Adenosine receptor antagonist compounds

Abstract

To provide adenosine receptor (e.g., A2A and / or A1 receptor) antagonist compounds and compositions comprising the compounds, and to provide methods of using the compounds and compositions for modulating (e.g., inhibiting or antagonizing) A2A and / or A1 receptor in a biological system.SOLUTION: An A2A and / or A1 antagonist compound of formula (I), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a composition comprising the compound are provided. Formula (I) (where, each symbol in the formula is as defined in the claims).SELECTED DRAWING: None

Description

[Technical Field]

[0001] Cross-references to related applications This application claims the benefit of priority from Korean Application No. 10-2019-0149117, filed November 19, 2019, which is incorporated herein by reference in its entirety. [Background technology]

[0002] Prologue Adenosine performs many biological functions through specific cellular receptors and is associated with a variety of biological activities, including immune function and inflammation.

[0003] There are four types of adenosine receptors (A1, A2A, A2B, and A3), which are coupled to heteromeric G proteins. A2A and A2B receptors are each connected to the Gs subtype of Gα protein. Stimulation of these receptors leads to a receptor format change, which induces shedding of activated Gs subunits by Gβρ dimers, resulting in the hydrolysis of intracellular adenosine triphosphate (ATP) and the generation of cyclic adenosine monophosphate (cAMP). cAMP synthesis activates protein kinase A (PKA) and phosphorylates other proteins. In T cells, type I PKA isoforms are primarily present around the T cell receptor (TCR). Increased cAMP levels, coupled to increased PKA activation, inhibits TCR signaling and contributes to the development of various diseases.

[0004] Cancer cells produce significantly more adenosine than normal cells. In cancer cells, high-density adenosine induces A2A receptor activation, thereby inhibiting the immune system and protecting the cells. In the tumor microenvironment, the adenosine concentration is 50 μM, which is higher than in normal cells and can lead to immunosuppression of T cell function and activation. A2A receptor antagonists can be used to modulate the inhibition of the immune system by cancer cells and induce anti-cancer effects.

[0005] A2A receptor antagonists are being developed for immuno-oncology treatment. A2A receptor antagonists can enhance antitumor immunity. A2A receptors are widely produced in leukocytes. Activation of A2A receptors on T cells reduces TCR-mediated cytotoxicity and cytokine production, suppresses T cell proliferation, and induces Treg cell proliferation. PD-1 or PD-L1 immune checkpoint inhibitors (e.g., antibody inhibitors) are widely used in immuno-oncology. However, statistically, only 20% to 30% of patients produce PD-1 / PD-L1, meaning many patients do not benefit from the efficacy of such inhibitors. Therapies involving immune checkpoint inhibitors in combination with A2A receptor antagonists are of interest.

[0006] Adenosine receptor regulation is important for the treatment of various indications: Modulating the activity of adenosine A1 receptors is important for the treatment of nervous system disorders, asthma, heart failure, and renal failure; antagonizing adenosine A2A receptors is important for the treatment of Parkinson's disease; modulating the activity of adenosine A2B receptors is important for the treatment of chronic lung disorders such as asthma, cancer, and immuno-oncology; and modulating adenosine A3 receptors is important for the treatment of asthma, chronic obstructive pulmonary disorder, glaucoma, cancer, and cerebral apoplexy. Summary of the Invention

[0007] overview The present disclosure provides adenosine receptor (e.g., A2A and / or A1 receptor) antagonist compounds and compositions comprising the compounds. The present disclosure also provides methods of using the compounds and compositions to modulate (e.g., inhibit or antagonize) A2A and / or A1 receptors in biological systems. The compounds and compositions are used in a variety of therapeutic applications, including the treatment of cancer, and immuno-oncology. The compounds and compositions are used in a variety of therapeutic applications, including the treatment of central nervous system or neurodegenerative diseases such as Parkinson's disease.

[0008] In a first aspect, the disclosure provides an A2A and / or A1 antagonist compound of formula (I), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000001.jpg2849(I) where Z 1 and Z 2 are independently selected from CR and N; Z 1 and Z 2 at least one of is N; R is H, (C1-C3) alkyl, or substituted (C1-C3) alkyl; Y 1 Or Y 4 is an independent CR 10 and N, and at least two of Y1 to Y4 are independently selected from CR 10 and; R 10 are each independently selected from H, (C-C)alkyl, substituted (C-C)alkyl, (C-C)alkenyl, substituted (C-C)alkenyl, (C-C)alkynyl, substituted (C-C)alkynyl, (C-C)haloalkyl, (C-C)alkoxy, substituted (C-C)alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol; R a and R b are each independently selected from H, F, (C-C) alkyl, and substituted (C-C) alkyl, or R a and R b are attached to a ring and together with the carbon atoms to which they are attached form a cyclopropyl or substituted cyclopropyl; A is phenyl, substituted phenyl, pyridyl, or substituted pyridyl.

[0009] In a second aspect, the present disclosure provides a pharmaceutical composition comprising a compound described herein, or a pharmaceutically acceptable salt thereof (e.g., a compound of Formula (I)-(XVIb)), and a pharmaceutically acceptable excipient.

[0010] In a third aspect, the present disclosure provides a method of modulating (e.g., inhibiting or antagonizing) adenosine A2A and / or A1 receptors, comprising contacting a sample or a cell or a biological system with an effective amount of a compound described herein (e.g., a compound of Formula (I)-(XVIb) or a pharmaceutically acceptable salt thereof).

[0011] Also provided are methods for treating cancer, comprising administering a therapeutically effective amount of an A2A and / or A1 receptor antagonist compound (e.g., as described herein) to a subject having cancer. In some embodiments, the method further comprises co-administering an additional active agent, such as an immune checkpoint inhibitor, to the subject. Also provided are methods for treating a central nervous system or neurodegenerative disease, comprising administering a therapeutically effective amount of an A2A and / or A1 receptor antagonist compound (e.g., a compound of Formula (I)-(XVIb), or a pharmaceutically acceptable salt thereof, as described herein) to a subject having or at risk of such a disease.

[0012] Detailed Description Adenosine receptor antagonist compounds As summarized above, the present disclosure provides A2A and / or A1 receptor antagonist compounds and compositions. These compounds can modulate adenosine A2A and / or A1 receptors in cells and biological systems of interest. These compounds have been used in a variety of therapeutic applications, including the treatment of cancer, immuno-oncology, and the treatment of central nervous system and neurodegenerative disorders such as Parkinson's disease.

[0013] The compounds of the present disclosure can be described as cyano-substituted fused pyrimidine compounds, comprising a core structure having a 2-amino-pyrimidine ring fused to a 5-membered heterocycle. The core structure itself can be further substituted with a benzonitrile substituent (e.g., 3-cyano-phenyl) or a derivative thereof (e.g., a cyano-pyridyl substituent). The fused 5-membered heterocycle can be further substituted with an optionally substituted benzyl group. In some cases, the compounds have a 9H-purin-2-amine or 1H-pyrazolo[3,4-d]pyrimidin-6-amine core structure that is further substituted as described herein.

[0014] In some embodiments, the compound is a benzonitrile-substituted fused pyrimidine compound of the formula: JPEG2025120182000002.jpg2841 or a derivative thereof (e.g., a derivative compound in which the 3-benzonitrile group is replaced with a cyano-pyridyl group). wherein Cy is a fused 5-membered heterocycle (e.g., a heteroaryl or heterocycloalkyl ring) containing at least one heteroatom selected from nitrogen, oxygen, and sulfur. R is an optionally substituted phenyl group. R 10 In some embodiments, R is independently selected from a halogen, a hydroxyl group, a thiol group, a carbonyl group, an amido group, a nitro group, an amino group, a substituted or unsubstituted (C-C) alkyl group, a substituted or unsubstituted (C-C) alkenyl group, a substituted or unsubstituted (C-C) alkynyl group, a substituted or unsubstituted (C-C) haloalkyl group, and a substituted or unsubstituted (C-C) aminoalkyl group or a substituted or unsubstituted (C-C) alkoxy group. 11 , R 12 , and / or R 13 is a phenyl substituted by a group, and R 10is one or more groups independently selected from hydrogen, halogen, hydroxyl group, thiol group, substituted or unsubstituted (C1-C5) alkyl group, substituted or unsubstituted (C2-C5) alkenyl group, substituted or unsubstituted (C2-C5) alkynyl group, substituted or unsubstituted (C1-C3) haloalkyl group, substituted or unsubstituted (C1-C5) alkoxy group, or cyano group.

[0015] In some embodiments, the aforementioned R 11 and R 13 are each independently hydrogen, halogen, a (C1-C5) alkyl group, or a (C1-C3) haloalkyl group, and 12 is hydrogen or an amino group, and each of the R 10 is independently hydrogen, halogen, a (C1-C5) alkyl group, or a cyano group. Further embodiments of the above formula are described herein.

[0016] Embodiments of the present disclosure include A2A and / or A1 antagonist compounds of formula (I), or solvates, hydrates, prodrugs, and / or stereoisomers thereof, or pharmaceutically acceptable salts thereof: JPEG2025120182000003.jpg2849(I) where: Z 1 and Z 2 are independently selected from CR and N; Z 1 and Z 2 at least one of is N; R is H, (C1-C3) alkyl, or substituted (C1-C3) alkyl; Y 1 Or Y 4 is an independent CR 10 and N are selected from Y 1 Or Y 4 At least two of the 10 and; R 10are each independently selected from H, (C-C)alkyl, substituted (C-C)alkyl, (C-C)alkenyl, substituted (C-C)alkenyl, (C-C)alkynyl, substituted (C-C)alkynyl, (C-C)haloalkyl, (C-C)alkoxy, substituted (C-C)alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol; R a and R b are each independently selected from H, F, (C-C) alkyl, and substituted (C-C) alkyl, or R a and R b are attached to a ring and together with the carbon atoms to which they are attached form a cyclopropyl or substituted cyclopropyl; A is phenyl, substituted phenyl, pyridyl, or substituted pyridyl.

[0017] In some embodiments of Formula (I), Y 1 Or Y 4 is CR 10 and N, where Y 1 Or Y 4 At least three of these independently 10 is.

[0018] In some embodiments of Formula (I), Z is a group such that the compound is a compound of Formula (Ia), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof. 1 is CR and Z 2 is N. JPEG2025120182000004.jpg3351(Ia)

[0019] In some embodiments of Formula (Ia), R is (C1-C3) alkyl. In some embodiments of Formula (Ia), R is H.

[0020] In some embodiments of Formula (I), Z is a group such that the compound is a compound of Formula (Ib), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof. 1 is CR and Z2 is N. JPEG2025120182000005.jpg3354(Ib)

[0021] In some embodiments of Formula (Ib), R is (C1-C3) alkyl. In some embodiments of Formula (Ib), R is H.

[0022] In some embodiments of Formula (I)-(Ib), A is phenyl. In some embodiments of Formula (I)-(Ib), A is substituted phenyl. In some embodiments of Formula (I)-(Ib), A is phenyl, or one, two, or three R 20 groups, and each R 20 are independently selected from (C-C) alkyl, substituted (C-C) alkyl, (C-C) alkenyl, substituted (C-C) alkenyl, (C-C) alkynyl, substituted (C-C) alkynyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol.

[0023] In some embodiments of Formula (I)-(Ib), A is pyridyl. In some embodiments of Formula (I)-(Ib), A is substituted pyridyl. A can be an optionally substituted pyridyl that is 2-pyridyl, 3-pyridyl, or 4-pyridyl. In some embodiments of Formula (I)-(Ib), A is pyridyl or one, two, or three R 20 pyridyl substituted with a group, and each R 20are independently selected from (C-C) alkyl, substituted (C-C) alkyl, (C-C) alkenyl, substituted (C-C) alkenyl, (C-C) alkynyl, substituted (C-C) alkynyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol.

[0024] In some embodiments of Formulas (I)-(Ib), R a and R b are H respectively.

[0025] In some embodiments of Formula (Ia)-(Ib), Y 1 Or Y 4 is CR 10 and N, where Y 1 Or Y 4 At least three of these independently 10 is.

[0026] In some embodiments of Formula (Ia)-(Ib), the compound includes compounds of Formula (IIa) or (IIb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000006.jpg42170 where R 1 Or R 9are independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) alkenyl, substituted (C-C) alkenyl, (C-C) alkynyl, substituted (C-C) alkynyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol.

[0027] In some embodiments of Formula (IIa)-(IIb), R 1 Or R 9 is independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, —NH, substituted amino, halogen, and hydroxyl. 1 Or R 9 are independently selected from H, NH2, F, CH3, and CF3.

[0028] In some embodiments of Formula (IIa)-(IIb), the compound includes compounds of Formula (IIIa) or (IIIb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000007.jpg46170 where R 21 and R 22 are independently H, (C1-C8) alkyl, substituted (C1-C8) alkyl, SO2R 30 , and C.O.R. 30 Selected from R 30 is (C1-C8) alkyl or substituted (C1-C8) alkyl.

[0029] In some embodiments of Formulas (IIIa)-(IIIb), R 21and R 22 are H respectively.

[0030] In some embodiments of Formula (IIIa)-(IIIb), R 5 , R 6 , R 8 and R 9 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. 5 , R 6 , R 8 and R 9 is independently selected from H, F, CH, and CF. In some embodiments of Formula (IIIa)-(IIIb), R 5 , R 6 , R 8 and R 9 are H respectively.

[0031] In some embodiments of Formula (IIIa)-(IIIb), R 2 Or R 4 are H and R respectively. 1 is selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. In some embodiments of Formula (IIIa)-(IIIb), R 1 is selected from H, F, CH3, and CF3.

[0032] In some embodiments of Formula (IIIa)-(IIIb), the compound is selected from the following: or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000008.jpg106170

[0033] In some embodiments of Formula (IIIa)-(IIIb), the compound is of Formula (IVa) or (IVb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000009.jpg41170

[0034] In some embodiments of Formula (IVa)-(IVb), R 6 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. 6 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, and (C1-C3) haloalkyl. In some embodiments of Formula (IVa)-(IVb), R 6 is CH3 or CF3.

[0035] In some embodiments of Formula (IVa)-(IVb), R 2 Or R 4 are H and R respectively. 1 is selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. In some embodiments of Formula (IVa)-(IVb), R 1 is selected from H, F, CH3, and CF3.

[0036] In some embodiments of Formula (IVa)-(IVb), the compound is selected from the following: or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000010.jpg143170

[0037] In some embodiments of Formula (IIIa)-(IIIb), the compound is of Formula (Va) or (Vb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000011.jpg41170

[0038] In some embodiments of Formulas (Va)-(Vb), R 5 and R 9 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. In some embodiments of Formula (Va)-(Vb), R 5 and R 9 is independently selected from H and halogen. In some embodiments of Formula (Va)-(Vb), R 5 is F. In some embodiments of Formulas (Va)-(Vb), R 9 is F. In some embodiments of Formulas (Va)-(Vb), R 5 and R 9 are each F. In some embodiments of Formulas (Va)-(Vb), R 5 is H and R 9 is F.

[0039] In some embodiments of Formula (Va)-(Vb), the compound is selected from the following: or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000012.jpg144170

[0040] In some embodiments of Formula (IIa)-(IIb), the compound is of Formula (VIa) or (VIb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000013.jpg41170

[0041] In some embodiments of Formulas (VIa)-(VIb), R 5 , R 6 and R 9 is independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, —NH, substituted amino, halogen, and hydroxyl. 6 is selected from H, (C-C) alkyl, substituted (C-C) alkyl, and (C-C) haloalkyl. In some embodiments of Formula (VIa)-(VIb), R 6 is CH3 or CF3. In some embodiments of Formula (VIa)-(VIb), R 6 is H.

[0042] In some embodiments of Formulas (VIa)-(VIb), R 5 and R 9 is independently selected from H and halogen. In some embodiments of Formula (VIa)-(VIb), R 5 is F. In some embodiments of Formulas (VIa)-(VIb), R 9 is F. In some embodiments of Formulas (VIa)-(VIb), R 5 and R 9 are each F. In some embodiments of Formulas (VIa)-(VIb), R 5 is H and R 9 is F.

[0043] In some embodiments of Formula (VIa)-(VIb), the compound is selected from the following: or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000014.jpg94170

[0044] In some embodiments of Formulas (IIa)-(VIb), R 1 Or R 4 is independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, —NH, substituted amino, halogen, and hydroxyl. 1 Or R 4 are independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, and halogen.

[0045] In some embodiments of Formulas (IIa)-(VIb), R 1 is H. In some embodiments of Formulas (IIa)-(VIb), R 1 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, and halogen. In some embodiments of Formula (IIa)-(VIb), R 1 is F, CH, or CF. In some embodiments of Formulas (IIa)-(VIb), R 2 , R 3 or R 4 is selected from (C-C) alkyl, substituted (C-C) alkyl, (C-C) haloalkyl, and halogen. In some embodiments of Formulas (IIa)-(VIb), R 2 , R 3 and R 4 are H respectively.

[0046] In some embodiments of Formula (Ia)-(Ib), the compound is of Formula (VIIa) or (VIIb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof. JPEG2025120182000015.jpg42170 where Y 1 Or Y 4 is CR 10and N are independently selected, and Y 1 Or Y 4 At least three of the 10 and; R 5 Or R 9 and each R 10 are independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) alkenyl, substituted (C-C) alkenyl, (C-C) alkynyl, substituted (C-C) alkynyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol.

[0047] In some embodiments of Formulas (VIIa)-(VIIb), Y 1 Or Y 4 One of them is N.

[0048] In some embodiments of Formulas (VIIa)-(VIIb), R 5 Or R 9 and each R 10 is independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, —NH, substituted amino, halogen, and hydroxyl. 5 Or R 9 and each R 10 are independently selected from H, NH2, F, CH3, and CF3.

[0049] In some embodiments of Formula (VIIa)-(VIIb), the compound is of Formula (VIIIa) or (VIIIb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000016.jpg46170 where, Y 1 Or Y 4 One of them is N; R 21 and R 22 is H, (C1-C8) alkyl, substituted (C1-C8) alkyl, SO2R 30 , and C.O.R. 30 are independently selected from R 30 is (C1-C8) alkyl or substituted (C1-C8) alkyl.

[0050] In some embodiments of Formulas (VIIIa)-(VIIIb), R 21 and R 22 are H respectively.

[0051] In some embodiments of Formulas (VIIIa)-(VIIIb), R 5 , R 6 , R 8 and R 9 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. 5 , R 6 , R 8 and R 9 is independently selected from H, F, CH, and CF. In some embodiments of Formula (VIIIa)-(VIIIb), R 5 , R 6 , R 8 and R 9 are H respectively.

[0052] In some embodiments of Formula (VIIIa)-(VIIIb), the compound is selected from the following: or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000017.jpg143170

[0053] In some embodiments of Formula (VIIIa)-(VIIIb), the compound is of Formula (IXa) or (IXb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000018.jpg41170

[0054] In some embodiments of Formulas (IXa)-(IXb), R 6 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. 6 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, and (C1-C3) haloalkyl. In some embodiments of Formula (IXa)-(IXb), R 6 is CH3 or CF3.

[0055] In some embodiments of Formula (IXa)-(IXb), the compound is selected from the following: or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000019.jpg219170 JPEG2025120182000020.jpg73170

[0056] In some embodiments of Formula (VIIIa)-(VIIIb), is Formula (Xa) or (Xb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof. JPEG2025120182000021.jpg41170

[0057] In some embodiments of Formulas (Xa)-(Xb), R 5 and R 9 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. In some embodiments of Formula (Xa)-(Xb), R 5 and R 9 is independently selected from H and halogen. In some embodiments of Formula (Xa)-(Xb), R 5 is F. In some embodiments of Formulas (Xa)-(Xb), R 9 is F. In some embodiments of Formulas (Xa)-(Xb), R 5 and R 9 are each F. In some embodiments of Formulas (Xa)-(Xb), R 5 is H and R 9 is F.

[0058] In some embodiments of Formula (Xa)-(Xb), the compound is selected from the following: or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000022.jpg213170 JPEG2025120182000023.jpg72170

[0059] In some embodiments of Formula (VIIa)-(VIIb), the compound is of Formula (XIa) or (XIb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000024.jpg41170

[0060] In some embodiments of Formulas (XIa)-(XIb), R 5 , R 6 and R 9is independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, —NH, substituted amino, halogen, and hydroxyl. 6 is selected from H, (C-C) alkyl, substituted (C-C) alkyl, and (C-C) haloalkyl. In some embodiments of Formula (XIa)-(XIb), R 6 is CH3 or CF3. In some embodiments of Formula (XIa)-(XIb), R 6 is H.

[0061] In some embodiments of Formulas (XIa)-(XIb), R 5 and R 9 is independently selected from H and halogen. In some embodiments of Formula (XIa)-(XIb), R 5 is F. In some embodiments of Formulas (XIa)-(XIb), R 9 is F. In some embodiments of Formulas (XIa)-(XIb), R 5 and R 9 are each F. In some embodiments of Formulas (XIa)-(XIb), R 5 is F and R 9 is H.

[0062] In some embodiments of Formula (XIa)-(XIb), the compound is selected from the following: or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000025.jpg124170

[0063] In some embodiments of Formula (VIIa)-(VIIb), the compound is of Formula (XIVa) or (XIVb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000026.jpg46170 where Y 4 is CR 4 or N; R 1 Or R 4 are independently selected from H, (C-C)alkyl, substituted (C-C)alkyl, (C-C)alkenyl, substituted (C-C)alkenyl, (C-C)alkynyl, substituted (C-C)alkynyl, (C-C)haloalkyl, (C-C)alkoxy, substituted (C-C)alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol; R 21 and R 22 is H, (C1-C8) alkyl, substituted (C1-C8) alkyl, SO2R 30 , and C.O.R. 30 are independently selected from R 30 is (C1-C8) alkyl, and substituted (C1-C8) alkyl.

[0064] In some embodiments of Formula (XIVa)-(XIVb), R 5 , R 6 , R 8 and R 9 are independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0065] In some embodiments of Formulas (XIVa)-(XIVb), R 21 and R 22 are H respectively.

[0066] In some embodiments of Formulas (XIVa)-(XIVb), R 2 Or R 4 are H and R respectively. 1is selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. In some embodiments of Formula (XIVa)-(XIVb), R 1 is selected from H, F, CH3, and CF3.

[0067] In some embodiments of Formula (XIVa)-(XIVb), the compound is of Formula (XVa) or (XVb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000027.jpg41170

[0068] In some embodiments of Formulas (XVa)-(XVb), R 5 and R 9 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. In some embodiments of Formula (XVa)-(XVb), R 5 and R 9 is independently selected from H and halogen. In some embodiments of Formula (XVa)-(XVb), R 5 is F. In some embodiments of Formulas (XVa)-(XVb), R 9 is F. In some embodiments of Formulas (XVa)-(XVb), R 5 and R 9 are each F. In some embodiments of Formulas (XVa)-(XVb), R 5 is H and R 9 is F.

[0069] In some embodiments of Formulas (XVa)-(XVb), R 1 teeth, In some embodiments of Formula (XVa)-(XVb), R is selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. 1 is selected from H, F, CH3, and CF3.

[0070] In some embodiments of Formulas (XVa)-(XVb), Y 4 In some embodiments of Formula (XVa)-(XVb), Y 4 is N.

[0071] In some embodiments of Formula (XVIa)-(XVIb), the compound is selected from the following: or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000028.jpg72170

[0072] In some embodiments of Formula (XIVa)-(XIVb), the compound is of Formula (XVIa) or (XVIb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000029.jpg41170

[0073] In some embodiments of Formulas (XVIa)-(XVIb), R 6 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. 6 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, and (C1-C3) haloalkyl. In some embodiments of Formula (XVIa)-(XVIb), R 6 is CH3 or CF3.

[0074] In some embodiments of Formulas (XVIa)-(XVIb), R 1 is selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl. 1 is selected from H, F, CH3, and CF3.

[0075] In some embodiments of Formulas (XVIa)-(XVIb), Y 4 In some embodiments of Formula (XVIa)-(XVIb), Y 4 is N.

[0076] In some embodiments of Formula (XVIa)-(XVIb), the compound is selected from the following, or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: JPEG2025120182000030.jpg75170

[0077] In some embodiments of Formula (I)-(Ib), the compound is of Formula (XIIa) or (XIIb). JPEG2025120182000031.jpg44170 where Y 5 Or Y 7 are each independently CR 20 or N and Y 5 Or Y 7 One of the is N;R 1 Or R 4 , R 8 , R 9 and each R 20are independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) alkenyl, substituted (C-C) alkenyl, (C-C) alkynyl, substituted (C-C) alkynyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol.

[0078] In some embodiments of Formulas (XIIa)-(XIIb), Y 5 is N. In some embodiments of Formulas (XIIa)-(XIIb), Y 6 is N. In some embodiments of Formulas (XIIa)-(XIIb), Y 7 is N.

[0079] In some embodiments of Formulas (XIIa)-(XIIb), R 8 , R 9 and each R 20 are independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, —NH2, substituted amino, halogen, and hydroxyl.

[0080] In some embodiments of Formulas (XIIa)-(XIIb), R 8 , R 9 and each R 20 are independently selected from H, NH2, F, CH3, and CF3.

[0081] In some embodiments of Formulas (XIIa)-(XIIb), R 1 Or R 4is independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, —NH, substituted amino, halogen, and hydroxyl. 1 Or R 4 are independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, and halogen.

[0082] In some embodiments of Formula (I)-(Ib), the compound is of Formula (XIIIa) or (XIIIb). JPEG2025120182000032.jpg39170 where Y 5 From Y 7 are each independently CR 20 or N and Y 5 From Y 7 One of them is N; R 8 , R 9 , each R 10 , and each R 20 are independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) alkenyl, substituted (C-C) alkenyl, (C-C) alkynyl, substituted (C-C) alkynyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol.

[0083] In some embodiments of Formulas (XIIIa)-(XIIIb), Y 5 is N. In some embodiments of Formulas (XIIIa)-(XIIIb), Y 6 is N. In some embodiments of Formulas (XIIIa)-(XIIIb), Y 7 is N.

[0084] In some embodiments of Formulas (XIIIa)-(XIIIb), R 8 , R 9 and each R 20 is independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, —NH, substituted amino, halogen, and hydroxyl. 8 , R 9 and each R 20 is independently selected from H, F, CH, and CF. In some embodiments of Formula (XIIIa)-(XIIIb), each R 10 are independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, —NH2, substituted amino, halogen, and hydroxyl.

[0085] In some embodiments of Formulas (XIIIa)-(XIIIb), Y 1 is CR 10 is.

[0086] In some embodiments of Formulas (XIIIa)-(XIIIb), R 10 is H. In some embodiments of Formulas (XIIIa)-(XIIIb), R 10 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, and halogen. In some embodiments of Formula (XIIIa)-(XIIIb), R 10 are H, F, CH3, and CF3.

[0087] In some embodiments of Formulas (XIIIa)-(XIIIb), Y 1 Or Y 4 are CR 10 is.

[0088] In some embodiments of Formulas (XIIIa)-(XIIIb), each R 10 are independently selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, and halogen.

[0089] In some embodiments of Formulas (XIIIa)-(XIIIb), Y 1 Or Y 4 In some embodiments of Formula (XIIIa)-(XIIIb), Y 1 Or Y 4 and only one of Y is N. In some embodiments of Formulas (XIIIa)-(XIIIb), Y 1 is N. In some embodiments of Formulas (XIIIa)-(XIIIb), Y 2 is N. In some embodiments of Formulas (XIIIa)-(XIIIb), Y 3 is N. In some embodiments of Formulas (XIIIa)-(XIIIb), Y 4 is N.

[0090] In some of the above embodiments, the compound is of Formula (Ia) to (XVIa) (e.g., a purine compound), or a salt thereof (e.g., a pharmaceutically acceptable salt). In some of the above embodiments, the compound is of Formula (Ib) to (XVIb) (e.g., a pyrazolopyrimidine compound), or a salt thereof (e.g., a pharmaceutically acceptable salt).

[0091] In some embodiments of Formula (I), the compound is selected from: 3-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo-[3,4-d]-pyrimidin-4-yl)-benzonitrile; 3-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo-[3,4-d]-pyrimidin-4-yl)-2-methylbenzonitrile; 3-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo-[3,4-d]-pyrimidin-4-yl)-2-fluorobenzonitrile; 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)-benzonitrile; 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile; 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)-2-methylbenzonitrile; 3-(2-amino-9-(4-amino-2,6-difluorobenzyl)-9H-purin-6-yl)-benzonitrile; 3-(2-amino-9-(4-amino-2,6-difluorobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile; 3-(2-amino-9-(4-amino-2-fluorobenzyl)-9H-purin-6-yl)-benzonitrile; 3-(2-amino-9-(4-amino-2-fluorobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile; 3-(2-amino-9-(4-amino-3-methylbenzyl)-9H-purin-6-yl)-benzonitrile; 3-(2-amino-9-(4-amino-3-methylbenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile; 3-(2-amino-9-(2,6-difluorobenzyl)-9H-purin-6-yl)benzonitrile; 3-(2-amino-9-(2,6-difluorobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile; and 3-(2-amino-9-(2,6-difluorobenzyl)-9H-purin-6-yl)-2-methylbenzonitrile.

[0092] All salts, and / or solvates, hydrates, prodrugs, and / or stereoisomers of the compounds described herein, e.g., compounds of Formula (I)-(XVIb), or compounds shown in Table 1, are included in the present disclosure. Thus, any of the compounds described herein may be referred to as a compound of Formula (I)-(XVIb), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0093] In some embodiments, the compound is represented by the structure of one of the compounds in Table 1, or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a salt thereof (e.g., a pharmaceutically acceptable salt). In some embodiments, the compound is represented by the structure of one of the compounds in Table 1 that is Formula (Ia) (e.g., a purine compound) or a salt thereof (e.g., a pharmaceutically acceptable salt). In some embodiments, the compound is represented by the structure of one of the compounds in Table 1 that is Formula (Ib) (e.g., a pyrazolo-pyrimidine compound) or a salt thereof (e.g., a pharmaceutically acceptable salt). The present disclosure is meant to include any one of the compounds in Table 1, or a salt thereof, and / or a solvate, hydrate, prodrug thereof, single stereoisomer, mixture of stereoisomers, and / or isotopically labeled form thereof. For example, solvates, hydrates, prodrugs and / or salts of stereoisomeric forms of any of the compounds described herein (e.g., of Formulas (I)-(XVIb)) or shown in Table 1 are meant to be included in the disclosure herein. JPEG2025120182000033.jpg250165 JPEG2025120182000034.jpg251167 JPEG2025120182000035.jpg245170 JPEG2025120182000036.jpg244170 JPEG2025120182000037.jpg225170 JPEG2025120182000038.jpg81170Isotopically labeled analogues

[0094] The present disclosure also includes isotopically labeled compounds identical to the compounds described herein, except that one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number normally found in nature ("isotopologues"). The compounds of the present disclosure may also contain unnatural proportions of atomic isotopes in one or more atoms that constitute such compounds. Examples of isotopes that can be incorporated into the compounds described herein include: 2 H("D"), 3 H, 13 C. 14 C. 15 N, 18 O. 17 O. 31 P, 32 P, 35 S, 18 F, and 36 Included are isotopes of each of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as Cl. For example, the compounds described herein may have one or more H atoms replaced with deuterium.

[0095] Generally, a reference or depiction of a particular element, such as hydrogen or H, is meant to include all isotopes of that element. For example, if an R group is defined to include hydrogen or H, it also includes deuterium and tritium. Thus, tritium, 14 C. 32 P and 35 Compounds containing radioisotopes such as S are within the scope of the present technology. Procedures for incorporating such labels into the compounds of the present technology will be readily apparent to those of skill in the art based on the disclosure herein.

[0096] Unless otherwise stated, the compounds described herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms, for example, the replacement of hydrogen by deuterium or tritium, or 13 C or 14 Compounds having the present structures except for the replacement of a carbon with a C-rich carbon are within the scope of this disclosure.

[0097] In some embodiments, 3 H and 14 Certain isotopically labeled compounds, such as those labeled with C, may be useful in compound and / or substrate tissue distribution assays. 3 H) and carbon-14 ( 14 C) Isotopes may be particularly preferred because they are easy to prepare and detect. Furthermore, substitution with heavier isotopes, such as deuterium, may offer certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced required dosage, and may therefore be preferred in some circumstances. Isotopically labeled compounds can generally be prepared by procedures similar to those disclosed herein, e.g., in the Examples section, by substituting isotopically labeled reagents for non-isotopically labeled reagents.

[0098] In some embodiments, the compounds disclosed herein are deuterated analogs of any of the compounds or salts thereof, as described herein. Deuterated analogs of compounds of Formulas (I)-(XVIb) are compounds in which one or more hydrogen atoms are replaced with deuterium. In some embodiments, deuterated analogs are compounds in which one or more hydrogen atoms are replaced with deuterium. a , R b , or R 1 Or R 20 A compound of formula (I) comprising a group:

[0099] Deuterium-substituted compounds are synthesized using a variety of methods, such as those described in: Dean, Dennis C.; Editor. Recent Advances in the Synthesis and Applications of Radiolabeled Compounds for Drug Discovery and Development. [In: Curr. Pharmac. Des., 2000;6(10)] 2000, 110 pp; George W.; Varma, Rajender S. Synthesis of Radiolabeled Compounds via Organometallic Intermediates, Tetrahedron, 1989, 45(21), 6601-21; and Evans, E. Anthony. Synthesis of Radiolabeled Compounds, J. Radioanal. Chem., 1981, 64(1-2), 9-32.

[0100] Deuterated starting materials are readily available and are amenable to the synthetic methods described herein to provide for the synthesis of deuterated compounds. Many deuterated reagents and building blocks are commercially available from chemical vendors such as Aldrich Chemical Co. Fluorinated Analogues

[0101] In some embodiments, the compounds disclosed herein are fluorinated analogs of any compound or salt thereof as described herein. Fluorinated analogs of compounds of Formula (I)-(XVIb) are compounds in which one or more hydrogen atoms or substituents are replaced with fluorine atoms. In some embodiments, fluorinated analogs are fluorinated R, R a , R b , or R 1 Or R 20

[0023] The compound of formula (I) comprises a group. In some embodiments of the fluorinated analog of the compound of formula (I), a hydrogen atom of an aliphatic or aromatic C-H bond is replaced with a fluorine atom. In some embodiments of the fluorinated analog of the compound of formula (I), at least one hydrogen of an optionally substituted aryl (e.g., phenyl) or an optionally substituted heteroaryl (e.g., pyridyl) is replaced with a fluorine atom. In some embodiments of the fluorinated analog of the compound of formula (I), a hydroxyl substituent (-OH) or an amino substituent (-NH2) is replaced with a fluorine atom. In some embodiments of the fluorinated analog of the compound, the compound comprises one or more substituents independently selected from -F, -CF3, -CF2CF3, -CHF2, -OCF3, -OCHF2, and -OCF2CF3. Isomers

[0102] As used herein, the term "compound" is meant to include all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted.

[0103] The compounds described herein may have asymmetric centers, geometric centers (e.g., double bonds), or both. All chiral, diastereomeric, racemic forms, and all geometric isomeric forms of a structure are intended unless a particular stereochemistry or isomeric form is specifically indicated. In some embodiments, the compounds described herein have one or more chiral centers. Unless the absolute stereochemistry is explicitly indicated, it is understood that each chiral center may independently be in the R or S configuration or a mixture thereof. Thus, the compounds described herein include optical isomers enriched or resolved at any or all asymmetric atoms, as is clear from the depiction. Racemic mixtures of R- and S-enantiomers, and enantio-enriched stereoisomeric mixtures containing R- and S-enantiomers, as well as individual optical isomers, can be isolated or synthesized to be substantially free of their enantiomeric or diastereomeric partners, and all of these stereoisomers are within the scope of the present technology.

[0104] Compounds of the present disclosure containing an asymmetrically substituted atom can be isolated in optically active or racemic forms. It is well known in the art how to prepare optically active forms, such as by resolution of racemic forms, by synthesis from optically active starting materials, or by use of chiral auxiliaries.

[0105] Geometric isomers resulting from the arrangement of substituents around a carbon-carbon double bond or the arrangement of substituents around a cycloalkyl or heterocyclic ring can also exist in the compounds of the present disclosure. Geometric isomers of olefins, C=N double bonds, or other types of double bonds can exist in the compounds described herein, and all such stable isomers are included in the present disclosure. Specifically, cis and trans geometric isomers of the compounds of the present disclosure can also exist and can be isolated as a mixture of isomers or as separate isomeric forms.

[0106] The compounds of the present disclosure also include tautomeric forms. Tautomers occur when a single bond swaps with an adjacent double bond, resulting in the concomitant migration of a proton. Tautomers include prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of protic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactimum pairs, amide-imidic acid pairs, enamine-imine pairs, and cyclic forms in which protons can occupy more than one position on a heterocyclic ring system, such as 1H- and 3H-imidazole, 1H-, 2H-, and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. Tautomeric forms may be in equilibrium or sterically locked into one form by appropriate substitution. Salts and other forms

[0107] In some embodiments, the compounds described herein are present in the form of a salt. In some embodiments, the compounds are provided in the form of a pharmaceutically acceptable salt.

[0108] The compounds contained in the present compositions that are basic in nature can form a wide variety of salts with various inorganic and organic acids. The acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, i.e., salts with pharmacologically acceptable anions, including, but not limited to, chloride.

[0109] Compounds containing amine functional groups or nitrogen-containing heteroaryl groups may be basic in nature and may react with various inorganic and organic acids to form corresponding salts. The compounds may be used in the form of pharmaceutically acceptable salts derived from inorganic or organic acids. In some embodiments, the pharmaceutically acceptable salts may be salts derived from hydrochloric acid (i.e., the hydrochloride salts of the compounds described herein), etc.

[0110] Pharmaceutically acceptable salts of the compounds of the present disclosure can be prepared by dissolving the compound in a water-miscible organic solvent such as acetone, methanol, ethanol, or acetonitrile, adding an excess amount of an aqueous organic or inorganic acid, and then precipitating or crystallizing. Additional salts can then be obtained by evaporating the solvent or excess acid from the mixture and drying it, or by filtering the extracted salt to produce the salt.

[0111] Other examples of salts include the anion of the disclosed compound combined with a suitable cation. For therapeutic use, the salts of the disclosed compounds may be pharmaceutically acceptable. However, non-pharmaceutical acceptable acid and base salts may also find use, for example, in the preparation or purification of pharmaceutically acceptable compounds.

[0112] Compounds included in the present compositions that are acidic in nature are capable of forming base salts with various pharmacologically acceptable cations. Examples of such salts include alkali metal or alkaline earth metal salts.

[0113] Compounds containing a basic or acidic moiety can also form pharmaceutically acceptable salts with various amino acids. The compounds of the present disclosure can contain both acidic and basic groups, for example, one amino group and one carboxylic acid group. In such cases, the compounds can exist as acid addition salts, zwitterions, or base salts.

[0114] The compounds described herein may exist in various forms, including crystalline, powdered, and amorphous forms of the compounds, such as pharmaceutically acceptable salts, including polymorphs, pseudopolymorphs, solvates, hydrates, nonsolvated polymorphs (including anhydrates), conformational polymorphs, and amorphous forms of the compounds, and mixtures thereof. It is understood that any one salt form of the subject compounds may exist in crystalline, powdered, or amorphous form.

[0115] The compounds described herein may exist as solvates, particularly hydrates, and all such solvates and hydrates are intended unless otherwise specified. The term "solvate" can refer to a compound of the present disclosure that contains stoichiometric or non-stoichiometric solvents combined by non-covalent intermolecular forces or salts thereof. Hydrates may be formed during the preparation of the compound or compositions containing the compound, or hydrates may form over time due to the hygroscopic nature of the compound. The compounds of the present technology may also exist as organic solvates, including, among others, dimethylformamide (DMF), ether, and alcohol solvates. The identification and preparation of specific solvates is within the skill of a synthetic organic or medicinal chemist.

[0116] In some embodiments, the compounds described herein exist in the form of a solvate. In some embodiments, the compounds described herein exist in the form of a hydrate when the solvent component of the solvate is water. Prodrug

[0117] In some embodiments, the compounds described herein exist in the form of prodrugs. Any convenient prodrug form of the subject compounds can be prepared, for example, according to the strategies and methods described by Rauchio et al. ("Prodrugs: Design and Clinical Applications," Nature Reviews Drug Discovery 7, 255-270 (February 2008)). compound synthesis

[0118] The compounds of the present disclosure can be synthesized according to standard methods known in the art [see, for example, "Organic Chemistry," Morrison and Boyd, 6th Edition, Prentice-Hall (1992)]. Some compounds and / or intermediates of the present disclosure are commercially available, known in the literature, or readily available to those of ordinary skill in the art using standard procedures. Some compounds of the present disclosure can be synthesized using the schemes, examples, or intermediates described herein. Where the synthesis of a compound, its intermediate, or variant is not fully described, one of ordinary skill in the art will recognize that reaction times, number of equivalents of reagents, and / or temperatures can be varied from the reactions described herein to prepare the compound or intermediate presented, or variants thereof, and that different workup and / or purification techniques may be necessary or desirable to prepare such compounds, intermediates, or variants.

[0119] The synthesized compounds may be verified for their appropriate structure by methods known to those skilled in the art, for example, by nuclear magnetic resonance (NMR) spectroscopy and / or mass spectrometry. Pharmaceutical Composition

[0120] The compounds of the present disclosure can be formulated into pharmaceutical compositions that can include one or more exemplary A2A and / or A1 receptor antagonist compounds (e.g., as described herein) and at least one excipient (e.g., a pharmaceutically acceptable excipient).

[0121] The compounds described herein may find use in pharmaceutical compositions for administration to subjects in need thereof in a variety of therapeutic applications in which inhibition or antagonism of the activity of A2A and / or A1 receptors is desirable.

[0122] Thus, in a second aspect, the present disclosure provides pharmaceutical compositions comprising at least one compound described herein, a pharmaceutically acceptable salt thereof, or a prodrug thereof, and at least one pharmaceutically acceptable excipient. The phrase "pharmaceutically acceptable excipient" refers to any ingredient other than the compounds of the present invention described herein (e.g., a vehicle capable of suspending or dissolving an active compound, or any other convenient pharmaceutically acceptable carrier, excipient, or additive) that is substantially non-toxic and non-inflammatory in patients. Excipients may include, for example, anti-adherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film-forming agents or coatings, flavors, fragrances, glidants (gliding agents), lubricants, preservatives, printing inks, adsorbents, dispensing or dispersing agents, sweeteners, and water of hydration. In some embodiments, the pharmaceutical compositions comprise a therapeutically effective amount of a compound described herein, a pharmaceutically acceptable salt thereof, or a prodrug thereof.

[0123] The pharmaceutical compositions can be formulated according to any convenient method and can be prepared in a variety of forms for oral administration, such as tablets, pills, powders, nanoparticles, capsules, syrups, suspensions, emulsions and microemulsions, or in forms for parenteral administration, such as preparations for intramuscular, intravenous or subcutaneous administration.

[0124] In certain instances, the pharmaceutical composition may include a pharmaceutically acceptable carrier, excipient, or additive. The pharmaceutical composition may be prepared as a pharmaceutical product by conventional methods, and may be prepared as various oral pharmaceutical products such as tablets, pills, powders, capsules, syrups, emulsions, microemulsions, etc., or as parenteral drugs such as intramuscular injections, intravenous injections, and subcutaneous injections.

[0125] When the pharmaceutical composition is prepared in the form of an oral medication, examples of the additives or carriers used may include cellulose, calcium silicate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, magnesium stearate, calcium stearate, gelatin, talc, surfactants, suspensions, emulsifiers, diluents, etc. When the pharmaceutical composition of the present disclosure is prepared in the form of an injection, the additives or carriers may include water, saline, aqueous glucose solution, similar sugar-soluble solutions, alcohols, glycols, ethers (e.g., polyethylene glycol 400), oils, fatty acids, fatty acid esters, glycerides, surfactants, suspensions, emulsifiers, etc.

[0126] In some embodiments, the pharmaceutical composition is formulated for parenteral administration to a subject in need thereof. In some parenteral embodiments, the pharmaceutical composition is formulated for intravenous administration to a subject in need thereof. In some parenteral embodiments, the pharmaceutical composition is formulated for subcutaneous administration to a subject in need thereof. Methods of Modulating A2A and / or A1 Receptors

[0127] Aspects of the present disclosure include methods for modulating the activity of A2A and / or A1 receptors in a biological system or sample. In some embodiments, modulating the activity of A2A and / or A1 receptors refers to inhibiting the A2A and / or A1 receptors in the sample. In some embodiments, modulating the activity of A2A and / or A1 receptors refers to antagonizing the activity of A2A and / or A1 receptors in a cell or biological system.

[0128] In some embodiments, the method includes contacting the sample with a compound of the present disclosure capable of modulating A2A and / or A1 receptor activity. In some embodiments, the method includes contacting a cell or biological system with a compound of the present disclosure capable of modulating A2A and / or A1 receptor activity.

[0129] The present disclosure provides compounds having A2A and / or A1 receptor modulating activity, e.g., A2A and / or A1 receptor inhibitory and / or antagonistic activity. In some embodiments, the compounds have A2A receptor inhibitory and / or antagonistic activity. In some embodiments, the compounds have A2A and A1 receptor inhibitory and / or antagonistic activity. In some embodiments, the compounds have A1 receptor inhibitory and / or antagonistic activity. The ability of a compound to modulate A2A and / or A1 receptor activity can be characterized using various assays, for example, A2A and / or A1 receptor binding assays and / or A2A receptor functional assays. For example, the experimental section describes A2A receptor binding assays. Exemplary compounds of the present disclosure were evaluated to determine K i IC values were determined to demonstrate that compounds of the present disclosure can provide specific binding to the human A2A receptor in an assay system. Additionally, the experimental section describes an A2A functional assay involving monitoring cAMP signals generated in a human recombinant A2A receptor stable cell line. Exemplary compounds of the present disclosure were evaluated and IC 50 values were obtained, indicating that the compounds are capable of antagonizing the human A2A receptor in the assay system.

[0130] Embodiments of the present disclosure include methods of modulating (e.g., inhibiting or antagonizing) A2A and / or A1 receptors using the compounds described herein. Such methods may include modulating A2A and / or A1 receptors in a biological system by contacting such a system with a compound (e.g., a compound having a structure according to Formulas (I)-(XVIb) or a compound in Table 1). Biological systems may include, but are not limited to, cells, tissues, organs, bodily fluids, organisms, non-mammalian subjects, and mammalian subjects (e.g., humans). Biological systems may include subjects (e.g., human subjects).

[0131] In some embodiments, the method for inhibiting or antagonizing A2A and / or A1 receptors comprises contacting a biological system or sample containing A2A and / or A1 receptors with an effective amount of any compound described herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein for inhibiting or antagonizing A2A and / or A1 receptors. In certain embodiments, the biological system or sample is in vitro. In other embodiments, the biological system or sample is in vivo. In some embodiments, the sample is a cell sample. In some embodiments, the A2A and / or A1 receptors are bound to a cell membrane.

[0132] Compounds of the present disclosure may antagonize the A2A receptor, e.g., as assessed by a human A2A functional assay, e.g., as described in Example 3. A2A receptor antagonist compounds by such methods have an IC50 of 10 μM or less, e.g., 2 μM or less, 1 μM or less, 500 nM or less, 300 nM or less, 100 nM or less, 30 nM or less, or 10 nM or less. 50 A2A receptor antagonist compounds by such methods may have a K value (e.g., as assessed by the cAMP assay of Example 3). The biological system may include a subject (e.g., a human subject). A2A receptor antagonist compounds by such methods may have a K value of 1 μM or less, e.g., 500 nM or less, 300 nM or less, 100 nM or less, 30 nM or less, 10 nM or less, 3 nM or less, or 1 nM or less. i The binding assay may have a value (e.g., as assessed by the binding assay of Example 2).

[0133] In some embodiments, the compound is an A2A receptor antagonist that exhibits a desirable level of inhibitory activity at the A1 receptor. In some embodiments, the compound is a dual antagonist of the A2A receptor and the A1 receptor. In some embodiments, the desirable level of A1 receptor inhibitory activity is a K1 at the human A1 receptor in a cellular model (e.g., as assessed in Example 4.1). iThe term "compounds" refers to compounds having a value of 1 mM or less, for example, 300 μM or less, 100 μM or less, 30 μM or less, 10 μM or less, 3 μM or less, or 1 μM or less. In some embodiments, compounds of the present disclosure exhibit inhibitory activity against A2A receptors and A1 receptors. Example 4, Table 5 in the Experimental Section provides A2A and A1 receptor inhibition data showing that compounds of the present disclosure can have potent activity as dual antagonists. In such cases, the compounds can be used to treat CNS and neurodegenerative diseases (e.g., compounds of Formulas (Ib) to (XVIb)). Treatment indications

[0134] Aspects of the present disclosure include methods of treating a subject for a therapeutic indication of interest using the compounds and / or compositions disclosed herein. The term "therapeutic indication" refers to a symptom, condition, disorder, or disease that may be alleviated, stabilized, ameliorated, cured, or otherwise addressed in a subject by some form of treatment or other therapeutic intervention (e.g., administration of an A2A and / or A1 receptor antagonist). Therapeutic indications related to modulation of the biological activity and / or dysfunction of A2A and / or A1 receptors are referred to herein as "adenosine receptor-related indications." In some embodiments, the methods of the present disclosure may include treating an adenosine receptor-related indication by administering an effective amount of a compound and / or composition disclosed herein (e.g., an A2A and / or A1 receptor antagonist compound) to a subject.

[0135] The term "treat" or "treatment" refers to inhibiting a disease, e.g., inhibiting a disease, condition, or disorder in a subject experiencing a symptom or condition of the disease, condition, or disorder, i.e., preventing further recurrence of the symptom and / or condition, or ameliorating a disease, e.g., ameliorating a disease, condition, or disorder in a subject experiencing a symptom or condition of the disease, condition, or disorder, i.e., reversing the symptom and / or condition, e.g., reducing the severity of the disease. In the context of this disclosure, in relation to any of the other conditions listed below, the terms "treat," "treatment," and the like refer to alleviating or alleviating at least one symptom associated with such condition, or to slowing or reversing the progression or expected progression of such condition.

[0136] The term "prevention" or "preventing" means preventing a disease, e.g., preventing a disease, condition, or disorder, in an entity that has not experienced or exhibited any pathology or symptoms of the disease, even though the entity may be predisposed to the disease, condition, or disorder.

[0137] The terms "individual," "patient," "subject," and "entity" are used interchangeably and refer to a human or non-human animal in need of treatment for a disease, more specifically a mammal such as a human, non-human primate, mouse, dog, cat, horse, cow, rabbit, rat, or other mammal. Cancer and Immuno-Oncology

[0138] The present disclosure provides methods for treating or preventing cancer in a subject using the subject compounds as therapeutic agents, and compositions containing the compounds. Any cancer for which an A2A receptor antagonist is believed by one skilled in the art to be useful is contemplated as a cancer treatable by this embodiment, either as monotherapy or in combination with other therapeutic agents discussed below.

[0139] "Cancer," a disease targeted for prevention or treatment by the aforementioned pharmaceutical compositions, refers collectively to diseases caused by cells with aggressive characteristics, such as the tendency for cells to divide and grow despite normal growth limitations, invasive characteristics that allow cells to invade surrounding tissues, and metastatic characteristics that allow cells to spread to other areas of the body. In some embodiments, the cancer is a solid tumor cancer. Cancers targeted for prevention or treatment using the compounds and pharmaceutical compositions of the present disclosure include, but are not limited to, lung cancer, breast cancer, prostate cancer, ovarian cancer, solenoma, cervical cancer, bladder cancer, head and neck cancer, renal cell carcinoma, esophageal cancer, pancreatic cancer, brain cancer, liver cancer, leukemia, lymphoma, melanoma, multiple myeloma, Ewing's sarcoma, osteosarcoma, colorectal neoplasm, bile duct cancer, choriocarcinoma, oral cancer, neuroblastoma, skin cancer, testicular cancer, stromal tumor, germ cell tumor, and thyroid cancer.

[0140] In some embodiments, the cancer is kidney, breast, lung, or liver cancer. In some embodiments, the cancer is a solid tumor cancer. In some embodiments, the liver cancer is hepatocellular carcinoma (HCC). In some embodiments, the lung cancer is non-small cell lung cancer (NSCLC). In some embodiments, the cancer is a solid tumor cancer.

[0141] In some embodiments, the method further comprises identifying a subject as suffering from or having cancer.

[0142] The compounds of the present disclosure can be administered alone or in combination with one or more additional agents described herein.Accordingly, an embodiment of the method includes co-administering an effective amount of a compound or composition of the present disclosure and an effective amount of an additional active agent to a subject.The compound and the additional active agent can be administered simultaneously or sequentially.

[0143] In some embodiments of the method, the additional active agent is an anti-cancer agent selected from an anti-angiogenic agent, an anti-inflammatory agent, an immune checkpoint inhibitor, a poly ADP-ribose polymerase (PARP) inhibitor, a chemotherapeutic agent, and an immunological anti-cancer agent.

[0144] Given the immunosuppressive role of adenosine, administration of an A2A receptor antagonist of the present disclosure (e.g., a compound of Formula (Ia) to (XVIa)) can enhance the efficacy of immunotherapy, such as immune checkpoint inhibitor therapy. In some embodiments, the additional active agent is an immune checkpoint inhibitor selected from a CTLA-4 inhibitor, a PD-1 inhibitor, and a PD-L1 inhibitor.

[0145] In some embodiments, the immune checkpoint inhibitor is an antibody or antibody fragment. In some embodiments, the additional therapeutic agent is an anti-PD-1 antibody. In other embodiments, the additional therapeutic agent is an anti-PD-L1 antibody. In some embodiments, the additional therapeutic agent is selected from pembrolizumab, nivolumab, atezolizumab, durvalumab, and avelumab. inflammatory diseases

[0146] The present disclosure provides methods for treating or preventing inflammatory diseases in a subject using the subject compounds as therapeutic agents, and compositions containing the compounds. Any inflammatory disease for which an A2A receptor antagonist is believed by one of skill in the art to be useful is contemplated as a disease treatable using the compounds of the present disclosure, either as monotherapy or in combination with other therapeutic agents (e.g., as described herein).

[0147] In some embodiments, the method includes administering a therapeutically effective amount of an A2A and / or A1 receptor antagonist compound (e.g., as described herein) to a subject having an inflammatory disease. In some embodiments, the inflammatory disease is a chronic inflammatory disease, such as rheumatoid arthritis (RA). In some embodiments, the inflammatory disease is an acute inflammatory disease.

[0148] In some embodiments, the method further comprises identifying a subject suffering from or at risk for an inflammatory disease.

[0149] In some embodiments, the method further comprises identifying an underlying disease or condition associated with the inflammatory disease. CNS and Neurodegenerative Diseases

[0150] The present disclosure provides methods for treating or preventing central nervous system (CNS) or neurodegenerative diseases or disorders in a subject using a compound of the present disclosure as a therapeutic agent or a pharmaceutical composition containing the compound. In some embodiments, the CNS disease is referred to as a neurodegenerative disease or disorder of the CNS. CNS and neurodegenerative diseases that can be prevented or treated using the compounds and pharmaceutical compositions of the present disclosure include, but are not limited to, Parkinson's disease (PD), Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), multiple sclerosis, Huntington's disease (HD), depression, schizophrenia, and epilepsy.

[0151] In some embodiments, the method is for treating or preventing Parkinson's disease, comprising administering to a subject having or at risk of Parkinson's disease an effective amount of a compound of the present disclosure. In some cases, the compound has dual inhibitory activity at both the A2A receptor and the A1 receptor (e.g., compounds of Formulas (Ib to XVIb)).

[0152] In some embodiments, the method further comprises identifying a subject suffering from or at risk for a CNS or neurodegenerative disease.

[0153] In some embodiments, the method further comprises identifying an underlying disease or condition associated with the CNS or neurodegenerative disease.

[0154] The amount of the aforementioned pharmaceutical composition administered is an amount effective for treating or preventing a disease in an entity or patient, and can be administered orally or parenterally depending on the purpose. For oral administration, the amount administered based on the active ingredient is 0.01-1,000 mg per kg of body weight, more specifically 0.1-300 mg per kg. For parenteral administration, 0.01-100 mg of the active ingredient per kg of body weight per day, more specifically 0.1-50 mg, is administered once or several times. The amount administered to a particular entity or patient can be determined based on many relevant factors, including the patient's weight, age, sex, health condition, diet, administration time, administration method, and severity of the disease. It is understood that dosages can be increased or decreased as appropriate by a specialist, and the aforementioned dosages are in no way intended to limit the scope of this disclosure. A physician or veterinarian with ordinary skill in the relevant art can determine and formulate the effective amount of the pharmaceutical composition. For example, a physician or veterinarian may start with an amount of a compound used in a pharmaceutical composition under the present disclosure that is less than the amount required to achieve the desired therapeutic effect, and gradually increase the amount administered until the desired effect is achieved.

[0155] The compounds and compositions of the present disclosure can be administered alone, in combination with a compound according to another example of the present disclosure, or in simultaneous, separate or sequential co-administration with at least one other therapeutic agent, such as another active pharmaceutical ingredient described herein.

[0156] In certain examples, pharmaceutical compositions of the present disclosure include within their scope pharmaceutical compositions containing an effective therapeutic amount of at least one compound according to the specific examples as an active ingredient, or in combination with a pharmaceutical carrier. Optionally, a compound according to an embodiment of the present disclosure can be administered independently, in combination with another compound according to the specific examples, or simultaneously with one or more other therapeutic agents, such as an anti-cancer agent (e.g., as described herein), or in simultaneous, separate, or sequential combination with an active pharmaceutical ingredient. definition

[0157] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.

[0158] It is understood that the definitions provided herein are not intended to be mutually exclusive, and thus some chemical moieties may be included in more than one term definition.

[0159] "C x -C y The term "(C alkyl)" when used in combination with a chemical moiety such as alkyl, alkenyl, or alkynyl, is meant to include groups containing x to y carbons in the chain. For example, the term "C alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups containing 1-6 carbons. In some embodiments, "(C x -C y The term "(C ) alkylene" refers to a substituted or unsubstituted alkylene chain having x to y carbon atoms in the alkylene chain. For example, "(C x -C y ) Alkylene may be selected from methylene, ethylene, propylene, butylene, pentylene, and hexylene, any one of which is optionally substituted.

[0160] The term "alkyl" refers to an unbranched or branched saturated hydrocarbon chain. In some embodiments, alkyl, as used herein, refers to an alkyl group having 1 to 20 carbon atoms (C-C 20 ) alkyl), 1 to 10 carbon atoms ((C1-C 10) alkyl), 1 to 8 carbon atoms ((C1-C8) alkyl), 1-6 carbon atoms ((C1-C6) alkyl), 1-5 carbon atoms ((C1-C5) alkyl), or 1-3 carbon atoms ((C1-C3) alkyl). Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, 2-pentyl, isopentyl, neopentyl, n-hexyl, 2-hexyl, 3-hexyl, and 3-methylpentyl. When an alkyl residue having a specific number of carbons is named, all geometric isomers having that number of carbons can be included. For example, "butyl" can include n-butyl, sec-butyl, isobutyl, and t-butyl, and "propyl" can include n-propyl and isopropyl. Unless otherwise stated herein, alkyl chains are optionally substituted with one or more substituents, such as those described herein.

[0161] The term "alkoxy" refers to an unbranched or branched alkyl group attached to an oxygen atom (alkyl-O-). In some embodiments, alkoxy as used herein refers to an alkyl group having 1 to 20 carbon atoms (C-C 20 )alkoxy), 1 to 10 carbon atoms ((C1-C 10 )alkoxy), 1 to 8 carbon atoms ((C1-C8)alkoxy), 1-6 carbon atoms ((C1-C6)alkoxy), 1-5 carbon atoms ((C1-C5)alkoxy), or 1-3 carbon atoms ((C1-C5)alkoxy). Examples include, but are not limited to, methoxy, ethoxy, n-propoxy, and butoxy. When an alkoxy residue having a specific number of carbons is named, all geometric isomers having that number of carbons can be encompassed, such as isopropoxy, isobutoxy, and t-butoxy. Unless otherwise stated herein, alkoxy chains are optionally substituted with one or more substituents, such as those described herein.

[0162] The term "alkylene" refers to an alkylene group in which the remainder of the molecule consists solely of carbon and hydrogen, contains no unsaturation, and preferably has from 1 to 20 carbon atoms (C-C 20 ) alkylene), 1-10 carbon atoms ((C1-C 10 (C-C) alkylene), 1-6 carbon atoms ((C-C) alkylene), or 1-5 carbon atoms ((C-C) alkylene). Examples include, but are not limited to, methylene, ethylene, propylene, butylene, etc. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group are through the terminal carbons, respectively. Unless stated otherwise herein, the alkylene chain is optionally substituted with one or more substituents, such as those described herein. Examples include methylene (-CH-), ethylene (-CHCH-), propylene (-CHCHCH-), 2-methylpropylene (-CH-CH(CH)-CH-), hexylene (-(CH)-), etc.

[0163] The term "alkenyl" refers to an aliphatic hydrocarbon group containing at least one carbon-carbon double bond, including straight-chain, branched-chain, and cyclic alkenyl groups. In some embodiments, an alkenyl group contains 2-10 carbon atoms (C2- 10 In another embodiment, the alkenyl group has 2-4 carbon atoms in the chain (C2-4 alkenyl). Exemplary alkenyl groups include, but are not limited to, ethenyl, propenyl, n-butenyl, i-butenyl, 3-methylbut-2-enyl, 3-methylbut-1-enyl, n-pentenyl, heptenyl, octenyl, cyclohexyl-butenyl, decenyl, and the like, and isomers thereof. An alkylalkenyl is an alkyl group, as defined herein, attached to an alkenyl group, as defined herein. An alkenyl group can be unsubstituted or substituted through available carbon atoms with one or more groups, as defined above for alkyl.

[0164] The term "alkynyl" refers to a straight-chain or branched monovalent hydrocarbyl group having 2 to 6 carbon atoms, preferably 2 to 3 carbon atoms, and having at least 1, and preferably 1 to 2, sites of acetylenic (C≡C-) unsaturation. Examples of such alkynyl groups include, but are not limited to, acetylenyl (C≡CH), propargyl (CHC≡CH), 1-propynyl, 1-butynyl, and 1-pentynyl.

[0165] The term "aminoalkyl" refers to a group formed by combining an alkyl and an amino group. Examples include, but are not limited to, -NHCH3, -NHCH2CH3-NHCHCH3), -N(CH3)2, -NCH3(CH2CH3)-NCH3(CH(CH3)2) and similar groups.

[0166] The term "aryl" refers to a monocyclic or polycyclic group having at least one hydrocarbon aromatic ring, where all ring atoms of at least one hydrocarbon aromatic ring are carbon. Aryl can include groups having a single aromatic ring (e.g., phenyl) and groups having multiple fused aromatic rings (e.g., naphthyl, anthryl). Aryl can also include groups having one or more aromatic hydrocarbon rings fused to one or more non-aromatic hydrocarbon rings (e.g., fluorene; 2,3-dihydro-1H-indene; 1,2,3,4-tetrahydronaphthalene). In certain embodiments, aryl includes groups having an aromatic hydrocarbon ring fused to a non-aromatic ring, where the non-aromatic ring contains at least one ring heteroatom independently selected from the group consisting of N, O, and S. For example, in some embodiments, aryl includes groups having a phenyl ring fused to a non-aromatic ring, where the non-aromatic ring contains at least one ring heteroatom independently selected from the group consisting of N, O, and S (e.g., chroman; thiochroman; 2,3-dihydrobenzofuran; indoline). In some embodiments, aryl, as used herein, refers to an alkyl group having 6 to 14 carbon atoms (C-C 14)aryl), or 6 to 10 carbon atoms ((C6-C 10 ) aryl). When the aryl contains a fused ring, the aryl can be attached to one or more substituents or moieties of the formulae described herein through any atom of the fused ring, where valence allows.

[0167] The term "cycloalkyl" refers to a monocyclic or polycyclic saturated hydrocarbon. In some embodiments, cycloalkyl is a cyclic group having from 3 to 20 carbon atoms (C-C 20 )cycloalkyl), 3 to 8 carbon atoms ((C3-C8)cycloalkyl), 3 to 6 carbon atoms ((C3-C6)cycloalkyl), or 3 to 5 carbon atoms ((C3-C5)cycloalkyl). In some embodiments, cycloalkyl has 3 to 8 carbon atoms with single or multiple cyclic rings, including fused, bridged, and spiro ring systems. Examples of suitable cycloalkyl groups include, but are not limited to, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, octahydropentalenyl, octahydro-1H-indene, decahydronaphthalene, cubane, bicyclo[3.1.0]hexane, bicyclo[1.1.1]pentane, and the like.

[0168] The term "carbocycle" refers to a saturated, unsaturated, or aromatic ring system in which each atom of the ring system is carbon. Carbocycles include 3-10 membered monocyclic rings, 6-12 membered bicyclic rings, and 6-12 membered bridged rings. Each ring of a bicyclic carbocycle can be selected from saturated, unsaturated, and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, can be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Bicyclic carbocycles include any combination of saturated, unsaturated, and aromatic bicyclic rings, as valence permits. Bicyclic carbocycles include any combination of ring sizes, such as 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems. Exemplary carbocycles include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl.

[0169] The term "haloalkyl" refers to a monohaloalkyl or polyhaloalkyl group which may be further substituted or unsubstituted, wherein the terms halogen and alkyl are as defined herein.

[0170] The term "heterocycle" refers to a saturated, unsaturated, or aromatic ring containing one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles include 3-10 membered monocyclic rings, 6-12 membered bicyclic rings, and 6-12 membered bridged rings. Bicyclic heterocycles include any combination of saturated, unsaturated, and aromatic bicyclic rings, valence permitting. In exemplary embodiments, an aromatic ring, such as pyridyl, can be fused to a saturated or unsaturated ring, such as cyclohexane, cyclopentane, morpholine, piperidine, or cyclohexene. Bicyclic heterocycles include any combination of ring sizes, such as 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems.

[0171] The term "heteroaryl" refers to a monocyclic or bicyclic or higher aromatic group, which may be substituted or unsubstituted, containing at least one heteroatom (e.g., a heteroatom selected from B, N, O, S, P(=O), Si, and P). In some embodiments, the term refers to an aromatic group of 4 to 10 carbon atoms and 1 to 4 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur in the ring. Such heteroaryl groups can have a single ring (i.e., pyridinyl or furyl) or multiple fused rings (i.e., indolizinyl or benzothienyl), where the fused rings may or may not be aromatic and / or contain heteroatoms if the point of attachment is through an atom of the aromatic heteroaryl group. In one embodiment, the nitrogen and / or sulfur ring atoms of the heteroaryl group are optionally oxidized to provide N-oxide (N→O), sulfinyl, or sulfonyl moieties. Examples of monocyclic heteroaryls include, but are not limited to, pyrazolyl, pyrrolyl, thiazolyl, oxazolyl, thiophenyl, furanyl, imidazolyl, isoxazolyl, triazolyl, thiadiazolyl, tetrazolyl, oxadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thiazolyl, and similar groups. Examples of bicyclic heteroaryls include, but are not limited to, indolyl, benzothiophenyl, benzofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzothiadiazole, benzotriazolyl, quinolinyl, isoquinolinyl, purinyl, furopyridinyl, oxochromene, dioxoisoindoline, pyrazolopyridinyl, pyrazolo[1,5-a]pyridinyl, and similar groups. Preferred heteroaryls include 5- or 6-membered heteroaryls such as pyridinyl, pyrrolyl, indolyl, thiophenyl, and furanyl.

[0172] The term "heterocycloalkyl" refers to a substituted or unsubstituted monocyclic alkyl containing one or more heteroatoms selected from B, N, O, S, P(=O), Si, and P. Examples include, but are not limited to, piperidinyl, piperazinyl, morpholinyl, pyrrolidinyl, thiomorpholinyl, imidazolidinyl, tetrahydrofurfuryl, and similar groups.

[0173] The term "heteroalkyl" refers to an alkyl substituent in which one or more carbon atoms and any associated hydrogen atoms are independently replaced with the same or different heteroatom groups. For example, one, two, or three carbon atoms can be independently replaced with the same or different heteroatom substituents.

[0174] The term "substituted" refers to moieties having substituents replacing a hydrogen on one or more carbon or substitutable heteroatoms, e.g., NH or NH, of a compound. "Substituted" or "substituted with" includes the implicit proviso that such substitution is subject to the allowed valences of the substituted atom and substituent, and it is understood that the substitution results in a stable compound. For example, stable compounds include, but are not limited to, compounds that do not undergo spontaneous transformation by rearrangement, cyclization, elimination, and the like. In certain embodiments, substituted refers to moieties having substituents replacing two hydrogen atoms on the same carbon atom, such as, for example, replacing two hydrogen atoms on a single carbon with an oxo, imino, or thioxo group. The term "substituted" is considered to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds.

[0175] Those skilled in the art will understand that, if necessary, the substituents themselves can be substituted.Unless specifically stated as "unsubstituted", reference to a chemical moiety herein is understood to include substituted and unsubstituted variants.For example, reference to a "heteroaryl" group or moiety, unless otherwise specified, implicitly includes both substituted and unsubstituted variants.

[0176] When referring to features of a compound, the phrase "optionally substituted" can be used interchangeably with the phrase "unsubstituted or substituted," and non-hydrogen substituents may or may not be present on a given atom or group; thus, the description includes structures in which non-hydrogen substituents are present and those in which they are absent. For example, "optionally substituted alkyl" encompasses both "alkyl" and "substituted alkyl" as defined herein. With respect to any group that contains one or more substituents, those of skill in the art understand that such groups are not intended to introduce substitutions or substitution patterns that are sterically impractical, synthetically impractical, and / or inherently unstable.

[0177] It will also be understood by those skilled in the art that when "optionally substituted" is used, any portion of the following term can be substituted.

[0178] The terms "linker," "bond," and "linking group" are used interchangeably and refer to a linking moiety that covalently bonds two or more substituents. A linking moiety may connect two groups, where the linker may be linear, branched, cyclic, or a single atom. In some embodiments, the linker is divalent. In some embodiments, the linker is a branched linker. In some embodiments, the two or more substituents covalently linked by the linking moiety are optionally substituted alkyl or alkoxy groups. In some embodiments, the linker is selected from -CO2-, -O-, -OCO-, -CONH-, -NHCO-, and -NH-.

[0179] In some embodiments, a substituent may be any substituent described herein, for example, halogen, hydroxy, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO), imino (=NH), oximo (=N-OH), hydrazino (=N-NH), -R b -OR a , -R b -℃(O)-R a , -R b -℃(O)-OR a , -R b -℃(O)-N(R a )2, -R b -N(R a )2, -R b -C(O)R a , -R b -C(O)OR a , -R b -C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(Ra)C(O)OR a , -R b -N(R a )C(O)R a , -R b N(R a )S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (t is 1 or 2), -R b -S(O) t OR a (t is 1 or 2), and -R b -S(O) t N(R a)2 (t is 1 or 2). In another exemplary embodiment, the substituents include alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, aralkynyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, and heteroarylalkyl, any of which can be selected from alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo, thioxo, cyano, nitro, imino, oximo, hydrazine, -R b -OR a , -R b -℃(O)-R a , -R b -℃(O)-OR a , -R b -℃(O)-N(R a )2, -R b -N(R a )2, -R b -C(O)R a , -R b -C(O)OR a , -R b -C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(Ra)C(O)OR a , -R b -N(R a )C(O)R a , -R b N(R a )S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (t is 1 or 2), -R b -S(O) t OR a (t is 1 or 2), and -R b -S(O) t N(R a )2 (t is 1 or 2) can be optionally substituted. Here, each R a , R b , and R cis independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, and heteroarylalkyl, where valences permit, each R a , R b , and R c is alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo, thioxo, cyano, nitro, imino, oximo, hydrazine, -R b -OR a , -R b -℃(O)-R a , -R b -℃(O)-OR a , -R b -℃(O)-N(R a )2, -R b -N(R a )2, -R b -C(O)R a , -R b -C(O)OR a , -R b -C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(Ra)C(O)OR a , -R b -N(R a )C(O)R a , -R b N(R a )S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (t is 1 or 2), -R b -S(O) t OR a (t is 1 or 2), and -R b -S(O) t N(R a )2(t is 1 or 2) can be substituted arbitrarily.

[0180] The term "isomer" or "three-dimensional isomer" refers to two or more compounds that contain the same number and types of atoms, groups, or components, but differ in the structural arrangement and connectivity of the atoms. This term can refer to compounds, or salts thereof, that are identical in terms of chemical equations or molecular equations, but differ optically or three-dimensionally, and can specifically be partial three-dimensional isomers, enantiomers, geometric isomers, or shape isomers.

[0181] The term "tautomer" refers to one of two or more structural isomers that exist in equilibrium and are readily converted from one isomeric form to another.

[0182] "Stereoisomers" refer to compounds made up of the same atoms joined by the same bonds, but with different, incompatible three-dimensional structures. The present disclosure contemplates various stereoisomers and mixtures thereof, and includes "enantiomers," which refer to two stereoisomers whose molecules are non-superimposable mirror images of one another.

[0183] Individual enantiomers and diastereomers of the compounds of the present disclosure can be prepared synthetically from commercially available starting materials containing asymmetric or stereogenic centers, or by preparation of racemic mixtures followed by resolution methods well known to those skilled in the art. These separation methods are exemplified by (1) attachment of a mixture of enantiomers to a chiral auxiliary, separation of the resulting mixture of diastereomers by recrystallization or chromatography, and liberation of the optically pure product from the auxiliary; (2) salt formation using an optically active resolving agent; (3) direct separation of a mixture of optical enantiomers on a chiral liquid chromatography column; or (4) kinetic separation using stereoselective chemical or enzymatic reagents. Racemic mixtures can also be resolved into their respective enantiomers by well-known methods such as chiral-phase gas chromatography or crystallization of compounds in chiral solvents. Stereoselective synthesis, chemical or enzymatic reactions in which a single reactant forms an unequal mixture of stereoisomers during the creation of a new stereocenter or the transformation of an existing stereocenter, is well known in the art. Stereoselective synthesis includes both enantioselective and diastereoselective transformations. See, for example, Carreira and Kvaerno, Classics in Stereoselective Synthesis, Wiley-VCH: Weinheim, 2009.

[0184] The symbol = denotes a bond that may be a single, double, or triple bond, as described herein. Substituents around a carbon-carbon double bond are designated as being in the "Z" or "E" configuration, and the terms "Z" and "E" are used in accordance with IUPAC standards. Unless otherwise specified, structures depicting double bonds include both the "E" and "Z" isomers.

[0185] Alternatively, substituents around a carbon-carbon double bond may be designated "cis" or "trans," with "cis" referring to substituents on the same side of the double bond and "trans" referring to substituents on opposite sides of the double bond. The arrangement of substituents around a carbocyclic ring may also be designated as "cis" or "trans." The term "cis" refers to substituents on the same side of the plane of the ring and the term "trans" refers to substituents on opposite sides of the plane of the ring. A mixture of compounds in which substituents are located on both the same and opposite sides of the plane of the ring is designated "cis / trans."

[0186] Singular articles such as "a," "an," "the," and similar referents in the context of describing elements are to be construed as covering both the singular and the plural unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, including the upper and lower limits of the range, unless otherwise stated herein, and each separate value is incorporated into the specification as if it were individually set forth herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or clearly contradicted by context. Any and all examples provided herein, or the use of exemplary language (i.e., "such as"), are intended merely to better clarify the embodiments and do not limit the scope of the claims unless otherwise expressly stated.

[0187] In some embodiments, when the term "about" is used before a quantitative value, the present disclosure also includes the specific quantitative value itself unless otherwise specified. As used herein, the term "about" refers to a ±10% variation from the nominal value unless otherwise indicated or inferred. When a percentage is provided regarding the amount of an ingredient or material in a composition, the percentage should be understood to be a percentage by weight unless otherwise specified or understood from the context.

[0188] Where molecular weights are provided and not absolute values, for example for polymers, the molecular weight should be understood to be an average molecular weight unless otherwise stated or understood from the context.

[0189] It should be understood that the order of steps or order for performing certain actions is immaterial so long as the present disclosure remains functional. Moreover, two or more steps or actions may be conducted simultaneously.

[0190] A dash ("-") symbol that is not between two letters or symbols indicates the point of attachment or bond for a substituent, for example, -NH2 is bonded through the nitrogen atom.

[0191] The term "pharmaceutically acceptable salt" refers to a salt that is acceptable for administration to a subject. It is understood that such salts, including counterions, have acceptable mammalian safety for a given administration regimen. Such salts can also be derived from pharmaceutically acceptable inorganic or organic bases and pharmaceutically acceptable inorganic or organic acids, and can include organic and inorganic counterions. The neutral form of the compounds described herein can be converted to the corresponding salt form by contacting the compound with a base or acid and isolating the resulting salt.

[0192] The terms "pharmaceutically acceptable excipient," "pharmaceutically acceptable diluent," "pharmaceutically acceptable carrier," and "pharmaceutically acceptable adjuvant" are used interchangeably and refer to an excipient, diluent, carrier, or adjuvant that is useful in preparing pharmaceutical compositions and is generally safe, non-toxic, and not biologically or otherwise undesirable. Also included are excipients, diluents, carriers, and adjuvants that can be used in veterinary and human medicines. The phrase "pharmaceutically acceptable excipient" includes both one and more such excipients, diluents, carriers, and / or adjuvants.

[0193] The term "pharmaceutical composition" is meant to encompass compositions suitable for administration to a subject, such as a mammal, particularly a human. Generally, a "pharmaceutical composition" is sterile and preferably free of contaminants that may elicit an undesirable response in the subject (i.e., the compounds in the pharmaceutical composition are of pharmaceutical grade). Pharmaceutical compositions can be designed to be administered to a subject or patient in need thereof via many different routes of administration, including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, intratracheal, intramuscular, subcutaneous, etc. Illustrative Embodiments

[0194] As described herein, the present invention will be described in various embodiments of the compounds, compositions, and methods of the present invention.The various embodiments described are intended to provide various illustrative examples and should not be interpreted as descriptions of alternative species.It should be noted that the descriptions of the various embodiments provided herein may overlap in scope.The embodiments discussed herein are merely illustrative and are not intended to limit the scope of the present technology.

[0195] Regardless of the scope of the appended claims, aspects of the present disclosure are set forth in the following paragraphs.

[0196] Item 1. A compound of formula (Ia) or formula (Ib), or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof. JPEG2025120182000039.jpg39170 where, R is H, (C1-C3) alkyl, or substituted (C1-C3) alkyl; Y 1 Or Y 4 is an independent CR 10 and N are selected from Y 1 Or Y 4 At least two of the 10 and; R 10are each independently selected from H, (C-C)alkyl, substituted (C-C)alkyl, (C-C)alkenyl, substituted (C-C)alkenyl, (C-C)alkynyl, substituted (C-C)alkynyl, (C-C)haloalkyl, (C-C)alkoxy, substituted (C-C)alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol; R a and R b are each independently selected from H, F, (C-C) alkyl, and substituted (C-C) alkyl, or R a and R b are attached to a ring and together with the carbon atoms to which they are attached form a cyclopropyl or substituted cyclopropyl; A is phenyl, substituted phenyl, pyridyl, or substituted pyridyl.

[0197] Item 2. A is phenyl or one, two, or three R 20 groups, and each R 20 are independently selected from (C-C) alkyl, substituted (C-C) alkyl, (C-C) alkenyl, substituted (C-C) alkenyl, (C-C) alkynyl, substituted (C-C) alkynyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol.

[0198] Item 3. A is pyridyl or one, two, or three R 20 pyridyl substituted with a group, and each R 20are independently selected from (C-C) alkyl, substituted (C-C) alkyl, (C-C) alkenyl, substituted (C-C) alkenyl, (C-C) alkynyl, substituted (C-C) alkynyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol.

[0199] Item 4. The compound according to any one of items 1-3, wherein R is H.

[0200] Item 5. R a and R b and each is H.

[0201] Item 6. The compound according to item 2, wherein the compound is of formula (IIa) or (IIb): JPEG2025120182000040.jpg41170 where R 1 Or R 9 are independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) alkenyl, substituted (C-C) alkenyl, (C-C) alkynyl, substituted (C-C) alkynyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol.

[0202] Item 7. R 1 Or R 9are independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, -NH2, substituted amino, halogen, and hydroxyl.

[0203] Item 8. R 1 Or R 9 is independently selected from H, NH2, F, CH3, and CF3.

[0204] Item 9. The compound according to item 6, wherein the compound is of formula (IIIa) or (IIIb): JPEG2025120182000041.jpg47170 where R 21 and R 22 are independently H, (C1-C8) alkyl, substituted (C1-C8) alkyl, SO2R 30 , and C.O.R. 30 Selected from R 30 is (C1-C8) alkyl or substituted (C1-C8) alkyl.

[0205] Item 10. R 21 and R 22 and each is H.

[0206] Item 11. R 5 , R 6 , R 8 and R 9 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0207] Item 12. R 5 , R 6 , R 8 and R 9is independently selected from H, F, CH3, and CF3.

[0208] Item 13. R 5 , R 6 , R 8 and R 9 Item 13. The compound according to item 12, wherein each is H.

[0209] Item 14. A compound according to any one of items 9 to 13, as described below: R 2 Or R 4 are H and R respectively. 1 is selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0210] Item 15. R 1 is selected from H, F, CH3, and CF3.

[0211] Item 16. The compound according to item 15, wherein the compound is selected from the following: JPEG2025120182000042.jpg108170

[0212] Item 17. The compound according to item 9, wherein the compound is of formula (IVa) or (IVb): JPEG2025120182000043.jpg41170

[0213] Item 18. R 6 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0214] Item 19. R 6is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, and (C1-C3) haloalkyl.

[0215] Item 20. R 6 Item 19. The compound according to item 19, wherein is CH3 or CF3.

[0216] Item 21. A compound according to any one of items 17 to 20, as described below. 2 Or R 4 are H and R respectively. 1 is selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0217] Item 22. R 1 22. The compound according to item 21, wherein is selected from H, F, CH3, and CF3.

[0218] Item 23. The compound according to any one of items 17 to 22, wherein the compound is selected from the following: JPEG2025120182000044.jpg143170

[0219] Item 24. The compound according to item 9, wherein the compound is of formula (Va) or (Vb): JPEG2025120182000045.jpg41170

[0220] Item 25. R 5 and R 9 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0221] Item 26. R 5 and R 926. The compound according to item 25, wherein is independently selected from H and halogen.

[0222] Item 27. R 5 27. The compound according to item 26, wherein is F.

[0223] Item 28. R 9 28. The compound according to item 26 or 27, wherein is F.

[0224] Item 29. The compound according to item 26, wherein the compound is selected from the following: JPEG2025120182000046.jpg149170

[0225] Item 30. The compound according to item 6, wherein the compound is of formula (VIa) or (VIb): JPEG2025120182000047.jpg41170

[0226] Item 31. R 5 , R 6 and R 9 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, -NH2, substituted amino, halogen, and hydroxyl.

[0227] Item 32. R 6 is selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, and (C1-C3) haloalkyl.

[0228] Item 33. R 6 33. The compound according to item 32, wherein is CH3 or CF3.

[0229] Item 34. R 6 33. The compound according to item 32, wherein is H.

[0230] Item 35. R 5 and R9 is independently selected from H and halogen.

[0231] Item 36. R 5 36. The compound according to item 35, wherein is F.

[0232] Item 37. R 9 The compound according to item 35 or 36, wherein is F.

[0233] Item 38. The compound according to item 35, wherein the compound is selected from the following: JPEG2025120182000048.jpg85170

[0234] Item 39. Each R 10 or R 1 or R 4 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, -NH2, substituted amino, halogen, and hydroxyl.

[0235] Item 40. Each R 10 or R 1 Or R 4 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl and halogen.

[0236] Item 41. R 1 is H.

[0237] Item 42. R 1 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl and halogen.

[0238] Item 43. R1 43. The compound according to item 42, wherein is F, CH3 or CF3.

[0239] Item 44. R 2 Or R 4 and each is H.

[0240] Item 45. The compound according to item 1 or 2, wherein the compound is of formula (VIIa) or (VIIb): JPEG2025120182000049.jpg42170 where R 5 Or R 9 and each R 10 are independently selected from H, (C-C) alkyl, substituted (C-C) alkyl, (C-C) alkenyl, substituted (C-C) alkenyl, (C-C) alkynyl, substituted (C-C) alkynyl, (C-C) haloalkyl, (C-C) alkoxy, substituted (C-C) alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol.

[0241] Item 46. Y 1 Or Y 4 46. The compound according to item 45, wherein one of

[0242] Item 47. R 5 Or R 9 and each R 10 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, -NH2, substituted amino, halogen, and hydroxyl.

[0243] Item 48. R 5 Or R 9 and each R 10is independently selected from H, NH2, F, CH3, and CF3.

[0244] Item 49. The compound according to item 46, wherein the compound is of formula (VIIIa) or (VIIIb): JPEG2025120182000050.jpg46170 where R 21 and R 22 is H, (C1-C8) alkyl, substituted (C1-C8) alkyl, SO2R 30 , and C.O.R. 30 are independently selected from R 30 is (C1-C8) alkyl or substituted (C1-C8) alkyl.

[0245] Item 50. R 21 and R 22 Item 49. The compound according to item 49, wherein each is H.

[0246] Item 51. R 5 , R 6 , R 8 and R 9 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0247] Item 52. R 5 , R 6 , R 8 and R 9 is independently selected from H, F, CH3, and CF3.

[0248] Item 53. R 5 , R 6 , R 8 and R 9 53. The compound according to item 52, wherein each is H.

[0249] Item 54. The compound according to item 53, wherein the compound is selected from the following: JPEG2025120182000051.jpg143170

[0250] Item 55. The compound according to item 49, wherein the compound is of formula (IXa) or (IXb): JPEG2025120182000052.jpg41170

[0251] Item 56. R 6 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0252] Item 57. R 6 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, and (C1-C3) haloalkyl.

[0253] Item 58. R 6 58. The compound according to item 57, wherein is CH3 or CF3.

[0254] Item 59. The compound according to item 58, wherein the compound is selected from the following: JPEG2025120182000053.jpg219170 JPEG2025120182000054.jpg73170

[0255] Item 60. The compound according to Item 49, wherein the compound is of formula (Xa) or (Xb): JPEG2025120182000055.jpg41170

[0256] Item 61. R 5 and R 9is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0257] Item 62. R 5 and R 9 is independently selected from H and halogen.

[0258] Item 63. R 5 63. The compound according to item 62, wherein is F.

[0259] Item 64. R 9 64. The compound according to item 62 or item 63, wherein is F.

[0260] Item 65. The compound according to item 62, wherein the compound is selected from the following: JPEG2025120182000056.jpg212170 JPEG2025120182000057.jpg72170

[0261] Item 66. The compound according to item 46, wherein the compound is of formula (XIa) or (XIb): JPEG2025120182000058.jpg41170

[0262] Item 67. R 5 , R 6 and R 9 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, -NH2, substituted amino, halogen, and hydroxyl.

[0263] Item 68. R 6is selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, and (C1-C3) haloalkyl.

[0264] Item 69. R 6 69. The compound according to item 68, wherein is CH3 or CF3.

[0265] Item 70. R 6 69. The compound according to item 68, wherein is H.

[0266] Item 71. R 5 and R 9 is independently selected from H and halogen.

[0267] Item 72. R 5 72. The compound according to item 71, wherein is F.

[0268] Item 73. R 9 73. The compound according to item 71 or item 72, wherein

[0269] Item 74. The compound according to item 71, wherein the compound is selected from the following: JPEG2025120182000059.jpg122170

[0270] Item 75. The compound according to item 45, wherein the compound is of formula (XIVa) or (XIVb): JPEG2025120182000060.jpg47170 where Y 4 is CR 4 or N; R 1 Or R 4are independently selected from H, (C-C)alkyl, substituted (C-C)alkyl, (C-C)alkenyl, substituted (C-C)alkenyl, (C-C)alkynyl, substituted (C-C)alkynyl, (C-C)haloalkyl, (C-C)alkoxy, substituted (C-C)alkoxy, -CONH, substituted amido, -NH, substituted amino, -COH, cyano, halogen, hydroxyl, -NO, -SOH, -SONH, substituted sulfonamide, and thiol; R 21 and R 22 is H, (C1-C8) alkyl, substituted (C1-C8) alkyl, SO2R 30 , and C.O.R. 30 are independently selected from R 30 is (C1-C8) alkyl, and substituted (C1-C8) alkyl.

[0271] Item 76. R 5 , R 6 , R 8 and R 9 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0272] Item 77. R 21 and R 22 and each is H.

[0273] Item 78. A compound according to any one of items 75 to 77, wherein: R 2 Or R 4 are H and R respectively. 1 is selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0274] Item 79. R 1 79. The compound according to item 78, wherein is selected from H, F, CH3, and CF3.

[0275] Item 80. The compound according to item 75, wherein the compound is of formula (XVa) or (XVb): JPEG2025120182000061.jpg41170

[0276] Item 81. R 5 and R 9 is independently selected from H, (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0277] Item 82. R 5 and R 9 82. The compound according to item 81, wherein is independently selected from H and halogen.

[0278] Item 83. R 5 83. The compound according to item 82, wherein is F.

[0279] Item 84. R 9 84. The compound according to item 82 or item 83, wherein is F.

[0280] Item 85. The compound according to item 75, wherein the compound is of formula (XVIa) or (XVIb): JPEG2025120182000062.jpg41170

[0281] Item 86. R 6 is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, substituted (C1-C5) alkoxy, halogen, and hydroxyl.

[0282] Item 87. R6 87. The compound according to item 86, wherein is selected from (C1-C5) alkyl, substituted (C1-C5) alkyl, and (C1-C3) haloalkyl.

[0283] Item 88. R 6 88. The compound according to item 87, wherein is CH3 or CF3.

[0284] Item 89. R 1 is selected from H, (C1-C5)alkyl, substituted (C1-C5)alkyl, (C1-C3)haloalkyl, (C1-C5)alkoxy, substituted (C1-C5)alkoxy, halogen, and hydroxyl.

[0285] Item 90. R 1 89. The compound according to item 89, wherein is selected from H, F, CH3, and CF3.

[0286] Item 91. Y 4 The compound according to any one of items 75 to 90, wherein is CH.

[0287] Item 92. Y 4 is N.

[0288] Item 93. The compound according to items 75, 80 or 85, wherein the compound is selected from the following: JPEG2025120182000063.jpg146170

[0289] Item 94. The compound according to item 1, wherein the compound is a compound of Table 1, or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0290] Item 95. A pharmaceutical composition comprising the compound according to any one of items 1 to 94, or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0291] Item 96. A compound for use in inhibiting or antagonizing adenosine A2A and / or A1 receptors, the compound being a compound according to any one of items 1-94, or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0292] Item 97. A pharmaceutical composition for use in inhibiting or antagonizing adenosine A2A and / or A1 receptors, the pharmaceutical composition being as described in item 95.

[0293] Item 98. A method for inhibiting adenosine A2A and / or A1 receptors, comprising contacting a sample containing adenosine A2A and / or A1 receptors with an effective amount of a compound according to any one of items 1-94, or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof, to inhibit adenosine A2A and / or A1 receptors.

[0294] Item 99. The method of item 98, wherein the sample is in vitro.

[0295] Item 100. A method for antagonizing adenosine A2A and / or A1 receptors, comprising: Cells containing adenosine A2A and / or A1 receptors are contacted with an effective amount of a compound according to any one of paragraphs 1-94, or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof, to antagonize the adenosine A2A and / or A1 receptors.

[0296] Item 101. The method of item 100, wherein the cells are contained in a sample of cells in vitro.

[0297] Item 102. The method of item 100, wherein the cell is contained in a biological system in vivo.

[0298] Item 103. A method of treating cancer, comprising administering to a subject having cancer a therapeutically effective amount of an A2A and / or A1 receptor antagonist compound according to any one of items 1-94, or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0299] Item 104. The method of item 103, wherein the cancer is a solid tumor cancer.

[0300] Item 105. The method of item 103, wherein the cancer is selected from lung cancer, breast cancer, prostate cancer, ovarian cancer, solenoma, cervical cancer, bladder cancer, head and neck cancer, renal cell carcinoma, esophageal cancer, pancreatic cancer, brain cancer, liver cancer, leukemia, lymphoma, melanoma, multiple myeloma, Ewing's sarcoma, osteosarcoma, colorectal tumor, bile duct cancer, choriocarcinoma, oral cancer, neuroblastoma, skin cancer, testicular cancer, stromal tumor, germ cell tumor, and thyroid cancer.

[0301] Item 106. The method of item 105, wherein the cancer is a liver cancer that is hepatocellular carcinoma (HCC).

[0302] Item 107. The method of item 105, wherein the cancer is a lung cancer that is non-small cell lung cancer (NSCLC).

[0303] Item 108. The method of any one of items 103 to 107, further comprising co-administering an additional active agent to the subject.

[0304] Item 109. The method of item 108, wherein the additional active agent is selected from an anti-angiogenic agent, an anti-inflammatory agent, an immune checkpoint inhibitor, a PARP inhibitor, a chemotherapeutic agent, and an immune anti-cancer agent.

[0305] Item 110. The method of item 109, wherein the additional active agent is an immune checkpoint inhibitor selected from a CTLA-4 inhibitor, a PD-1 inhibitor, and a PD-L1 inhibitor.

[0306] Item 111. The method of item 110, wherein the immune checkpoint inhibitor is an antibody or antibody fragment.

[0307] Item 112. A method of treating an inflammatory disease, comprising administering to a subject having an inflammatory disease a therapeutically effective amount of an A2A and / or A1 receptor antagonist compound according to any one of items 1-94, or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0308] Item 113. The method of item 112, wherein the inflammatory disease is a chronic inflammatory disease.

[0309] Item 114. The method of item 112, wherein the inflammatory disease is an acute inflammatory disease.

[0310] Item 115. A method of treating a central nervous system disorder, comprising administering to a subject having or at risk for a central nervous system disorder a therapeutically effective amount of an A2A and / or A1 receptor antagonist compound according to any one of items 1-94, or a solvate, hydrate, prodrug, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0311] Item 116. The method of item 115, wherein the central nervous system disease is selected from Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, multiple sclerosis, Huntington's disease, depression, schizophrenia, and epilepsy.

[0312] Item 117. The method of item 116, wherein the central nervous system disease is Parkinson's disease.

[0313] Item 118. The compound according to any one of items 1 to 94 or the pharmaceutical composition according to item 95 for use in treating a disease selected from cancer, an inflammatory disease and a central nervous system disease.

[0314] Item 119. Use of a compound according to any one of items 1 to 94 or a pharmaceutical composition according to item 95 in the manufacture of a medicament for treating a disease selected from cancer, an inflammatory disease, and a central nervous system disease.

[0315] Item 120. Compound of the formula: JPEG2025120182000064.jpg2842 or a solvate, three-dimensional isomer, or pharmaceutically acceptable salt thereof, wherein Cy is a fused 5-membered heterocycle (e.g., a heteroaryl or heterocycloalkyl ring) containing at least one heteroatom selected from nitrogen, oxygen, and sulfur; R is R 11 , R 12 , and / or R 13 is a phenyl group substituted with; R 11 , R 12 , and / or R 13 are each independently a halogen, a hydroxyl group, a thiol group, a carbonyl group, an amido group, a nitro group, an amino group, a substituted or unsubstituted (C1-C5) alkyl group, a substituted or unsubstituted (C2-C5) alkenyl group, a substituted or unsubstituted (C2-C5) alkynyl group, a substituted or unsubstituted (C1-C3) haloalkyl group, and a substituted or unsubstituted (C1-C3) aminoalkyl group or a substituted or unsubstituted (C1-C5) alkoxy group, R 10 is one or more groups independently selected from hydrogen, halogen, hydroxyl group, thiol group, substituted or unsubstituted (C1-C5) alkyl group, substituted or unsubstituted (C2-C5) alkenyl group, substituted or unsubstituted (C2-C5) alkynyl group, substituted or unsubstituted (C1-C3) haloalkyl group, substituted or unsubstituted (C1-C5) alkoxy group, or cyano.

[0316] Item 121. Cy contains at least one heteroatom selected from nitrogen, oxygen, and sulfur to form a 5-membered heteroaryl ring fused with the atoms of the pyrimidine ring, and R is independently selected from hydrogen, halogen, amino group, hydroxyl group, thiol group, (C1-C5) alkyl group, (C2-C5) alkenyl group, (C2-C5) alkynyl group, (C1-C3) haloalkyl group, (C1-C3) aminoalkyl group, or (C1-C5) alkoxy group. 11 , R 12 , and / or R 13 and each R is independently selected from hydrogen, halogen, hydroxyl, thiol, (C-C) alkyl, (C-C) alkenyl, (C-C) alkynyl, (C-C) haloalkyl, (C-C) alkoxy, and cyano; or a solvate, a steric isomer, or a pharmaceutically acceptable salt thereof.

[0317] Item 122. Cy contains at least one nitrogen atom and forms a 5-membered heteroaryl ring fused with an atom of the pyrimidine ring, and R is a group having the structure JPEG2025120182000065.jpg2323, which is bonded to the nitrogen atom of the Cy ring, and R 11 , R 12 , and / or R 13 are each hydrogen, halogen, an amino group, a (C1-C5) alkyl group, a (C2-C5) alkenyl group, a (C2-C5) alkynyl group, a (C1-C3) haloalkyl group, a (C1-C3) aminoalkyl group, or a (C1-C5) alkoxy group, and each R 10 are independently selected from hydrogen, halogen, hydroxyl, thiol, (C1-C5) alkyl, (C2-C5) alkenyl, (C2-C5) alkynyl, (C1-C3) haloalkyl, (C1-C5) alkoxy, and cyano.

[0318] Item 123. Cy contains at least one nitrogen atom and forms a 5-membered heteroaryl ring with the atoms of the pyrimidine ring, and R is JPEG2025120182000066.jpg2323, which is bonded to the nitrogen atom of the Cy ring, R 11 and R 13 are each hydrogen, halogen, a (C1-C5) alkyl group, a (C2-C5) alkenyl group, a (C2-C5) alkynyl group, a (C1-C3) haloalkyl group, or a (C1-C5) alkoxy group; R 12 teeth hydrogen or an amino group, and each R 10 is independently selected from hydrogen, halogen, a (C1-C5) alkyl group, a (C2-C5) alkenyl group, a (C2-C5) alkynyl group, a (C1-C3) haloalkyl group, a (C1-C5) alkoxy group, and cyano, or a solvate, a three-dimensional isomer, or a pharmaceutically acceptable salt thereof.

[0319] Item 124. R 11 and R 13 are each hydrogen, halogen, a (C1-C5) alkyl group, a (C2-C5) alkenyl group, a (C2-C5) alkynyl group, a (C1-C3) haloalkyl group, or a (C1-C5) alkoxy group; R 12 is hydrogen or amino group, R 10 is independently selected from hydrogen, halogen, a (C1-C5) alkyl group, a (C2-C5) alkenyl group, a (C2-C5) alkynyl group, a (C1-C3) haloalkyl group, or a (C1-C5) alkoxy group, or cyano, or a solvate, a three-dimensional isomer, or a pharmaceutically acceptable salt thereof.

[0320] Item 125. R 11 and R 13 are each hydrogen, halogen, a C1-C5 alkyl group, or a C1-C3 haloalkyl group, and R 12 is hydrogen or an amino group, and R 10125. The compound or solvate, tertiary isomer or pharmaceutically acceptable salt of item 124, wherein is independently selected from hydrogen, halogen, a C1-C5 alkyl group, or cyano.

[0321] Item 126. Cy forms a pyrazolopyrimidine or purine with a pyrimidine chain bound to Cy, R is JPEG2025120182000067.jpg2828, and bonded to the nitrogen atom of Cy, R 11 and R 13 are each hydrogen, halogen, a C1-C5 alkyl group, a C2-C5 alkenyl group, a C2-C5 alkynyl group, a C1-C3 haloalkyl group, or a C1-C5 alkoxy group; R 2 is hydrogen or an amino group, R 10 is hydrogen, halogen, a C1-C5 alkyl group, a C2-C5 alkenyl group, a C2-C5 alkynyl group, a C1-C3 haloalkyl group, a C1-C5 alkoxy group, or a cyano group, or a solvate, a three-dimensional isomer, or a pharmaceutically acceptable salt thereof.

[0322] Item 127. R 11 and R 13 are each hydrogen, halogen, a C1-C5 alkyl group, a C2-C5 alkenyl group, a C2-C5 alkynyl group, a C1-C3 haloalkyl group, or a C1-C5 alkoxy group; R 2 is hydrogen or an amino group, R 10 is hydrogen, halogen, a C1-C5 alkyl group, a C2-C5 alkenyl, a C2-C5 alkynyl, a C1-C3 haloalkyl group, a C1-C5 alkoxy or cyano, or a solvate, a three-dimensional isomer or a pharmaceutically acceptable salt thereof.

[0323] Item 128. The compound of item 120, or a solvate, a three-dimensional isomer, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of: 3-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo-[3,4-d]-pyrimidin-4-yl)-benzonitrile; 3-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo-[3,4-d]-pyrimidin-4-yl)-2-methylbenzonitrile; 3-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo-[3,4-d]-pyrimidin-4-yl)-2-fluorobenzonitrile; 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)-benzonitrile; 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile; 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)-2-methylbenzonitrile; 3-(2-amino-9-(4-amino-2,6-difluorobenzyl)-9H-purin-6-yl)-benzonitrile; 3-(2-amino-9-(4-amino-2,6-difluorobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile; 3-(2-amino-9-(4-amino-2-fluorobenzyl)-9H-purin-6-yl)-benzonitrile; 3-(2-amino-9-(4-amino-2-fluorobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile; 3-(2-amino-9-(4-amino-3-methylbenzyl)-9H-purin-6-yl)-benzonitrile; 3-(2-amino-9-(4-amino-3-methylbenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile; 3-(2-amino-9-(2,6-difluorobenzyl)-9H-purin-6-yl)-benzonitrile; 3-(2-amino-9-(2,6-difluorobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile; and 3-(2-amino-9-(2,6-difluorobenzyl)-9H-purin-6-yl)-2-methylbenzonitrile.

[0324] Item 129. A pharmaceutical composition for preventing or treating cancer, comprising an effective amount of a compound of any one of items 119 to 127, or a solvate, three-dimensional isomer, or pharmaceutically acceptable salt thereof.

[0325] Item 130. The pharmaceutical composition according to item 129, wherein the composition has an antagonistic effect on adenosine A2A receptors.

[0326] Item 131. The pharmaceutical composition of item 129, wherein the cancer is lung cancer, breast cancer, prostate cancer, ovarian cancer, solenoma, cervical cancer, bladder cancer, head and neck cancer, renal cell carcinoma, esophageal cancer, pancreatic cancer, brain cancer, liver cancer, leukemia, lymphoma, melanoma, multiple myeloma, Ewing's sarcoma, osteosarcoma, colorectal tumor, bile duct cancer, choriocarcinoma, oral cancer, neuroblastoma, skin cancer, testicular cancer, stromal tumor, germ cell tumor, or thyroid cancer.

[0327] Item 132. The pharmaceutical composition of item 129, wherein the composition is administered simultaneously, separately, or sequentially with another anticancer agent.

[0328] Item 133. The pharmaceutical composition of item 129, wherein the composition is manufactured in a pharmaceutically acceptable form, which is a tablet, pill, powder, capsule, syrup, emulsion, or microemulsion. [Example]

[0329] The following examples are provided to illustrate the present disclosure and should not be construed as limiting the scope of the present technology in any way. All functionally equivalent methods are within the scope of the present technology. In addition to those described herein, various modifications of the present technology will be apparent to those skilled in the art from the foregoing description. Such modifications are within the scope of the appended claims.

[0330] All temperatures are in degrees Celsius unless otherwise noted. Efforts have been made to ensure accuracy with respect to numbers used (amounts, temperatures, etc.) but some experimental errors and deviations should be allowed for.

[0331] If an abbreviation is not defined, it has its generally accepted meaning. General synthesis method

[0332] The final compound was confirmed by high performance liquid chromatography / mass spectrometry (HPLC / MS) analysis and was determined to be greater than 90% by weight pure. 1 H nuclear magnetic resonance (NMR) spectra were recorded in CDCl3 (residual internal standard CHCl3 = δ 7.26), dimethyl sulfoxide (DMSO)-d6 (residual internal standard CD3S°C D2H = δ 2.50), methanol-d4 (residual internal standard CD2HOD = δ 3.30), or acetone-d6 (residual internal standard CD3C°C D2H = δ 2.05). Reported chemical shifts (δ) are given in parts per million (ppm), and coupling constants (J) are given in hertz (Hz). Spin multiplicities are: s = singlet, bs = broad singlet, bm = broad multiplet, d = doublet, t = triplet, q = quartet, p = quintet, dd = doublet of doublets, ddd = doublet of doublets, dt = doublet of triplets, td = triplet of doublets, tt = triplet of triplets, and m = multiplet.

[0333] HPLC-MS analysis was performed using gradient elution. Medium pressure liquid chromatography (MPLC) was performed using both normal and reverse phase silica gel columns. Example 1: Preparation of Compounds

[0334] The compounds of the present disclosure are produced by adapting well-known chemical transformations according to the methods illustrated by Schemes 1 or 2 below. The synthesis of some exemplary compounds in Table 1 is described below in Examples 1.1 to 1.69. Several other compounds in Table 1 were prepared by adapting the methods described herein and evaluated according to the biological assays described in Examples 2-4.

[0335] General Scheme 1 An exemplary synthesis method for JPEG2025120182000068.jpg79161 Scheme 1 is described in detail below.

[0336] To explain in more detail with reference to the above-mentioned Scheme 1, the first step is to dissolve the A-2 compound in tetrahydrofuran and water, and then to produce the compound of Scheme A-3 by chain reaction of the solution with hydrazine monohydrate at 25°C to 60°C for 24 hours; the second step is to dissolve the A-3 compound in N,N-dimethylformamide, and then to produce the compound of Scheme A-4 by nucleophilic substitution at 25°C to 60°C for 24 hours under the condition that potassium carbonate or cesium carbonate is provided, to produce the compound of Scheme A-5. It is possible to prepare the compound shown by the above-mentioned Scheme 1 through a series of processes including two steps; and a third step in which the A-5 compound is dissolved in 1,4-dioxane and distilled water, and the resulting solution is subjected to Suzuki coupling reaction with the compound of Scheme A-6 at 100 to 115°C for 2 to 24 hours under conditions in which 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile and its derivatives, tetrakis(triphenylphosphine)palladium, and sodium carbonate are provided, or the solution undergoes Suzuki coupling reaction under the same conditions, the solution is dissolved in an ethanol solvent, and the solution is subjected to reduction reaction under conditions in which tin(II) chloride dihydrate and acid are used to produce the A compound. Here, R in the above-mentioned Scheme 1 is a1 , R a2 , R a3 , R a4 , and R a5 is a substituent of the formulae described herein, for example, R of formula (IIb) 1 Or R 9 and a cyano group.

[0337] General Scheme 2 An exemplary synthesis method for JPEG2025120182000069.jpg74161 Scheme 2 is described in detail below.

[0338] The compounds shown by the above schemes can be produced through a series of processes including: a first step in which compound B-1 is dissolved in N,N-dimethylformamide and the solution is reacted in a nucleophilic substitution reaction with the compound of Scheme A-4 at 25 to 60°C for 24 hours under conditions of providing potassium carbonate or cesium carbonate, to produce compound B-2; and a second step in which compound B-2 is dissolved in 1,4-dioxane and distilled water and the solution is reacted in a Suzuki coupling reaction with the compound of Scheme A-6 at 100 to 115°C for 2 to 24 hours under conditions of providing 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile and its derivatives, tetrakis(triphenylphosphine)palladium, and sodium carbonate, or the solution is subjected to a Suzuki coupling reaction under the same conditions, the solution is dissolved in ethanol, and the solution is reduced with tin(II) chloride dihydrate and acid to produce compound B. Here, R in the above scheme 2 is a1 , R a2 , R a3 , R a4 , and R a5 is a substituent of the formulae described herein, for example, R of formula (IIa) 1 Or R 9 and a cyano group.

[0339] Example 1.1: Preparation of 3-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (Compound 201)

[0340] 1.1.1. Preparation of 4-chloro-(4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine JPEG2025120182000070.jpg36130 Reagents and conditions: (a) hydrazine, TEA, THF:H2O (3:1), room temperature to 50°C, 3 hours, quantitative; (b) 4-nitrobenzyl bromide, K2CO3, DMF, 0°C to room temperature, 12 hours, 58%.

[0341] Step 1: Preparation of 4-chloro-1H-pyrazolo[3,4-d]pyrimidin-6-amine

[0342] 2-Amino-4,6-dichloropyrimidine-5-carbaldehyde (S1, 4 g, 20.83 mmol) was dissolved in tetrahydrofuran (THF) and HO, and hydrazine monohydrate (0.78 mL, 24.99 mmol) and triethylamine (TEA, 3.51 mL, 24.99 mmol) were added. The reaction mixture was then heated to 50 °C, stirred for 3 hours, and concentrated. The concentrate was then washed and purified with distilled water to obtain the intermediate compound (S2, 3.45 g, 100%). 1 H-NMR (DMSO-d6, 400MHz): δ13.25 (s, 1H), 8.02 (s, 1H), 7.14 (s, 2H).

[0343] Step 2: Preparation of 4-chloro-1-(4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine

[0344] The intermediate compound (S2, 3.0 g, 18.12 mmol) produced in Step 1 above was dissolved in dimethylformamide (DMF) and 4-nitrobenzyl bromide (3.92 g, 18.12 mmol), and potassium carbonate (K2CO3, 3.75 g, 27.18 mmol) was added to the solution. This was then stirred at room temperature for 12 hours. The reaction mixture was diluted with ethyl acetate (EA) and washed with distillation. It was then dried over magnesium sulfate, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give the desired compound (S3, 3.2 g, 58%). 1 H-NMR (DMSO-d6, 400MHz): δ8.19 (d, 2H), 8.09 (s, 1H), 7.40-7.38 (m, 4H), 5.56 (s, 2H).

[0345] 1.1.2. Preparation of 3-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (Compound 201). JPEG2025120182000071.jpg36161 Reagents and conditions: (c) 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile, Pd(PPh3)4, Na2CO3, dioxane:H2O (8:1), room temperature to 105°C, 12 hours, 70%; (d) SnCl2.2H2O, concentrated HCl, EtOH, room temperature to 50°C, 3 hours, 45%.

[0346] Step 1: Preparation of 3-(6-amino-1-(4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile

[0347] The compound of Example 1-1 (S3, 1.0 g, 3.28 mmol) was dissolved in 1,4-dioxane and HO, and 3-(4,4,5,5-tetramethyl-1,3,2-dioxaneboron-2-yl)benzonitrile (1.12 g, 4.92 mmol), tetrakis(triphenylphosphine)palladium ((0)(Pd(PPh)), 758 mg, 0.65 mmol), and sodium carbonate (NaCO, 695 mg, 6.56 mmol) were added thereto. The reaction mixture was then stirred at 105 °C for 12 hours in a sealed tube, diluted with dichloromethane (DCM), and washed with distilled water. The mixture was then dried over magnesium sulfate, filtered, and concentrated. The concentrate was washed with ethyl acetate / hexane (EA / Hex) to obtain the intermediate compound (S4, 852 mg, 70%). 1 H-NMR (DMSO-d6, 400MHz): δ8.54(s, 1H), 8.51-8.49(m, 2H), 8.08(dd, 1H), 7.80(t, 1H), 7.42(d, 2H), 7.15(s, 2H), 5.62(s, 2H).

[0348] Step 2: Preparation of 3-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile

[0349] The intermediate compound (S4, 400 mg, 1.07 mmol) produced in Step 1 above was dissolved in ethanol (EtOH), and then tin(II) chloride dihydrate (SnCl2H2O, 730 mg, 3.23 mmol) and concentrated HCl (aq) (1.8 mL, 21.54 mmol) were added. The reaction mixture was then stirred at 50 °C for 3 hours, diluted with dichloromethane (DCM), and washed with distilled water. It was then dried over magnesium sulfate, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to yield the desired compound (201, 161 mg, 45%). 1 H-NMR (DMSO-d6, 400MHz): δ8.50(s, 1H), 8.48-8.45(m, 1H), 8.35(s, 1H), 8.07-8.04( m, 1H), 7.79(t, 1H), 7.07(s, 2H), 6.95(d, 2H), 6.47(d, 2H), 5.23(s, 2H), 5.04(s, 2H).

[0350] Example 1.2: Preparation of 3-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-methylbenzonitrile (Compound 205) JPEG2025120182000072.jpg36161 Reagents and conditions: (a) 2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile, Pd(PPh3)4, Na2CO3, dioxane:H2O (8:1), room temperature to 105°C, 12 hours, 81%; (b) SnCl2.2H2O, concentrated HCl, EtOH, room temperature to 50°C, 3 hours, 74%.

[0351] Step 1: Preparation of 3-(6-amino-1-(4-nitrobenzyl[3,4-d]pyrimidin-4-yl)-2-methylbenzonitrile

[0352] The same process as in Step 1 of the above Example 1.1.2 was used, except that in Step 1 of the above Example 1.1.2, 2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxyboron-2-yl)benzonitrile (120 mg, 0.49 mmol) was used instead of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (1.12 g, 4.92 mmol) to give intermediate compound S5 (103 mg, 81%). 1 H-NMR (DMSO-d6, 400MHz): δ8.21(d, 2H), 7.97(d, 1H), 7.90(s, 1H), 7.82(d, 1H), 7.56(t, 1H), 7.46(d, 2H), 7.13(s, 2H), 5.59(s, 2H), 2.52(s, 3H)

[0353] Step 2: Preparation of 3-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-methylbenzonitrile

[0354] The same process as in Step 2 of Example 1.1.2 above was carried out on the intermediate compound (S5, 103 mg, 0.26 mmol) from Step 1 above, except that the amounts of tin(II) chloride dihydrate (SnCl.2HO, 180 mg, 0.80 mmol) and concentrated HCl (aq) (0.45 mL, 5.34 mmol) were changed to give the desired compound (205.71 mg, 74%). 1 H-NMR (DMSO-d6, 400MHz): δ7.95(d, 1H), 7.79(d, 1H), 7.77(s, 1H), 7.54(t, 1H) , 7.05(s, 2H), 6.98(d, 2H), 6.48(d, 2H), 5.21(s, 2H), 5.04(s, 2H), 2.49(s, 3H).

[0355] Example 1.3: Preparation of 3-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile (Compound 202) JPEG2025120182000073.jpg36161 Reagents and conditions: (a) 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile, Pd(PPh3)4, Na2CO3, dioxane:H2O (8:1), room temperature to 105°C, 12 hours, 90%; (b) SnCl2.2H2O, concentrated HCl, EtOH, room temperature to 50°C, 3 hours, 92%.

[0356] Step 1: Preparation of 3-(6-amino-1-(4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile The same process as in Step 1 of the above Example 1.1.2 was carried out to give intermediate compound S6 (1.15 g, 90%), except that in Step 1 of the above Example 1.1.2, 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (1.21 g, 4.92 mmol) was used instead of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (1.12 g, 4.92 mmol), and the amounts of tetrakis(triphenylphosphine)palladium(0) (Pd(PPh), 758 mg, 0.65 mmol) and sodium carbonate (NaCO, 695 mg, 6.56 mmol) were changed. 1 H-NMR (DMSO-d6, 400MHz): δ8.22-8.15 (m, 4H), 8.10 (d, 1H), 7.61 (t, 1H), 7.43 (d, 2H), 7.21 (s, 2H), 5.60 (s, 2H).

[0357] Step 2: Preparation of 3-(6-amino-1-4(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile

[0358] The same process as in Step 2 of Example 1.1.2 above was carried out on the intermediate compound (S6, 144 mg, 0.37 mmol) from Step 1 above, except that the amounts of tin(II) chloride dihydrate (SnCl.2HO, 250 mg, 1.11 mol) and concentrated HCl(aq) (0.62 ml, 7.40 mmol) were changed to give the desired compound (202, 131 mg, 92%). 1 H-NMR (DMSO-d6, 400MHz): δ8.14(t, 2H) 7.96(d, 1H), 7.11(s, 2H), 6.96(d, 2H), 6.48(d, 2H), 5.22(s, 2H), 5.04(s, 2H).

[0359] Example 1.4: Preparation of 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)benzonitrile (Compound 101)

[0360] 1.4.1. Preparation of 6-chloro-9(4-nitrobenzyl)-9H-purin-2-amine JPEG2025120182000074.jpg3687Reagents and conditions: (a) 4-nitrobenzyl bromide, K2CO3, DMF, room temperature.

[0361] 6-Chloro-9H-purin-2-amine (S12, 3 g, 18.12 mmol) was dissolved in N,N-dimethylformamide (DMF) and 4-nitrobenzyl bromide (4.1 g, 19.30 mmol), and potassium carbonate (K2CO3, 3.76 g, 27.18 mmol) was added. The mixture was stirred at room temperature. The reaction mixture was then diluted with dichloromethane (DCM) and washed with distilled water. It was then dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the intermediate compound (S8, 395 mg, 42%). 1 H-NMR (DMSO-d6, 400MHz): δ8.27(s, 1H), 8.22-8.20(m, 2H), 7.47-7.45(m, 2H), 6.96(s, 2H), 5.46(s, 2H).

[0362] 1.4.2. Preparation of 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)benzonitrile JPEG2025120182000075.jpg28170 Reagents and conditions: (a) Pd(PPh3)4, Na2CO3 in H2O, 1,4-dioxane, microwave, 105 °C, 30 min. (b) SnCl22H2O, concentrated HCl(aq), EtOH, 55 °C, 4 h.

[0363] Step 1: Preparation of 3-(2-amino-9-(4-nitrobenzyl)-9H-purin-6-yl)benzonitrile

[0364] The intermediate compound (S8, 200 mg, 0.652 mmol) was dissolved in 1,4-dioxane, and 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (224 mg, 0.978 mmol), tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4, 151 mg, 0.131 mmol), and sodium carbonate (Na2CO3, 139 mg, 1.304 mmol) were added. The solution was heated to 105 °C using a microwave and stirred for 30 min. The reaction mixture was then diluted with dichloromethane (DCM) and washed with distilled water. It was then dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the intermediate compound (S9, 395 mg, 54%). 1 H-NMR (DMSO-d6, 400MHz): δ9.10-9.08(m, 1H), 9.05-9.02(m, 1H), 8.39(s, 1H), 8.22( d, 2H), 8.03-8.01(m, 1H), 7.80(t, 1H), 7.51-7.49(m, 2H), 6.73(s, 2H), 5.53(s, 2H).

[0365] Step 2: Preparation of 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)benzonitrile

[0366] The intermediate compound (S9, 244 mg, 0.624 mmol) produced in Step 1 above was dissolved in ethanol (EtOH), and tin(II) chloride dihydrate (SnCl2.2HO, 423 mg, 1.872 mmol) and concentrated HCl (aq) (1.1 mL, 12.48 mmol) were added. The solution was stirred at 55 °C for 4 h. The reaction mixture was then diluted with dichloromethane (DCM) and washed with distilled water. The solution was then dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the desired compound (101, 41 mg, 38%). 1 H-NMR (DMSO-d6, 400MHz): δ9.07(s, 1H), 9.02(d, 1H), 8.24(s, 1H), 8.00(d, 1H) , 7.78(t, 1H), 7.05-7.03(m, 2H), 6.69(br, 2H), 6.52-6.50(m, 2H), 5.11(d, 4H).

[0367] Example 1.5: Preparation of 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile (Compound 102) JPEG2025120182000076.jpg28161Reagents and conditions: (a) Pd(PPh3)4, Na2CO3 in H2O, 1,4-dioxane, microwave, 105 °C, 1 h. (b) SnCl2.2H2O, concentrated HCl(aq), EtOH, 55 °C, 4 h.

[0368] Step 1: Preparation of 3-(2-amino-9-(4-nitrobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile

[0369] The same process as in Step 1 of the above-mentioned Example 1.1 was carried out, except that in Step 1 of the above-mentioned Example 1.1, 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (224 mg, 0.978 mmol) was used instead of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (224 mg, 0.978 mmol) to give the intermediate compound (S10, 395 mg, 54%).1 H-NMR (DMSO-d6, 400MHz): δ8.29(s, 1H), 8.27-8.17(m, 2H), 8.13-8.07(m, 1H), 7.65-7.50(m, 4H), 6.78(d, 2H), 5.51(d, 2H).

[0370] Step 2: Preparation of 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile

[0371] The same process as in Step 2 of Example 1.1 above was carried out on the intermediate compound of Step 1 above (S10, 244 mg, 0.624 mmol) to give the desired compound (102, 52 mg, 48%). 1 H-NMR (DMSO-d6, 400MHz): δ8.22-8.13(m, 2H), 8.13-8.07(m, 1H), 7.57(t, 1H), 7.05(d, 2H), 6.73(s, 2H), 6.51(d, 2H), 5.11(d, 4H).

[0372] Example 1.6: Preparation of 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)-2-methylbenzonitrile (Compound 105) JPEG2025120182000077.jpg28161 Reagents and conditions: (a) Pd(PPh3)4, Na2CO3 in H2O, 1,4-dioxane, microwave, set at T = 105 °C, 1 h. (b) SnCl2.2H2O, concentrated HCl(aq), EtOH, 55 °C, 4 h.

[0373] Step 1: Preparation of 3-(2-amino-9-(4-nitrobenzyl)-9H-purin-6-yl)-2-methylbenzonitrile

[0374] The same process as in Step 1 of Example 1.1 above was carried out, except that 2-methyl-3-(4,4,5,5,5,-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (224 mg, 0.978 mmol) was used instead of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (224 mg, 0.978 mmol) in Step 1 of Example 1.1 above, to obtain intermediate compound (S11, 395 mg, 54%). 1 H-NMR (DMSO-d6, 400MHz): δ8.27-8.22(m, 2H), 7.83(d, 1H), 7.88(d, 1H), 7.65-7.52(m, 4H), 6.71(s, 2H), 5.50(s, 2H).

[0375] Step 2: Preparation of 3-(2-amino-9-(4-aminobenzyl)-9H-purin-6-yl)-2-methylbenzonitrile

[0376] The same process as in Step 2 of Example 1.1 above was carried out on the intermediate compound of Step 1 above (S11, 244 mg, 0.624 mmol) to give the desired compound (105, 52 mg, 48%). 1 H-NMR (DMSO-d6, 400MHz): δ8.10(d, 1H), 7.91(d, 1H), 7.83(d, 1H), 7.51(t, 1H), 7.06(d, 2H), 6.66(s, 2H), 6.55-6.46(m, 2H), 5.10(d, 4H), 3.32(s, 3H).

[0377] Example 1.7: Preparation of 3-(2-amino-9-(4-amino-2,6-difluorobenzyl)-9H-purin-6-yl)benzonitrile (Compound 107) JPEG2025120182000078.jpg36117 JPEG2025120182000079.jpg36117 Reagents and conditions: (a) 2-(bromomethyl)-1,3-difluoro-5-nitrobenzene, K2CO3, DMF, room temperature, overnight; (b) Pd(PPh3)4, K2CO3, 1,4-dioxane, H2O, 115 °C, 4 h; (c) SnCl2.2H2O, concentrated HCl(aq), EtOH, 55 °C, 4 h.

[0378] Step 1: Preparation of 6-chloro-9-(2,6-difluoro-4-nitrobenzyl)-9H-purin-2-amine

[0379] 6-Chloro-9H-purin-2-amine (S12, 500 mg, 2.95 mmol) was dissolved in dimethylformamide (DMF) and 2-(bromomethyl)-1,3-difluoro-5-nitrobenzene (817.4 mg, 3.24 mmol). Potassium carbonate (KCO, 611.6 mg, 4.43 mmol) was added, and the solution was stirred at room temperature for 1 day. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the intermediate compound (S13, 300 mg, 33%). 1 H-NMR (DMSO-d6, 400MHz): δ8.68 (s, 1H), 7.55 (d, 2H), 6.93 (s, 2H), 4.99 (s, 2H).

[0380] Step 2: Preparation of 3-(2-amino-9-(2,6-difluoro-4-nitrobenzyl)-9H-purin-6-yl)benzonitrile

[0381] The intermediate compound (S13, 150 mg, 0.44 mmol) produced in Step 1 above was dissolved in 1,4-dioxane, and 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (151 mg, 0.66 mmol), tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4, 25.4 mg, 0.022 mmol), and potassium carbonate (K2CO3, 121.6 mg, 0.88 mmol) were added. The mixture was heated to 115 °C and stirred for 4 h. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the intermediate compound (S14, 95.5 mg, 53.3%). 1 H-NMR (methanol-d4, 400 MHz): δ 8.98 (s, 1H), 8.93 (dd, 1H), 8.18 (s, 1H), 8.01-7.96 (m, 2H), 7.86 (dd, 1H), 7.70 (t, 1H), 5.56 (s, 2H).

[0382] Step 3: Preparation of 3-(2-amino-9-(4-amino-2,6-difluorobenzyl)-9H-purin-6-yl)benzonitrile

[0383] The intermediate compound (S14, 95.1 mg, 0.23 mmol) produced in Step 2 above was dissolved in ethanol (EtOH), and tin(II) chloride dihydrate (SnCl.2HO, 263.4 mg, 1.17 mmol) was added thereto, and the solution was stirred at 55° C. for 4 hours. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the desired compound (107, 38.7 mg, 44.6%). 1 H-NMR (DMSO-d6, 400MHz): δ9.06(s, 1H), 8.99(dd, 1H), 8.05(s, 1H), 7.98(dd , 1H), 7.78(t, 1H), 6.67(s, 2H), 6.25-6.18(m, 2H), 5.89(s, 2H), 5.17(s, 2H).

[0384] Example 1.8: Preparation of 3-(2-amino-9-(4-amino-2,6-difluorobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile (Compound 108) JPEG2025120182000080.jpg28161 Reagents and conditions: (a) Pd(PPh3)4, K2CO3 1,4-dioxane, H2O, 115 °C, overnight; (b) SnCl2.2H2O, concentrated HCl(aq), EtOH, 55 °C, 3 h.

[0385] Step 1: Preparation of 3-(2-amino-9-(2,6-difluoro-4-nitrobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile

[0386] The same process as in Step 2 of Example 1.4 above was carried out, except that 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (130.5 mg, 0.53) was used instead of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (151 mg, 0.66 mmol), to give intermediate compound S15 (100 mg, 53.4%). 1 H-NMR (CDCl3, 400MHz): δ8.15-8.13(m, 1H), 7.90-7.87(m, 3H), 7.75-7.74(m, 1H), 7.40(t, 1H), 5.54(s, 2H), 5.12(s, 2H).

[0387] Step 2: Preparation of 3-(2-amino-9-(4-amino-2,6-difluorobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile

[0388] The intermediate compound (S15, 100 mg, 0.24 mmol) produced in Step 1 was dissolved in ethanol (EtOH), and tin(II) chloride dihydrate (SnCl.2HO, 265 mg, 1.18 mmol) was added to the solution, which was then stirred at 50° C. for 3 hours. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the desired compound (108, 32.9 mg, 34.7%). 1 H-NMR (methanol-d4, 400 MHz): δ 8.12-8.08 (m, 1H), 7.96-7.91 (m, 2H), 7.53 (t, 1H), 6.26 (d, 2H), 5.29 (s, 2H).

[0389] Example 1.9: Preparation of 3-(2-amino-9-(4-amino-2-fluorobenzyl)-9H-purin-6-yl)benzonitrile (Compound 109) JPEG2025120182000081.jpg36115 JPEG2025120182000082.jpg33110 Reagents and conditions: (a) 1-(bromomethyl)-2-fluoro-4-nitrobenzene, K2CO3, DMF, room temperature, overnight; (b) Pd(PPh3)4, K2CO3, 1,4-dioxane, H2O, 115°C, 5 hours; (c) SnCl2.2H2O, EtOH, 50°C, 1 hour.

[0390] Step 1: Preparation of 6-chloro-9-(2-fluoro-4-nitrobenzyl)-9H-purin-2-amine

[0391] 6-Chloro-9H-purin-2-amine (S12, 500 mg, 2.95 mmol) was dissolved in dimethylformamide (DMF), and 1-(bromomethyl)-2-fluoro-4-nitrobenzene (758.2 mg, 3.24 mmol) and potassium carbonate (KCO, 611.6 mg, 4.43 mmol) were added to the solution, which was stirred for 1 day. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the intermediate compound (S16, 742 mg, 77%). 1 H-NMR (DMSO-d6, 400MHz): δ8.23(s, 1H), 8.18(dd, 1H), 8.03(dd, 1H), 7.30(t, 1H), 6.97(s, 2H), 5.49(s, 2H).

[0392] Step 2: Preparation of 3-(2-amino-9-(2-fluoro-4-nitrobenzyl)-9H-purin-6-yl)-benzonitrile

[0393] The intermediate compound (S16, 300 mg, 0.93 mmol) produced in Step 1 was dissolved in 1,4-dioxane and HO. 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-benzonitrile (318.4 mg, 1.39 mmol), tetrakis(triphenylphosphine)palladium ((0)Pd(PPh3)4, 53.7 mg, 0.05 mmol), and potassium carbonate (K2CO3, 257 mg, 1.86 mmol) were added to the solution, which was then heated to 115 °C and stirred for 5 h. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the intermediate compound (S17, 162 mg, 44.8%). 1H-NMR (DMSO-d6, 400MHz): δ9.09(s, 1H), 9.02(dd, 1H), 8.35(s, 1H), 8.19(dd , 1H), 8.06-8.01(m, 2H), 7.81(t, 1H), 7.31(t, 1H), 6.75(s, 2H), 5.55(s, 2H).

[0394] Step 3: Preparation of 3-(2-amino-9-(4-amino-2-fluorobenzyl)-9H-purin-6-yl)benzonitrile

[0395] The intermediate compound (S17, 89 mg, 0.23 mmol) produced in the second step was dissolved in ethanol (EtOH), and then tin(II) chloride dihydrate (SnCl2.2HO, 257 mg, 1.14 mmol) was added to the solution, and the solution was stirred at 50 °C for 1 hour. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the desired compound (109, 19 mg, 23%). 1 H-NMR (DMSO-d6, 400MHz): δ9.06(s, 1H), 9.01(d, 1H), 8.14(s, 1H), 8.00(d, 1H) , 7.79(t, 1H), 6.98(t, 1H), 6.69(s, 2H), 6.32(d, 2H), 5.48(s, 2H), 5.16(s, 2H).

[0396] Example 1.10: Preparation of 3-(2-amino-9-(4-amino-2-fluorobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile (Compound 103) JPEG2025120182000083.jpg28161Reagents and conditions: (a) Pd(PPh3)4, K2CO3 1,4-dioxane, H2O, 115 °C, overnight; (b) SnCl2.2H2O, EtOH, 50 °C, 3 h.

[0397] Step 1: Preparation of 3-(2-amino-9-(2-fluoro-4-nitrobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile

[0398] The same process as in Step 2 of Example 1.6 above was carried out, except that 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (138 mg, 0.56 mmol) was used instead of 3-(4,4,5,5-teremethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (318 mg, 1.39 mmol), to give intermediate compound S18 (80 mg, 42.7%). 1 H-NMR (methanol-d4, 400 MHz): δ 8.21 (s, 1H), 8.15-8.07 (m, 3H), 7.98-7.93 (m, 1H), 7.59-7.52 (m, 2H), 5.60 (s, 2H).

[0399] Step 2: Preparation of 3-(2-amino-9-(4-amino-2-fluorobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile

[0400] The same process as in Step 3 of Example 1.6 above was carried out on the intermediate compound (S18, 65 mg, 0.16 mmol) from Step 1 above, except that the amount of tin(II) chloride dihydrate (SnCl.2HO, 180 mg, 0.80 mmol) was changed to give the target compound (103, 14.4 mg, 24%). 1 H-NMR (DMSO-d6, 400MHz): δ8.17(t, 1H), 8.10(t, 1H), 8.05(s, 1H), 7.58(t, 1H), 7.01(t, 1H), 6.74(s, 2H), 6.36-6.32(m, 2H), 5.50(s, 2H), 5.15(s, 2H).

[0401] Example 1.11: Preparation of 3-(2-amino-9-(4-amino-3-methylbenzyl)-9H-purin-6-yl)benzonitrile (Compound 104) JPEG2025120182000084.jpg36115 JPEG2025120182000085.jpg33110 Reagents and conditions: (a) 4-(bromomethyl)-2-methyl-1-nitrobenzene, K2CO3, DMF, room temperature, overnight; (b) Pd(PPh3)4, K2CO3, 1,4-dioxane, H2O, 115°C, 2 hours; (c) SnCl2.2H2O, EtOH, 55°C, overnight.

[0402] Step 1: Preparation of 6-chloro-9-(3-methyl-4-nitrobenzyl)-9H-purin-2-amine

[0403] 6-Chloro-9H-purin-2-amine (S12, 846 mg, 4.99 mmol) was dissolved in N,N-dimethylformamide (DMF), 4-(bromomethyl)-2-methyl-1-nitrobenzene (1.263 g, 5.49 mmol) and potassium carbonate (KCO, 1.03 g, 7.49 mmol) were added, and the solution was stirred at room temperature for 1 day. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the intermediate compound (S19, 1.28 g, 80.5%). 1 H-NMR (CDCl3, 400MHz): δ7.97 (d, 1H), 7.77 (s, 1H), 7.22-7.18 (m, 2H), 5.30 (s, 2H), 2.59 (s, 3H).

[0404] Step 2: Preparation of 3-(2-amino-9-(3-methyl-4-nitrobenzyl)-9H-purin-6-yl)benzonitrile

[0405] The intermediate compound (S19, 278 mg, 0.87 mmol) produced in Step 1 was dissolved in 1,4-dioxane and HO. 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-benzonitrile (300 mg, 1.31 mmol), tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4, 50.3 mg, 0.043 mmol), and potassium carbonate (K2CO3, 240.4 mg, 1.74 mmol) were added to the solution. The solution was heated to 105 °C and stirred for 2 h. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the intermediate compound (S20, 143 mg, 42.7%). 1 H-NMR (CDCl3, 400MHz): δ9.07(s, 1H), 9.02(d, 1H), 7.98(d, 1H), 7.84(s, 1H), 7.78( d, 1H), 7.59-7.51(m, 1H), 7.50-7.44(m, 2H), 5.36(s, 2H), 5.08(s, 2H), 2.59(s, 3H).

[0406] Step 3: Preparation of 3-(2-amino-9-(4-amino-3-methylbenzyl)-9H-purin-6-yl)benzonitrile

[0407] The intermediate compound (S20, 143 mg, 0.37 mmol) produced in Step 2 above was dissolved in ethanol (EtOH), tin(II) chloride dihydrate (SnCl2.2HO, 418.6 mg, 1.86 mmol) was added, and the solution was stirred at 55 °C for 1 day. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the desired compound (104, 23.6 mg, 18%). 1H-NMR (DMSO-d6, 400MHz): δ9.08(s, 1H), 9.02(d, 1H), 8.23(s, 1H), 7.99(d, 1H), 7.79(t, 1H) , 6.96(s, 1H), 6.91(d, 1H), 6.86(s, 2H), 6.55(d, 1H), 5.11(s, 2H), 4.87(s, 2H), 2.01(s, 3H).

[0408] Example 1.12: Preparation of 3-(2-amino-9-(4-amino-3-methylbenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile (Compound 106) JPEG2025120182000086.jpg28158 Reagents and conditions: (a) Pd(PPh3)4, K2CO3, 1,4-dioxane, H2O, 115°C, 2 hours; (c) SnCl2.2H2O, EtOH, 55°C, overnight

[0409] Step 1: Preparation of 3-(2-amino-9-(3-methyl-4-nitrobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile

[0410] The intermediate compound (S19, 300 mg, 0.94 mmol) was dissolved in 1,4-dioxane and HO. 2-Fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (349 mg, 1.41 mmol), tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4, 54.3 mg, 0.05 mmol), and potassium carbonate (K2CO3, 260 mg, 1.88 mmol) were added to the solution. The solution was heated to 115 °C and stirred for 2 h. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the intermediate compound (S21, 298.2 mg, 78.6%). 1H-NMR (CDCl3, 400MHz): δ8.22-8.16(m, 1H), 7.99(d, 1H), 7.83(s, 1H), 7.79(t , 1H), 7.70-7.64(m, 1H), 7.43(t, 1H), 7.25(s, 1H), 5.13(s, 2H), 2.60(s, 3H).

[0411] Step 2: Preparation of 3-(2-amino-9-(4-amino-3-methylbenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile

[0412] The intermediate compound (S21, 298.2 mg, 0.74 mmol) produced in Step 1 above was dissolved in ethanol (EtOH), and then tin(II) chloride dihydrate (SnCl2.2HO, 835 mg, 3.7 mmol) was added to the solution, which was stirred at 55 °C for 1 day. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the desired compound (106, 98 mg, 35.5%). 1 H-NMR (DMSO-d6, 400MHz): δ8.19-8.14(m, 2H), 8.10-8.07(m, 1H), 7.57(t, 1H), 6.97( s, 1H), 6.92(d, 1H), 6.73(s, 2H), 6.55(d, 1H), 5.09(s, 2H), 4.88(s, 2H), 2.01(s, 3H).

[0413] Example 1.13: Preparation of 3-(2-amino-9-(2,6-difluorobenzyl)-9H-purin-6-yl)benzonitrile (Compound 112) JPEG2025120182000087.jpg30153 Reagents and conditions: (a) 2,6-difluorobenzyl bromide, K2CO3, DMF, room temperature, 4 hours; (b) Pd(PPh3)4, K2CO3, 1,4-dioxane, H2O, 115 °C, overnight.

[0414] Step 1: Preparation of 6-chloro-9-(2,6-difluorobenzyl)-9H-purin-2-amine

[0415] 6-Chloro-9H-purin-2-amine (S12, 800 mg, 4.72 mmol) was dissolved in dimethylformamide (DMF), and 2,6-difluorobenzyl bromide (1.46 g, 7.08 mmol) and potassium carbonate (KCO, 1.95 g, 14.16 mmol) were added to the solution. The solution was stirred at room temperature for 4 hours. The reaction mixture was then filtered through a Celite pad, and the remaining liquid was diluted with ethyl acetate (EA). The solution was then washed with distilled water, dried over sodium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the intermediate compound (S22, 850 mg, 58%). 1 H-NMR (DMSO-d6, 400MHz): δ8.10 (s, 1H), 7.49-7.46 (m, 1H), 7.15 (t, 2H), 6.68 (s, 2H), 5.35 (s, 2H).

[0416] Step 2: Preparation of 3-(2-amino-9-(2,6-difluorobenzyl)-9H-purin-6-yl)benzonitrile

[0417] The intermediate compound (S22, 100 mg, 0.31 mmol) produced in Step 1 above was dissolved in 1,4-dioxane and HO. The solution was added to a solution of 3-(4,4,5,5-tetramethyl-1,3,2)-dioxaborolan-2-yl)benzonitrile (85.2 mg, 0.37 mmol), tetrakis(triphenylphosphine)palladium(0) (Pd(PPh) 18 mg, 0.02 mmol), and potassium carbonate (KCO 85.7 mg, 0.62 mmol). The solution was heated to 115 °C and stirred for 1 day. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the desired compound (S22, 81 mg, 89%). 1H-NMR (CDCl3, 400MHz): δ9.02(s, 1H), 8.98(dd, 1H), 7.87(s, 1H), 7.75(dd, 1H), 7.61(t, 1H), 7.39-7.32(m, 1H), 6.97(t, 2H), 5.39(s, 2H), 5.07(s, 2H).

[0418] Example 1.14: Preparation of 3-(2-amino-9-(2,6-difluorobenzyl)-9H-purin-6-yl)-2-fluorobenzonitrile (Compound 113) JPEG2025120182000088.jpg36137 Reagents and conditions: (a) Pd(PPh3)4, K2CO3, 1,4-dioxane, H2O, 115 °C, overnight.

[0419] The intermediate compound S22 (50 mg, 0.17 mmol) produced in Step 1 of Example 1.10 above was dissolved in 1,4-dioxane and HO. 2-Fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaneboron-2-yl)benzonitrile (49.4 mg, 0.20 mmol), tetrakis(triphenylphosphine)palladium(0) (Pd(PPh) (8.7 mg, 0.008 mmol), and potassium carbonate (KCO) (47 mg, 0.34 mmol) were added to the solution. The solution was heated to 115 °C and stirred for 1 day. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the desired compound (113, 45 mg, 70%). 1 H-NMR (DMSO-d6, 400MHz): δ8.18-8.11(m, 1H), 8.10-8.08(m, 2H), 7.57(t, 1H), 7.51-7.46(m, 1H), 7.16(t, 2H), 6.73(s, 2H), 5.39(s, 2H).

[0420] Example 1.15: Preparation of 3-(2-amino-9-(2,6-difluorobenzyl)-9H-purin-6-yl)-2-methylbenzonitrile (Compound 114) JPEG2025120182000089.jpg36140Reagents and conditions: (a) Pd(PPh3)4, K2CO3, 1,4-dioxane, H2O, 115 °C, 4 h.

[0421] The intermediate compound S22 (50 mg, 0.17 mmol) produced in Step 1 of Example 1.10 above was dissolved in 1,4-dioxane and HO. 2-Methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaneboron-2-yl)benzonitrile (48.6 mg, 0.20 mmol), tetrakis(triphenylphosphine)palladium(0) (Pd(PPh) (8.7 mg, 0.008 mmol), and potassium carbonate (KCO) (47 mg, 0.34 mmol) were added, and the solution was heated to 115 °C and stirred for 4 hours. The reaction mixture was then diluted with dichloromethane (DCM), washed with distilled water, dried over magnesium sulfate, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the desired compound S114 (47.6 mg, 74%). 1 H-NMR (methanol-d4, 400 MHz): δ 8.07 (s, 1H), 7.83 (dd, 1H), 7.77 (dd, 1H), 7.54-7.46 (m, 2H), 7.07 (t, 2H), 5.50 (s, 2H), 2.51 (s, 3H).

[0422] Example 1.16: Preparation of 3-[2-amino-9-[[4-amino-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]benzonitrile (Compound 110) JPEG2025120182000090.jpg69150Reagents and conditions: (a) K2CO3, DMAc, 25°C, 3 hours; (b) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110°C, 15 hours; (c) Fe, NH4Cl, EtOH / H2O, 80°C, 2 hours.

[0423] Step 1: Preparation of 6-chloro-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-2-amine.

[0424] To a solution of 4-(bromomethyl)-1-nitro-2-(trifluoromethyl)benzene (3 g, 10.56 mmol) in DMAc (50 mL) was added intermediate S12, 6-chloro-9H-purin-2-amine (1.79 g, 10.56 mmol), and K2CO3 (2.92 g, 21.12 mmol). The mixture was then stirred at 25 °C for 3 hours. After that, water (80 mL) was added to the mixture and extracted with ethyl acetate (80 mL × 3). The combined organic phases were washed with brine (150 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (80 g Sepa Flash® silica flash column, eluent: 20-100% ethyl acetate / petroleum ether gradient @ 60 mL / min). Compound S23, 6-chloro-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-2-amine (2.5 g, 6.62 mmol, 63% yield, 98.7% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6): δ8.27(s, 1H), 8.12(d, J=8.3Hz, 1H), 8.05(s, 1H), 7.67(dd, J=8.3Hz, 1H), 6.97(s, 2H), 5.50(s, 2H). MS: m / z=373.0 (M+1, ESI+).

[0425] Step 2: Preparation of 3-[2-amino-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]benzonitrile

[0426] To a solution of compound S23, 6-chloro-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-2-amine (2.5 g, 6.71 mmol, 1 equiv.) in dioxane (40 mL) and water (10 mL) was added (3-cyanophenyl)boronic acid (1.97 g, 13.42 mmol, 2 equiv.), Pd(dppf)Cl (490 mg, 670.80 μmol, 0.1 equiv.), and KCO (1.85 g, 13.42 mmol, 2 equiv.). The mixture was then stirred at 110° C. for 15 hours. The mixture was concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (40 g Sepa Flash® silica flash column, eluent: 20–60% ethyl acetate / petroleum ether gradient @ 50 mL / min). Compound S24, 3-[2-amino-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]benzonitrile (2.3 g, 4.56 mmol, 68% yield, 87% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6): δ9.08(t, J=1.5Hz, 1H), 9.02(td, J=1.4, 8.0Hz, 1H), 8.38(s, 1H), 8.13(d, J=8.4Hz, 1H), 8.08(d, J=1.2Hz, 1H), 8.01(td, J=1.3, 7.8Hz, 1H), 7.79(t, J=7.8Hz, 1H), 7.71(dd, J=1.5, 8.4Hz, 1H), 6.73(s, 2H), 5.56(s, 2H). MS: m / z=440.0 (M+1, ESI+).

[0427] Step 3: Preparation of 3-[2-amino-9-[[4-amino-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]benzonitrile To a solution of 3-[2-amino-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]benzonitrile (2.3 g, 4.55 mmol, 87% purity, 1 equiv.) in ethanol (30 mL) and water (6 mL), iron powder (1.27 g, 22.77 mmol, 5 equiv.) and NHCl (1.95 g, 36.44 mmol, 8 equiv.) were added. The mixture was then stirred at 80 °C for 2 h. The reaction mixture was filtered, and the filtrate was concentrated in vacuo. Water (80 mL) was added to the mixture, which was then extracted with ethyl acetate (80 mL × 3). The combined organic phase was filtered and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Welch Ultimate XB-CN 250*70*10 μm; mobile phase: [hexane-EtOH (0.1% ammonium hydroxide)]; B%: 30%-70%, 15 min) to give the product. The product was purified by recrystallization from THF (20 mL) at 70 °C to give the crude product. The crude product was purified by preparative HPLC (column: Welch Ultimate XB-SiOH 250*50*10 μm; mobile phase: [hexane-EtOH (0.1% ammonium hydroxide)]; B%: 5%-45%, 15 min). Compound 110, or 3-[2-amino-9-[[4-amino-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]benzonitrile (602.34 mg, 1.44 mmol, 32% yield, 97.62% purity), was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6)δ=9.07(s, 1H), 9.01(brd, J=7.9Hz, 1H), 8.30(s, 1H), 8.00(brd, J=7.7Hz, 1H), 7.78(t, J= 7.9Hz, 1H), 7.41(s, 1H), 7.29(brd, J=8.6Hz, 1H), 6.79(d, J=8.6Hz, 1H), 6.71(s, 2H), 5.65(s, 2H), 5.19(s, 2H). HRMS-TOF:410.1347

[0428] Example 1.17: Preparation of 3-[2-amino-9-[[4-amino-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]2-fluorobenzonitrile (Compound 111) JPEG2025120182000091.jpg44154 Reagents and conditions: (a) Pd(PPh3)4, K2CO3, dioxane / H2O, 110°C, 15 h; (b) Fe, NH4Cl, EtOH / H2O, 60°C, 1 h.

[0429] Step 1: Preparation of 3-[2-amino-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]-2-fluorobenzonitrile

[0430] A mixture of (3-cyano-2-fluorophenyl)boronic acid (265 mg, 1.61 mmol, 1.5 equiv.), intermediate compound S23 or 6-chloro-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-2-amine (400 mg, 1.07 mmol), Pd(PPh3)4 (124 mg, 107.00 μmol), and K2CO3 (296 mg, 2.14 mmol) in dioxane (10 mL) and water (1 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred under a nitrogen atmosphere at 100 °C for 15 h. The reaction mixture was concentrated under reduced pressure to remove dioxane and water, yielding a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g Sepa Flash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient @ 35 mL / min). The intermediate compound 3-[2-amino-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]-2-fluorobenzonitrile (200 mg, 437.31 μmol, 41% yield) was obtained as a yellow solid. MS: m / z=440.3 (M+1, ESI+).

[0431] Step 2: Preparation of 3-[2-amino-9-[[4-amino-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]2-fluorobenzonitrile.

[0432] To a solution of 3-[2-amino-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]-2-fluorobenzonitrile (300 mg, 655.97 μmol, 1 equiv.) in ethanol (15 mL) and water (5 mL) was added NH4Cl (292 mg, 5.25 mmol, 8 equiv.) and iron powder (183 mg, 3.28 mmol, 5 equiv.). The mixture was stirred at 60 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove ethanol, yielding a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 40%-70%, 10 min). Compound 111 or 3-[2-amino-9-[[4-amino-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]benzonitrile (75.32 mg, 175.81 μmol, 27% yield, 99.75% purity) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6): δ8.21(s, 1H), 8.17(t, 1H), 8.09(t, 1H), 7.57(t, 1H), 7 .41(s, 1H), 7.30(brd, 1H), 6.80(d, 1H), 6.75(s, 2H), 5.65(s, 2H), 5.17(s, 2H). HRMS-TOF:428.1244.

[0433] Example 1.18: Preparation of 6-[2-amino-9-[(4-aminophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (Compound 115) JPEG2025120182000092.jpg39153Reagents and conditions: (a) XPhos-Pd-G3, THF, 90°C, 15 hours. (b) Fe, NH4Cl, THF / H2O, 80°C, 2 hours.

[0434] Step 1: Preparation of 6-[2-amino-9-[(4-nitrophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0435] To a solution of intermediate compound S8, 6-chloro-9-[(4-nitrophenyl)methyl]purin-2-amine (800 mg, 2.63 mmol, 1 equiv.) in THF (20 mL) was added 6-tributylstannylpyridine-2-carbonitrile (2.06 g, 5.25 mmol, 2 equiv.) and XPhos-Pd-G3 (222 mg, 262.56 μmol, 0.1 equiv.). The mixture was then stirred at 90° C. for 15 hours. Water (30 mL) was added to the mixture, which was then extracted with ethyl acetate (30 mL×3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (12 g SepaFlash® silica flash column, 0-100% ethyl acetate / petroleum ether gradient @ 35 mL / min). The compound 6-[2-amino-9-[(4-nitrophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (167 mg, 372.27 μmol, 14% yield, 83% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6): δ8.91(dd, J=0.9, 8.1Hz, 1H), 8.37(s, 1H), 8.31-8.25(m, 1H), 8.22(d, J=8.7Hz, 2H), 8.18(dd, J=0.9, 7.6Hz, 1H), 7.50(d, J=8.7Hz, 2H), 6.85(brs, 1H), 5.53(s, 2H). MS: m / z=373.1 (M+1, ESI+).

[0436] Step 2: Preparation of 6-[2-amino-9-[(4-aminophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0437] To a solution of 6-[2-amino-9-[(4-nitrophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (160 mg, 429.72 μmol, 1 equiv.) in water (3 mL) and THF (9 mL) was added NH4Cl (184 mg, 3.44 mmol, 8 equiv.) and iron powder (120 mg, 2.15 mmol, 5 equiv.). The mixture was then stirred at 80 °C for 2 h. The reaction mixture was filtered, and saturated NaHCO3 solution (30 mL) was added to the filtrate, followed by extraction with ethyl acetate (80 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Waters Xbridge C18 150*50 mm*10 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 5%-41%, 10 min). Compound 115 or 6-[2-amino-9-[(4-aminophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (18.59 mg, 53.55 μmol, 12% yield, 95.94% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.92(brd, J=7.9Hz, 1H), 8.34-8.22(m, 2H), 8.16(brd, J=7.5Hz, 1H ), 7.05 (brd, J=8.0Hz, 2H), 6.81 (brs, 2H), 6.52 (brd, J=8.0Hz, 2H), 5.13 (brd, J=7.3Hz, 4H). HRMS-TOF:343.1415.

[0438] Example 1.19: Preparation of 6-[2-amino-9-[(4-amino-2-fluorophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (Compound 119) JPEG2025120182000093.jpg38153Reagents and conditions: (a) XPhos-Pd-G3, THF, 90 °C, 15 h. (b) Fe, NH4Cl, THF / H2O, 80 °C, 1.5 h.

[0439] Step 1: Preparation of 6-[2-amino-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile.

[0440] To a mixture of intermediate compound S16 or 6-chloro-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-2-amine (0.8 g, 2.48 mmol, 1 equiv.) in THF (10 mL) was added 6-tributylstannylpyridine-2-carbonitrile (1.8 g, 4.58 mmol, 1.85 equiv.) and XPhos-Pd-G3 (210 mg, 247.92 μmol, 0.1 equiv.). The mixture was then stirred at 90 °C for 15 h. Saturated aqueous potassium fluoride solution (30 mL) was added to the mixture, which was then extracted with ethyl acetate (30 mL × 3). The organic phases were combined and concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (25 g SepaFlash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient 50 mL / min). The compound 6-[2-amino-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (260 mg, 652.78 μmol, 26% yield, 98.0% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.90(d, J=7.9Hz, 1H), 8.33(s, 1H), 8.28(t, J=7.9Hz, 1H), 8.21-8 .15(m, 2H), 8.05(dd, J=1.8, 8.5Hz, 1H), 7.33(t, J=8.1Hz, 1H), 6.84(brs, 2H), 5.55(s, 2H). MS: m / z=391.0 (M+1, ESI+).

[0441] Step 2: Preparation of 6-[2-amino-9-[(4-amino-2-fluorophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0442] To a solution of 6-[2-amino-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (150 mg, 384.29 μmol, 1 equiv.) in water (1.5 mL) and THF (6 mL) was added iron powder (107 mg, 1.92 mmol, 5 equiv.) and NH4Cl (164 mg, 3.07 mmol, 8 equiv.). The mixture was then stirred at 80 °C for 1.5 h. The reaction mixture was filtered. Saturated NaHCO3 solution (30 mL) was added to the filtrate and extracted with ethyl acetate (30 mL × 2). The combined organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini NX-C18 (75*30 mm*3 μm); mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 13%-40%, 7 min). Compound 119 or 6-[2-amino-9-[(4-amino-2-fluoro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (21.61 mg, 58.63 μmol, 15% yield, 98.70% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.91(dd, J=1.0, 8.0Hz, 1H), 8.30-8.24(m, 1H), 8.16(d, J=7.8Hz, 1H), 8.14(s, 1H), 6.99(t, J=8.5Hz, 1H), 6.81(s, 2H), 6.41-6.28(m, 2H), 5.49(brs, 2H), 5.18(s, 2H). HRMS-TOF:361.1321.

[0443] Example 1.20: Preparation of 6-[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (Compound 123) JPEG2025120182000094.jpg38158Reagents and conditions: (a) XPhos-Pd-G3, THF, 90 °C, 15 h. (b) Fe, NH4Cl, THF / H2O, 80 °C, 1.5 h.

[0444] Step 1: Preparation of 6-[2-amino-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0445] A mixture of intermediate compound S13 or 6-chloro-9-[(2,6-difluoro-4-nitrophenyl)methyl]purin-2-amine (1 g, 2.94 mmol, 1 equiv.), 6-tributylstannylpyridine-2-carbonitrile (2.31 g, 5.87 mmol, 2 equiv.), and XPhos-Pd-G3 (250 mg, 293.54 μmol, 0.1 equiv.) in THF (15 mL) was degassed with nitrogen and heated at 90 °C for 15 h. Water (30 mL) was added to the mixture, and the mixture was extracted with ethyl acetate (30 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1). The compound 6-[2-amino-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (500 mg, 1.05 mmol, 36% yield, 85% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.86(d, J=8.0Hz, 1H), 8.31-8.23(m, 2H), 8.15(d, J=7.6Hz, 1H), 8.09(d, J=7.4Hz, 2H), 6.76(brs, 2H), 5.52(s, 2H) MS: m / z = 409.1 (M+1, ESI+).

[0446] Step 2: Preparation of 6-[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0447] To a solution of 6-[2-amino-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (200 mg, 489.81 μmol, 1 equiv.) in THF (8 mL) and water (2 mL) was added iron powder (137 mg, 2.45 mmol, 5 equiv.) and NH4Cl (210 mg, 3.92 mmol, 8 equiv.). The mixture was then stirred at 80 °C for 1.5 h. The reaction mixture was filtered, and the filtrate was added with saturated aqueous NaHCO3 (30 mL) and extracted with ethyl acetate (80 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Waters Xbridge 150*25 mm*5 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 18%-38%, 9 min). Compound 123 or 6-[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (31.07 mg, 79.80 μmol, 16% yield, 99.33% purity) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6) δ=8.88(dd, J=1.1, 8.1Hz, 1H), 8.29-8.22(m, 1H), 8.15(dd, J=1.0 , 7.8Hz, 1H), 8.03(s, 1H), 6.77(s, 2H), 6.22(d, J=10.5Hz, 2H), 5.88(s, 2H), 5.17(s, 2H). HRMS-TOF:379.1235.

[0448] Example 1.21: Preparation of 6-[2-amino-9-[(2,6-difluorophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (Compound 131) JPEG2025120182000095.jpg36146Reagents and conditions: (a) XPhos-Pd-G3, THF, 90°C, 15 hours.

[0449] To a solution of intermediate compound S22 or 6-chloro-9-[(2,6-difluorophenyl)methyl]purin-2-amine (200 mg, 676.42 μmol, 1 equiv.) and 6-tributylstannylpyridine-2-carbonitrile (266 mg, 676.42 μmol, 1 equiv.) in THF (10 mL) was added XPhos-Pd-G3 (57.26 mg, 67.64 μmol, 0.1 equiv.). The mixture was then stirred at 90 °C for 15 h. Aqueous potassium fluoride solution (30 mL) was added to the mixture, and the mixture was extracted with ethyl acetate (30 mL × 3). The combined organic phases were dried over anhydrous NaSO, filtered, and concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (12 g SepaFlash® silica flash column, 0-100% ethyl acetate / petroleum ether gradient @ 35 mL / min). Compound 133 or 6-[2-amino-9-[(2,6-difluorophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (40.26 mg, 110.81 μmol, 16% yield, 96.84% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.87(dd, J=1.0, 8.1Hz, 1H), 8.29-8.22(m, 1H), 8.18(s, 1H), 8.15 (dd, J=1.0, 7.7Hz, 1H), 7.58-7.39(m, 1H), 7.16(t, J=8.1Hz, 2H), 6.77(s, 2H), 5.42(s, 2H). HRMS-TOF:364.1121.

[0450] Example 1.22: Preparation of 6-[2-amino-9-[(4-amino-3-methyl-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (Compound 127) JPEG2025120182000096.jpg38155Reagents and conditions: (a) XPhos-Pd-G3, THF, 90°C, 15 hours. (b) Fe, NH4Cl, THF / H2O, 80°C, 1 hour.

[0451] Step 1: Preparation of 6-[2-amino-9-[(3-methyl-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0452] A mixture of intermediate compound S19 or 6-chloro-9-[(3-methyl-4-nitrophenyl)methyl]purin-2-amine (0.8 g, 2.51 mmol, 1 equiv.), 6-tributylstannylpyridine-2-carbonitrile (1.80 g, 4.58 mmol, 1.82 equiv.), and XPhos-Pd-G3 (212 mg, 251.01 μmol, 0.1 equiv.) in THF (10 mL) was degassed and heated to 90 °C under nitrogen for 15 h. Saturated aqueous potassium fluoride solution (30 mL) was added to the mixture, and the mixture was extracted with ethyl acetate (30 mL × 2). The combined organic phases were dried over anhydrous NaSO, filtered, and concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (25 g SepaFlash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient @ 50 mL / min). The compound 6-[2-amino-9-[(3-methyl-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (350 mg, 900.89 μmol, 35% yield, 99.45% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.91(dd, J=0.8, 8.0Hz, 1H), 8.36(s, 1H), 8.32-8.24(m, 1H), 8.17(dd, J=0. 9, 7.7Hz, 1H), 7.98(d, J=8.4Hz, 1H), 7.41(s, 1H), 7.28(brd, J=8.4Hz, 1H), 6.83(s, 2H), 5.45(s, 2H). MS: m / z=387.1 (M+1, ESI+).

[0453] Step 2: Preparation of 6-[2-amino-9-[(4-amino-3-methyl-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0454] To a solution of 6-[2-amino-9-[(3-methyl-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (150 mg, 388.23 μmol, 1 equiv.) in water (1 mL) and THF (4 mL) was added iron powder (108 mg, 1.94 mmol, 5 equiv.) and NH4Cl (166 mg, 3.11 mmol, 8 equiv.). The mixture was then stirred at 80 °C for 1 h. The reaction mixture was filtered. Saturated NaHCO3 solution (80 mL) was added to the filtrate and extracted with ethyl acetate (80 mL × 3). The combined organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini NX-C18 75*30 mm*3 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 10%-40%, 8 min). Compound 127 or 6-[2-amino-9-[(4-amino-3-methyl-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (37.55 mg, 101.86 μmol, 26% yield, >99% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6)δ=8.90(dd, J=1.0, 8.1Hz, 1H), 8.28-8.23(m, 1H), 8.22(s, 1H), 8.15(dd, J=1.1, 7.7Hz, 1H), 6.9 5(s, 1H), 6.91(dd, J=1.9, 8.1Hz, 1H), 6.79(s, 2H), 6.55(d, J=8.1Hz, 1H), 5.11(s, 2H), 4.89-4.83(m, 2H), 2.01(s, 3H). HRMS-TOF:357.1571.

[0455] Example 1.23: Preparation of 2-[2-amino-9-[(4-aminophenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (Compound 116) JPEG2025120182000097.jpg69153 Reagents and conditions: (a) CuI, DMSO, 110°C, 15 hours; (b) NaOMe, MeOH, 25°C, 0.5 hours; (c) Pd(OAc)2, Cy3P, K2CO3, dioxane, 130°C, 10 hours, MW; (d) Fe, NH4Cl, EtOH / H2O, 60°C, 1 hour.

[0456] Step 1: Preparation of methyl 3-((4-cyanopyridin-2-yl)sulfonyl)propanoate

[0457] A mixture of 2-bromopyridine-4-carbonitrile (4.4 g, 24.04 mmol, 1 equiv.), sodium 3-methoxy-3-oxo-propane-1-sulfinate (5.02 g, 28.85 mmol, 1.2 equiv.), and CuI (5.49 g, 28.85 mmol, 1.2 equiv.) in DMSO (50 mL) was degassed and purged with nitrogen three times, and the mixture was then stirred at 110 °C under a nitrogen atmosphere for 15 h. The mixture was then cooled to room temperature, diluted with ethyl acetate (50 mL), and filtered through a pad of silica. The filtrate was washed with water (2 × 20 mL), brine (20 mL), dried over anhydrous sodium sulfate, filtered through a pad of silica, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (ISCO®; 20 g Sepa Flash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient @ 35 mL / min). The compound methyl 3-[(4-cyano-2-pyridyl)sulfonyl]propanoate (3 g, 11.80 mmol, 49% yield) was obtained as a white solid. MS: m / z = 254.9 (M+1, ESI+).

[0458] Step 2: Preparation of (4-cyano-2-pyridyl)sulfinyloxysodium

[0459] To a solution of methyl 3-[(4-cyano-2-pyridyl)sulfonyl]propanoate (1.1 g, 4.33 mmol, 1 equiv.) in methanol (10 mL) was added sodium methoxide (233 mg, 4.33 mmol, 1 equiv.). The mixture was stirred at 25 °C for 0.5 h. The reaction mixture was concentrated under reduced pressure to remove methanol and give a residue. The crude product of intermediate compound S25 or (4-cyano-2-pyridyl)sulfinyloxysodium (700 mg, crude) was obtained as a yellow solid and used in the next step without further purification. MS: m / z = 167.0 (M+1, ESI+).

[0460] Step 3: Preparation of 2-[2-amino-9-[(4-nitrophenyl)methyl]purin-6-yl]pyridine-4-carbonitrile

[0461] A mixture of intermediate compound S25 or (4-cyano-2-pyridyl)sulfinyloxysodium (374 mg, 1.97 mmol, 1.2 equiv.), intermediate compound S8 or 6-chloro-9-[(4-nitrophenyl)methyl]purin-2-amine (500 mg, 1.64 mmol, 1 equiv.), tricyclohexylphosphine (92 mg, 328.20 μmol, 0.2 equiv.), KCO (453 mg, 3.28 mmol, 2 equiv.), and Pd(OAc) (36 mg, 164.10 μmol, 0.1 equiv.) in dioxane (2 mL) was placed in a microwave tube. The sealed tube was heated at 130 °C in a microwave oven for 10 h. The reaction mixture was concentrated under reduced pressure to remove dioxane and give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g Sepa Flash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient @ 40 mL / min). The compound 2-[2-amino-9-[(4-nitrophenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (360 mg, 734.81 μmol, 45% yield, 76% purity) was obtained as a yellow solid. MS: m / z = 373.1 (M+1, ESI+).

[0462] Step 4: Preparation of 2-[2-amino-9-[(4-aminophenyl)methyl]purin-6-yl]pyridine-4-carbonitrile

[0463] To a solution of intermediate compound S26 or 2-[2-amino-9-[(4-nitrophenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (150 mg, 402.86 μmol, 1 equiv.) and NHCl (172 mg, 3.22 mmol, 8 equiv.) in ethanol (9 mL) and water (3 mL), iron powder (112 mg, 2.01 mmol, 5 equiv.) was added. The mixture was stirred at 60 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to remove ethanol and water, yielding a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 5%-25%, 10 min). Compound 116 or 2-[2-amino-9-[(4-aminophenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (22.85 mg, 64.35 μmol, 16% yield, 96.90% purity, 0.2 FA) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ = 9.01 (dd, 1H), 8.94 (s, 1H), 8.22 (s, 1H), 7.98 (dd, 1H), 7.04 (d, 2H), 6.74 (s, 2H), 6.51 (d, 2H), 5.13 (s.4H). HRMS-TOF:343.1415.

[0464] Example 1.24: Preparation of 2-[2-amino-9-[(4-amino-2-fluoro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (Compound 120) JPEG2025120182000098.jpg38154 Reagents and conditions: (a) Pd(OAc)2, Cy3P, K2CO3, dioxane, 130 °C, 10 h, MW; (b) Fe, NH4Cl, EtOH / H2O, 60 °C, 1 h.

[0465] Step 1: Preparation of 2-[2-amino-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile.

[0466] A mixture of intermediate compound S25 or (4-cyano-2-pyridyl)sulfinyloxysodium (424 mg, 2.23 mmol, 1.2 equiv.), intermediate compound S16 or 6-chloro-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-2-amine (600 mg, 1.86 mmol, 1 equiv.), tricyclohexylphosphine (104 mg, 371.88 μmol, 0.2 equiv.), KCO (513 mg, 3.72 mmol, 2 equiv.), and palladium acetate (42 mg, 185.94 μmol, 0.1 equiv.) in dioxane (4 mL) was placed in a microwave tube. The sealed tube was heated at 130 °C for 10 h in a microwave oven. The reaction mixture was concentrated under reduced pressure to remove dioxane and give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g Sepa Flash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient @ 35 mL / min). The compound 2-[2-amino-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (300 mg, 768.58 μmol, 41% yield) was obtained as a white solid. MS: m / z = 391.1 (M+1, ESI+).

[0467] Step 2: Preparation of 2-[2-amino-9-[(4-amino-2-fluoro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile

[0468] To a solution of 2-[2-amino-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (150 mg, 230.57 μmol, 1 equiv.) and NH₄Cl (98 mg, 1.84 mmol, 8 equiv.) in ethanol (15 mL) and water (5 mL) was added iron powder (64 mg, 1.15 mmol, 5 equiv.). The mixture was stirred at 60°C for 1 hour. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Waters Xbridge 150*25 mm*5 μm; mobile phase: [water (10 mM NH₄HCO₃)-ACN]; B%: 10%-40%, 9 min) to give the product in 87% purity. Then, it was purified again by preparative HPLC (column: Waters Xbridge C18 150*50 mm*10 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 8%-38%, 10 min). Finally, compound 120 or 2-[2-amino-9-[(4-amino-2-fluoro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (9.83 mg, 26.40 μmol, 11% yield, 97.25% purity) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6) δ = 9.02(d, 1H), 8.89(s, 1H), 8.13(s, 1H), 7.99(d, 1H), 6.98(t, 1H), 6.75(s, 2H), 6.33(d, 2H), 5.48(s, 2H), 5.17(s2H). HRMS-TOF:361.1319.

[0469] Example 1.25: Preparation of -[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (Compound 124) JPEG2025120182000099.jpg38154 Reagents and conditions: (a) Pd(OAc)2, Cy3P, K2CO3, dioxane, 130 °C, 10 h, MW; (b) Fe, NH4Cl, EtOH / H2O, 60 °C, 1 h. Step 1: 2-[2-amino-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (4-Cyano-2-pyridyl)sulfinyloxysodium (419 mg, 2.20 mmol, 1.5 equiv.), 6-chloro-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-2-amine (500 mg, 1.47 mmol, 1 equiv.), tricyclohexylphosphine (123 mg, 440.31 μmol, 0.3 equiv.), KCO (405 mg, 2.94 mmol, 2 equiv.), and palladium acetate (50 mg, 223.09 μmol, 0.15 equiv.) were taken in a microwave tube in dioxane (10 mL). The sealed tube was heated at 130 °C in a microwave for 10 h. The reaction mixture was concentrated under reduced pressure to remove dioxane and give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150*25mm*10μm; mobile phase: [water (0.225% FA)-ACN]; B%: 23%-53%, 10 min). The compound 2-[2-amino-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (120 mg, 293.89 μmol, 20% yield) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6) δ = 9.02 (d, 1H), 8.85 (s, 1H), 8.28 (s, 1H), 8.11-8.07 (m, 2H), 7.99 (d, 1H), 6.72 (s, 2H), 5.52 (s, 2H). MS:m / z=409.1(M+1, ESI+) Step 2: 2-[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile To a solution of 2-[2-amino-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (100 mg, 244.91 μmol, 1 equiv.) and NHCl (104 mg, 1.96 mmol, 8 equiv.) in ethanol (9 mL) and water (3 mL) was added iron powder (68 mg, 1.22 mmol, 5 equiv.). The mixture was stirred at 60° C. for 1 hour. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 13%-43%, 10 min) to give the product with 90% purity. Purification by a second preparative HPLC (column: Waters Xbridge 150*25 mm*5 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 22%-52%, 9 min) gave the compound 2-[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (34.57 mg, 89.87 μmol, 37% yield, 97.96% purity) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ = 9.01(dd, 1H), 8.87(s, 1H), 8.03(s, 1H), 7.99(dd, 1H), 6.72(s, 2H), 6.22(d, 2H), 5.88(s, 2H), 5.17(s, 2H). HRMS(TOF):379.1229

[0470] Example 1.26: Preparation of 2-[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (Compound 132) JPEG2025120182000100.jpg39158Reagents and conditions: (a) Pd(OAc)2, Cy3P, K2CO3, dioxane, 130 °C, 10 h, MW.

[0471] A mixture of intermediate compound S25 or (4-cyano-2-pyridyl)sulfinyloxysodium (231 mg, 1.22 mmol, 1.2 equiv.), intermediate compound S22 or 6-chloro-9-[(2,6-difluorophenyl)methyl]purin-2-amine (300 mg, 1.01 mmol, 1 equiv.), palladium acetate (22 mg, 101.46 μmol, 0.1 equiv.), tricyclohexylphosphine (56 mg, 202.93 μmol, 0.2 equiv.), and KCO (280 mg, 2.03 mmol, 2 equiv.) in dioxane (6 mL) was degassed and purged with nitrogen three times. The mixture was then stirred at 130 °C for 10 h under microwave conditions. The mixture was concentrated under reduced pressure to remove dioxane and give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30 mm*3 μm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 15%-45%, 7 min). Compound 132 or 2-[2-amino-9-[(2,6-difluorophenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (49.39 mg, 130.41 μmol, 13% yield, 93.37% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ = 9.01(dd, 1H), 8.87(s, 1H), 8.18(s, 1H), 7.99(dd, 1H), 7.54-7.43(m, 1H), 7.21-7.12(m, 2H), 6.73(s, 2H), 5.41(s, 2H). HRMS-TOF:364.1121.

[0472] Example 1.27: Preparation of 2-[2-amino-9-[(4-amino-3-methyl-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (Compound 128) JPEG2025120182000101.jpg38155Reagents and conditions: (a) Pd(OAc)2, Cy3P, K2CO3, dioxane, 130 °C, 10 h, MW; (b) Fe, NH4Cl, EtOH / H2O, 60 °C, 1 h.

[0473] Step 1: Preparation of 2-[2-amino-9-[(3-methyl-4-nitro-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile

[0474] A mixture of intermediate compound S25 or sodium 4-cyanopyridine-2-sulfinate (357 mg, 1.88 mmol, 1.2 equiv.), intermediate compound S19 or 6-chloro-9-[(3-methyl-4-nitrophenyl)methyl]purin-2-amine (500 mg, 1.57 mmol, 1 equiv.), palladium acetate (35 mg, 157.00 μmol, 0.1 equiv.), tricyclohexylphosphine (87 mg, 314.00 μmol, 0.2 equiv.), and KCO (433 mg, 3.14 mmol, 2 equiv.) in dioxane (2 mL) was degassed and purged with nitrogen three times. The mixture was then stirred at 130 °C for 10 h under microwave conditions. The reaction mixture was concentrated under reduced pressure to remove dioxane and give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g Sepa Flash® silica flash column, eluent: 0-10% ethyl acetate / petroleum ether gradient @ 30 mL / min). The compound 2-[2-amino-9-[(3-methyl-4-nitrophenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (300 mg, 535.76 μmol, 34% yield, 69% purity) was obtained as a yellow solid. MS: m / z = 387.3 (M+1, ESI+).

[0475] Step 2: Preparation of 2-[2-amino-9-[(4-amino-3-methyl-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile To a solution of 2-[2-amino-9-[(3-methyl-4-nitrophenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (150 mg, 388.23 μmol, 1 equiv.) and NHCl (7 M, 444 μL, 8 equiv.) in water (5 mL) and ethanol (15 mL), iron powder (108 mg, 1.94 mmol, 5 equiv.) was added. The mixture was stirred at 60 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove ethanol and water, yielding a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 8%-28%, 9 min). Compound 128 or 2-[2-amino-9-[(4-amino-3-methyl-phenyl)methyl]purin-6-yl]pyridine-4-carbonitrile (21.82 mg, 58.15 μmol, 15% yield, 96.70% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.02(d, 1H), 8.90(s, 1H), 8.23(s, 1H), 7.99(d, 1H), 6.95(s, 1) H), 6.93-6.90(m, 1H), 6.75(s, 2H), 6.55(d, 1H), 5.12(s, 2H), 4.87(s, 2H), 2.06(s, 3H). HRMS-TOF:357.1569.

[0476] Example 1.28: Preparation of 2-[2-amino-9-[[4-amino-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]pyridine-4-carbonitrile (Compound 135) JPEG2025120182000102.jpg42155Reagents and conditions: (a) Pd(OAc)2, Cy3P, K2CO3, dioxane, 130 °C, 10 h, MW; (b) Fe, NH4Cl, EtOH / H2O, 60 °C, 1 h.

[0477] Step 1: Preparation of 2-[2-amino-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]pyridine-4-carbonitrile.

[0478] A mixture of intermediate compound S25 or sodium 4-cyanopyridine-2-sulfinate (230 mg, 1.21 mmol, 1.5 equiv.), intermediate compound S23 or 6-chloro-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-2-amine (300 mg, 804.96 μmol, 1 equiv.), potassium carbonate (222 mg, 1.61 mmol, 2 equiv.), and Cy3P-Pd-G3 (52 mg, 80.50 μmol, 0.1 equiv.) in dioxane (5 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred under a nitrogen atmosphere at 120° C. for 16 hours. The reaction mixture was concentrated under reduced pressure to remove dioxane and give a residue. The residue was purified by preparative HPLC (column: Shimpak C18 150*25*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 34%-54%, 10 min). Compound 135 or 2-[2-amino-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]pyridine-4-carbonitrile (300 mg, 647.23 μmol, 80% yield, 95% purity) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6) δ=9.04(dd, J=0.7, 4.9Hz, 1H), 8.90(s, 1H), 8.37(s, 1H), 8.13(s, 1H), 8 .08(s, 1H), 8.01(dd, J=1.5, 5.0Hz, 1H), 7.71(dd, J=1.2, 8.4Hz, 1H), 6.80(s, 2H), 5.57(s, 2H).

[0479] Step 2: Preparation of 2-[2-amino-9-[[4-amino-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]pyridine-4-carbonitrile.

[0480] To a solution of 2-[2-amino-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]pyridine-4-carbonitrile (300 mg, 681.29 μmol, 1 equiv.) in ethanol (15 mL) and water (5 mL), iron powder (190 mg, 3.41 mmol, 5 equiv.) and NHCl (292 mg, 5.45 mmol, 8 equiv.) were added. The mixture was stirred at 60°C for 1 hour. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Shimpak C18 150*25*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 17%-47%, 10 min). Compound 135 or 2-[2-amino-9-[[4-amino-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]pyridine-4-carbonitrile (40.01 mg, 96.51 μmol, 14% yield, 98.98% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.02(d, 1H), 8.89(s, 1H), 8.29(s, 1H), 7.99(dd, 1H), 7.40(d , 1H), 7.29(dd, 1H), 6.82-6.74(m, 3H), 5.64(s, 2H), 5.20(s, 2H)HRMS-TOF:411.1281.

[0481] Example 1.29: Preparation of 5-[2-amino-9-[(4-aminophenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (Compound 117) JPEG2025120182000103.jpg42158Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110°C, 15 hours. (b) Fe, NH4Cl, EtOH / H2O, 80°C, 1 hour.

[0482] Step 1: Preparation of 5-[2-amino-9-[(4-nitrophenyl)methyl]purin-6-yl]pyridine-3-carbonitrile.

[0483] A mixture of intermediate compound S8 or 6-chloro-9-[(4-nitrophenyl)methyl]purin-2-amine (200 mg, 656.40 μmol, 1 equiv.), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-3-carbonitrile (181 mg, 787.68 μmol, 1.2 equiv.), Pd(dppf)Cl (48 mg, 65.64 μmol, 0.1 equiv.), and KCO (181 mg, 1.31 mmol, 2 equiv.) in dioxane (10 mL) and water (0.5 mL) was degassed and then heated to 110 °C under nitrogen for 15 h. Water (130 mL) was added to the mixture, and the mixture was extracted with ethyl acetate (130 mL × 2). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative TLC (SiO, PE:EA=1:1). The compound 5-[2-amino-9-[(4-nitrophenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (150 mg, 330.34 μmol, 50% yield, 82% purity) was obtained as a yellow solid. MS: m / z=373 (M+1, ESI+).

[0484] Step 2: Preparation of 5-[2-amino-9-[(4-aminophenyl)methyl]purin-6-yl]pyridine-3-carbonitrile

[0485] To a solution of 5-[2-amino-9-[(4-nitrophenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (150 mg, 330.34 μmol, 1 equiv.) in ethanol (9 mL) and water (3 mL) was added iron powder (92 mg, 1.65 mmol, 5 equiv.) and NH4Cl (141 mg, 2.64 mmol, 8 equiv.). The mixture was then stirred at 80 °C for 1 h. The reaction mixture was filtered, and the filtrate was concentrated in vacuo. Water (50 mL) was then added to the mixture, and it was extracted with ethyl acetate (80 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Waters Xbridge 150*25 mm*5 μm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 14%-44%, 10 min). Compound 117 or 5-[2-amino-9-[(4-aminophenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (20.59 mg, 58.88 μmol, 18% yield, 98.45% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=10.01(d, J=1.8Hz, 1H), 9.30(s, 1H), 9.16(d, J=1.8Hz, 1H), 8.29(s , 1H), 7.05(brd, J=8.3Hz, 2H), 6.77(s, 2H), 6.52(d, J=8.3Hz, 2H), 5.14(s, 2H), 5.11(s, 2H). HRMS-TOF:343.1410.

[0486] Example 1.30: Preparation of 5-[2-amino-9-[(4-amino-2-fluoro-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (Compound 121) JPEG2025120182000104.jpg39158 Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110°C, 15 hours. (b) Fe, NH4Cl, EtOH / H2O, 80°C, 1 hour.

[0487] Step 1: Preparation of 5-[2-amino-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile.

[0488] A mixture of intermediate compound S16 or 6-chloro-9-[(2-fluoro-4-nitrophenyl)methyl]purin-2-amine (300 mg, 929.71 μmol, 1 equiv.), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-3-carbonitrile (320 mg, 1.39 mmol, 1.5 equiv.), KCO (257 mg, 1.86 mmol, 2 equiv.), and Pd(dppf)Cl (68 mg, 92.97 μmol, 0.1 equiv.) in dioxane (10 mL) and water (0.5 mL) was degassed and heated to 110 °C under nitrogen for 15 h. Water (30 mL) was added to the mixture, which was then extracted with ethyl acetate (30 mL × 3). The combined organic phase was washed with brine (50 mL x 2), dried over anhydrous NaSO, filtered, and concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (20 g SepaFlash® silica flash column, 0-80% ethyl acetate / petroleum ether gradient @ 35 mL / min). Compound 121, or 5-[2-amino-9-[(2-fluoro-4-nitrophenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (200 mg, 502.14 μmol, 54% yield, 98% purity), was obtained as a yellow solid. MS: m / z = 391.2 (M+1, ESI+).

[0489] Step 2: Preparation of 5-[2-amino-9-[(4-amino-2-fluoro-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile.

[0490] To a solution of 5-[2-amino-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (200 mg, 512.39 μmol, 1 equiv.) and NH₄Cl (220 mg, 4.10 mmol, 8 equiv.) in ethanol (9 mL) and water (3 mL) was added iron powder (143 mg, 2.56 mmol, 5 equiv.). The mixture was then stirred at 80°C for 1 hour. The mixture was concentrated in vacuo to give a residue. Water (30 mL) was added to the residue and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, and concentrated in vacuo to give a residue. The crude product was triturated with methanol (10 mL) and THF (1 mL) at 60°C for 30 minutes. Compound 121 or 5-[2-amino-9-[(4-amino-2-fluoro-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (60 mg, 166.51 μmol, 32% yield, 95.08% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=10.01(d, J=2.0Hz, 1H), 9.30(t, J=2.1Hz, 1H), 9.17(d, J=2.0Hz, 1H), 8.20(s, 1H), 6.99(t, J=8.4Hz, 1H), 6.79(s, 2H), 6.44-6.28(m, 2H), 5.50(s, 2H), 5.18(s, 2H). HRMS-TOF:361.1319.

[0491] Example 1.31: Preparation of 5-[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (Compound 125) JPEG2025120182000105.jpg39158 Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110°C, 2 h; (b) Fe, NH4Cl, EtOH / H2O, 80°C, 1 h.

[0492] Step 1: Preparation of 5-[2-amino-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile.

[0493] A mixture of intermediate compound S13 or 6-chloro-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-2-amine (200 mg, 587.07 μmol, 1 equiv.), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-3-carbonitrile (270 mg, 1.17 mmol, 2 equiv.), Pd(dppf)Cl (43 mg, 58.71 μmol, 0.1 equiv.), and KCO (162 mg, 1.17 mmol, 2 equiv.) in dioxane (10 mL) and water (2 mL) was placed in a microwave tube. The sealed tube was heated at 110 °C for 2 h under microwave conditions. Then, water (130 mL) was added to the mixture, and it was extracted with ethyl acetate (130 mL × 2). The combined organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g Sepa Flash® silica flash column, 0-100% ethyl acetate / petroleum ether gradient @ 40 mL / min). Compound 125, or 5-[2-amino-9-[(2,6-difluoro-4-nitrophenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (200 mg, 426.14 μmol, 72% yield, 87% purity), was obtained as a yellow solid. MS: m / z = 408.9 (M+1, ESI+).

[0494] Step 2: Preparation of 5-[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile.

[0495] To a solution of 5-[2-amino-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (200 mg, 489.81 μmol, 1 equiv.) in ethanol (8 mL) and water (2 mL), iron powder (137 mg, 2.45 mmol, 5 equiv.) and NH4Cl (210 mg, 3.92 mmol, 8 equiv.) were added. The mixture was then stirred at 80 °C for 1 h. The reaction mixture was then filtered, the filtrate was concentrated in vacuo, and the residue was then added to water (50 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (Column: Waters Vinridis Silica 2-EP OBD 50*150 mm*5 μm; Mobile phase: [Heptane-EtOH (0.1% NH₃·H₂O)]; B%: 5%-45%, 10 min). Compound 125 or 5-[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (16.22 mg, 40.31 μmol, 8% yield, 96.64% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.99(d, J=2.0Hz, 1H), 9.28(t, J=2.1Hz, 1H), 9.15(d, J=2.1 Hz, 1H), 8.09(s, 1H), 6.74(s, 2H), 6.22(d, J=10.4Hz, 2H), 5.88(s, 2H), 5.17(s, 2H). HRMS-TOF:379.1229.

[0496] Example 1.32: Preparation of 5-[2-amino-9-[(2,6-difluorophenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (Compound 133) JPEG2025120182000106.jpg38150Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110 °C, 15 h.

[0497] A mixture of intermediate compound S22 or 6-chloro-9-[(2,6-difluorophenyl)methyl]purin-2-amine (150 mg, 507.31 μmol, 1 equiv.), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-3-carbonitrile (140 mg, 608.78 μmol, 1.2 equiv.), Pd(dppf)Cl (37 mg, 50.73 μmol, 0.1 equiv.), and KCO (140 mg, 1.01 mmol, 2 equiv.) in dioxane (10 mL) and water (1 mL) was degassed and heated to 110 °C under nitrogen for 15 h. The mixture was concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Xtimate C18 150*40 mm*10 μm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 30%-60%, 10 min). Compound 133 or 5-[2-amino-9-[(2,6-difluorophenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (60.03 mg, 161.42 μmol, 32% yield, 96.75% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.98(d, J=1.7Hz, 1H), 9.27(s, 1H), 9.16(d, J=1.7Hz, 1H) , 8.24(s, 1H), 7.55-7.42(m, 1H), 7.16(t, J=8.1Hz, 2H), 6.74(s, 2H), 5.42(s, 2H). HRMS-TOF:364.1121.

[0498] Example 1.33: Preparation of 5-[2-amino-9-[(4-amino-3-methyl-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (Compound 129) JPEG2025120182000107.jpg39158Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110°C, 15 hours. (b) Fe, NH4Cl, EtOH / H2O, 80°C, 1 hour.

[0499] Step 1: Preparation of 5-[2-amino-9-[(3-methyl-4-nitro-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile

[0500] A mixture of intermediate compound S19 or 6-chloro-9-[(3-methyl-4-nitrophenyl)methyl]purin-2-amine (300 mg, 941.27 μmol, 1 equiv.), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-3-carbonitrile (325 mg, 1.41 mmol, 1.5 equiv.), Pd(dppf)Cl (69 mg, 94.13 μmol, 0.1 equiv.), and KCO (260 mg, 1.88 mmol, 2 equiv.) in dioxane (10 mL) and water (0.5 mL) was degassed and heated to 110 °C under nitrogen for 15 h. Water (80 mL) was then added to the mixture, which was then extracted with ethyl acetate (80 mL × 3). The combined organic phases were dried over anhydrous NaSO, filtered, and concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (12 g SepaFlash® silica flash column, eluent: 0-80% ethyl acetate / petroleum ether gradient @ 30 mL / min) to give the compound 5-[2-amino-9-[(3-methyl-4-nitro-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (300 mg, 750.84 μmol, 79% yield, 96.7% purity) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=10.02(d, J=2.0Hz, 1H), 9.31(t, J=2.1Hz, 1H), 9.17(d, J=2.0Hz, 1H), 8.41(s, 1H) , 7.97(d, J=8.4Hz, 1H), 7.41(s, 1H), 7.28(dd, J=1.5, 8.4Hz, 1H), 6.80(s, 2H), 5.45(s, 2H), 2.49(brs, 3H). MS: m / z=387 (M+1, ESI+).

[0501] Step 2: Preparation of 5-[2-amino-9-[(4-amino-3-methyl-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile

[0502] To a solution of 5-[2-amino-9-[(3-methyl-4-nitrophenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (300 mg, 722.1 μmol, 1 equiv.) in ethanol (9 mL) and water (3 mL) was added iron powder (202 mg, 3.61 mmol, 5 equiv.) and NH4Cl (310 mg, 5.78 mmol, 8 equiv.). The mixture was then stirred at 80 °C for 1 h. The reaction mixture was filtered, and the filtrate was concentrated in vacuo. Water (50 mL) was then added to the residue, which was then extracted with ethyl acetate (50 mL × 3). The combined organic phases were dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give a residue. The crude product was triturated with methanol (8 mL) at 25 °C for 30 min, then filtered, and the solid was dried under reduced pressure. Compound 129 or 5-[2-amino-9-[(4-amino-3-methyl-phenyl)methyl]purin-6-yl]pyridine-3-carbonitrile (36.5 mg, 99.14 μmol, 13% yield, 96.26% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=10.01(d, J=2.0Hz, 1H), 9.29(t, J=2.0Hz, 1H), 9.15(d, J=2.0Hz, 1H), 8.28(s, 1H) ), 6.96(s, 1H), 6.92(dd, J=1.7, 8.1Hz, 1H), 6.76(s, 2H), 6.56(d, J=8.1Hz, 1H), 5.12(s, 2H), 2.01(s, 3H). HRMS-TOF:357.1555.

[0503] Example 1.34: Preparation of 4-[2-amino-9-[(4-aminophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (Compound 118) JPEG2025120182000108.jpg39158 Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110 °C, 2 h, MW. (b) Fe, NH4Cl, EtOH / H2O, 80 °C, 1 h.

[0504] Step 1: Preparation of 4-[2-amino-9-[(4-nitrophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile.

[0505] A mixture of intermediate compound S8 or 6-chloro-9-[(4-nitrophenyl)methyl]purin-2-amine (200 mg, 656.40 μmol, 1 equiv.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonitrile (755 mg, 3.28 mmol, 5 equiv.), K2CO3 (181 mg, 1.31 mmol, 2 equiv.), and Pd(dppf)Cl2 (48 mg, 65.64 μmol, 0.1 equiv.) in dioxane (10 mL) and water (2 mL) was placed in a microwave tube. The sealed tube was heated at 110 °C for 2 h under microwave conditions. Water (130 mL) was then added to the mixture, which was then extracted with ethyl acetate (130 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (12 g SepaFlash® silica flash column, eluent: 0-80% ethyl acetate / petroleum ether gradient @ 35 mL / min). The compound 4-[2-amino-9-[(4-nitrophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (200 mg, 504.92 μmol, 77% yield, 94% purity) was obtained as a yellow solid. MS: m / z = 373.1 (M+1, ESI+).

[0506] Step 2: Preparation of 4-[2-amino-9-[(4-aminophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0507] To a solution of 4-[2-amino-9-[(4-nitrophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (200 mg, 537.14 μmol, 1 equiv.) and NHCl (230 mg, 4.30 mmol, 8 equiv.) in ethanol (8 mL) and water (2 mL) was added iron powder (150 mg, 2.69 mmol, 5 equiv.). The mixture was then stirred at 80° C. for 1 h. The reaction mixture was filtered, the filtrate was concentrated in vacuo, and the residue was then added to water (50 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phase was concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Waters Vinridis Silica 2-EP OBD 50*150 mm*5 μm; mobile phase: [Heptane-EtOH (0.1% NH3HO)]; B%: 5%-45%, 10 min). Compound 118 or 4-[2-amino-9-[(4-aminophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (38.42 mg, 107.37 μmol, 20% yield, 92.28% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6)δ=9.05(s, 1H), 8.98(d, J=5.3Hz, 1H), 8.89(dd, J=1.4, 5.2Hz, 1H), 8.33 (s, 1H), 7.05(brd, J=8.3Hz, 2H), 6.82(s, 2H), 6.52(d, J=8.4Hz, 2H), 5.14(s, 2H), 5.11(s, 2H). HRMS-TOF:343.1413.

[0508] Example 1.35: Preparation of 4-[2-amino-9-[(4-amino-2-fluoro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (Compound 122) JPEG2025120182000109.jpg39158 Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110 °C, 2 h, MW. (b) Fe, NH4Cl, THF / H2O, 80 °C, 1 h.

[0509] Step 1: Preparation of 4-[2-amino-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0510] A mixture of intermediate compound S16 or 6-chloro-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-2-amine (300 mg, 929.71 μmol, 1 equiv.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonitrile (428 mg, 1.86 mmol, 2 equiv.), KCO (257 mg, 1.86 mmol, 2 equiv.), and Pd(dppf)Cl (68 mg, 92.97 μmol, 0.1 equiv.) in dioxane (8 mL) and water (2 mL) was placed in a microwave tube. The sealed tube was heated at 110 °C for 2 h under microwave conditions. The mixture was concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (12 g SepaFlash® silica flash column, eluent: 0-50% ethyl acetate / petroleum ether gradient @ 60 mL / min) to give the compound 4-[2-amino-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (200 mg, 478.83 μmol, 52% yield, 93.45% purity) as a yellow solid. 1 H NMR (400MHz, DMSO-d6)δ=9.04(s, 1H), 8.99(d, J=4.6Hz, 1H), 8.88(dd, J=1.5, 5.1Hz, 1H), 8.41(s, 1H), 8 .18(dd, J=2.1, 9.9Hz, 1H), 8.04(dd, J=1.9, 8.6Hz, 1H), 7.35(t, J=8.0Hz, 1H), 6.86(s, 2H), 5.56(s, 2H). MS: m / z=391.1 (M+1, ESI+).

[0511] Step 2: Preparation of 4-[2-amino-9-[(4-amino-2-fluoro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0512] To a solution of 4-[2-amino-9-[(2-fluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (200 mg, 512.39 μmol, 1 equiv.) in ethanol (9 mL) and water (3 mL) was added iron powder (143 mg, 2.56 mmol, 5 equiv.) and NH4Cl (219 mg, 4.10 mmol, 8 equiv.). The mixture was then stirred at 80 °C for 1 hour. The reaction mixture was filtered, and the filtrate was concentrated in vacuo. Water (80 mL) was added to the residue, which was then extracted with ethyl acetate (80 mL × 3). The combined organic phase was washed with brine (100 mL × 2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini NX-C18 (75*30 mm*3 μm); mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 18%-48%, 8 min). Compound 122 or 4-[2-amino-9-[(4-amino-2-fluoro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (20.14 mg, 54.73 μmol, 11% yield, 98.80% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6)δ=9.04(s, 1H), 8.97(d, J=5.1Hz, 1H), 8.87(dd, J=1.5, 5.1Hz, 1H), 8.22(s, 1H) , 6.99(t, J=8.5Hz, 1H), 6.81(s, 2H), 6.34(s, 1H), 6.32(brd, J=1.2Hz, 1H), 5.48(s, 2H), 5.18(s, 2H). HRMS-TOF:361.1317.

[0513] Example 1.36: Preparation of 4-[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (Compound 126) JPEG2025120182000110.jpg39158 Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110°C, 2 hours. (b) Fe, NH4Cl, THF / H2O, 80°C, 0.5 hours.

[0514] Step 1: Preparation of 4-[2-amino-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0515] To a solution of intermediate compound S13 or 6-chloro-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-2-amine (160 mg, 469.66 μmol, 1 equiv.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonitrile (540 mg, 2.35 mmol, 5 equiv.) in dioxane (14 mL) and water (1.4 mL), K2CO3 (130 mg, 939.32 μmol, 2 equiv.) and Pd(dppf)Cl2 (34 mg, 46.97 μmol, 0.1 equiv.) were added. The mixture was stirred at 110 °C for 2 hours. Water (30 mL) was added to the mixture, which was then extracted with ethyl acetate (30 mL × 3). The combined organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g Sepa Flash® silica flash column, eluent: 0-42% ethyl acetate / petroleum ether gradient @ 40 mL / min). The compound 4-[2-amino-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (75 mg, 174.50 μmol, 37% yield, 95% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.02-9.00(m, 1H), 8.97(dd, J=0.6, 5.1Hz, 1H), 8.85(dd , J=1.6, 5.1Hz, 1H), 8.38(s, 1H), 8.10(d, J=7.5Hz, 2H), 6.79(s, 2H), 5.53(s, 2H). MS:m / z=409.1(M+1, ESI+)

[0516] Step 2: Preparation of 4-[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0517] To a solution of 4-[2-amino-9-[(2,6-difluoro-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (92 mg, 225.31 μmol, 1 equiv.) in ethanol (8 mL) and water (2 mL), iron powder (63 mg, 1.13 mmol, 5 equiv.) and NH4Cl (96 mg, 1.80 mmol, 8 equiv.) were added. The mixture was stirred at 80°C for 0.5 hours. The mixture was filtered, and the filtrate was concentrated in vacuo to give a residue. Water (40 mL) was added to the residue, which was then extracted with ethyl acetate (30 mL x 3). The combined organic layers were concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Welch Ultimate XB-CN 250*25*10 μm; mobile phase: [heptane-EtOH (0.1% NH₃·H₂O)]; B%: 25%-65%, 15 min). Compound 126 or 4-[2-amino-9-[(4-amino-2,6-difluoro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (38 mg, 99.43 μmol, 44% yield, 97.67% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.03(dd, J=0.8, 1.6Hz, 1H), 8.97(dd, J=0.9, 5.1Hz, 1H), 8.86(dd, J =1.6, 5.1Hz, 1H), 8.13(s, 1H), 6.79(s, 2H), 6.21(d, J=10.5Hz, 2H), 5.88(s, 2H), 5.18(s, 2H). HRMS-TOF:379.1231.

[0518] Example 1.37: Preparation of 4-[2-amino-9-[(2,6-difluorophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (Compound 134) JPEG2025120182000111.jpg37150Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110 °C, 15 h.

[0519] A mixture of intermediate compound S22 or 6-chloro-9-[(2,6-difluorophenyl)methyl]purin-2-amine (100 mg, 338.21 μmol, 1 equiv.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonitrile (117 mg, 507.31 μmol, 1.5 equiv.), KCO (94 mg, 676.42 μmol, 2 equiv.), and Pd(dppf)Cl (25 mg, 33.82 μmol, 0.1 equiv.) in dioxane (5 mL) and water (0.5 mL) was degassed and heated to 110 °C under nitrogen for 15 hours. Water (20 mL) was added to the mixture, and it was extracted with ethyl acetate (30 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (4 g SepaFlash® silica flash column, 0-100% ethyl acetate / petroleum ether gradient @ 30 mL / min) to give an impure product. The impure product was purified by preparative HPLC (column: Xtimate C18 150*40 mm*10 μm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 30%-60%, 10 min). Compound 134, or 4-[2-amino-9-[(2,6-difluorophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (18.9 mg, 50.98 μmol, 15% yield, 98.03% purity), was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.03(s, 1H), 8.98(dd, J=0.6, 5.1Hz, 1H), 8.86(dd, J=1.6, 5.1H z, 1H), 8.29(s, 1H), 7.54-7.43(m, 1H), 7.17(t, J=8.1Hz, 2H), 6.80(s, 2H), 5.44(s, 2H). HRMS-TOF:364.1120.

[0520] Example 1.38: Preparation of 4-[2-amino-9-[(4-amino-3-methyl-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (Compound 130) JPEG2025120182000112.jpg40155Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110 °C, 2 h, MW. (b) Fe, NH4Cl, THF / H2O, 80 °C, 1 h.

[0521] Step 1: Preparation of 4-[2-amino-9-[(3-methyl-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0522] A mixture of intermediate compound S19 or 6-chloro-9-[(3-methyl-4-nitrophenyl)methyl]purin-2-amine (100 mg, 313.76 μmol, 1 equiv.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonitrile (144 mg, 627.51 μmol, 2 equiv.), Pd(dppf)Cl (23 mg, 31.38 μmol, 0.1 equiv.), and KCO (87 mg, 627.51 μmol, 2 equiv.) in dioxane (3 mL) and water (0.3 mL) was degassed and purged with nitrogen three times, then stirred under microwave irradiation at 110 °C for 2 h. Water (30 mL) was added to the mixture, and the mixture was extracted with ethyl acetate (30 mL × 3). The combined organic phase was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g Sepa Flash® silica flash column, 0-80% ethyl acetate / petroleum ether gradient @ 35 mL / min). The compound 4-[2-amino-9-[(3-methyl-4-nitrophenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (104 mg, 242.26 μmol, 77% yield, 90% purity) was obtained as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ=9.05(s, 1H), 8.99(d, J=5.1Hz, 1H), 8.89(dd, J=1.6, 5.1Hz, 1H), 8.44(s, 1H), 7.97(d, J=8.4Hz, 1H), 7.41(s, 1H), 7.28(dd, J=1.4, 8.5Hz, 1H), 6.85(s, 2H), 5.45(s, 2H), 2.51(s, 3H). MS:m / z=387.1(M+1, ESI+)

[0523] Step 2: Preparation of 4-[2-amino-9-[(4-amino-3-methyl-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile

[0524] To a solution of 4-[2-amino-9-[(3-methyl-4-nitro-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (200 mg, 517.64 μmol, 1 equiv.) in ethanol (6 mL) and water (2 mL), iron powder (145 mg, 2.59 mmol, 5 equiv.) and NH4Cl (221 mg, 4.14 mmol, 8 equiv.) were added. The mixture was stirred at 80 °C for 1 h. The mixture was filtered, and the filtrate was concentrated in vacuo to give a residue. Water (40 mL) was added to the residue, which was then extracted with ethyl acetate (30 mL × 3). The combined organic layers were concentrated under reduced pressure to give a residue. The crude product was triturated with THF at 70 °C for 30 min. Compound 130 or 4-[2-amino-9-[(4-amino-3-methyl-phenyl)methyl]purin-6-yl]pyridine-2-carbonitrile (62 mg, 167.01 μmol, 32% yield, 96.73% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.04(s, 1H), 8.97(d, J=5.1Hz, 1H), 8.87(dd, J=1.5, 5.1Hz, 1H), 8.32(s, 1H), 6.99-6.88(m, 2H), 6.85(s, 2H), 6.54(d, J=8.0Hz, 1H), 5.12(s, 2H), 4.87(s, 2H), 2.00(s, 3H). HRMS-TOF:357.1568.

[0525] Example 1.39: Preparation of 4-[2-amino-9-[[4-amino-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]pyridine-2-carbonitrile (Compound 136) JPEG2025120182000113.jpg42155Reagents and conditions: (a) APhos-Pd-G3, K3PO4, DMAc, 60°C, 16 hours (b) Fe, NH4Cl, EtOH / H2O, 60°C, 1 hour.

[0526] Step 1: Preparation of 4-[2-amino-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]pyridine-2-carbonitrile

[0527] A mixture of intermediate compound S23 or 6-chloro-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-2-amine (50 mg, 134.16 μmol, 1 equiv.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonitrile (34 mg, 147.58 μmol, 1.1 equiv.), KPO (1.5 M aqueous solution, 90 μL, 1 equiv.), and APhos-Pd-G (8.52 mg, 13.42 μmol, 0.1 equiv.) in DMAc (2 mL) was degassed and purged with nitrogen three times. The mixture was then stirred under a nitrogen atmosphere at 60 °C for 16 h. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by preparative TLC (SiO, ethyl acetate / petroleum ether = 1:1). The compound 4-[2-amino-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]pyridine-2-carbonitrile (40 mg, 90.84 μmol, 68% yield) was obtained as a yellow oil. MS: m / z=441.1 (M+1, ESI+).

[0528] Step 2: Preparation of 4-[2-amino-9-[[4-amino-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]pyridine-2-carbonitrile To a solution of 4-[2-amino-9-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]pyridine-2-carbonitrile (40 mg, 90.84 μmol, 1 equiv.) in ethanol (15 mL) and water (5 mL), iron powder (25 mg, 454.2 μmol, 5 equiv.) and NH4Cl (39 mg, 726.72 μmol, 8 equiv.) were added. The mixture was stirred at 60 °C for 1 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Welch Ultimate XB-SiOH 250*50*10 μm; mobile phase: [hexane-EtOH (0.1% ammonium hydroxide)]; B%: 10%-50%, 15 min). Compound 136 or 4-[2-amino-9-[[4-amino-3-(trifluoromethyl)phenyl]methyl]purin-6-yl]pyridine-2-carbonitrile (8.9 mg, 20.71 μmol, 23% yield, 95.51% purity) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6)δ=9.03(s, 1H), 8.97(d, J=5.1Hz, 1H), 8.87(dd, J=1.3, 5.1Hz, 1H), 8.38(s, 1H) , 7.41(s, 1H), 7.29(brd, J=8.4Hz, 1H), 6.84(s, 2H), 6.79(d, J=8.5Hz, 1H), 5.66(s, 2H), 5.20(s, 2H). HRMS-TOF:411.1305.

[0529] Example 1.40: Preparation of 3-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (Compound 204)

[0530] 1.40.1. Preparation of 3-(6-amino-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (Intermediate Compound S107) JPEG2025120182000114.jpg33122Reagents and conditions: (a) Pd(PPh3)4, Na2CO3, dioxane / H2O, 100 °C, 16 h.

[0531] A mixture of 4-chloro-1H-pyrazolo[3,4-d]pyrimidin-6-amine (500 mg, 1 equiv.), (3-cyanophenyl)boronic acid (519 mg, 1.2 equiv.), Pd(PPh3)4 (340 mg, 0.1 equiv.), and Na2CO3 (625 mg, 2 equiv.) in dioxane (20 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred at 100 °C under a nitrogen atmosphere for 16 h. The reaction mixture was partitioned between ethyl acetate (100 mL) and water (30 mL). The organic phase was separated, washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The crude product, 3-(6-amino-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (500 mg, crude), was obtained as a yellow solid and used in the next step without further purification. MS: m / z = 237.1 (M+1, ESI+).

[0532] 1.40.2. Preparation of 3-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (Compound 204). JPEG2025120182000115.jpg36148Reagents and conditions: (a) K2CO3, DMF, 80°C, 16 h; (b) Fe, NH4Cl, EtOH / H2O, 60°C, 1 h.

[0533] Step 1: Preparation of 3-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile

[0534] To a solution of 4-(chloromethyl)-2-methyl-1-nitrobenzene (500 mg, 1.27 equiv.) and intermediate compound S107 or 3-(6-amino-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (500 mg, 1 equiv.) in DMF (10 mL) was added K2CO3 (585 mg, 2 equiv.). The mixture was stirred at 80 °C for 16 h. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 40%-70%, 10 min). The compound 3-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (200 mg, 24% yield) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6)δ=8.56-8.42(m, 3H), 8.10-8.06(m, 1H), 8.00-7.92(m, 1H), 7.8 6-7.76(m, 1H), 7.40-7.30(m, 1H), 7.24-7.08(m, 3H), 5.65-5.43(m, 2H), 2.49(s, 3H). MS: m / z=386.0 (M+1, ESI+).

[0535] Step 2: Preparation of 3-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (Compound 204)

[0536] To a solution of 3-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (150 mg, 1 equiv.) in ethanol (12 mL) and water (4 mL), iron powder (108 mg, 1.95 mmol, 5 equiv.) and NH4Cl (166 mg, 8 equiv.) were added. The mixture was stirred at 60°C for 1 hour. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 23%-53%, 10 min). Compound 204 or 3-[6-amino-1-[(4-amino-3-methyl-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (54.26 mg, 38% yield, 98.27% purity) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6) δ=8.54-8.49(m, 1H), 8.49-8.45(m, 1H), 8.37-8.31(m, 1H), 8.09-8.01(m, 1H), 7.84-7.70(m, 1H), 7.16-6 .99(m, 2H), 6.91-6.86(m, 1H), 6.86-6.80(m, 1H), 6.58-6.45(m, 1H), 5.33-5.14(m, 2H)), 4.98-4.57(m, 2H), 2.02-1.96(m, 3H).

[0537] Example 1.41: Preparation of 3-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile (Compound 206)

[0538] 1.41.1. Preparation of 4-chloro-1-(3-methyl-4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Intermediate Compound S108) JPEG2025120182000116.jpg42120Reagents and conditions: (a) K2CO3, DMAc, 80°C, 16 hours.

[0539] To a solution of 4-chloro-1H-pyrazolo[3,4-d]pyrimidin-6-amine (0.8 g, 1 eq.) and 4-(chloromethyl)-2-methyl-1-nitrobenzene (720 mg, 0.8) in DMAc (50 mL) was added K2CO3 (1.30 g, 2 eq.). The mixture was stirred at 80 °C for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The crude product, 4-chloro-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (800 mg, crude), was obtained as a yellow solid and used in the next step without further purification. MS: m / z = 354.9 (M+1, ESI+).

[0540] 1.41.2. Preparation of 3-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile (Compound 206) JPEG2025120182000117.jpg39153 Reagents and conditions: (a) Pd(PPh3)4, K2CO3, dioxane / H2O, 100°C, 16 h; (b) Fe, NH4Cl, EtOH / H2O, 60°C, 1 h.

[0541] Step 1: Preparation of 3-(6-amino-1-(3-methyl-4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile

[0542] A mixture of (3-cyano-2-fluorophenyl)boronic acid (434 mg, 1.2 equiv.), intermediate compound S108 or 4-chloro-1-[(3-methyl-4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (700 mg, 1 equiv.), Pd(PPh3)4 (253 mg, 219.63 μmol, 0.1 equiv.), and Na2CO3 (465 mg, 2 equiv.) in dioxane (10 mL) and water (2 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred at 110 °C under a nitrogen atmosphere for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g Sepa Flash® silica flash column, eluent: 0-80% ethyl acetate / petroleum ether gradient @ 40 mL / min). The compound 3-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (800 mg, 90% yield) was obtained as a yellow solid. MS: m / z=386.0 (M+1, ESI+).

[0543] Step 2: Preparation of 3-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile (Compound 206)

[0544] To a solution of 3-[6-amino-1-[(3-methyl-4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (200 mg, 1 eq.) in water (4 mL) and ethanol (12 mL), iron powder (138 mg, 5 eq.) and NH4Cl (212 mg, 8 eq.) were added. The mixture was stirred at 60 °C for 1 hour. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Synergi C18 150*25 mm*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 16%-46%, 11 min), affording a compound with 89% purity after the first purification. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150*25mm*10μm; mobile phase: [water (0.225% FA)-ACN]; B%: 18%-48%, 10 min). Compound 206 or 3-[6-amino-1-[(4-amino-3-methyl-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (111.35 mg, 59% yield, 97.78% purity) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6) δ=8.21-8.10(m, 2H), 8.00-7.94(m, 1H), 7.66-7.52(m, 1H), 7.16-7.07(m, 2 H), 6.92-6.80(m, 2H), 6.56-6.46(m, 1H), 5.34-5.15(m, 2H), 4.93-4.67(m, 2H), 2.06-1.88(m, 3H).

[0545] Example 1.42: Preparation of 3-(6-amino-1-(4-amino-3-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (Compound 210)

[0546] 1.42.1. Preparation of 4-chloro-1-(4-nitro-3-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Intermediate Compound S109) JPEG2025120182000118.jpg41132Reagents and conditions: (a) K2CO3, DMAc, 80°C, 16 hours.

[0547] To a solution of 4-(chloromethyl)-1-nitro-2-(trifluoromethyl)benzene (3.39 g, 1.2 equiv.) and 4-chloro-1H-pyrazolo[3,4-d]pyrimidin-6-amine (2 g, 1 equiv.) in DMF (20 mL) was added K2CO3 (3.26 g, 2 equiv.). The mixture was stirred at 80 °C for 16 h. The reaction mixture was concentrated under reduced pressure to remove DMF, leaving a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g Sepa Flash® silica flash column, 0-100% ethyl acetate / petroleum ether gradient @ 45 mL / min). The compound 4-chloro-1-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]pyrazolo[3,4-d]pyrimidin-6-amine (2 g, 45% yield) was obtained as a yellow solid. MS: m / z = 373.3 (M+1, ESI+).

[0548] 1.42.2. Preparation of 3-(6-amino-1-(4-amino-3-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (Compound 210) JPEG2025120182000119.jpg42160Reagents and conditions: (a) Pd(PPh3)4, K2CO3, dioxane / H2O, 100°C, 15 h; (b) Fe, NH4Cl, EtOH / H2O, 60°C, 2 h.

[0549] Step 1: Preparation of 3-(6-amino-1-(4-nitro-3-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile

[0550] A mixture of (3-cyanophenyl)boronic acid (236 mg, 1.5 equiv.), intermediate compound S109 or 4-chloro-1-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]pyrazolo[3,4-d]pyrimidin-6-amine (400 mg, 1 equiv.), Pd(PPh3)4 (124 mg, 0.1 equiv.), and K2CO3 (296 mg, 2 equiv.) in dioxane (10 mL) and water (1 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred at 100 °C under a nitrogen atmosphere for 15 h. The reaction mixture was concentrated under reduced pressure to remove dioxane and water, yielding a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g Sepa Flash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient @ 35 mL / min). The compound 3-[6-amino-1-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (300 mg, 63% yield) was obtained as a yellow oil. MS: m / z=440.1 (M+1, ESI+).

[0551] Step 2: Preparation of 3-(6-amino-1-(4-amino-3-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (Compound 210)

[0552] To a solution of 3-[6-amino-1-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (290 mg, 1 equiv.) in water (5 mL) and ethanol (15 mL) was added iron powder (190 mg, 5 equiv.) and NH4Cl (292 mg, 8 equiv.). The mixture was stirred at 60°C for 2 hours. The reaction mixture was concentrated under reduced pressure to remove ethanol, yielding a residue. The residue was purified by preparative HPLC (column: Unisil 3-100 C18 Ultra 150*50 mm*3 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 43%-63%, 10 min). Compound 210 or 3-[6-amino-1-[[4-amino-3-(trifluoromethyl)phenyl]methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (59.98 mg, 21% yield, 99.14% purity) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6) δ=8.52(s, 1H), 8.47(d, J=8.1Hz, 1H), 8.38(s, 1H), 8.06(d, J=7.7Hz, 1H), 7.79(t, J=7.8 Hz, 1H), 7.30(d, J=1.5Hz, 1H), 7.23-7.17(m, 1H), 7.10(s, 2H), 6.78(d, J=8.4Hz, 1H), 5.59(s, 2H), 5.30(s, 2H).

[0553] Example 1.43: Preparation of 3-(6-amino-1-(4-amino-3-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile (Compound 211) JPEG2025120182000120.jpg42161Reagents and conditions: (a) Pd(PPh3)4, K2CO3, dioxane / H2O, 100°C, 15 h; (b) Fe, NH4Cl, EtOH / H2O, 60°C, 2 h.

[0554] Step 1: Preparation of 3-(6-amino-1-(4-nitro-3-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile

[0555] A mixture of (3-cyano-2-fluorophenyl)boronic acid (265 mg, 1.5 equiv.), intermediate compound S109 or 4-chloro-1-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]pyrazolo[3,4-d]pyrimidin-6-amine (400 mg, 1 equiv.), Pd(PPh3)4 (124 mg, 0.1 equiv.), and K2CO3 (296 mg, 2 equiv.) in dioxane (10 mL) and water (1 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred under a nitrogen atmosphere at 100 °C for 15 h. The reaction mixture was concentrated under reduced pressure to remove dioxane and water, yielding a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g Sepa Flash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient @ 35 mL / min). The compound 3-[6-amino-1-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (350 mg, 71% yield) was obtained as a yellow oil. MS: m / z=458.3 (M+1, ESI+).

[0556] Step 2: Preparation of 3-(6-amino-1-(4-amino-3-(trifluoromethyl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile (Compound 211)

[0557] To a solution of 3-[6-amino-1-[[4-nitro-3-(trifluoromethyl)phenyl]methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (300 mg, 1 equiv.) in ethanol (15 mL) and water (5 mL), iron powder (183 mg, 5 equiv.) and NH4Cl (280 mg, 8 equiv.) were added. The mixture was stirred at 60°C for 2 hours. The reaction mixture was concentrated under reduced pressure to remove ethanol, leaving a residue. The residue was purified by preparative HPLC (column: Unisil 3-100 C18 Ultra 150*50 mm*3 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 43%-63%, 10 min). Compound 211 or 3-[6-amino-1-[[4-amino-3-(trifluoromethyl)phenyl]methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (41.58 mg, 15% yield, 98.23% purity) was obtained as a pale yellow solid. 1 H NMR (400MHz, DMSO-d6)δ=8.20-8.11(m, 2H), 8.00(d, J=3.5Hz, 1H), 7.60(t, J=7.8Hz, 1H), 7.30(d , J=1.6Hz, 1H), 7.23-7.17(m, 1H), 7.15(s, 2H), 6.78(d, J=8.4Hz, 1H), 5.60(s, 2H), 5.29(s, 2H).

[0558] Example 1.44: Preparation of 3-(6-amino-1-(2,6-difluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile (Compound 213) JPEG2025120182000121.jpg40164Reagents and conditions: (a) K2CO3, DMAc, 80 °C, 16 h; (b) Pd(PPh3)4, K2CO3, dioxane / H2O, 80 °C, 16 h.

[0559] Step 1: Preparation of 4-chloro-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (intermediate compound S110)

[0560] To a solution of 4-chloro-1H-pyrazolo[3,4-d]pyrimidin-6-amine (8 g, 47.18 mmol, 1 equiv.) and 2-(bromomethyl)-1,3-difluorobenzene (11.72 g, 56.61 mmol, 1.2 equiv.) in DMAc (100 mL) was added KCO (13.04 g, 94.36 mmol, 2 equiv.). The mixture was stirred at 80 °C for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g Sepa Flash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient @ 60 mL / min). Intermediate compound S110 or 4-chloro-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (4 g, 13.53 mmol, 29% yield) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6) δ = 8.35-7.77 (m, 1H), 7.53-7.41 (m, 1H), 7.40-7.31 (m, 2H), 7.19-6.98 (m, 2H), 5.60-5.20 (m, 2H).

[0561] Step 2: Preparation of 3-(6-amino-1-(2,6-difluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile (Compound 213)

[0562] A mixture of (3-cyano-2-fluorophenyl)boronic acid (251 mg, 1.5 equiv.), intermediate compound S110 or 4-chloro-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (300 mg, 1 equiv.), Pd(PPh3)4 (117 mg, 0.1 equiv.), and K2CO3 (280 mg, 2 equiv.) in dioxane (10 mL) and water (2 mL) was degassed and purged with nitrogen three times. The mixture was then stirred at 80 °C under a nitrogen atmosphere for 16 hours. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Unisil 3-100 C18 Ultra 150 * 50 mm * 3 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 45%-65%, 10 min). Compound 213 or 3-[6-amino-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (118.51 mg, 30% yield, 98.93% purity) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6) δ=8.22-8.09(m, 2H), 7.95(d, J=3.5Hz, 1H), 7.60(t, J=7 .8Hz, 1H), 7.52-7.39(m, 1H), 7.19(s, 2H), 7.13(t, J=8.0Hz, 2H), 5.45(s, 2H).

[0563] Example 1.45: Preparation of 3-(6-amino-1-(4-amino-2-fluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (Compound 209) JPEG2025120182000122.jpg33160Reagents and conditions: (a) K2CO3, DMAc, 60°C, 16 hours; (b) Pd(PPh3)4, K2CO3, dioxane / H2O, 80°C, 16 hours; (c) Fe, NH4Cl, EtOH / H2O, 80°C, 3 hours.

[0564] Step 1: Preparation of 4-chloro-1-(2-fluoro-4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (intermediate compound S111)

[0565] To a solution of 4-chloro-1H-pyrazolo[3,4-d]pyrimidin-6-amine (5.1 g, 1 equiv.) and 1-(bromomethyl)-2-fluoro-4-nitrobenzene (7 g, 1 equiv.) in DMF (20 mL) was added KCO (8.3 g, 2 equiv.). The mixture was stirred at 60 °C for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g Sepa Flash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient @ 40 mL / min). Intermediate compound S111, or 4-chloro-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (5 g, 52% yield), was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ = 8.15-8.11 (m, 1H), 8.08-8.06 (m, 1H), 8.05-8.01 (m, 1H), 7.47-7.38 (m, 2H), 7.33 (t, J = 8.0Hz, 1H), 5.57 (s, 2H). MS: m / z=323.3 (M+1, ESI+).

[0566] Step 2: Preparation of 3-(6-amino-1-(2-fluoro-4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile

[0567] A mixture of (3-cyanophenyl)boronic acid (341 mg, 1.5 equiv.), intermediate compound S111 or 4-chloro-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (500 mg, 1 equiv.), Pd(PPh3)4 (179 mg, 0.1 equiv.), and K2CO3 (428 mg, 2 equiv.) in dioxane (10 mL) and water (2 mL) was degassed and purged with nitrogen three times, after which the mixture was stirred under a nitrogen atmosphere at 100 °C for 16 h. The reaction mixture was concentrated under reduced pressure to remove dioxane and give a residue. The residue was purified by preparative HPLC (column: Unisil 3-100 C18 Ultra 150*50mm*3μm; mobile phase: [water (0.225% FA)-ACN]; B%: 45%-65%, 10 min). The compound 3-[6-amino-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (200 mg, 33% yield, 99% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6)δ=8.55-8.51(m, 1H), 8.49-8.46(m, 1H), 8.31-8.26(m , 1H), 8.18-8.01(m, 4H), 7.38-7.28(m, 1H), 7.22-7.05(m, 2H), 5.62(s, 2H). MS: m / z=389.9 (M+1, ESI+).

[0568] Step 3: Preparation of 3-(6-amino-1-(4-amino-2-fluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (Compound 209)

[0569] To a solution of 3-[6-amino-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (200 mg, 513.69 μmol, 1 equiv.) in ethanol (15 mL) and water (5 mL), iron powder (143 mg, 5 equiv.) and NH4Cl (219 mg, 8 equiv.) were added. The mixture was stirred at 80°C for 3 hours. The reaction mixture was concentrated under reduced pressure to remove ethanol and water, yielding a residue. The residue was purified by preparative HPLC (column: Unisil 3-100 C18 Ultra 150*50 mm*3 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 30%-50%, 10 min). Compound 209 or 3-[6-amino-1-[(4-amino-2-fluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (36.42 mg, 19% yield, 97.66% purity) was obtained as an off-white solid. 1 H NMR (400MHz, DMSO-d6)δ=8.51(s, 1H), 8.49-8.44(m, 1H), 8.35(s, 1H), 8.08-8.01(m, 1H), 7.79(t , J=7.8Hz, 1H), 7.07(s, 2H), 6.88(t, J=8.5Hz, 1H), 6.36-6.22(m, 2H), 5.40(s, 2H), 5.28(s, 2H).

[0570] Example 1.46: Preparation of 3-(6-amino-1-(4-amino-2,6-difluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)benzonitrile (Compound 207) JPEG2025120182000123.jpg59161 Reagents and conditions: (a1) NBS, AIBN, CCl4, 90°C, 15 hours; (a2) K2CO3, DMAc, 80°C, 3 hours; (b) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110°C, 2 hours, MW; (c) Fe, NH4Cl, EtOH / H2O, 80°C, 1 hour.

[0571] Step 1: Preparation of 4-chloro-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (intermediate compound S112)

[0572] To a solution of 1,3-difluoro-2-methyl-5-nitrobenzene (2 g, 11.55 mmol, 1 equiv.) in tetrachloromethane (50 mL), NBS (3.08 g, 17.33 mmol, 1.5 equiv.) and AIBN (190 mg, 1.16 mmol, 0.1 equiv.) were added. The mixture was then stirred at 90 °C for 15 hours. The reaction mixture was concentrated in vacuo to give a residue. To the residue, DMAc (50 mL), 4-chloro-1H-pyrazolo[3,4-d]pyrimidin-6-amine (1.96 g, 11.55 mmol, 1 equiv.), and KCO (3.19 g, 23.11 mmol, 2 equiv.) were added. The mixture was then stirred at 80 °C for 3 hours. Water (130 mL) and ethyl acetate (130 mL) were added to the mixture, and the layers were separated. The aqueous was extracted with ethyl acetate (130 mL × 2). The combined organic phases were washed with brine (150 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (40 g Sepa Flash® silica flash column, eluent: 0-30% ethyl acetate / petroleum ether gradient @ 60 mL / min). Intermediate compound S112, or 4-chloro-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (1.5 g, 4.26 mmol, 37% yield, 96.7% purity), was obtained as a yellow solid.

[0573] Step 2: Preparation of 3-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile

[0574] Intermediate compound S112 or 4-chloro-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (500 mg, 1.47 mmol, 1 equiv.), (3-cyanophenyl)boronic acid (323 mg, 2.20 mmol, 1.5 equiv.), K2CO3 (405 mg, 2.94 mmol, 2 equiv.), and Pd(dppf)Cl2 (107 mg, 146.77 μmol, 0.1 equiv.) were placed in a microwave tube in dioxane (6 mL) and water (2 mL). The sealed tube was heated in a microwave at 110 °C for 2 h. Then, water (80 mL) and ethyl acetate (80 mL) were added to the mixture, and the layers were separated. The aqueous was extracted with ethyl acetate (80 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (20 g SepaFlash® silica flash column, 0-100% ethyl acetate / petroleum ether gradient @ 50 mL / min eluent). The compound 3-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (200 mg, 441.90 μmol, 30% yield, 90% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.49(d, J=1.2Hz, 1H), 8.47-8.41(m, 1H), 8.35(s, 1H), 8.16-8. 07(m, 2H), 8.06(td, J=1.3, 7.8Hz, 1H), 7.79(t, J=7.9Hz, 1H), 7.16(s, 2H), 5.55(s, 2H). MS: m / z=408.0 (M+1, ESI+).

[0575] Step 3: Preparation of 3-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (Compound 207)

[0576] To a solution of 3-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (200 mg, 491.00 μmol, 1 equiv.) in ethanol (8 mL) and water (2 mL), iron powder (137 mg, 2.45 mmol, 5 equiv.) and NH4Cl (210 mg, 3.93 mmol, 8 equiv.) were added. The mixture was then stirred at 80 °C for 1 h. The reaction mixture was filtered, and the filtrate was concentrated in vacuo. Water (30 mL) was then added to the mixture, which was then extracted with ethyl acetate (30 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Welch Ultimate XB-CN 250*70*10 μm; mobile phase: [hexane-EtOH (0.1% ammonium hydroxide)]; B%: 30%-70%, 15 min). Compound 207 or 3-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]benzonitrile (62.95 mg, 163.33 μmol, 33% yield, 97.91% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.49(t, J=1.4Hz, 1H), 8.45(td, J=1.4, 7.9Hz, 1H), 8.29(s, 1H), 8.05(td, J=1.3, 7.7Hz, 1H), 7.79(t, J=7.8Hz, 1H), 7.07(s, 2H), 6.30-6.06(m, 2H), 5.80(s, 2H), 5.25(s, 2H).

[0577] Example 1.47: Preparation of 3-(6-amino-1-(4-amino-2-fluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile (Compound 203) JPEG2025120182000124.jpg39160Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110°C, 16 h; (b) Fe, NH4Cl, EtOH / H2O, 80°C, 3 h.

[0578] Step 1: Preparation of 3-(6-amino-1-(2-fluoro-4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile

[0579] A mixture of (3-cyano-2-fluorophenyl)boronic acid (383 mg, 2.33 mmol, 1.5 equiv.), intermediate compound S111 or 4-chloro-1-[(2-fluoro-4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (500 mg, 1 equiv.), Pd(PPh3)4 (179 mg, 0.1 equiv.), and K2CO3 (428 mg, 2 equiv.) in dioxane (10 mL) and water (2 mL) was degassed and purged with nitrogen three times. The mixture was then stirred under a nitrogen atmosphere at 100 °C for 16 h. The reaction mixture was concentrated under reduced pressure to remove dioxane and yield a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g Sepa Flash® silica flash column, eluent: 0-80% ethyl acetate / petroleum ether gradient @ 30 mL / min). The compound 3-[6-amino-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (300 mg, 48% yield) was obtained as a yellow solid. MS: m / z=407.9 (M+1, ESI+).

[0580] Step 2: Preparation of 3-(6-amino-1-(4-amino-2-fluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile (Compound 203)

[0581] To a solution of 3-[6-amino-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (209 mg, 1 eq.) in ethanol (15 mL) and water (5 mL), iron powder (143 mg, 5 eq.) and NH4Cl (219 mg, 8 eq.) were added. The mixture was stirred at 80°C for 3 hours. The reaction mixture was concentrated under reduced pressure to remove ethanol, leaving a residue. The residue was purified by preparative HPLC (column: Unisil 3-100 C18 Ultra 150*50 mm*3 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 33%-53%, 10 min). After obtaining the target compound with 80% purity, the residue was purified by a second preparative HPLC (column: Phenomenex Gemini-NX C18 75*30 mm*3 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 22%-52%, 7 min). Compound 203 or 3-[6-amino-1-[(4-amino-2-fluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (62.37 mg, 31% yield, 97.40% purity) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6)δ=8.21-8.10(m, 2H), 7.98(d, J=3.7Hz, 1H), 7.61(t, J=7.8Hz, 1H), 7.14(s, 2H), 6.97-6.84(m, 1H), 6.32(s, 1H), 6.31-6.28(m, 1H), 5.42(s, 2H), 5.27(s, 2H).

[0582] Example 1.48: Preparation of 3-(6-amino-1-(4-amino-2,6-difluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile (Compound 208) JPEG2025120182000125.jpg41160Reagents and conditions: (a) Pd(dppf)Cl4, K2CO3, dioxane / H2O, 110°C, 15 hours. (b) Fe, NH4Cl, EtOH / H2O, 80°C, 1 hour.

[0583] Step 1: Preparation of 3-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile

[0584] To a solution of intermediate compound S112 or 4-chloro-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (400 mg, 1.17 mmol, 1 equiv.) in water (2 mL) and dioxane (8 mL), (3-cyano-2-fluoro-phenyl)boronic acid (300 mg, 1.82 mmol, 1.55 equiv.), Pd(dppf)Cl (86 mg, 117.41 μmol, 0.1 equiv.), and KCO (325 mg, 2.35 mmol, 2 equiv.) were added. The mixture was then stirred at 110° C. for 15 hours. The mixture was then concentrated in vacuo to give a residue that was purified by flash silica gel chromatography (12 g SepaFlash® silica flash column, 20-100% ethyl acetate / petroleum ether gradient @ 50 mL / min). The compound 3-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (140 mg, 322.25 μmol, 27% yield, 97.9% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.20-8.12(m, 2H), 8.12-8.05(m, 2H), 7.99(d, J=3.6Hz, 1H), 7.60(t, J=7.8Hz, 1H), 7.22(s, 2H), 5.54(s, 2H). MS: m / z=426.1 (M+1, ESI+).

[0585] Step 2: Preparation of 3-(6-amino-1-(4-amino-2,6-difluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-2-fluorobenzonitrile (Compound 208)

[0586] To a solution of 3-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (140 mg, 329.16 μmol, 1 equiv.) in ethanol (8 mL) and water (2 mL), iron powder (92 mg, 1.65 mmol, 5 equiv.) and NH4Cl (140 mg, 2.63 mmol, 8 equiv.) were added. The mixture was then stirred at 80 °C for 1 h. The reaction mixture was filtered, and the filtrate was concentrated in vacuo. Water (50 mL) was added to the mixture, which was then extracted with ethyl acetate (30 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Welch Ultimate XB-CN 250*70*10 μm; mobile phase: [hexane-EtOH (0.1% NH₃·H₂O)]; B%: 35%-75%, 15 min). Compound 208, or 3-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-2-fluoro-benzonitrile (44.01 mg, 109.41 μmol, 33% yield, 98.28% purity), was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6)δ=8.14(dd, J=6.7, 7.7Hz, 2H), 7.93(d, J=3.6Hz, 1H), 7. 60(t, J=7.8Hz, 1H), 7.13(s, 2H), 6.27-6.07(m, 2H), 5.80(s, 2H), 5.24(s, 2H).

[0587] Example 1.49: Preparation of 4-[6-amino-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (Compound 234) JPEG2025120182000126.jpg36161Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 80 °C, 16 h.

[0588] A mixture of intermediate compound S110 or 4-chloro-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (300 mg, 1 equiv.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonitrile (279 mg, 1.2 equiv.), KCO (279 mg, 2 equiv.), and Pd(dppf)Cl (74 mg, 0.1 equiv.) in dioxane (10 mL) and water (2 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred under a nitrogen atmosphere at 80 °C for 16 h. The reaction mixture was concentrated under reduced pressure to remove dioxane and give a residue. The residue was purified by preparative HPLC (column: Unisil 3-100 C18 Ultra 150*50mm*3μm; mobile phase: [water (0.225% FA)-ACN]; B%: 38%-58%, 10 min). Compound 234 or 4-[6-amino-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (124.39 mg, 34% yield, 99.96% purity) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6) δ=8.96(d, J=5.1Hz, 1H), 8.58(d, J=0.6Hz, 1H), 8.41-8.39(m, 1 H), 8.39-8.36(m, 1H), 7.51-7.41(m, 1H), 7.25(s, 2H), 7.17-7.08(m, 2H), 5.47(s, 2H).

[0589] Example 1.50: Preparation of 4-(6-amino-1-(4-amino-2,6-difluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)picolinonitrile (Compound 226) JPEG2025120182000127.jpg38160Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 80°C, 16 h; (b) Fe, NH4Cl, EtOH / H2O, 80°C, 2 h.

[0590] Step 1: Preparation of 4-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile

[0591] To a solution of intermediate compound S112 or 4-chloro-1-[(2,6-difluoro-4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (400 mg, 1.17 mmol, 1 equiv.) in dioxane (10 mL) and water (2 mL) was added 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonitrile (540 mg, 2.35 mmol, 2 equiv.), Pd(dppf)Cl (86 mg, 117.41 μmol, 0.1 equiv.), and KCO (325 mg, 2.35 mmol, 2 equiv.). The mixture was then stirred at 100° C. for 15 hours. The mixture was concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (20 g Sepa Flash® silica flash column, eluent: 0-60% ethyl acetate / petroleum ether gradient @ 50 mL / min) to give the compound 4-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (300 mg, 625.68 μmol, 53% yield, 85% purity) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.96(d, J=5.1Hz, 1H), 8.58(s, 1H), 8.43(s, 1H), 8.3 8(dd, J=1.7, 5.1Hz, 1H), 8.09(brd, J=7.1Hz, 2H), 7.28(brs, 2H), 5.56(s, 2H). MS: m / z=409.1 (M+1, ESI+).

[0592] Step 2: Preparation of 4-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (Compound 226) To a mixture of 4-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (300 mg, 734.72 μmol, 1 equiv.) in water (3 mL) and ethanol (9 mL), iron powder (205 mg, 3.67 mmol, 5 equiv.) and NH4Cl (314 mg, 5.88 mmol, 8 equiv.) were added. The mixture was then stirred at 80 °C for 2 h. The reaction mixture was filtered, and the filtrate was concentrated in vacuo. Water (30 mL) was added to the mixture, and the mixture was extracted with ethyl acetate (30 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Welch Ultimate XB-CN 250*70*10 μm; mobile phase: [hexane-ethanol (0.1% NH₃·H₂O)]; B%: 30%-70%, 15 min). Compound 226 or 4-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (70.80 mg, 178.73 μmol, 24% yield, 95.51% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.96(dd, J=0.8, 5.1Hz, 1H), 8.58(dd, J=0.8, 1.6Hz, 1H ), 8.46-8.24(m, 2H), 7.19(s, 2H), 6.29-6.08(m, 2H), 5.80(s, 2H), 5.26(s, 2H).

[0593] Example 1.51: Preparation of 4-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)picolinonitrile (Compound 230) JPEG2025120182000128.jpg36160Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110°C, 16 h; (b) Fe, NH4Cl, EtOH / H2O, 60°C, 3 h.

[0594] Step 1: Preparation of 4-(6-amino-1-(3-methyl-4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)picolinonitrile

[0595] A mixture of intermediate compound S108 or 4-chloro-1-[(3-methyl-4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (500 mg, 1 equiv.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonitrile (541 mg, 1.5 equiv.), KCO (433 mg, 2 equiv.), and Pd(dppf)Cl (114 mg, 0.1 equiv.) in dioxane (10 mL) and water (2 mL) was degassed and purged with N three times, then the mixture was stirred under a N atmosphere at 110 °C for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g Sepa Flash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient @ 30 mL / min). The compound 4-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (500 mg, 82.49% yield) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.99(dd, J=0.7, 5.1Hz, 1H), 8.56(s, 1H), 8.43(dd, J=1.7, 5.1Hz, 1H), 7.98-7.91(m, 1H), 7.39-7.31(m, 1H), 7.30-7.07(m, 3H), 5.68-5.49(m, 2H), 2.49-2.47(m, 3H). MS: m / z=387.1 (M+1, ESI+).

[0596] Step 2: Preparation of 4-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)picolinonitrile (Compound 230)

[0597] To a solution of 4-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (400 mg, 1 eq.) in ethanol (15 mL) and water (5 mL) was added iron powder (289 mg, 5 eq.) and NH4Cl (443 mg, 8 eq.). The mixture was stirred at 60°C for 3 hours. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30 mm*3 μm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 15%-45%, 11.5 min). Compound 230 or 4-[6-amino-1-[(4-amino-3-methyl-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (41.4 mg, 7% yield, 95.38% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.97(dd, J=0.6, 5.1Hz, 1H), 8.60(d, J=0.9Hz, 1H), 8.44(s, 1H), 8.40(dd, J=1.7, 5.1Hz, 1H), 7.17(s, 2H), 6.93-6.79(m, 2H), 6.52(d, J=8.1Hz, 1H), 5.24(s, 2H), 4.81(s, 2H), 1.99(s, 3H).

[0598] Example 1.52: Preparation of 6-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)picolinonitrile (Compound 215) JPEG2025120182000129.jpg54161Reagents and conditions: (a) NaOMe, MeOH, 25°C, 0.5 h; (b) Pd(OAc)2, K2CO3, Cy3P, dioxane, 120°C, 16 h; (c) Fe, NH4Cl, EtOH / H2O, 60°C, 3 h.

[0599] Step 1: Preparation of (6-cyano-2-pyridyl)sulfinyloxysodium (intermediate compound S113)

[0600] To a solution of sodium methoxide (212 mg, 1 eq.) in methanol (10 mL) was added methyl 3-[(6-cyano-2-pyridyl)sulfonyl]propanoate (1 g, 1 eq.). The mixture was stirred at 25° C. for 0.5 h. The reaction mixture was concentrated under reduced pressure to remove methanol and give a residue. The crude product of intermediate compound S113 or (6-cyano-2-pyridyl)sulfinyloxysodium (1 g, crude) was obtained as a yellow solid and used in the next step without further purification. MS: m / z=167.0 (M+1, ESI+).

[0601] Step 2: Preparation of 6-[6-amino-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile

[0602] A mixture of intermediate compound S113 or 4-chloro-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (500 mg, 1 equiv.), (6-cyano-2-pyridyl)sulfinyloxysodium (374 mg, 1.2 equiv.), palladium acetate (36 mg, 0.1 equiv.), Cy3P (92 mg, 320.2 equiv.), and K2CO3 (453 mg, 2 equiv.) in dioxane (5 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred under a nitrogen atmosphere at 120 °C for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g Sepa Flash® silica flash column, eluent: 70–100% ethyl acetate / petroleum ether gradient @ 35 mL / min). The compound 6-[6-amino-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (200 mg, 504.92 μmol, 31% yield, 94% purity) was obtained as a white solid. 1H NMR (400MHz, DMSO-d6)δ=8.66(dd, J=1.3, 8.0Hz, 1H), 8.50(s, 1H), 8.35-8.30(m, 1H), 8. 28-8.24(m, 1H), 8.20(d, J=8.9Hz, 2H), 7.41(d, J=8.9Hz, 2H), 7.20(s, 2H), 5.64(s, 2H). MS: m / z=373.1 (M+1, ESI+).

[0603] Step 3: Preparation of 6-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)picolinonitrile (Compound 215)

[0604] To a solution of 6-[6-amino-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (200 mg, 1 equiv.) in water (5 mL) and ethanol (15 mL) was added iron powder (150 mg, 5 equiv.) and NH4Cl (230 mg, 8 equiv.). The mixture was stirred at 60°C for 3 hours. The reaction mixture was concentrated under reduced pressure to remove ethanol and water to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30 mm*3 μm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 10%-40%, 11.5 min) to give the desired compound with 89% purity. The residue was then re-purified by preparative HPLC (column: Welch Ultimate XB-CN 250*50*10 μm; mobile phase: [hexane-EtOH (0.1% NH3 HO)]; B%: 20%-60%, 15 min) to give the desired compound with 91% purity. The residue was then purified by preparative HPLC (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 16%-46%, 10 min). Compound 215 or 6-[6-amino-1-[(4-aminophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (21.55 mg, 11% yield, 98.25% purity) was obtained as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ=8.65(d, J=7.6Hz, 1H), 8.39(s, 1H), 8.35-8.27(m, 1H), 8.26-8.22(m, 1H), 7.12(s, 2H), 6.94(brd, J=8.3Hz, 2H), 6.48(brd, J=8.3Hz, 2H), 5.25(s, 2H), 5.03(s, 2H).

[0605] Example 1.53: Preparation of 6-[6-amino-1-[(4-amino-2-fluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (Compound 219) JPEG2025120182000130.jpg36160Reagents and conditions: (a) Pd(OAc)2, K2CO3, Cy3P, dioxane, 120°C, 16 hours; (c) Fe, NH4Cl, THF / H2O, 80°C, 4 hours.

[0606] Step 1: Preparation of 6-[6-amino-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile

[0607] A mixture of intermediate compound S113 or (6-cyano-2-pyridyl)sulfinyloxysodium (495.01 mg, 2.60 mmol, 1.2 equiv.), intermediate compound S111 or 4-chloro-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (700 mg, 2.17 mmol, 1 equiv.), palladium acetate (48.70 mg, 216.93 μmol, 0.1 equiv.), Cy3P (121.67 mg, 433.86 μmol, 140.66 μL, 0.2 equiv.), and K2CO3 (599.64 mg, 4.34 mmol, 2 equiv.) in dioxane (5 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred at 120 °C under a nitrogen atmosphere for 16 h. The mixture was concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (12 g SepaFlash® silica flash column, 10-50% ethyl acetate / petroleum ether gradient @ 40 mL / min eluent). The compound 6-[6-amino-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (220 mg, 503.88 μmol, 26% yield, 89.4% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.66(dd, J=1.1, 7.9Hz, 1H), 8.49(s, 1H), 8.36-8.29(m, 1H), 8.28-8.23(m, 1H), 8.16(dd, J=2.3, 9.8Hz, 1H), 8.04(dd, J=1.8, 8.6Hz, 1H), 7.29(t, J=8.0Hz, 1H), 7.22(s, 2H), 5.64(s, 2H).

[0608] Step 2: Preparation of 6-[6-amino-1-[(4-amino-2-fluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (Compound 219)

[0609] To a mixture of 6-[6-amino-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (200 mg, 512.39 μmol, 1 equiv.) in THF (10 mL) and water (2 mL), iron powder (143 mg, 2.56 mmol, 5 equiv.) and NH4Cl (219.27 mg, 4.10 mmol, 8 equiv.) were added. The mixture was then stirred at 80 °C for 4 h. The reaction mixture was filtered, and 50 mL of saturated aqueous NaHCO3 was added to the filtrate, followed by extraction with ethyl acetate (50 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Welch Ultimate XB-CN 250*70*10 μm; mobile phase: [hexane-EtOH (0.1% NH₃·H₂O)]; B%: 35%-75%, 15 min). Compound 219, or 6-[6-amino-1-[(4-amino-2-fluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (56.69 mg, 151.63 μmol, 30% yield, 96.38% purity), was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.65(dd, J=1.2, 8.0Hz, 1H), 8.39(s, 1H), 8.34-8.29(m, 1H), 8.26-8.22(m, 1H) ), 7.13(s, 2H), 6.95-6.79(m, 1H), 6.31(dd, J=1.8, 5.4Hz, 1H), 6.28(s, 1H), 5.40(s, 2H), 5.29(s, 2H).

[0610] Example 1.54: Preparation of 6-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (Compound 223) JPEG2025120182000131.jpg38160Reagents and conditions: (a) Pd(OAc)2, K2CO3, Cy3P, dioxane, 120°C, 16 hours. (c) Fe, NH4Cl, THF / H2O, 80°C, 5 hours.

[0611] Step 1: Preparation of 6-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile

[0612] A mixture of intermediate compound S113 or sodium 6-cyanopyridine-2-sulfinate (530 mg, 2.79 mmol, 2 equiv.), intermediate compound S112 or 4-chloro-1-[(2,6-difluoro-4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (470 mg, 1.38 mmol, 0.99 equiv.), palladium acetate (31 mg, 139.36 μmol, 0.1 equiv.), Cy3P (78 mg, 278.72 μmol, 0.2 equiv.), and K2CO3 (385 mg, 2.79 mmol, 2 equiv.) in dioxane (15 mL) was degassed and purged with nitrogen three times. The mixture was then stirred under a nitrogen atmosphere at 120 °C for 16 h. The mixture was concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (20 g Sepa Flash® silica flash column, eluent: 30-60% ethyl acetate / petroleum ether gradient @ 50 mL / min) to give the compound 6-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (180 mg, 290.95 μmol, 21% yield, 66% purity) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.65(dd, J=1.1, 8.0Hz, 1H), 8.38(s, 1H), 8.35-8. 33(m, 1H), 8.26-8.21(m, 1H), 8.12-8.06(m, 2H), 7.22(s, 2H), 5.57(s, 2H). MS: m / z=409.0 (M+1, ESI+).

[0613] Step 2: Preparation of 6-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (Compound 223)

[0614] To a mixture of 6-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (180 mg, 290.95 μmol, 66% purity, 1 equiv.) in THF (10 mL) and water (3 mL) was added iron powder (81 mg, 1.45 mmol, 5 equiv.) and NH4Cl (124 mg, 2.33 mmol, 8 equiv.). The mixture was then stirred at 80 °C for 5 h. The reaction mixture was filtered, and saturated aqueous NaHCO3 solution (50 mL) was added to the filtrate, followed by extraction with ethyl acetate (80 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Welch Ultimate XB-CN 250*70*10 μm; mobile phase: [hexane-EtOH (0.1% ammonium hydroxide)]; B%: 40%-80%, 10 min). Compound 223 or 6-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (24.24 mg, 60.94 μmol, 21% yield, 95.11% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.65(dd, J=0.9, 8.0Hz, 1H), 8.35(s, 1H), 8.34-8.29(m, 1H) ), 8.27-8.21(m, 1H), 7.13(s, 2H), 6.18(d, J=10.3Hz, 2H), 5.80(s, 2H), 5.27(s, 2H).

[0615] Example 1.55: Preparation of 6-(6-amino-1-(2,6-difluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)picolinonitrile (Compound 231) JPEG2025120182000132.jpg37160Reagents and conditions: (a) Pd(OAc)2, K2CO3, Cy3P, dioxane, 120 °C, 10 h, MW.

[0616] A mixture of (6-cyano-2-pyridyl)sulfinyloxysodium (231 mg, 1.2 equiv.), intermediate compound S110 or 4-chloro-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (300 mg, 1 equiv.), palladium acetate (23 mg, 0.1 equiv.), Cy3P (57 mg, 0.2 equiv.), and K2CO3 (280 mg, 2 equiv.) in dioxane (2 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred at 120 °C for 10 h under microwave conditions. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150*25mm*10μm; mobile phase: [water (0.225% FA)-ACN]; B%: 37%-67%, 11 min). Compound 231 or 6-[6-amino-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (70.52 mg, 18% yield, 95.65% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.65(dd, J=1.0, 8.0Hz, 1H), 8.36(s, 1H), 8.33-8.27(m, 1H), 8.26-8.20(m, 1H), 7.51-7.41(m, 1H), 7.18(s, 2H), 7.12(t, J=8.0Hz, 2H), 5.48(s, 2H).

[0617] Example 1.56: Preparation of 6-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)picolinonitrile (Compound 227) JPEG2025120182000133.jpg38160Reagents and conditions: (a) Pd(OAc)2, K2CO3, Cy3P, dioxane, 120°C, 16 hours. (b) Fe, NH4Cl, EtOH / H2O, 60°C, 2 hours.

[0618] Step 1: Preparation of 6-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile

[0619] A mixture of intermediate compound S113 or (6-cyano-2-pyridyl)sulfinyloxysodium (356 mg, 1.2 equiv.), intermediate compound S108 or 4-chloro-1-[(3-methyl-4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (500 mg, 1 equiv.), palladium acetate (35 mg, 0.1 equiv.), Cy3P (88 mg, 0.2 equiv.), and K2CO3 (433 mg, 2 equiv.) in dioxane (5 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred at 120 °C under a nitrogen atmosphere for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 24 g Sepa Flash® silica flash column, eluent: 50–80% ethyl acetate / petroleum ether gradient @ 35 mL / min). The compound 6-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (300 mg, 40% yield, 81% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.66(dd, J=1.1, 7.9Hz, 1H), 8.48(s, 1H), 8.37-8.29(m, 1H), 8.28-8 .22(m, 1H), 7.95(d, J=8.5Hz, 1H), 7.32(s, 1H), 7.24-7.14(m, 3H), 5.55(s, 2H), 2.47(s, 3H). MS: m / z=387.1 (M+1, ESI+).

[0620] Step 2: Preparation of 6-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)picolinonitrile (Compound 227)

[0621] To a solution of 6-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (311 mg, 1 equiv.) in water (5 mL) and ethanol (15 mL) was added iron powder (225 mg, 5 equiv.) and NH4Cl (344 mg, 8 equiv.). The mixture was stirred at 60°C for 2 hours. The reaction mixture was concentrated under reduced pressure to remove ethanol and water to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30mm*3μm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 10%-40%, 11.5min) to give the desired compound with 88% purity, and the residue was re-purified by preparative HPLC (column: Welch Ultimate XB-SiOH 250*50*10μm; mobile phase: [hexane-EtOH (0.1% NH3·H2O)]; B%: 15%-55%, 15min) to give the desired compound with 90% purity. The residue was then purified by preparative HPLC (column: Welch Ultimate XB-SiOH 250*50*10 μm; mobile phase: [hexane-EtOH (0.1% NH₃·HO)]; B%: 15%-55%, 15 min), affording the desired compound with 91% purity. Finally, the residue was purified by preparative HPLC (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 21%-51%, 10 min). Compound 227, or 6-[6-amino-1-[(4-amino-3-methyl-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (30.69 mg, 10% yield, 98.87% purity), was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.65(dd, J=1.1, 8.0Hz, 1H), 8.39(s, 1H), 8.34-8.27(m, 1H), 8.26-8.20(m, 1H), 7.11( s, 2H), 6.86(s, 1H), 6.82(dd, J=1.9, 8.1Hz, 1H), 6.52(d, J=8.0Hz, 1H), 5.24(s, 2H), 4.80(s, 2H), 1.99(s, 3H).

[0622] Example 1.57: Preparation of 2-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)isonicotinonitrile (Compound 216) JPEG2025120182000134.jpg36161Reagents and conditions: (a) Pd(OAc)2, K2CO3, Cy3P, dioxane, 120°C, 16 hours. (b) Fe, NH4Cl, EtOH / H2O, 60°C, 2 hours.

[0623] Step 1: Preparation of 2-(6-amino-1-(4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)isonicotinonitrile

[0624] A mixture of sodium 4-cyanopyridine-2-sulfinate (298 mg, 1.2 equiv.), intermediate compound S3 or 4-chloro-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (400 mg, 1 equiv.), palladium acetate (29 mg, 0.1 equiv.), Cy3P (73 mg, 0.2 equiv.), and K2CO3 (362 mg, 2 equiv.) in dioxane (5 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred under a nitrogen atmosphere at 120 °C for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g Sepa Flash® silica flash column, eluent: 70–90% ethyl acetate / petroleum ether gradient @ 30 mL / min). The compound 2-[6-amino-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (300 mg, 55% yield, 89% purity) was obtained as a yellow solid. MS: m / z=373.1 (M+1, ESI+).

[0625] Step 2: Preparation of 2-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)isonicotinonitrile (Compound 216)

[0626] To a solution of 2-[6-amino-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (300 mg, 1 equiv.) in water (5 mL) and ethanol (15 mL), iron powder (225 mg, 5 equiv.) and NH₄Cl (345 mg, 8 equiv.) were added. The mixture was stirred at 60°C for 2 h. The reaction mixture was concentrated under reduced pressure to remove ethanol and water, yielding a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30 mm*3 μm; mobile phase: [water (10 mM NH₄HCO₃)-ACN]; B%: 18%-48%, 8 min) to give the desired compound in 80% purity. The residue was then purified by a second run of preparative HPLC (column: Welch Ultimate XB-SiOH 250*50*10 μm; mobile phase: [hexane-EtOH (0.1% NH₃·HO)]; B%: 15%-55%, 15 min), affording the desired compound with 80% purity. Finally, the residue was purified by a third run of preparative HPLC (column: Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 17%-47%, 10 min). Compound 216, or 2-[6-amino-1-[(4-aminophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (23.18 mg, 8% yield, 95.02% purity), was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.54(d, J=2.1Hz, 1H), 9.22(d, J=2.0Hz, 1H), 8.92(t, J=2.1Hz, 1H), 8.45(s, 1H), 7.13(s, 2H), 6.96(d, J=8.4Hz, 2H), 6.63-6.37(m, 2H), 5.25(s, 2H), 5.04(s, 2H). HRMS-TOF:343.1408.

[0627] Example 1.58: Preparation of 2-(6-amino-1-(4-amino-2-fluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)isonicotinonitrile (Compound 220) JPEG2025120182000135.jpg36160 Reagents and conditions: (a) Pd(OAc)2, K2CO3, Cy3P, dioxane, 120°C, 16 hours. (b) Fe, NH4Cl, EtOH / H2O, 60°C, 1 hour.

[0628] Step 1: Preparation of 2-(6-amino-1-(2-fluoro-4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)isonicotinonitrile

[0629] A mixture of sodium 4-cyanopyridine-2-sulfinate (424 mg, 1.2 equiv.), intermediate compound S111 or 4-chloro-1-[(2-fluoro-4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (600 mg, 1 equiv.), palladium acetate (41 mg, 0.1 equiv.), Cy3P (104 mg, 0.2 equiv.), and K2CO3 (513 mg, 2 equiv.) in dioxane (5 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred at 120 °C under a nitrogen atmosphere for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g Sepa Flash® silica flash column, eluent: 70–90% ethyl acetate / petroleum ether gradient @ 30 mL / min). The compound 2-[6-amino-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (300 mg, 41% yield, 99% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.13-9.06(m, 1H), 8.68(dd, J=0.9, 1.5Hz, 1H), 8.55(s, 1H), 8.16(dd, J=2.3, 9.9Hz, 1H), 8.09(dd, J=1.6, 4.9Hz, 1H), 8.03(dd, J=2.1, 8.5Hz, 1H), 7.30(t, J=8.0Hz, 1H), 7.21(s, 2H), 5.63(s, 2H).

[0630] Step 2: Preparation of 2-(6-amino-1-(4-amino-2-fluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)isonicotinonitrile (Compound 220)

[0631] To a solution of 2-[6-amino-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (300 mg, 1 equiv.) in water (5 mL) and ethanol (15 mL) was added iron powder (214 mg, 5 equiv.) and NH4Cl (328 mg, 8 equiv.). The mixture was stirred at 60°C for 1 hour. The reaction mixture was concentrated under reduced pressure to remove ethanol and water to give a residue. The residue was purified by preparative HPLC (column: Welch Ultimate XB-CN 250*70*10 μm; mobile phase: [Hexane-EtOH (0.1% NH₃·H₂O)]; B%: 15%-55%, 15 min (0.1% NH₃·H₂O)]; B%: 30%-70%, 15 min). After preparative HPLC, the desired compound was obtained with 90% purity. The residue was purified by a second preparative HPLC (column: Welch Ultimate XB-CN 250*70*10 μm; mobile phase: [Hexane-EtOH (0.1% aqueous ammonia)]; B%: 30%-70%, 15 min). Compound 220 or 2-[6-amino-1-[(4-amino-2-fluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (33.67 mg, 89.81 μmol, purity 96.12%) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.08(dd, J=0.7, 5.0Hz, 1H), 8.68(d, J=1.3Hz, 1H), 8.46(s, 1H), 8.08(dd, J=1.6, 5.0Hz, 1H), 7.13(s, 2H), 6.88(t, J=8.4Hz, 1H), 6.33-6.30(m, 1H), 6.29(s, 1H), 5.40(s, 2H), 5.29(s, 2H).

[0632] Example 1.59: Preparation of 2-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (Compound 224) JPEG2025120182000136.jpg90118Reagents and conditions: (a) Fe, NH4Cl, EtOH / H2O, 80°C, 3 hours. (b) Pd(OAc)2, K2CO3, PCy3, dioxane, 120°C, 16 hours.

[0633] Step 1: Preparation of 1-(4-amino-2,6-difluorobenzyl)-4-chloro-1H-pyrazolo[3,4-d]pyrimidin-6-amine (intermediate compound S114)

[0634] To a mixture of intermediate compound S112 or 4-chloro-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (700 mg, 1.38 mmol, 67% purity, 1 equiv.) in water (3 mL) and ethanol (10 mL), iron powder (385 mg, 6.88 mmol, 5 equiv.) and NH4Cl (590 mg, 11.01 mmol, 8 equiv.) were added. The mixture was then stirred at 80 °C for 3 h. The reaction mixture was filtered, and the filtrate was concentrated in vacuo. Water (30 mL) was added to the mixture, which was then extracted with ethyl acetate (30 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by preparative HPLC (column: Welch Ultimate XB-CN 250*70*10 μm; mobile phase: [hexane-EtOH (0.1% NH₃·H₂O)]; B%: 20%-60%, 15 min). Intermediate compound S114 or 1-(4-amino-2,6-difluorobenzyl)-4-chloro-1H-pyrazolo[3,4-d]pyrimidin-6-amine (400 mg, 1.03 mmol, 75% yield, 88% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ = 7.92 (s, 1H), 7.32 (s, 2H), 6.25-6.09 (m, 2H), 5.80 (s, 2H), 5.18 (s, 2H).

[0635] Step 2: Preparation of 2-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (Compound 224)

[0636] A mixture of (4-cyano-2-pyridyl)sulfinyloxysodium (550 mg, 2.89 mmol, 2.55 equiv.), intermediate compound S114 or 1-[(4-amino-2,6-difluorophenyl)methyl]-4-chloropyrazolo[3,4-d]pyrimidin-6-amine (400 mg, 1.13 mmol, 88% purity, 1 equiv.), palladium acetate (25 mg, 113.30 μmol, 0.1 equiv.), Cy3P (63 mg, 226.59 μmol, 73.46 μL, 0.2 equiv.), and K2CO3 (313 mg, 2.27 mmol, 2 equiv.) in dioxane (10 mL) was degassed and purged with nitrogen three times. The mixture was then stirred at 120 °C under a nitrogen atmosphere for 16 h. The reaction mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by flash silica gel chromatography (20 g Sepa Flash® silica flash column, eluent: 20-40% ethyl acetate / petroleum ether gradient @ 40 mL / min). Compound 224 or 2-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (70 mg, 181.87 μmol, 16% yield, 98.30% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.07(dd, J=0.8, 5.0Hz, 1H), 8.67(dd, J=0.9, 1.5Hz, 1H), 8.41(s, 1H), 8.06(dd, J=1.7, 5.0Hz, 1H), 7.11(s, 2H), 6.27-6.11(m, 2H), 5.79(s, 2H), 5.26(s, 2H).

[0637] Example 1.60: Preparation of 2-(6-amino-1-(2,6-difluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)isonicotinonitrile (Compound 232) JPEG2025120182000137.jpg38160Reagents and conditions: (a) Pd(OAc)2, K2CO3, Cy3P, dioxane, 120°C, 16 hours.

[0638] A mixture of (4-cyano-2-pyridyl)sulfinyloxysodium (232 mg, 1.2 equiv.), intermediate compound S110 or 4-chloro-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (300 mg, 1 equiv.), palladium acetate (23 mg, 0.1 equiv.), Cy3P (57 mg, 0.2 equiv.), and K2CO3 (281 mg, 2 equiv.) in dioxane (5 mL) was degassed and purged with nitrogen three times, then the mixture was stirred under a nitrogen atmosphere at 120 °C for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Unisil 3-100 C18 Ultra 150*50mm*3μm; mobile phase: [water (0.225% FA)-ACN]; B%: 43%-63%, 10 min). Compound 232 or 2-[6-amino-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (66.33 mg, 18% yield, 99.34% purity) was obtained as a white solid. 1 H NMR (400MHz, DMSO-d6) δ=9.06(d, J=5.0Hz, 1H), 8.67(s, 1H), 8.44-8.38(m, 1H), 8.09 -8.03(m, 1H), 7.51-7.39(m, 1H), 7.22-7.15(m, 2H), 7.14-7.09(m, 2H), 5.47(s, 2H).

[0639] Example 1.61: 2-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)isonicotinonitrile (Compound 228) JPEG2025120182000138.jpg36160Reagents and conditions: (a) Pd(OAc)2, K2CO3, Cy3P, dioxane, 120°C, 16 hours. (b) Fe, NH4Cl, EtOH / H2O, 60°C, 2 hours.

[0640] Step 1: Preparation of 2-(6-amino-1-(3-methyl-4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)isonicotinonitrile

[0641] A mixture of (4-cyano-2-pyridyl)sulfinyloxysodium (215 mg, 1.2 equiv.), intermediate compound S108 or 4-chloro-1-[(3-methyl-4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (300 mg, 1 equiv.), palladium acetate (21 mg, 0.1 equiv.), Cy3P (52 mg, 0.2 equiv.), and K2CO3 (260 mg, 2 equiv.) in dioxane (5 mL) was degassed and purged with nitrogen three times. The mixture was then stirred at 120 °C under a nitrogen atmosphere for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 24 g Sepa Flash® silica flash column, eluent: 50–80% ethyl acetate / petroleum ether gradient @ 35 mL / min). The compound 2-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (200 mg, 55% yield) was obtained as a yellow solid. MS: m / z=387.1 (M+1, ESI+).

[0642] Step 2: Preparation of 2-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)isonicotinonitrile (Compound 228)

[0643] To a solution of 2-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (200 mg, 1 equiv.) in ethanol (15 mL) and water (5 mL) was added iron powder (145 mg, 5 equiv.) and NH4Cl (221 mg, 8 equiv.). The mixture was stirred at 60°C for 2 hours. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30 mm*3 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 20%-50%, 8 min). After preparative HPLC, the compound was obtained with 80% purity. The residue was purified by a second preparative HPLC (column: Shim-pack C18 150*25*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 16%-46%, 10 min). Compound 228, or 2-[6-amino-1-[(4-amino-3-methyl-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-4-carbonitrile (19.65 mg, 10% yield, 96.15% purity), was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.08(dd, J=0.8, 4.9Hz, 1H), 8.67(dd, J=0.9, 1.6Hz, 1H), 8.45(s, 1H), 8.07(dd, J=1.6, 5.0Hz, 1 H), 7.10(s, 2H), 6.86(s, 1H), 6.82(dd, J=1.9, 8.1Hz, 1H), 6.51(d, J=8.0Hz, 1H), 5.23(s, 2H), 4.80(s, 2H), 1.98(s, 3H).

[0644] Example 1.62: Preparation of 5-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)nicotinonitrile (Compound 217) JPEG2025120182000139.jpg36168Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110°C, 16 h; (b) Fe, NH4Cl, THF / H2O, 60°C, 3 h.

[0645] Step 1: Preparation of 5-(6-amino-1-(4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)nicotinonitrile

[0646] A mixture of (5-cyano-3-pyridyl)boronic acid (583 mg, 1.5 equiv.), intermediate compound S3 or 4-chloro-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (800 mg, 1 equiv.), KCO (726 mg, 2 equiv.), and Pd(dppf)Cl (192 mg, 0.1 equiv.) in dioxane (10 mL) and water (2 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred at 110 °C under a nitrogen atmosphere for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 24 g Sepa Flash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient @ 30 mL / min). The compound 5-[6-amino-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (800 mg, 82% yield) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ = 9.63-9.52(m, 1H), 9.37-9.31(m, 2H), 9.25-9.23(m, 1H), 9.15-9.12(m, 2H), 7.46-7.42(m, 2H), 5.65-5.60(m, 2H). MS: m / z=373.1 (M+1, ESI+).

[0647] Step 2: Preparation of 5-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)nicotinonitrile (Compound 217)

[0648] To a solution of 5-[6-amino-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (300 mg, 1 equiv.) in THF (15 mL) and water (5 mL) was added iron powder (224 mg, 5 equiv.) and NH4Cl (344 mg, 8 equiv.). The mixture was stirred at 60 °C for 3 h. The reaction mixture was concentrated under reduced pressure to remove THF and water to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30 mm*3 μm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 10%-40%, 11.5 min). Compound 217 or 5-[6-amino-1-[(4-aminophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (27.63 mg, 7% yield, 98.17% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.54(d, J=2.1Hz, 1H), 9.22(d, J=2.0Hz, 1H), 8.92(t, J=2.1Hz, 1H), 8.45(s, 1H), 7.13(s, 2H), 6.96(d, J=8.4Hz, 2H), 6.63-6.37(m, 2H), 5.25(s, 2H), 5.04(s, 2H).

[0649] Example 1.63: Preparation of 5-[6-amino-1-[(4-amino-2-fluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (Compound 221) JPEG2025120182000140.jpg33161Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110°C, 16 h; (b) Fe, NH4Cl, EtOH / H2O, 60°C, 2 h.

[0650] Step 1: Preparation of 5-(6-amino-1-(2-fluoro-4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)nicotinonitrile

[0651] A mixture of (5-cyano-3-pyridyl)boronic acid (343 mg, 1.5 equiv.), intermediate compound S111 or 4-chloro-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (500 mg, 1 equiv.), KCO (428 mg, 2 equiv.), and Pd(dppf)Cl (113 mg, 0.1 equiv.) in dioxane (10 mL) and water (2 mL) was degassed and purged with nitrogen three times. The mixture was then stirred at 110 °C under a nitrogen atmosphere for 16 h. The reaction mixture was concentrated under reduced pressure to remove dioxane and yield a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g Sepa Flash® silica flash column, eluent: 0–80% ethyl acetate / petroleum ether gradient @ 30 mL / min). The compound 5-[6-amino-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (300 mg, 50% yield) was obtained as a yellow solid. MS: m / z=390.9 (M+1, ESI+).

[0652] Step 2: Preparation of 5-(6-amino-1-(4-amino-2-fluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)nicotinonitrile (Compound 221)

[0653] To a solution of 5-[6-amino-1-[(2-fluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (300 mg, 1 eq.) in ethanol (15 mL) and water (5 mL) was added iron powder (214 mg, 5 eq.) and NH4Cl (328 mg, 8 eq.). The mixture was stirred at 60°C for 2 hours. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30 mm*3 μm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 13%-43%, 11.5 min). Compound 221 or 5-[6-amino-1-[(4-amino-2-fluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (37.61 mg, 13% yield, 94.57% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6)δ=9.54(d, J=2.1Hz, 1H), 9.23(d, J=2.0Hz, 1H), 8.91(t, J=2.0Hz, 1H), 8. 45(s, 1H), 7.14(s, 2H), 6.90(t, J=8.4Hz, 1H), 6.31(d, J=10.8Hz, 2H), 5.41(s, 2H), 5.29(s, 2H).

[0654] Example 1.64: Preparation of 5-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (Compound 225) JPEG2025120182000141.jpg39160Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 100°C, 15 h; (b) Fe, NH4Cl, EtOH / H2O, 80°C, 2 h.

[0655] Step 1: Preparation of 5-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile

[0656] To a solution of intermediate compound S112 or 4-chloro-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (400 mg, 1.17 mmol, 1 equiv.) in dioxane (10 mL) and water (2 mL), (5-cyano-3-pyridyl)boronic acid (346 mg, 2.34 mmol, 2 equiv.), Pd(dppf)Cl (86 mg, 117.00 μmol, 0.1 equiv.), and KCO (323 mg, 2.34 mmol, 2 equiv.) were added. The mixture was then stirred at 100° C. for 15 hours. The mixture was concentrated in vacuo to give a residue. The residue was purified by flash silica gel chromatography (20 g Sepa Flash® silica flash column, eluent: 0-60% ethyl acetate / petroleum ether gradient @ 50 mL / min). The compound 5-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (400 mg, 852.27 μmol, 73% yield, 87% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.51(d, J=2.0Hz, 1H), 9.21(d, J=1.9Hz, 1H), 8.89(t , J=2.1Hz, 1H), 8.44(s, 1H), 8.09(d, J=7.3Hz, 2H), 7.23(s, 2H), 5.55(s, 2H). MS: m / z=409.0 (M+1, ESI+).

[0657] Step 2: Preparation of 5-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (Compound 225)

[0658] To a mixture of 5-[6-amino-1-[(2,6-difluoro-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (400 mg, 979.62 μmol, 1 equiv.) in water (3 mL) and ethanol (9 mL), iron powder (274 mg, 4.90 mmol, 5 equiv.) and NH4Cl (420 mg, 7.84 mmol, 8 equiv.) were added. The mixture was then stirred at 80 °C for 2 h. The reaction mixture was filtered, and the filtrate was concentrated in vacuo. Water (30 mL) was then added to the mixture, which was then extracted with ethyl acetate (30 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Welch Ultimate XB-CN 250*70*10 μm; mobile phase: [hexane-EtOH (0.1% NH₃·H₂O)]; B%: 30%-70%, 15 min) to give an impure product, which was repurified by preparative HPLC (column: Welch Ultimate XB-CN 250*50*10 μm; mobile phase: [hexane-EtOH (0.1% NH₃·H₂O)]; B%: 25%-65%, 15 min). Compound 225 or 5-[6-amino-1-[(4-amino-2,6-difluoro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (113.05 mg, 297.07 μmol, 30% yield, 99.42% purity) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.52(d, J=2.1Hz, 1H), 9.21(d, J=1.8Hz, 1H), 8.89(t , J=2.1Hz, 1H), 8.38(s, 1H), 7.13(s, 2H), 6.18(d, J=10.1Hz, 2H), 5.25(s, 2H).

[0659] Example 1.65: Preparation of 5-(6-amino-1-(2,6-difluorobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)nicotinonitrile (Compound 233) JPEG2025120182000142.jpg39160Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110 °C, 16 h.

[0660] A mixture of intermediate compound S110 or 4-chloro-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (400 mg, 1 equiv.), (5-cyano-3-pyridyl)boronic acid (300 mg, 1.5 equiv.), KCO (373 mg, 2 equiv.), and Pd(dppf)Cl (99 mg, 0.1 equiv.) in dioxane (10 mL) and water (2 mL) was degassed and purged with nitrogen three times, then the mixture was stirred under a nitrogen atmosphere at 110° C. for 16 hours. The reaction mixture was concentrated under reduced pressure to remove dioxane and provide a residue. The residue was purified by preparative HPLC (column: Unisil 3-100 C18 Ultra 150*50mm*3μm; mobile phase: [water (0.225% FA)-ACN]; B%: 45%-65%, 10 min) to give the desired compound with 80% purity, followed by a second preparative HPLC (column: Phenomenex Luna C18 150*25mm*10μm; mobile phase: [water (0.225% FA)-ACN]; B%: 31%-61%, 10 min). Compound 233, or 5-[6-amino-1-[(2,6-difluorophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (69.15 mg, 14% yield, 97.76% purity), was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=8.96(d, J=5.1Hz, 1H), 8.58(d, J=0.6Hz, 1H), 8.41-8.39(m, 1 H), 8.39-8.36(m, 1H), 7.51-7.41(m, 1H), 7.25(s, 2H), 7.17-7.08(m, 2H), 5.47(s, 2H).

[0661] Example 1.66: Preparation of 5-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)nicotinonitrile (Compound 229) JPEG2025120182000143.jpg36161Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane, 110°C, 16 h; (b) Fe, NH4Cl, EtOH / H2O, 60°C, 2 h.

[0662] Step 1: Preparation of 5-(6-amino-1-(3-methyl-4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)nicotinonitrile

[0663] A mixture of (5-cyano-3-pyridyl)boronic acid (208 mg, 1.5 equiv.), intermediate compound S108 or 4-chloro-1-[(3-methyl-4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (300 mg, 1 equiv.), KCO (260 mg, 2 equiv.), and Pd(dppf)Cl (68.87 mg, 0.1 equiv.) in dioxane (10 mL) was degassed and purged with nitrogen three times, and then the mixture was stirred at 110 °C under a nitrogen atmosphere for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 24 g Sepa Flash® silica flash column, eluent: 0-100% ethyl acetate / petroleum ether gradient @ 35 mL / min). The compound 5-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (300 mg, 82% yield) was obtained as a yellow solid. MS: m / z=386.9 (M+1, ESI+).

[0664] Step 2: Preparation of 5-(6-amino-1-(4-amino-3-methylbenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)nicotinonitrile (Compound 229)

[0665] To a solution of 5-[6-amino-1-[(3-methyl-4-nitro-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (400 mg, 1 equiv.) in ethanol (15 mL) and water (5 mL) was added iron powder (289 mg, 5 equiv.) and NH4Cl (443 mg, 8 equiv.). The mixture was stirred at 60°C for 2 hours. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (Shim-pack C18 150*25*10 μm column; mobile phase: [water (0.225% FA)-ACN]; B%: 12%-42%, 10 min) to obtain the desired compound with a purity of 92%. The mixture was then purified by a second preparative HPLC (Phenomenex Gemini-NX C18 75*30 mm*3 μm column; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 14%-44%, 11.5 min). Compound 229, or 5-[6-amino-1-[(4-amino-3-methyl-phenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-3-carbonitrile (58.12 mg, 15% yield, 97.57% purity), was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.53(d, J=2.1Hz, 1H), 9.22(d, J=2.1Hz, 1H), 8.91(t, J=2.1Hz, 1H), 8.43(s , 1H), 7.12(s, 2H), 6.91-6.77(m, 2H), 6.52(d, J=8.1Hz, 1H), 5.23(s, 2H), 4.80(s, 2H), 1.99(s, 3H).

[0666] Example 1.67: Preparation of 4-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)picolinonitrile (Compound 218) JPEG2025120182000144.jpg36160Reagents and conditions: (a) Pd(dppf)Cl2, K2CO3, dioxane / H2O, 110°C, 16 h; (b) Fe, NH4Cl, EtOH / H2O, 60°C, 2 h.

[0667] Step 1: Preparation of 4-(6-amino-1-(4-nitrobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)picolinonitrile

[0668] A mixture of intermediate compound S3 or 4-chloro-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-6-amine (800 mg, 1 equiv.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbonitrile (906 mg, 1.5 equiv.), KCO (725 mg, 2 equiv.), and Pd(dppf)Cl (192 mg, 0.1 equiv.) in dioxane (10 mL) and water (2 mL) was degassed and purged with nitrogen three times. The mixture was then stirred under a nitrogen atmosphere at 110 °C for 16 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g Sepa Flash® silica flash column, eluting with a 30–80% ethyl acetate / petroleum ether gradient at 35 mL / min). The compound 4-[6-amino-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (500 mg, 51% yield) was obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ=9.05-8.94(m, 1H), 8.84-8.71(m, 2H), 8.58(s, 1H), 8.43(dd, J=1. 8, 5.1Hz, 1H), 8.09-8.07(m, 1H), 7.46-7.41(m, 2H), 7.31-7.24(m, 2H), 5.67-5.61(m, 2H). MS: m / z=372.9 (M+1, ESI+).

[0669] Step 2: Preparation of 4-(6-amino-1-(4-aminobenzyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl)picolinonitrile (Compound 218)

[0670] To a solution of 4-[6-amino-1-[(4-nitrophenyl)methyl]pyrazolo[3,4-d]pyrimidin-4-yl]pyridine-2-carbonitrile (300 mg, 1 equiv.) in water (5 mL) and ethanol (15 mL) was added iron powder (224 mg, 5 equiv.) and NH4Cl (344 mg, 8 equiv.). The mixture was stirred at 60 °C for 2 h. The reaction mixture was then concentrated under reduced pressure to remove ethanol and water, yielding a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX C1...

Claims

1. A compound of formula (Ib) or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof: (Ib) R is H, (C 1 -C 3 ) alkyl, or substituted (C 1 -C 3 ) alkyl; Y 1 is CH; Y 2 Or Y 4 is independently, CR 10 and N, and Y 2 Or Y 4 At least one of the following is independently CR 10 and R 10 are each independently H, (C 1 -C 8 ) alkyl, substituted (C 1 -C 8 ) alkyl, (C 2 -C 8 ) alkenyl, substituted (C 2 -C 8 ) alkenyl, (C 2 -C 8 ) alkynyl, substituted (C 2 -C 8 ) alkynyl, (C 1 -C 3 ) haloalkyl, (C 1 -C 8 ) alkoxy, substituted (C 1 -C 8 ) alkoxy, -CONH 2 , substituted amide, —NH 2 , substituted amino, —CO 2 H, cyano, halogen, hydroxyl, -NO 2 , -SO 3 H, -SO 2 NH 2 , substituted sulfonamide, and thiol; R a and R b are each independently H, F, (C 1 -C 3 ) alkyl, and substituted (C 1 -C 3 ) alkyl, or R a and R b are attached to a ring and together with the carbon atoms to which they are attached form a cyclopropyl or substituted cyclopropyl; A is phenyl, substituted phenyl, pyridyl, or substituted pyridyl.

2. i) A is phenyl or one, two or three R 20 phenyl substituted with R 20 are each independently 1 -C 8 ) alkyl, substituted (C 1 -C 8 ) alkyl, (C 2 -C 8 ) alkenyl, substituted (C 2 -C 8 ) alkenyl, (C 2 -C 8 ) alkynyl, substituted (C 2 -C 8 ) alkynyl, (C 1 -C 3 ) haloalkyl, (C 1 -C 8 ) alkoxy, substituted (C 1 -C 8 ) alkoxy, -CONH 2 , substituted amide, —NH 2 , substituted amino, —CO 2 H, cyano, halogen, hydroxyl, -NO 2 , -SO 3 H, -SO 2 NH 2 , substituted sulfonamides, and thiols; or ii) A is pyridyl or one, two, or three R 20 pyridyl substituted with R 20 are each independently 1 -C 8 ) alkyl, substituted (C 1 -C 8 ) alkyl, (C 2 -C 8 ) alkenyl, substituted (C 2 -C 8 ) alkenyl, (C 2 -C 8 ) alkynyl, substituted (C 2 -C 8 ) alkynyl, (C 1 -C 3 ) haloalkyl, (C 1 -C 8 ) alkoxy, substituted (C 1 -C 8 ) alkoxy, -CONH 2 , substituted amide, —NH 2 , substituted amino, —CO 2 H, cyano, halogen, hydroxyl, -NO 2 , -SO 3 H, -SO 2 NH 2 , a substituted sulfonamide, and a thiol, or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

3. R a and R b 10. The compound of claim 1, wherein each is H, or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

4. A is an unsubstituted or substituted phenyl represented by the formula: Or, the following group but, and In the formula, R 2 Or R 9 are independently H, (C 1 -C 8 ) alkyl, substituted (C 1 -C 8 ) alkyl, (C 2 -C 8 ) alkenyl, substituted (C 2 -C 8 ) alkenyl, (C 2 -C 8 ) alkynyl, substituted (C 2 -C 8 ) alkynyl, (C 1 -C 3 ) haloalkyl, (C 1 -C 8 ) alkoxy, substituted (C 1 -C 8 ) alkoxy, -CONH 2 , substituted amide, —NH 2 , substituted amino, —CO 2 H, cyano, halogen, hydroxyl, -NO 2 , -SO 3 H, -SO 2 NH 2 4. The compound of claim 3, wherein the compound is selected from the group consisting of aryl, arylsulfonamides ...

5. R 2 Or R 9 are independently H, NH 2 , F, C.H. 3 , and C.F. 3 5. The compound of claim 4, selected from: or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

6. R 7 NR 21 R 22 and R 21 and R 22 are independently H, (C 1 -C 8 ) alkyl, substituted (C 1 -C 8 ) alkyl, SO 2 R 30 , and COR 30 Selected from, and R 30 is (C 1 -C 8 ) alkyl, or substituted (C 1 -C 8 ) alkyl; 5. A compound according to claim 4, or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof. where:

7. R 21 and R 22 and R are each H, or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

8. R 5 , R 6 , R 8 and R 9 are independently H, (C 1 -C 5 ) alkyl, substituted (C 1 -C 5 ) alkyl, (C 1 -C 3 ) haloalkyl, (C 1 -C 5 ) alkoxy, substituted (C 1 -C 5 7. The compound of claim 6, or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

9. R 5 , R 6 , R 8 and R 9 However, independently, H, F, CH 3 , and C.F. 3 selected from, and R 2 Or R 4 9. The compound of claim 8, wherein each is H, or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

10. R 2 Or R 4 are H, 5. A compound according to claim 4, or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

11. 2. The compound of claim 1, wherein the compound of formula (Ib) is selected from the following: or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

12. A pharmaceutical composition comprising a compound according to any one of claims 1 to 11 or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable excipient.

13. 1. An in vitro method for inhibiting adenosine A2A and / or A1 receptors, comprising: A sample containing adenosine A2A and / or A1 receptors is contacted with an effective amount of a compound according to any one of claims 1-11, or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 12, to inhibit adenosine A2A and / or A1 receptors.

14. 1. An in vitro method of antagonizing adenosine A2A and / or A1 receptors, comprising: A cell containing adenosine A2A and / or A1 receptors is contacted with an effective amount of a compound according to any one of claims 1-11, or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 12, to antagonize the adenosine A2A and / or A1 receptors.

15. A pharmaceutical composition for treating cancer, comprising as an active ingredient an A2A and / or A1 receptor antagonist compound according to any one of claims 1 to 11, or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

16. The pharmaceutical composition of claim 15, wherein the cancer is a solid tumor cancer.

17. 16. The pharmaceutical composition of claim 15, wherein the cancer is selected from the group consisting of lung cancer, breast cancer, prostate cancer, ovarian cancer, solenoma, cervical cancer, bladder cancer, head and neck cancer, renal cell carcinoma, esophageal cancer, pancreatic cancer, brain cancer, liver cancer, leukemia, lymphoma, melanoma, multiple myeloma, Ewing's sarcoma, osteosarcoma, colorectal tumor, bile duct cancer, choriocarcinoma, oral cancer, neuroblastoma, skin cancer, testicular cancer, stromal tumor, germ cell tumor, and thyroid cancer.

18. 16. The pharmaceutical composition of claim 15, wherein the composition is administered in combination with an additional active agent.

19. 20. The pharmaceutical composition of claim 18, wherein the additional active agent is selected from an anti-angiogenic agent, an anti-inflammatory agent, an immune checkpoint inhibitor, a PARP inhibitor, a chemotherapeutic agent, and an immune anti-cancer agent.

20. 19. The pharmaceutical composition of claim 18, wherein the additional active agent is an immune checkpoint inhibitor selected from a CTLA-4 inhibitor, a PD-1 inhibitor, and a PD-L1 inhibitor.

21. 21. The pharmaceutical composition of claim 20, wherein the immune checkpoint inhibitor is an antibody or antibody fragment.

22. A pharmaceutical composition for treating an inflammatory disease, comprising an A2A and / or A1 receptor antagonist compound according to any one of claims 1 to 11, or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

23. Use of an A2A and / or A1 receptor antagonist compound, or a solvate, hydrate, and / or stereoisomer thereof, or a pharmaceutically acceptable salt thereof, according to any one of claims 1-11, in the manufacture of a pharmaceutical composition according to any one of claims 15-22.

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