (2 mL) and DMF (4 mL) was added t-BuXphos (93 mg, 0.22 mmol) and Zn(CN)2 (154 mg, 1.3 mmol). Under N2 atmosphere, the resulting mixture was stirred at 70 °C for 12 h. The resulting mixture was diluted with H2O (50 mL). The resulting mixture was extracted with EtOAc (3x50 mL). The combined organic layers were washed with brine (1x50 mL) and then dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (petroleum ether / ethyl acetate = 1:1) to give methyl 2-((1-(2-(5-cyanoisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (350 mg, 65% yield) as a solid. MS: (ES + )m / z=494.3[M+H] + .

[0351] Step 2 : Preparation of 2-((1-(2-(5-cyanoisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomers 1 and 2.

[0352] The title compound was prepared using methods analogous to those described in Step 5 of Example 7 to provide Example 17: 2-((1-(2-(5-cyanoisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (6.4 mg, 3% yield, 99% ee), 1 H NMR (methanol-d4, 300 MHz): δ7.91–7.82 (m, 1H), 7.82–7.72 (m, 2H), 7.72–7.64 (m, 1H), 7.60–7.45 (m, 2H), 7.14–7.02 (m, 1H), 6.53–6.39 (m, 2H), 5.55–5.42 (m, 1H), 5.22–5.00 (m, 4H), 3.71 (s, 3H), 2.34 (s, 3H), 1.62 (d, J=6.7 Hz, 3H), MS: (ES + )m / z=480.1[M+H] +; and Example 18: 2-((1-(2-(5-cyanoisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (6.3 mg, 3% yield, >95% ee), 1 H NMR (methanol-d4, 300 MHz): δ7.91–7.82 (m, 1H), 7.82–7.72 (m, 2H), 7.72–7.64 (m, 1H), 7.60–7.45 (m, 2H), 7.14–7.04 (m, 1H), 6.54–6.40 (m, 2H), 5.55–5.42 (m, 1H), 5.22–5.01 (m, 4H), 3.71 (s, 3H), 2.35 (s, 3H), 1.62 (d, J=6.7 Hz, 3H), MS: (ES + )m / z=480.1[M+H] + .

[0353] Example 19: 2-((1-(2-(5-Carbamoylindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid.

[0354]

[0355] Step 1 : Preparation of 2-((1-(2-(5-carbamoylindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid.

[0356]

[0357] To a stirred solution of methyl 2-((1-(2-(5-cyanoisoindolin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (from Example 18, Step 1) (100 mg, 0.2 mmol) in MeOH (1 mL) and H2O (0.5 mL) was added NaOH (162 mg, 4.1 mmol). The resulting mixture was stirred at 50 °C for 6 h. The resulting mixture was diluted with H2O (30 mL). The resulting mixture was extracted with DCM (3x30 mL). The combined organic layers were washed with brine (1x30 mL) and then dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC (eluted with PE:EA=1:1) and preparative HPLC using the following conditions (column: XBridge Prep OBD C18 column, 30*150 mm, 5 μm; mobile phase A: 10 mmol NH4HCO3+0.05% NH4OH, mobile phase B: ACN; gradient: 15% B to 40% B in 9 min, flow rate: 60 mL / min) to give Example 19: 2-((1-(2-(5-carbamoylindololin-2-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (10.2 mg, 10% yield). 1 H NMR (methanol-d4, 300MHz): δ8.70(s,1H),7.97(s,1H),7.88(d,J=1.6Hz,1H),7.86–7.73(m,2H),7.68(d,J=2.0Hz,1H),7.49–7.39(m,2H),7.36(s, 1H),7.13–7.02(m,1H),6.49–6.34(m,2H),5.40(d,J=7.0Hz,1H),5.18–4.99(m,4H),3.61(s,3H),2.29(s,3H),1.55(d,J=6.5Hz,3H),MS:(ES + )m / z=498.2[M+H] - .

[0358] Example 20: 2-((1-(7-fluoro-3,6-dimethyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0359] Example 21: 2-((1-(7-fluoro-3,6-dimethyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0360]

[0361] Step 1 : Preparation of 6-amino-3-bromo-2-fluoro-N-methylbenzamide.

[0362] 6-amino-3-bromo-2-fluorobenzoic acid (20 g, 21.4 mmol), methylamine hydrochloride (8.66 g, 32 mmol), DCM (200 mL), DIEA (44.6 mL, 64.1 mmol) and HATU (48.8 g, 32 mmol) were added to a 1 L round-bottom flask at room temperature. The mixture was stirred at room temperature for 1 h. The resulting mixture was extracted with CH2Cl2 (3x200 mL). The combined organic layers were washed with brine (3x200 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE:EA=1:1) to give 6-amino-3-bromo-2-fluoro-N-methylbenzamide (19.3 g, 92% yield) as a light yellow oil. MS: (ES + )m / z=249.0[M+H] + .

[0363] Step 2 : Preparation of 6-amino-2-fluoro-N,3-dimethylbenzamide.

[0364] 6-amino-3-bromo-2-fluoro-N-methylbenzamide (19.3 g, 19.5 mmol), dioxane (360 mL), H2O (40 mL), methylboric acid (8 g, 33.4 mmol), K3PO4 (49.8 g, 58.7 mmol) and Pd(dppf)Cl2 (2.86 g, 0.98 mmol) were added to a 1000 mL round-bottom flask. Under N2 atmosphere, the mixture was stirred at 110 °C for 2 h. The reaction was quenched with water (400 mL) at room temperature. The resulting mixture was extracted with CH2Cl2 (3x200 mL). The combined organic layer was washed with brine (3x200 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE:EA=1:1) to give 6-amino-2-fluoro-N,3-dimethylbenzamide (8.40 g, 59% yield) as a white solid. MS: (ES + )m / z=183.1[M+H] + .

[0365] Step 3 : Preparation of 2-amino-3-bromo-4-fluoro-N,5-dimethylbenzamide.

[0366] 2-Amino-4-fluoro-N,5-dimethylbenzamide (8.4 g, 46.1 mmol), MeCN (100 mL) and NBS (8.21 g, 46.1 mol) were added to a 500 mL round-bottom flask. The mixture was stirred at 75 ° C for 4 h. The reaction was quenched with water (50 mL) at room temperature. The resulting mixture was extracted with CH2Cl2 (3x200 mL). The combined organic layers were washed with brine (3x100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE:EA=1:1) to give 2-amino-3-bromo-4-fluoro-N,5-dimethylbenzamide (14.6 g, >95% crude yield) as a light yellow solid. MS: (ES + )m / z=263.0[M+H] + .

[0367] Step 4 : Preparation of 2-((1-(7-fluoro-3,6-dimethyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomers 1 and 2).

[0368] Using procedures analogous to those described in Examples 7 and 8, the title compound was prepared from 2-amino-3-bromo-4-fluoro-N,5-dimethylbenzamide utilizing 8-bromo-2-chloro-7-fluoro-3,6-dimethylquinazolin-4(3H)-one, which was provided from 2-amino-3-bromo-4-fluoro-N-,5-dimethylbenzamide via procedures analogous to those described in Intermediate 1 (Steps 2 and 3) to afford Example 20: 2-((1-(7-fluoro-3,6-dimethyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1) (15.6 mg, 7% yield, >99% ee), 1 H NMR (400MHz, methanol-d4) δ7.89–7.82(m,2H),7.21–7.17(m,1H),6.83(d,J=8.5Hz,1H),6.52–6.44(m,1H),5.68(d,J=7.3Hz,1H) ,3.58(s,3H),3.52–3.43(m,4H),3.25–3.15(m,2H),3.00–2.94(m,4H),2.33(d,J=2.4Hz,3H),1.76(d,J=6.9Hz,3H),MS:(ES + )m / z=522.4[M+H] +; and Example 21: 2-((1-(7-fluoro-3,6-dimethyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2) (18.9 mg, 9% yield, 98.4% ee), 1 H NMR (400MHz, methanol-d4) δ7.89–7.82(m,2H),7.21–7.17(m,1H),6.83d,J=8.5Hz,1H),6.52–6.44(m,1H),5.68(m,1H),3.58 (s,3H),3.52–3.44(m,4H),3.25–3.15(m,2H),3.00–2.94(m,4H),2.33(d,J=2.1Hz,3H),1.76(d,J=6.9Hz,3H),MS:(ES + )m / z=522.4[M+H] + .

[0369] Example 22: 2-((1-(6-chloro-2-(4,4-dimethylpiperidin-1-yl)-3-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0370] Example 23: 2-((1-(6-Chloro-2-(4,4-dimethylpiperidin-1-yl)-3-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0371]

[0372] The title compound was prepared using Intermediate 5 using methods analogous to those described in Examples 7 and 8 to afford Example 22: 2-((1-(6-chloro-2-(4,4-dimethylpiperidin-1-yl)-3-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1) (3.2 mg, 21% yield, 99% ee) as a white solid. 1 HNMR(400MHz,DMSO-d6)δ8.70–8.65(m,1H),7.85–7.74(m,2H),7.60(s,1H),7.22–7.18(m,1H),6.55–6.50(m,1H ),6.50–6.45(m,1H),5.45–5.35(m,1H),3.50(s,3H),3.22–3.18(m,4H),2.55–3.45(m,7H),0.95(s,6H),MS:(ES +)m / z=469.1[M+H] + ; and Example 23: 2-((1-(6-chloro-2-(4,4-dimethylpiperidin-1-yl)-3-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2) (3.8 mg, 25% yield, 99% ee) as a white solid. 1 HNMR(400MHz,DMSO-d6)δ8.70–8.65(m,1H),7.85–7.78(m,2H),7.60(s,1H),7.22–7.18(m,1H),6.55–6.50(m,1H ),6.50–6.45(m,1H),5.45–5.35(m,1H),3.50(s,3H),3.22–3.18(m,4H),2.55–3.45(m,7H),0.95(s,6H),MS:(ES + )m / z=469.1[M+H] + .

[0373] Example 24: 2-((1-(6-chloro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)-5-fluorobenzoic acid (Enantiomer 1); and

[0374] Example 25: 2-((1-(6-Chloro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)-5-fluorobenzoic acid (Enantiomer 2).

[0375]

[0376] The title compound was prepared using intermediate 5 using methods analogous to those described in Examples 7 and 8 to provide Example 24: 2-((1-(6-chloro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)-5-fluorobenzoic acid, Enantiomer 1 (10.4 mg, 18% yield, 99% ee). 1HNMR(DMSO-d6,400MHz): δ13.10(s,1H),8.26(s,1H),7.84(s,1H),7.59(s1H),7.55–7.50(m,1H),7.18–7.10(m,1H),6 .44–4.40(m,1H),5.40–5.30(m,1H),3.48(s,3H),3.40–3.20(m,6H),2.83–2.60(m,4H),1.59(d,J=6.6Hz,3H),MS:(ES + )m / z=542.1[M+H] + ; and Example 25: 2-((1-(6-chloro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)-5-fluorobenzoic acid, Enantiomer 2 (10.9 mg, 19% yield, 99% ee) as a white solid. 1 HNMR(DMSO-d6,400MHz): δ13.10(s,1H),8.26(s,1H),7.84(s,1H),7.59(s 1H),7.55–7.50(m,1H),7.18–7.10(m,1H),6.44–4.40(m,1H),5.40–5.30(m,1H), 3.48(s,3H),3.40–3.20(m,6H),2.83–2.60(m,4H),1.59(d,J=6.6Hz,3H),MS:(ES + )m / z=542.1[M+H] + .

[0377] Example 26: 2-((1-(6-chloro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0378] Example 27: 2-((1-(6-Chloro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0379]

[0380] The title compound was prepared using Intermediate 5 using methods analogous to those described in Examples 7 and 8 to give Example 26 as a white solid: 2-((1-(6-chloro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1) (31.4 mg, 18% yield, 99% ee). 1 HNMR (400MHz, DMSO-d6) δ12.70(s,1H),8.40(s,1H),7.84(s,1H),7.79(d,J=7.8Hz,1H),7.58(s,1H),7.25–7.20(m,1H),6.52–6. 48(m,1H),6.44–6.40(m,1H),5.38–5.25(m,1H),3.48(s,3H),3.30–3.20(m,6H),2.90–2.85(m,4H),1.59(d,J=6.5Hz,3H),MS:(ES + )m / z=524.1[M+H] + ; and Example 27: 2-((1-(6-chloro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (36.6 mg, 21% yield, 99% ee) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ12.70(s,1H),8.40(s,1H),7.84(s,1H),7.79(d,J=7.8Hz,1H),7.58(s,1H),7.25–7.20(m,1H),6.52–6.4 8(m,1H),6.44–6.40(m,1H),5.38–5.25(m,1H),3.48(s,3H),3.30–3.18(m,6H),2.90–2.85(m,4H),1.59(d,J=6.5Hz,3H),MS:(ES + )m / z=524.1[M+H] + .

[0381] Example 28: 2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)-6-fluorobenzoic acid.

[0382]

[0383] Example 29: 2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)-5-fluorobenzoic acid (Enantiomer 1); and

[0384] Example 30: 2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)-5-fluorobenzoic acid (Enantiomer 2).

[0385]

[0386] The title compound was prepared using methods analogous to those described in Examples 7 and 8 to provide Example 29: 2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)-5-fluorobenzoic acid, Enantiomer 1 (51 mg, 27% yield, 99% ee). 1 H NMR(CD3OD-d4,300MHz):7.84(s,1H),7.58–7.55(m,1H),7.50(s,1H),6.90–6.85(m,1H),6.44–4.40(m,1H ),5.40–5.30(m,1H),3.60(s,3H),3.40–3.20(m,4H),2.35(s,3H),1.70–1.55(m,7H),1.10(s,6H),MS:(ES + )m / z=467.1[M+H] + ; and Example 30: 2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)-5-fluorobenzoic acid, Enantiomer 2 (59 mg, 31% yield, 98% ee) as a white solid. 1 H NMR(CD3OD-d4,300MHz):7.84(s,1H),7.58–7.55(m,1H),7.50(s,1H),6.90–6.85(m,1H),6.44–4.40(m,1H ),5.40–5.30(m,1H),3.60(s,3H),3.40–3.20(m,4H),2.35(s,3H),1.70–1.55(m,7H),1.10(s,6H),MS:(ES + )m / z=467.1[M+H] + .

[0387] Example 31: 2-((1-(2-((4-chloro-3-(trifluoromethoxy)benzyl)thio)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, (Enantiomer 1); and

[0388] Example 32: 2-((1-(2-((4-chloro-3-(trifluoromethoxy)benzyl)thio)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, (Enantiomer 2).

[0389]

[0390] Step 1 : Preparation of 8-bromo-3,6-dimethyl-2-thioxo-2,3-dihydroquinazolin-4(1H)-one.

[0391] 2-Amino-3-bromo-5-methylbenzoic acid (20 g, 86.9 mmol) and EtOH (200 mL) were added to a 1000 mL round-bottom flask at room temperature. Methyl isothiocyanate (12.7 g, 174 mmol) and Et3N (110 g, 1090 mmol) were added to the above mixture in batches at room temperature. The resulting mixture was stirred for another 4 h at 80 ° C. The precipitated solid was collected by filtration and washed with Et2O (3x10 mL). The resulting solid was dried under vacuum. This gave 8-bromo-3,6-dimethyl-2-thionyl-2,3-dihydroquinazoline-4 (1H) -one (20.4 g, 82% yield) as an off-white solid. MS: (ES + )m / z=286.0[M+H] + .

[0392] Step 2 : Preparation of 8-acetyl-3,6-dimethyl-2-thionyl-2,3-dihydroquinazolin-4(1H)-one.

[0393] At room temperature, under an argon atmosphere, a solution of 8-bromo-3,6-dimethyl-2-thione-2,3-dihydroquinazoline-4 (1H) -one (10g, 35.1mmol) in dioxane (100mL) was treated with tributyl (1-ethoxyvinyl) stannane (25.4g, 70.1mmol) for 3min, and then Pd (PPh3) 4 (8.1g, 7mmol) was added in batches at room temperature. Under an argon atmosphere, the resulting mixture was stirred at 100°C overnight. At room temperature, 1N HCl aqueous solution (100mL) was added dropwise to the above mixture within 1min. The resulting mixture was stirred at 50°C for 1h. The resulting mixture was concentrated under reduced pressure. The crude product was used directly in the next step without further purification. This gave 8-acetyl-3,6-dimethyl-2-thione-2,3-dihydroquinazoline-4 (1H) -one (2g, 23% yield) as a pale yellow solid. MS:(ES + )m / z=249.2[M+H] + .

[0394] Step 3 : Preparation of 8-acetyl-2-((4-chloro-3-(trifluoromethoxy)benzyl)thio)-3,6-dimethylquinazolin-4(3H)-one.

[0395] A solution of 8-acetyl-3,6-dimethyl-2-thionyl-2,3-dihydroquinazolin-4(1H)-one (2 g, 8.1 mmol) in THF (100 mL) was treated with 4-(bromomethyl)-1-chloro-2-(trifluoromethoxy)benzene (2.45 g, 8.5 mmol) at room temperature under argon atmosphere for 1 min, and then Cs2CO3 (5.27 g, 16.1 mmol) was added in portions at room temperature. Under argon atmosphere, the resulting mixture was stirred at 80 ° C for 2 h. After filtration, the resulting mixture was concentrated under vacuum to remove solids. The residue was purified by silica gel column chromatography (eluted with CH2Cl2) to give 8-acetyl-2-((4-chloro-3-(trifluoromethoxy)benzyl)thio)-3,6-dimethylquinazolin-4(3H)-one (0.85 g, 23% yield) as a light yellow solid. MS: (ES + )m / z=457.1[M+H] + .

[0396] Step 4 : Preparation of 2-((1-(2-((4-chloro-3-(trifluoromethoxy)benzyl)thio)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomers 1 and 2.

[0397]

[0398] The title compound was prepared using methods analogous to those described in Examples 7 and 8 to afford Example 31: 2-((1-(2-((4-chloro-3-(trifluoromethoxy)benzyl)thio)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (62.7 mg, 42% yield, 99% ee) as a white solid. 1 H NMR (methanol-d4, 400 MHz): δ7.89–7.80 (m, 2H), 7.57 (m, 1H), 7.52 (m, 2H), 7.45 (m, 1H), 7.02 (m, 1H), 6.47 (m, 1H), 6.16–6.09 (m, 1H), 5.35 (m, 1H), 4.66 (m, 2H), 3.66 (s, 3H), 2.34 (s, 3H), 1.47 (m, 3H), MS: (ES + )m / z=578.1[M+H] + ; and Example 32: 2-((1-(2-((4-chloro-3-(trifluoromethoxy)benzyl)thio)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (54.8 mg, 37% yield, 96% ee) as a white solid. 1 H NMR (methanol-d4, 400 MHz): δ7.89–7.80 (m, 2H), 7.57 (m, 1H), 7.52 (m, 2H), 7.45 (m, 1H), 7.02 (m, 1H), 6.47 (m, 1H), 6.13 (m, 1H), 5.35 (m, 1H), 4.70 (m, 1H), 4.62 (m, 1H), 3.66 (m, 3H), 2.35 (s, 3H), 1.47 (m, 3H), MS: (ES + )m / z=578.1[M+H] + .

[0399] Example 33: 5-fluoro-2-((1-(6-fluoro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0400] Example 34: 5-Fluoro-2-((1-(6-fluoro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0401]

[0402] The title compound was prepared using methods analogous to those described in Examples 7, 8, 11 and 12 to provide Example 33: 5-fluoro-2-((1-(6-fluoro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (31.6 mg, 20% yield, 99% ee), 1 H NMR (400 MHz, methanol-d4) δ7.68–7.53 (m, 2H), 7.47–7.38 (m, 1H), 7.04–6.92 (m, 1H), 6.45–6.37 (m, 1H), 5.57–5.46 (m, 1H), 3.62 (s, 3H), 3.43–3.37 (m, 4H), 3.24–3.17 (m, 2H), 2.99–2.89 (m, 4H), 1.65 (s, 3H), MS: (ES + )m / z=526.4[M+H] + ; and Example 34: 5-fluoro-2-((1-(6-fluoro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2) (26.0 mg, 16% yield, 99% ee), 1 H NMR (400 MHz, methanol-d4) δ7.68–7.57 (m, 2H), 7.47–7.37 (m, 1H), 7.02–6.92 (m, 1H), 6.47–6.38 (m, 1H), 5.55–5.46 (m, 1H), 3.63 (s, 3H), 3.44–3.37 (m, 4H), 3.24–3.17 (m, 2H), 2.99–2.88 (m, 4H), 1.65 (s, 3H), MS: (ES + )m / z=526.4[M+H] + .

[0403] Example 35: 2-((1-(6-fluoro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0404] Example 36: 2-((1-(6-Fluoro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0405]

[0406] Step 1 : Preparation of methyl 2-((1-(6-fluoro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazine-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0407] 6-fluoro-8-(1-hydroxyethyl)-3-methyl-2-(4-(2,2,2-trifluoroethyl)piperazine-1-yl)quinazoline-4(3H)-one (50 mg, 0.13 mmol) (prepared using a method similar to those described in Example 7) and CH2Cl2 (2 mL) were added to a 250 mL round-bottom flask. PBr3 (87 mg, 0.32 mmol) was added dropwise to the mixture at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at room temperature overnight under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The crude product was dissolved in CH2Cl2 (50 mL). The mixture was alkalized to pH 7 by adding a saturated NaHCO3 aqueous solution. The resulting mixture was extracted with DCM (2x50 mL). The combined organic layer was washed with brine (1x50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was dissolved in ACN (10 mL). Methyl anthranilate (489 mg, 0.32 mmol) was added to the mixture at room temperature. The resulting mixture was stirred at 80 ° C for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE: EA = 2: 1) to give methyl 2-((1-(6-fluoro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazine-1-yl)-3,4-dihydroquinazoline-8-yl)ethyl)amino)benzoate (75 mg, crude yield> 95%) as a light yellow solid. MS: (ES + )m / z=522.3[M+H] + .

[0408] Step 2 : Preparation of 2-((1-(6-fluoro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, enantiomers 1 and 2.

[0409]

[0410] Using procedures analogous to those described in Examples 7 and 8, the title compound was prepared using Intermediate 4 to give Example 35: 2-((1-(6-fluoro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (24 mg, 21% yield, 99% ee), 1H NMR (400MHz, methanol-d4) δ7.96-7.85(m,1H),7.68-7.57(m,1H),7.49-7.37(m,1H),7.21-7.08(m,1H),6.61-6.47(m,1H),6.47–6.35 (m,1H),5.60-5.44(m,1H),3.62(s,3H),3.46-3.35(m,4H),3.25-3.11(m,2H),3.02-2.81(m,4H),1.64(d,J=6.7Hz,3H),MS:(ES + )m / z=508.3[M+H] + ; and Example 36: 2-((1-(6-fluoro-3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (11.5 mg, 10% yield, 99% ee), 1 H NMR (400MHz, methanol-d4) δ7.95-7.86(m,1H),7.66-7.57(m,1H),7.47-7.37(m,1H),7.19-7.07(m,1H),6.60-6.48(m,1H),6.47–6.35 (m,1H),5.58-5.44(m,1H),3.62(s,3H),3.47-3.35(m,4H),3.25-3.11(m,2H),3.05-2.77(m,4H),1.64(d,J=6.7Hz,3H),MS:(ES + )m / z=508.3[M+H] + .

[0411] Example 37: 2-((1-(3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-6-(trifluoromethyl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0412] Example 38: 2-((1-(3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-6-(trifluoromethyl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0413]

[0414] The title compound was prepared using intermediate 6 using methods analogous to those described in Examples 35 and 36 to give Example 37 as a white solid: 2-((1-(3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-6-(trifluoromethyl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (30 mg, 25% yield, 99% ee). 1 H NMR (400MHz, DMSO-d6) δ12.68(s,1H),8.45(d,J=6.9Hz,1H),8.14(d,J=2.2Hz,1H),7.84(d,J=2.2Hz,1H),7.81–7.75(m,1H),7.24–7.14(m,1H) ,6.52-6.46(m,2H),5.48–5.35(m,1H),3.48(s,3H),3.44–3.36(m,4H), 3.29(d,J=10.2Hz,2H),2.90–2.75(m,4H),1.61(d,J=6.6Hz,3H),MS:(ES - )m / z=556.3[MH]; and Example 38 as a white solid: 2-((1-(3-methyl-4-oxo-2-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)-6-(trifluoromethyl)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (30 mg, 25% yield, 99% ee). 1 H NMR(400MHz, DMSO-d6)δ12.73(s,1H),8.51–8.45(m,1H),8.14(d,J=2.2Hz,1H),7.84(d,J=2.2Hz,1H),7.81–7.74(m,1H),7.26–7.13(m,1H),6 .52-6.45(m,2H),5.46–5.38(m,1H),3.48(s,3H),3.45–3.37(m,4H),3.29(d,J=10.2Hz,2H),2.90–2.75(m,4H),1.61(d,J=6.6Hz,3H),MS:(ES - )m / z=556.4[MH].

[0415] Example 39: 2-((1-(2-(isobutylthio)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0416] Example 40: 2-((1-(2-(Isobutylthio)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0417]

[0418] The title compound was prepared using methods analogous to those described in Examples 31 and 32 to afford Example 39: 2-((1-(2-(isobutylthio)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (6.3 mg, 5% yield, 99% ee) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ8.60(s,1H),7.84–7.74(m,2H),7.51(s,1H),7.15-7.11(m,1H),6.51-6.47( m,1H),6.31(d,J=8.5Hz,1H),5.50(d,J=7.2Hz,1H),3.55(s,3H),3.24(d,J=6.7Hz,2H),2.33(s,3H) , 2.13-2.06 (m, 1H), 1.54 (d, J = 6.6 Hz, 3H), 1.07-1.01 (m, 6H), LCMS (ESI) m / z = 426.0 (M+H), and Example 40 as a white solid: 2-((1-(2-(isobutylthio)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (3.8 mg, 2% yield, 99% ee). 1H NMR(400MHz,DMSO-d6)δ8.66(s,1H),7.82-7.79(m,1H),7.77-7.75(m,1H),7.51 (d,J=2.1Hz,1H),7.14-7.09(m,1H),6.53–6.44(m,1H),6.30(d,J=8.4Hz,1H),5 .50(d,J=6.9Hz,1H),3.55(s,3H),3.24(d,J=6.7Hz,2H),2.33(s,3H),2.15-2.0 4 (m, 1H), 1.54 (d, J = 6.7 Hz, 3H), 1.07-1.01 (m, 6H), LCMS (ESI) m / z = 426.0 (M + H).

[0419] Example 41: 2-((1-(2-((4-chlorophenyl)amino)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0420] Example 42: 2-((1-(2-((4-chlorophenyl)amino)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0421]

[0422] Step 1 : Preparation of 8-bromo-3,6-dimethyl-2-thionyl-2,3-dihydroquinazolin-4(1H)-one.

[0423] 2-amino-3-bromo-5-methylbenzoic acid (30.0 g, 130 mmol), TEA (45 mL, 326 mmol) and methyl isothiocyanate (19.0 g, 261 mmol) in EtOH (500 mL) were added to a 1000 mL round-bottom flask at room temperature. Under an argon atmosphere, the resulting mixture was stirred at 100 ° C for 4 h. The resulting mixture was concentrated under reduced pressure. The residue was dissolved in Et2O (500 mL). The precipitated solid was collected by filtration and washed with Et2O (2x300 mL). This obtained 8-bromo-3,6-dimethyl-2-sulfinyl (sulfanylidene) -1H-quinazoline-4-one (31.8 g, 86% yield) as a brown solid. 1 H NMR (400MHz, DMSO-d6) δ10.74 (s, 1H), 7.76 (d, J = 2.0Hz, 1H), 7.73-7.70 (m, 1H), 3.66 (s, 3H), 2.33 (s, 3H).

[0424] Step 2 : Preparation of 8-bromo-3,6-dimethyl-2-(methylthio)quinazolin-4(3H)-one.

[0425] At room temperature, dimethyl sulfate (5.9 g, 47.3 mmol) was added in batches to a stirred mixture of 8-bromo-3,6-dimethyl-2-sulfinyl-1H-quinazoline-4-one (9.0 g, 31.5 mmol) and NaOH (2.52 g, 63.1 mmol) in DMF (90 mL) under an argon atmosphere. The resulting mixture was stirred for 2 h at room temperature under an argon atmosphere. The mixture was cooled to 0 ° C. The reaction was quenched with ice water (50 mL) at 0 ° C. The resulting solid was washed with water five times (10 mL each time) and then dried under reduced pressure. This obtained 8-bromo-3,6-dimethyl-2-(methylthio)quinazoline-4 (3H)-one (8.0 g, 85% yield) as a white solid. 1 H NMR (300 MHz, chloroform-d) δ 7.99-7.91 (m, 1H), 7.83-7.76 (m, 1H), 3.60 (s, 3H), 2.73 (s, 3H), 2.46-2.40 (m, 3H).

[0426] Step 3 : Preparation of 8-acetyl-3,6-dimethyl-2-(methylthio)quinazolin-4(3H)-one.

[0427] At room temperature, tetrakis(triphenylphosphine)palladium(2.7g, 2.4mmol) was added in batches to a stirred mixture of 8-bromo-3,6-dimethyl-2-(methylthio)quinazoline-4(3H)-one(7.1g, 23.7mmol) and tributyl(1-ethoxyvinyl)stannane(10.3g, 28.5mmol) in 1,4-dioxane(100mL) under an argon atmosphere. The resulting mixture was stirred overnight at 100°C. The mixture was cooled to room temperature. At 0°C, 1N HCl aqueous solution(14.4mL) was added to the mixture in batches within 5min. The resulting mixture was stirred at room temperature for 1h. The reaction was quenched with ice water(50mL) at room temperature. The aqueous layer was extracted with EtOAc(3x50mL). The resulting mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE:EA=1:0 and increased to PE:EA=5:1 within 30 min) to give 8-acetyl-3,6-dimethyl-2-(methylthio)quinazolin-4(3H)-one (4.0 g, 64% yield) as a white solid. 1H NMR (300 MHz, CHLOROFORM-d) δ 8.22-8.15 (m, 1H), 7.86-7.79 (m, 1H), 3.64 (s, 3H), 2.87 (s, 3H), 2.66 (s, 3H), 2.51-2.44 (m, 3H).

[0428] Step 4 : Preparation of 8-(1-hydroxyethyl)-3,6-dimethyl-2-(methylthio)quinazolin-4(3H)-one.

[0429] Under nitrogen atmosphere, a solution of 8-acetyl-3,6-dimethyl-2-(methylthio)quinazolin-4(3H)-one (2.9 g, 11.1 mmol) in MeOH (20 mL) was treated with NaBH4 (836 mg, 22 mmol) at 0°C for 5 min. The resulting mixture was stirred at room temperature for 1 h. The resulting mixture was concentrated under reduced pressure and then quenched with ice water (50 mL) at 0°C. The resulting mixture was extracted with CH2Cl2 (2x50 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE:EA=1:0, increased to PE:EA=5:1 in 30 min) to give 8-(1-hydroxyethyl)-3,6-dimethyl-2-(methylthio)quinazolin-4(3H)-one (2.3 g, 79% yield) as a white solid. LCMS (ESI) m / z = 265.2 (M+H).

[0430] Step 5 : Preparation of methyl 2-((1-(3,6-dimethyl-2-(methylthio)-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0431] Under nitrogen atmosphere, a solution of 8-(1-hydroxyethyl)-3,6-dimethyl-2-(methylthio)quinazoline-4(3H)-one (2.5g, 9.4mmol) in DCM (30mL) was treated with PBr3 (25.6g, 94.5mmol) at 0°C for 5min. The resulting mixture was stirred at room temperature for 3h. The reaction was quenched with ice water (100mL). The resulting mixture was extracted with CH2Cl2 (2x100mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was suspended in MeCN (30mL), and K2CO3 (2.6g, 18.9mmol) and methyl anthranilate (4.3g, 28.3mmol) were added in batches at room temperature. The resulting mixture was stirred at 80°C for 2h. The reaction was then cooled to room temperature and quenched with ice water (50mL). The resulting mixture was extracted with CH2Cl2 (2x50mL). The combined organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE:EA=1:0, increased to PE:EA=5:1 in 30min), to give methyl 2-((1-(3,6-dimethyl-2-(methylthio)-4-oxo-3,4-dihydroquinazoline-8-yl)ethyl)amino)benzoate (2.5g, 67% yield) as a white solid. LCMS(ESI)m / z=397.9(M+H).

[0432] Step 6 : Preparation of methyl 2-((1-(3,6-dimethyl-2-(methylsulfonyl)-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0433] Under nitrogen atmosphere, a solution of methyl 2-((1-(3,6-dimethyl-2-(methylthio)-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (1.5 g, 3.7 mmol) in DMF (15 mL) was treated with m-CPBA (1.9 g, 11.3 mmol) at 0 ° C for 5 min. The resulting mixture was stirred at room temperature overnight. The reaction was quenched with saturated NaHCO3 solution (50 mL) at 0 ° C. The resulting mixture was extracted with CH2Cl2 (2x50 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE:EA=1:0, increasing to PE:EA=5:1 within 30 min) to give methyl 2-((1-(3,6-dimethyl-2-(methylsulfonyl)-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (911 mg, 56% yield) as an off-white solid. LCMS (ESI) m / z=430.0 (M+H).

[0434] Step 7 : Preparation of methyl 2-((1-(2-((4-chlorophenyl)amino)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate.

[0435] Under nitrogen atmosphere, to a mixture of methyl 2-((1-(3,6-dimethyl-2-(methylsulfonyl)-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (120 mg, 0.3 mmol) and 4-chloroaniline (107 mg, 0.8 mmol) in THF (4.0 mL) was added dropwise LiHMDS (2M, 1.2 mL) in THF at 0 ° C. The resulting mixture was then warmed to room temperature and stirred for 30 min. The reaction was quenched with ice water (20 mL). The resulting mixture was extracted with EtOAc (2x20 mL). The combined organic layer was washed with brine, dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluted with PE:EA=1:0, increasing to PE:EA=1:1 over 30 min) to give methyl 2-((1-(2-((4-chlorophenyl)amino)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoate (125 mg, 94% yield) as a white solid. 1 H NMR (300MHz, DMSO-d6) δ8.82(s,1H),8.14(d,J=6.3Hz,1H),7.79(d,J=9.0Hz,3H),7.70(d,J=1.8Hz,1H),7.38(d,J=8.9Hz,3H),7.22-7 .11(m,1H),6.57-6.46(m,1H),6.29(d,J=8.6Hz,1H),5.32-5.22(m,1H),3.83(s,3H),3.63(s,3H),2.29(s,3H),1.50(d,J=6.6Hz,3H).

[0436] Step 8 : Preparation of 2-((1-(2-((4-chlorophenyl)amino)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomers 1 and 2).

[0437]

[0438] The title compound was prepared using methods analogous to those described in Examples 7 and 8 to afford Example 41: 2-((1-(2-((4-chlorophenyl)amino)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (29 mg, 36% yield, 99% ee) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ12.65(s,1H),8.82(s,1H),8.32(s,1H),8.03–7.60(m,4H),7.39(d,J =6.7Hz,3H),7.17-7.09(m,1H),6.53-6.45(m,1H),6.24(d,J=8.6Hz,1H),5.26(s,1H),3.63( s, 3H), 2.29 (s, 3H), 1.49 (d, J = 6.6 Hz, 3H). LCMS (ESI) m / z = 463.2 (M+H), and Example 42 as a white solid: 2-((1-(2-((4-chlorophenyl)amino)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (29 mg, 36% yield, 99% ee). 1 H NMR(400MHz, DMSO-d6)δ8.82(s,1H),8.54(s,1H),7.85–7.75(m,3H),7.69(d,J=2.0Hz,1H),7.43–7.35(m,3H),7.12-7.03(m,1H),6 .50-6.41(m,1H),6.20(d,J=8.5Hz,1H),5.24(s,1H),3.63(s,3H),2.29(s,3H),1.48(d,J=6.6Hz,3H), LCMS(ESI)m / z=463.2(M+H).

[0439] Example 43: 2-((1-(2-((4-chlorobenzyl)amino)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0440] Example 44: 2-((1-(2-((4-chlorobenzyl)amino)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0441]

[0442] The title compound was prepared using methods analogous to those described in Examples 41 and 42 to afford Example 43: 2-((1-(2-((4-chlorobenzyl)amino)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (32 mg, 39% yield, 95% ee) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.44(s,1H),7.85-7.78(m,1H),7.75(d,J=7.9Hz,1H),7.58(s,1H),7.44(d,J=8.1Hz,2H),7.32( d,J=8.2Hz,2H),7.26(d,J=2.2Hz,1H),7.06-6.97(m,1H),6.48-6.40(m,1H),6.11(d,J=8.5Hz,1H),5.19(d,J=6.4Hz,1H ), 4.74-4.64 (m, 1H), 4.60-4.51 (m, 1H), 3.50 (s, 3H), 2.22 (s, 3H), 1.25 (d, J = 6.6 Hz, 3H). LCMS (ESI) m / z = 476.9 / 479.0 (M+H), and Example 44 as a white solid: 2-((1-(2-((4-chlorobenzyl)amino)-3,6-dimethyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (30 mg, 36% yield, 95% ee). 1 H NMR(400MHz,DMSO-d6)δ8.40(s,1H),7.85–7.78(m,1H),7.77-7.71(m,1H),7.58(d,J=2.2Hz ,1H),7.44(d,J=8.2Hz,2H),7.35–7.29(m,2H),7.26(d,J=2.2Hz,1H),7.07-6.99(m,1H),6. 48-6.40(m,1H),6.12(d,J=8.5Hz,1H),5.19(d,J=6.4Hz,1H),4.74-4.64(m,1H),4.60-4.50 (m, 1H), 3.50 (s, 3H), 2.22 (s, 3H), 1.26 (d, J = 6.6Hz, 3H). LCMS (ESI) m / z = 477.0 / 479.0 (M+H).

[0443] Example 45: 2-((1-(3,6-dimethyl-4-oxo-2-(((R)-2-phenylpropyl)amino)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0444] Example 46: 2-((1-(3,6-dimethyl-4-oxo-2-(((R)-2-phenylpropyl)amino)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0445]

[0446] The title compound was prepared using methods analogous to those described in Examples 41 and 42 to afford Example 45: 2-((1-(3,6-dimethyl-4-oxo-2-(((R)-2-phenylpropyl)amino)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (25 mg, 36% yield, 99% ee) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ12.66(s,1H),8.41(s,1H),7.82-7.75(m,1H),7.63-7.58(m,1H),7.33(d,J=2.2Hz,1H),7.26 (d,J=4.3Hz,4H),7.22–7.13(m,3H),6.54-6.45(m,1H),6.42(d,J=8.5Hz,1H),5.46(m,1H),3.75-3.62(m,1H),3.54- 3.43 (m, 1H), 3.37 (m, 4H), 2.25 (s, 3H), 1.54 (d, J = 6.6 Hz, 3H), 1.29 (d, J = 6.9 Hz, 3H), LCMS (ESI) m / z = 471.1 (M+H); and Example 46 as a white solid: 2-((1-(3,6-dimethyl-4-oxo-2-(((R)-2-phenylpropyl)amino)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (16 mg, 23% yield, 99% ee). 1 H NMR (400MHz, DMSO-d6) δ12.66(s,1H),8.44(s,1H),7.80-7.73(m,1H),7.61(d,J=1.8H z,1H),7.36–7.26(m,5H),7.26-7.18(m,2H),7.15-7.07(m,1H),6.50-6.42(m,1H),6. 39(d,J=8.6Hz,1H),5.46(m,1H),3.69–3.61(m,1H),3.61–3.54(m,1H),3.38(m,4H),2 .26 (s, 3H), 1.56 (d, J = 6.6Hz, 3H), 1.25 (d, J = 10.5Hz, 3H), LCMS (ESI) m / z = 471.1 (M + H).

[0447] Example 47: 2-((1-(3,6-dimethyl-4-oxo-2-(((S)-2-phenylpropyl)amino)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0448] Example 48: 2-((1-(3,6-dimethyl-4-oxo-2-(((S)-2-phenylpropyl)amino)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0449]

[0450] The title compound was prepared using methods analogous to those described in Examples 41 and 42 to afford Example 47: 2-((1-(3,6-dimethyl-4-oxo-2-(((S)-2-phenylpropyl)amino)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (30 mg, 56% yield, 99% ee) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ12.64(s,1H),8.41(s,1H),7.79(d,J=8.0,1.7Hz,1H),7.61(m,1H),7.33(d,J=2.2Hz,1H),7. 26(d,J=4.4Hz,4H),7.22–7.13(m,3H),6.55–6.45(m,1H),6.42(d,J=8.4Hz,1H),5.46(m,1H),3.74-3.64(m,1H),3.54 –3.43 (m, 1H), 3.37 (m, 4H), 2.25 (s, 3H), 1.54 (d, J = 6.7 Hz, 3H), 1.29 (d, J = 7.2 Hz, 3H), LCMS (ESI) m / z = 471.2 (M+H); and Example 48 as a yellow solid: 2-((1-(3,6-dimethyl-4-oxo-2-(((S)-2-phenylpropyl)amino)-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid enantiomer 2 (12 mg, 24% yield, 99% ee). 1H NMR(400MHz,DMSO-d6)δ12.62(s,1H),8.40(s,1H),7.80-7.73(m,1H),7.61(d,J =2.1Hz,1H),7.37–7.25(m,5H),7.27-7.18(m,2H),7.15-7.10(m,1H),6.55-6.3 6(m,2H),5.45(m,1H),3.70-3.59(m,1H),3.57-3.46(m,1H),3.38(m,4H)2.26(s , 3H), 1.56 (d, J = 6.7Hz, 3H), 1.26 (d, J = 7.2Hz, 3H), LCMS (ESI) m / z = 471.2 (M + H).

[0451] Example 49: 2-((1-(3-cyclopropyl-2-(4,4-dimethylpiperidin-1-yl)-6-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1), and

[0452] Example 50: 2-((1-(3-Cyclopropyl-2-(4,4-dimethylpiperidin-1-yl)-6-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0453]

[0454] The title compound was prepared from Intermediate 7 using methods analogous to those described in Examples 7 and 8 to afford Example 49 as a white solid: 2-((1-(3-cyclopropyl-2-(4,4-dimethylpiperidin-1-yl)-6-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (20 mg, 99% ee). 1H NMR (400MHz, DMSO-d6) δ12.61(s,1H),8.38(s,1H),7.80-7.73(m,1H),7.64(d,J=2.4Hz,1H),7.39(d,J=2.1Hz,1H ),7.22-7.13(m,1H),6.53-6.44(m,2H),5.35(m,1H),3.52–3.36(m,4H),3.22-3.13(m,1H),2.28(s,3H),1.58-1. 41 (m, 7H), 1.20–1.10 (m, 2H), 1.00 (s, 6H), 0.81-0.73 (m, 1H), 0.73–0.62 (m, 1H), LCMS (ESI) m / z=475.3 (M+H); and Example 50 as a white solid: 2-((1-(3-cyclopropyl-2-(4,4-dimethylpiperidin-1-yl)-6-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (19 mg, 99% ee). 1 H NMR(400MHz,DMSO-d6)δ12.61(s,1H),8.38(s,1H),7.80-7.73(m,1H),7.66-7.60( m,1H),7.39(d,J=2.1Hz,1H),7.22-7.13(m,1H),6.53-6.44(m,2H),5.35(s,1H),3 .48–3.41(m,4H),3.22-3.13(m,1H),2.28(s,3H),1.56-1.42(m,7H),1.19–1.13(m , 2H), 1.00 (s, 6H), 0.75 (s, 1H), 0.67 (d, J = 7.9Hz, 1H), LCMS (ESI) m / z = 475.2 (M + H).

[0455] Example 51: 2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3-(fluoromethyl)-6-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 1); and

[0456] Example 52: 2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3-(fluoromethyl)-6-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomer 2).

[0457]

[0458] Step 1: Preparation of 8-bromo-2-(4,4-dimethylpiperidin-1-yl)-6-methyl-3H-quinazolin-4-one.

[0459] To a stirred solution of 8-bromo-2-chloro-6-methyl-3H-quinazolin-4-one (7.0 g, 25.6 mmol) and 4,4-dimethylpiperidine (2.9 g, 25.6 mmol) in NMP (100 mL) was added K2CO3 (14.2 g, 102 mmol) at room temperature. The resulting mixture was stirred at 110 ° C overnight. The reaction was cooled to room temperature and quenched with water (150 mL). The aqueous layer was extracted with EtOAc (3x150 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SO4. The resulting mixture was concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN (0.1% FA) in water, 10% to 95% gradient in 30 min; detector, UV 254 nm. This afforded 8-bromo-2-(4,4-dimethylpiperidin-1-yl)-6-methyl-3H-quinazolin-4-one (6.1 g, 68% yield) as a yellow solid. LCMS (ESI) m / z = 350.1 / 352.3 (M+H).

[0460] Step 2 : Preparation of 8-bromo-2-(4,4-dimethylpiperidin-1-yl)-3-(fluoromethyl)-6-methylquinazolin-4-one.

[0461] Under nitrogen atmosphere, a solution of 8-bromo-2-(4,4-dimethylpiperidin-1-yl)-6-methyl-3H-quinazolin-4-one (1.2 g, 3.4 mmol) in DMF (15 mL) was treated with NaH (411 mg, 17.1 mmol, 60% oil solution) at 0 ° C for 10 min, and then bromofluoromethane (1.9 g, 17.1 mmol) was added dropwise. The resulting mixture was stirred at 60 ° C for 4 h. The reaction was cooled to room temperature and quenched with water (50 mL). The aqueous layer was extracted with EtOAc (3x50 mL). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by reverse phase flash chromatography using the following conditions: column, C18 silica gel; mobile phase, MeCN in water, 40% to 100% gradient in 20 min. This afforded 8-bromo-2-(4,4-dimethylpiperidin-1-yl)-3-(fluoromethyl)-6-methylquinazolin-4-one (1.0 g, 76% yield) as a yellow solid. LCMS (ESI) m / z = 382.1 / 384.3 (M+H).

[0462] Step 3: Preparation of 2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3-(fluoromethyl)-6-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid (Enantiomers 1 and 2).

[0463]

[0464] The title compound was prepared using methods analogous to those described in Examples 7 and 8 to give Example 51 as a white solid: 2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3-(fluoromethyl)-6-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 1 (11 mg, 99% ee). 1 H NMR (400MHz, DMSO-d6) δ12.63(s,1H),8.46(s,1H),7.80-7.75(m,1H),7.58(d,J=2.1Hz,1H),7.48(d,J=2. 0Hz,1H),7.18-7.12(m,1H),6.51–6.40(m,2H),6.33(m,1H),6.20(m,1H),5.43(m,1H),3.9-3.83(m,4H),2 .31 (s, 3H), 1.57 (d, J = 6.6 Hz, 3H), 1.45-1.36 (m, 4H), 1.00 (s, 6H), LCMS (ESI) m / z = 467.2 (M+H); and Example 52 as a white solid: 2-((1-(2-(4,4-dimethylpiperidin-1-yl)-3-(fluoromethyl)-6-methyl-4-oxo-3,4-dihydroquinazolin-8-yl)ethyl)amino)benzoic acid, Enantiomer 2 (9 mg, 99% ee).

[0465] 1 H NMR(400MHz,DMSO-d6)δ12.63(s,1H),8.46(s,1H),7.80-7.75(m,1H),7.58( d,J=2.1Hz,1H),7.48(d,J=2.0Hz,1H),7.18-7.12(m,1H),6.51–6.40(m,2H), 6.33(m,1H),6.20(m,1H),5.43(m,1H),3.9-3.83(m,4H),2.31(s,3H),1.57( d, J=6.6Hz, 3H), 1.45-1.36 (m, 4H), 1.00 (s, 6H); LCMS (ESI) m / z=467.2 (M+H).

[0466] Assays and compound testing

[0467] In vitro cell proliferation: T47D cells expressing mutant PI3Ka (H1047R) and cells expressing WT PI3Ka were measured. EC50 values ​​for proliferation inhibition of SKBR3 cells.

[0468] T47D or SKBR3 cells were trypsinized, resuspended in culture medium, and plated onto 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 plated at a density of 1,500 cells / well and dispensed into 384-well assay-ready plates (Corning, 89089-790) at 50uL. The assay-ready plates were previously printed with 10-point dilutions of the target compound and controls. Echo655 was used to stamp the plates under 40nL of compound or DMSO. Cells were grown at 37°C and 5% CO2 for 72 hours. After 72 hours, the cells were equilibrated at room temperature for 15 minutes. 30uL of CellTiter-Glo reagent was added to the plate, which was then shaken at 300-500rpm for 30 minutes at temperature. The cells were then read on an Envision plate reader. The percentage of proliferation inhibition was calculated using the following formula: % inhibition = 100 x (Lum D -Lum 样品 ) / (Lum D -Lum inh ), where D is obtained from cells treated with 0.1% DMSO only; Inh is obtained from cells treated with 10uM apellis. The effective concentration (EC50) to achieve 50% inhibition of proliferation was calculated by fitting the curve using Xlfit (v5.3.1.3), equation 201: Y = bottom + (top - bottom) / (1 + 10^((LogEC50-X) * slope)).

[0469] Reagent List:

[0470]

[0471] In vitro cell pAKT: T47D cells expressing mutant PI3Ka (H1047R) and cells expressing WT PI3Ka were measured IC50 values ​​for inhibition of AKT phosphorylation (pAKT) in SKBR3 cells.

[0472] After trypsinizing T47D or SKBR3 cells, they were resuspended in the medium and seeded onto assay preparation plates. The T47D medium consisted of RPMI, 10% FBS, and insulin (0.2 units / mL). The SKBR3 medium consisted of McCoy's 5a and 10% FBS. The cells were seeded at a density of 5000 cells / well and dispensed at 12.5 uL into 384-well assay preparation plates (Perkin-Elmer, 6008238). The assay preparation plates had previously been printed with 10-point dilutions of the target compounds and controls. The Echo 655 was used to stamp the plates with 12.5 nL of compound or DMSO. The cells were grown at 37 °C and 5% CO2 for 6 hours. After 6 hours, 4 uL of lysis buffer reagent was added to the plates, and then the plates were centrifuged at 1000 rpm for 1 minute. Then the plates were incubated at room temperature for 30 minutes. After 30 minutes, 4 uL of antibody mixture containing Eu Cryptate, d2-Cryptate, and detection buffer was added to the plates. The plates were centrifuged at 1000 rpm for 1 minute and then incubated overnight at room temperature. The plates were read on an Envision plate reader using the HTRF protocol. The percent inhibition of AKT phosphorylation was calculated using the following formula: % Inhibition = 100 x (pAKT HC - pAKT Sample) / (pAKT HC - pAKT LC), where pAKT HC was obtained from cells treated with only 0.1% DMSO; pAKT LC was obtained from cells treated with 10 uM alpelisib. The IC50 (the concentration at which 50% inhibition of pAKT was achieved) was calculated by fitting a curve using Xlfit (v5.3.1.3), equation 201: Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X) * Slope)).

[0473] Reagent List:

[0474] Reagents supplier Part Number Gibco RPMI 1640 Medium, without phenol red Fisher 11835030 Gibco RPMI 1640 Medium Fisher 11875093 McCoy's 5A (modified) medium, HEPES Fisher 12330031 DMEM, high glucose, HEPES, without phenol red Fisher 21063045 Gibco Trypsin-EDTA (0.5%), without phenol red Fisher 15400054 PBS Solarbio P1020 Trypan blue solution 0.4% Fisher T10282 Fetal Bovine Serum Australian Source Sigma 82051-458 ProxiPlate-384Plus, white, TC treated, box of 160 PerkinElmer 6008238 pAKT(Ser473)HTRF Cisbio 64AKSPEH

[0475] For the EC50 values shown in Table 1, "A" refers to 1 nM < EC50 < 500 nM; "B" refers to 500 nM < EC50 < 2 μM; "C" refers to 2 μM < EC50 < 15 μM; and "D" refers to EC50 > 15 μM.

[0476] Table 1. Cell Proliferation Data

[0477]

[0478]

[0479] For the IC50 values shown in Table 2, "A" means 1 nM < IC50 < 1 μM; "B" means 1 μM < IC50 < 5 μM; "C" means 5 μM < IC50 < 15 μM; and "D" means IC50 > 15 μM.

[0480] Table 2. Cellular pAKT Data

[0481]

[0482]

[0483] CD1 mice received a single IV or PO dose, and then plasma was sampled continuously at 0.0833 (IV only), 0.25, 0.5, 1, 2, 4, 8, 24 hours post-dose. The desired series of working solution concentrations were achieved by diluting the analyte stock solution with 50% aqueous acetonitrile. 10 μL of the working solution (0.5, 1, 2, 5, 10, 50, 100, 500, 1000 ng / mL) was added to 10 μL of blank female CD1 mouse plasma to achieve calibration standards of 0.5 - 1000 ng / mL with a total volume of 20 μL. Five plasma quality control samples at 1 ng / mL, 2 ng / mL, 5 ng / mL, 50 ng / mL, and 800 ng / mL were prepared independently of the samples used for 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 standards, 20 μL of the QC samples, and 20 μL of the unknown samples (10 μL plasma and 10 μL blank solution) were added separately to 200 μL of an acetonitrile-containing IS mixture to precipitate proteins. Then, after centrifugation at 4°C and 4000 rpm for 15 min, the samples were vortexed for 30 seconds. The supernatant was diluted with water at a ratio of 1:2 (V / V, 1:2), and then 5 μL of the diluted supernatant was injected into the LC / MS / MS system for quantitative analysis. The results are shown in Table 3.

[0484] Table 3: Mouse Pharmacokinetic Data

[0485]

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Claims

1. Compound of formula (1) or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, in: R1 is H, C1-C4 alkyl or C3-C7 cycloalkyl; Each R2 is independently H, C1-C4 alkyl, C3-C7 cycloalkyl, R 14 -C≡C-, halogen, CN, CF3, OCF3, CFH2 or CF2H; R3 is H, C1-C4 alkyl, C3-C7 cycloalkyl, CF3, CFH2, or CF2H, and when R3 is not H, the carbon atom attached to R3 is a chiral center and exists as a (R)- and (S)-racemic mixture or as an (R)- or (S)-enantiomer; R4 is H or C1-C4 alkyl; R6 is H, C1-C4 alkyl, C3-C7 cycloalkyl, heteroaryl, CF3, CFH2 or CF2H; R7 is H, C1-C4 alkyl, C3-C7 cycloalkyl, halogen, CN, CF3, OCF3, OCH3, CFH2 or CF2H; R8 is H, C1-C4 alkyl, C3-C7 cycloalkyl, halogen, CN, CF3, OCF3, OCH3, CFH2 or CF2H; R5 is -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; or <h2 style=";text-align:left;direction:ltr">-(NR<h2 style=";text-align:left;direction:ltr"> 10 <h2 style=";text-align:left;direction:ltr"> )-L1-L2-L3-L4-L5-L6-L7-R9, in: Each of L1, L2, L3, L6 and L7 is independently (CHR 11 )、(CHR 11 -O)、(CHR 11 -S), (C3-C7 cycloalkyl) or a bond; L4 is C=O, C=S or a bond; L5 is NR 10 , S, O or key; R9 is H, C(=O)R 12 、C(=O)NR 12 R 13 、C(=O)OR 12 , C1-C6 alkyl, C1-C6 fluoroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each of C1-C6 alkyl, C1-C6 fluoroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is unsubstituted or substituted, or when NR 10 When present, R9 and R 10 Together with the attached nitrogen atom, may form a 4-7 membered non-aromatic heterocyclic ring, wherein the ring may contain additional heteroatoms and be substituted or unsubstituted; R 10 and R 11 Each of is H or C1-C4 alkyl, wherein C1-C4 alkyl is unsubstituted or substituted; and R 12 and R 13 Each of R is independently H or C1-C6 alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein each of C1-C6 alkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl is unsubstituted or substituted, or R 12 and R 13 Together with the attached nitrogen atom, may form a 4-7 membered non-aromatic heterocyclic ring, wherein the ring may contain additional heteroatoms and be substituted or unsubstituted; Each R 14 are independently H, C1-C3 alkyl or C3-C7 cycloalkyl; Or R5 is Nonaromatic N-linked heterocycles wherein the heterocycle is substituted or unsubstituted, optionally contains one or more additional atoms selected from N (wherein N is substituted or unsubstituted), O, Si (wherein Si is substituted or unsubstituted) and S (wherein S is oxidized or unoxidized), and is optionally part of a bridged, fused or spiro ring system.

2. The compound according to claim 1, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R5 is -(NR 10 )-L1-L2-L3-L4-L5-L6-L7-R9, where L1 to L7, R9 and R 10 As defined.

3. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R5 is -O-L1-L2-L3-L4-L5-L6-L7-R9, wherein L1 to L7 and R9 are as defined.

4. The compound according to claim 1 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein 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; wherein L1 to L7 and R9 are as defined.

5. The compound according to claim 1, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R5 is an N-linked non-aromatic heterocyclic ring wherein the heterocycle is substituted or unsubstituted, optionally contains one or more additional atoms selected from N (wherein N is substituted or unsubstituted), O, Si (wherein Si is substituted or unsubstituted) and S (wherein S is oxidized or unoxidized), and is optionally part of a bridged, fused or spiro ring system.

6. A compound according to claim 5, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein the heterocycle is substituted or unsubstituted, optionally contains one or more additional heteroatoms selected from N, O, Si and S, and is not part of a bridged, fused or spiro ring system.

7. A compound according to claim 5, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein the heterocycle is substituted or unsubstituted, optionally contains one or more additional heteroatoms selected from N, O, Si and S, and is part of a bridged, fused or spiro ring system.

8. The compound according to claim 5, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein the heterocycle is substituted or unsubstituted, contains no additional heteroatoms, and is not part of a bridged, fused or spirocyclic system.

9. The compound according to claim 5, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein the heterocycle is substituted or unsubstituted, does not contain additional heteroatoms, and is part of a bridged, fused or spiro ring system.

10. The compound according to claim 5, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein the heterocycle is substituted or unsubstituted, contains at least one sulfur ring atom, and is not part of a bridged, fused or spiro ring system.

11. The compound according to claim 5, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein the heterocycle is substituted or unsubstituted, contains at least one sulfur ring atom, and is part of a bridged, fused or spiro ring system.

12. The compound according to claim 5, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein the heterocycle is substituted or unsubstituted, contains at least one oxygen ring atom, and is not part of a bridged, fused or spiro ring system.

13. The compound according to claim 5, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein the heterocycle is substituted or unsubstituted, contains at least one oxygen ring atom, and is part of a bridged, fused or spiro ring system.

14. A compound according to claim 5, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein the heterocycle is substituted or unsubstituted, contains at least one additional nitrogen ring atom, and is not part of a bridged, fused or spiro ring system.

15. A compound according to claim 5, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein the heterocycle is substituted or unsubstituted, contains at least one additional nitrogen ring atom, and is part of a bridged, fused or spiro ring system.

16. The compound according to claim 1 or its solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein the compound of formula (1) is selected from 17. A compound according to any one of claims 1 to 15, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R5 is selected from in: A is O, S, S(O) or S(O)2; Each R h and R i Independently selected from H, CH3, c-Pr, c-Bu, CF3 and OH; Each R j is independently selected from CF3, CH2CF3, CH2CF2H, OCH3, OCH2CF3, OCF3, Oc-Pr, aryl, heteroaryl, COCH3 and CO2CH3; and Each R k and R m Independently selected from H, C1-C3 alkyl and acetyl (COCH3).

18. The compound according to claim 1, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R1 is H.

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

20. The compound according to claim 1, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R3 is CH3.

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

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

23. The compound according to claim 1, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R7 is CH3 or F.

24. The compound according to claim 1, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R8 is H.

25. The compound according to claim 1, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R1, each R2 and R4 is H.

26. The compound according to claim 1, wherein the compound of formula (1) is a compound of formula (2) Or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R3 is CH3, CF3, CFH2 or CF2H; R5 is as defined in the compound of formula (1); R7 is CH3 or F; and the carbon marked with * is a chiral center and exists as a (R)- and (S)-racemic mixture or a (R)- or (S)-enantiomer.

27. The compound according to claim 1, wherein the compound of formula (1) is a compound of formula (3) or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R2 is as defined in the compound of formula (1); R3 is CH3, CF3, CFH2 or CF2H; R7 is CH3 or F; R 15 is OCH3, OCH2CH3, OCH2CF3, O-cyclopropyl, CH2CF3, CH2CF2H, aryl or heteroaryl; each R 16 are independently H or C1-C3 alkyl; and the carbon marked with an * is a chiral center and exists as a (R)- and (S)-racemic mixture or as a (R)- or (S)-enantiomer.

28. The compound according to claim 1, wherein the compound of formula (1) is a compound of formula (4) or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R2 is as defined in the compound of formula (1); R3 is CH3, CF3, CFH2 or CF2H; R7 is CH3 or F; R 15 is OCH3, OCH2CH3, OCH2CF3, O-cyclopropyl, CH2CF3, CH2CF2H, aryl or heteroaryl; and the carbon marked with * is a chiral center and exists as a (R)- and (S)-racemic mixture or as a (R)- or (S)-enantiomer.

29. The compound according to claim 1, wherein the compound of formula (1) is a compound of formula (5) or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, wherein R2 is as defined in the compound of formula (1); R3 is CH3, CF3, CFH2 or CF2H; R7 is CH3 or F; R 15 is OCH3, OCH2CH3, OCH2CF3, O-cyclopropyl, CH2CF3, CH2CF2H, aryl or heteroaryl; and the carbon marked with * is a chiral center and exists as a (R)- and (S)-racemic mixture or as a (R)- or (S)-enantiomer.

30. The compound according to claim 1 or its solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound, or a pharmaceutically acceptable salt thereof, wherein the compound of formula (1) is selected from one or more of the following 31. A pharmaceutical composition comprising a compound according to any one of claims 1 to 30 or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotope-labeled compound thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

32. The pharmaceutical composition of claim 31 further comprising one or more anticancer agents.

33. The pharmaceutical composition of claim 32, wherein the one or more anticancer agents are selected from 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 and a proteasome inhibitor.

34. A method for treating a disease in which PI3K activity is associated with a subject in need of such treatment, the method comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 30, or a solvate, enantiomer, diastereomer, tautomer, polymorph or isotopically labeled compound thereof, or a pharmaceutically acceptable salt thereof.

35. The method of claim 34, wherein the disease is cancer.

36. The method of claim 34, wherein the disease is selected from congenital lipomatous overgrowth, vascular malformations, epidermal nevus, scoliosis / skeletal and spine syndrome (CLOVES), mosaic tissue overgrowth syndrome, venous malformations and brain malformations associated with severe epilepsy, or PIK3CA-associated overgrowth syndrome.

37. The method of claim 34, wherein the disease is cancer carrying a PI3Kα H1047R mutation.

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