EA049302B1 - TRIAZOLOPYRIMIDINES AS A2A/A2B INHIBITORS - Google Patents

Abstract

This invention relates to compounds of formula (I): or pharmaceutically acceptable salts or stereoisomers thereof, which modulate the activity of adenosine receptors, such as A2A and A2B receptor subtypes, and can be used in the treatment of diseases associated with adenosine receptor activity, including, for example, cancer, inflammatory diseases, cardiovascular diseases and neurodegenerative diseases.

Description

Cross-reference to related applications

This application claims priority to U.S. Provisional Patent Application No. 62/891,685, filed August 26, 2019, the disclosure of which is incorporated herein by reference in its entirety.

Field of technology

The present invention relates to triazolopyrimidine compounds that modulate the activity of adenosine receptors, such as the A2A and A2B subtypes, and are useful for the treatment of diseases associated with adenosine receptor activity, including, for example, cancer, inflammatory diseases, cardiovascular diseases and neurodegenerative diseases.

State of the art

Adenosine is an extracellular signaling molecule that can modulate the immune response through many types of immune cells. Adenosine was first recognized as a physiological regulator of coronary vascular tone by Drury and Szent-Gyorgyu (Sachdeva, S. and Gupta, M. Saudi Pharmaceutical Journal, 2013, 21, 245-253), but it was not until 1970 that Sattin and Rall showed that adenosine regulates cell function by binding to specific receptors on the cell surface (Sattin, A., and Rall, TW, 1970. Mol. Pharmacol. 6,13-23; Hasko', G., at al., 2007, Pharmacol. Tep. 113,264-275).

Adenosine plays a vital role in various other physiological functions. It is involved in the synthesis of nucleic acids when linked to three phosphate groups; it forms ATP, an essential component of the cellular energy system. Adenosine can be formed by enzymatic cleavage of extracellular ATP or can also be released from damaged neurons and glial cells by passing through the damaged plasma membrane (Tautenhahn, M. et al. Neuropharmacology, 2012, 62, 1756-1766). Adenosine exerts various pharmacological effects in both the periphery and the central nervous system through its action on specific receptors localized on cell membranes (Matsumoto, T. et al. Pharmacol. Res., 2012, 65, 81-90). Alternative pathways for the formation of extracellular adenosine have been described. These pathways involve the production of adenosine from nicotinamide dinucleotide (NAD) instead of ATP by the combined action of CD38, CD203a, and CD73. CD73-independent adenosine production can also occur via other phosphates, such as alkaline phosphatase or prostate-specific phosphatase.

In humans, four subtypes of adenosine receptors are known, including the A1, A2A, A2B, and A3 receptors. A1 and A2A are high-affinity receptors, whereas A2B and A3 are low-affinity receptors. Adenosine and its agonists can act through one or more of these receptors and can modulate the activity of adenylate cyclase, the enzyme responsible for increasing cyclic AMP (cAMP). Different receptors have different stimulatory and inhibitory effects on this enzyme. Increased intracellular cAMP concentrations can suppress the activity of immune and inflammatory cells (Livingston, M. et al., InflaMM. Res., 2004, 53, 171-178).

The adenosine receptor A2A can transmit signals to the periphery and the CNS, with agonists being investigated as anti-inflammatory drugs and antagonists being investigated in neurodegenerative diseases (Carlsson, J. et al., J. Med. Chern., 2010, 53, 3748-3755). In most cell types, the A2A subtype inhibits intracellular calcium levels, while A2B potentiates them. The A2A receptor, in general, inhibits the inflammatory response of immune cells (Borrmann, T. et al., J. Med. Chem., 2009, 52(13), 3994-4006).

A2B receptors are highly expressed in the gastrointestinal tract, bladder, lungs, and on mast cells (Antonioli, L. et al., Nature Reviews Cancer, 2013, 13, 842-857). The A2B receptor, although structurally closely related to the A2A receptor and capable of activating adenylate cyclase, is functionally distinct from it. It has been suggested that this subtype may utilize signal transduction systems distinct from adenylate cyclase (Livingston, M. et al., InflaMM. Res., 2004, 53, 171-178). Among all adenosine receptors, the A2B adenosine receptor is a low-affinity receptor that is thought to be inactive under physiological conditions and is activated by increased extracellular adenosine levels (Ryzhov, S. et al. Neoplasia, 2008, 10, 987-995). Activation of the A2B adenosine receptor can stimulate adenylate cyclase and phospholipase C through activation of Gs and Gq proteins, respectively. Binding to mitogen-activated protein kinases has also been described (Borrmann, T. et al., J. Med. Chem., 2009, 52(13), 3994-4006).

In the immune system, engagement of adenosine signaling may be a critical regulatory mechanism protecting tissues from excessive immune responses. Adenosine can negatively modulate immune responses through many types of immune cells, including T cells, natural killer cells, macrophages, dendritic cells, mast cells, and myeloid-derived suppressor cells (Allard, B. et al. Current Opinion in Pharmacology, 2016, 29, 7-16).

In tumors, this pathway is hijacked by the tumor microenvironment and sabotages the immune system's antitumor ability, promoting cancer progression. In the tumor microenvironment

- 1 049302 Adenosine was mainly generated from extracellular ATP via CD39 and CD73. Several cell types can generate adenosine by expressing CD39 and CD73. This includes tumor cells, T effector cells, T regulatory cells, tumor-associated macrophages, myeloid-derived suppressor cells (MDSCs), endothelial cells, cancer-associated fibroblasts (CAFs), and mesenchymal stromal/stem cells (MSCs). Hypoxia, inflammation, and other immunosuppressive signals in the tumor microenvironment can induce CD39, CD73 expression and subsequent adenosine production. As a result, adenosine levels in solid tumors are unusually high compared to normal physiological conditions.

A2A are primarily expressed on cells of lymphoid origin, including T effector cells, T regulatory cells, and natural killer cells. Blocking the A2A receptor can prevent subsequent immunosuppressive signals that transiently inactivate T cells. A2B receptors are primarily expressed on cells of monocyte origin, including dendritic cells, tumor-associated macrophages, myeloid-derived suppressor cells (MDSCs), and mesenchymal stromal/stem cells (MSCs). Blocking the A2B receptor in preclinical models can suppress tumor growth, block metastasis, and enhance tumor antigen presentation.

Regarding the safety profile of ADORA2A/ADORA2B (A2A/A2B) blockade, all A2A and A2B receptor knockout mice are viable, do not exhibit growth abnormalities, and are fertile (Allard, B. et al. Current Opinion in Pharmacology, 2016, 29, 7-16). A2A KO mice showed elevated levels of proinflammatory cytokines only after LPS challenge and no signs of inflammation at baseline (Antonioli, L. et al., Nature Reviews Cancer, 2013, 13, 842-857). A2B KO mice exhibited normal platelet, red blood cell, and white blood cell counts but increased inflammation at baseline (TNF-alpha, IL-6) compared to naïve A2B KO mice (Antonioli, L. et al., Nature Reviews Cancer, 2013, 13, 842-857). Exaggerated TNF-alpha and IL-6 production was detected after LPS treatment. A2B KO mice also exhibited increases in vascular adhesion molecules that mediate inflammation as well as leukocyte adhesion/rolling; enhanced mast cell activation; increased susceptibility to IgE-mediated anaphylaxis; and increased vascular leakage and neutrophil influx during hypoxia (Antonioli, L. et al., Nature Reviews Cancer, 2013, 13, 842-857).

Thus, there is a need to develop new ligands selective for adenosine receptors, such as the A2A and A2B subtypes, for the treatment of diseases such as cancer, inflammatory diseases, cardiovascular diseases and neurodegenerative diseases. This application addresses this need and others.

The essence of the invention

The present invention relates, inter alia, to compounds of formula (I):

Su ²

Nhb(I) or pharmaceutically acceptable salts thereof, the constituent elements being as defined herein.

The present invention also provides pharmaceutical compositions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

The present invention also provides methods for inhibiting adenosine receptor activity comprising contacting the receptor with a compound of formula (I) or a pharmaceutically acceptable salt thereof.

The present invention also provides methods for treating a disease or disorder associated with abnormal expression of adenosine receptors, comprising administering to said patient a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

The present invention also provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in any of the methods described herein.

The present invention also provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the preparation of a medicament for use in any of the methods described herein.

- 2 049302

Detailed description

Connections.

The present invention relates, inter alia, to compounds of formula (I):

Oh ²

NH, (I) or their pharmaceutically acceptable salts; where

R ² is selected from LW and L-W'-Z;

wherein L is selected from C _1-3 alkyl, -C _1-3 alkyl-O-, -OC _1-3 alkyl-, -C _1-3 alkyl-NH-, -NH-C _1-3 alkyl-, -NH-C _1-3 alkyl-NH- and -N(C _1-3 alkyl)-;

wherein W is 5-6 membered heteroaryl optionally substituted with 1, 2 or 3 groups each independently selected from cyano, halogen and C _1-3 alkyl;

wherein W' is phenyl or a 5-6-membered heteroaryl, wherein each of the phenyl or 5-6-membered heteroaryl W' is optionally substituted with 1, 2 or 3 groups each independently selected from cyano, halogen and C _1-3 alkyl; and wherein Z is phenyl or a 5-6-membered heteroaryl, wherein each of the phenyl or 5-6-membered heteroaryl Z is optionally substituted with 1, 2 or 3 groups each independently selected from cyano, halogen, C _1-3 alkyl, amine and C _1-3 alkoxy;

Cy ¹ is selected from cyanophenyl and cyanofluorophenyl; and

Cy ² is a 5-6 membered heteroaryl optionally substituted with 1, 2 or 3 groups each independently selected from C _1-3 alkyl, C _1-3 alkoxy, NH ₂ , NH(C _1-3 alkyl) and N(C _1-3 alkyl) ₂ , and wherein the ring-forming carbon atom of Cy ² is optionally substituted with an oxo group.

In some embodiments, R ² is LW.

In some embodiments, R ² is L-W'-Z.

In some embodiments, L is selected from -CH2-, -NH-CH2-, -O-CH2-, -NH-, -CH2-O-, -NH-CH(CH3)-, and -NH-C(CH3)2-.

In some embodiments, W is pyridinyl, which is optionally substituted with 1 or 2 groups each independently selected from methyl, fluoro, and cyano.

In some embodiments, W' is selected from tetrazolyl and phenyl, wherein phenyl is optionally substituted with fluoro.

In some embodiments, Z is selected from phenyl, pyridinyl, pyrazolyl, thiazolyl, pyrimidinyl, and pyrazinyl, each of which is optionally substituted with 1 or 2 groups independently selected from cyano, halogen, methyl, amino, and methoxy.

In some embodiments, Cy ¹ is 3-cyanophenyl, optionally substituted with fluorine.

In some embodiments, Cy ² is selected from pyrimidinyl, 1-methyl-6-oxo-1,6-dihydropyridinyl, 1-methyl-6-oxo-1,6-dihydropyridazinyl, pyridinyl, dimethylaminopyridinyl, aminopyridinyl, 1-ethyl-6-oxo-1,6-dihydropyridinyl, methylpyridinyl, dimethylpyridinyl, and methoxymethylpyridinyl.

In some embodiments of the invention, the compound of formula (I) is selected from:

3-(5-amino-2-((5-(3-aminophenyl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(6-methylpyridin-2-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(6-methoxypyridin-2-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(pyridin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

-(2-((5-(1H-pyrazol-1-yl)-1H-tetrazol-1-yl)methyl)-5-amino-8-(1-methyl-6-oxo-1,6dihydropyridin-3-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(thiazol-4-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(pyrimidin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(pyrazin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-((5-(pyridin-2-yl)-1H-tetrazole-1

- 3 049302 (yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(pyridin-2-yl)-1H-tetrazol-1-yl)methyl)-8-(pyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2-(dimethylamino)pyridin-4-yl)-2-((5-(pyridin-2-yl)-1H-tetrazol-1-yl)methyl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2-aminopyridin-4-yl)-2-((5-(pyridin-2-yl)-III-tetrazol-1-yl)methyl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-(2-fluoro-6-(pyridin-4-yl)benzyl)-8-(pyridin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile;

3-(5-amino-8-(2-aminopyridin-4-yl)-2-(2-(2-aminopyridin-4-yl)-6-fluorobenzyl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-ethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((3-methylpyridin-2-yl)methoxy)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-((3-methylpyridin-2-yl)methoxy)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3-methylpyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-((3-methylpyridin-2-yl)methoxy)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-ethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((3-methylpyridin-2-yl)methoxy)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile;

3-(5-amino-8-(1-ethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile;

3-(5-amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(pyridin-2-ylmethyl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile;

3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-((3-fluoropyridin-2-yl)methoxy)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(2-methoxy-6-methylpyridin-4-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-(pyridin-2-ylamino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(pyridin-2-ylamino)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(pyridin-2-ylamino)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(3-methylpyridin-4-yl)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile;

3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile;

3-(5-amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((6-methylpyridin-2-yl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin- 4 049302

7-yl)benzonitrile;

3-(5-amino-2-((pyridin-2-yloxy)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile;

2-((5-amino-7-(3-cyanophenyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-2yl)methoxy)nicotinonitrile;

2-((5-amino-7-(3-cyanophenyl)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile;

2-((5-amino-7-(3-cyanophenyl)-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile;

2-((5-amino-7-(3-cyanophenyl)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-2yl)methoxy)nicotinonitrile;

2-((5-amino-7-(3-cyanophenyl)-8-(2,6-dimethylpyridin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-2yl)methoxy)nicotinonitrile;

2-((5-amino-7-(3-cyanophenyl)-8-(2-methoxy-6-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile;

3-(5-amino-2-((1-(pyridin-2-yl)ethyl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile;

3-(5-amino-2-((2-(pyridin-2-yl)propan-2-yl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(pyridin-2-yl)-2H-tetrazol-2-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(pyrimidin-2-yl)-III-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(pyrimidin-4-yl)-2-((5-(pyrimidin-4-yl)-III-tetrazol-1-yl)methyl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(pyridin-3-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(pyridin-4-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((6-methylpyridin-2-yl)methoxy)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-((6-methylpyridin-2-yl)methoxy)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((6-methylpyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-((6-methylpyridin-2-yl)methoxy)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-((6-methylpyridin-2-yl)methoxy)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(2,6-dimethylpyridin-4-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(2-methoxy-6-methylpyridin-4-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-(((6-methylpyridin-2-yl)methyl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((6-methylpyridin-2-yl)methyl)amino)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(((6-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-(((6-methylpyridin-2-yl)methyl)amino)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-(((6-methylpyridin-2-yl)methyl)amino)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; and

3-(5-amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-(((6-methylpyridin-2-yl)methyl)amino)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

or a pharmaceutically acceptable salt thereof.

- 5 049302

In some embodiments of the invention, the compound of formula I is selected from:

3-(5-amino-2-((5-(3-aminophenyl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(6-methylpyridin-2-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(6-methoxypyridin-2-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(pyridin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

-(2-((5-(1H-pyrazol-1-yl)-1H-tetrazol-1-yl)methyl)-5-amino-8-(1-methyl-6-oxo-1,6dihydropyridin-3-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(thiazol-4-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(pyrimidin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(pyrazin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-((5-(pyridin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(pyridin-2-yl)-III-tetrazol-1-yl)methyl)-8-(pyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2-(dimethylamino)pyridin-4-yl)-2-((5-(pyridin-2-yl)-III-tetrazol-1-yl)methyl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2-aminopyridin-4-yl)-2-((5-(pyridin-2-yl)-III-tetrazol-1-yl)methyl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-(2-fluoro-6-(pyridin-4-yl)benzyl)-8-(pyridin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile;

3-(5-amino-8-(2-aminopyridin-4-yl)-2-(2-(2-aminopyridin-4-yl)-6-fluorobenzyl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-ethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((3-methylpyridin-2-yl)methoxy)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-((3-methylpyridin-2-yl)methoxy)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3-methylpyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-((3-methylpyridin-2-yl)methoxy)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-ethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((3-methylpyridin-2-yl)methoxy)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile;

3-(5-amino-8-(1-ethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile;

3-(5-amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(pyridin-2-ylmethyl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile;

3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-((3-fluoropyridin-2-yl)methoxy)-[1,2,4]triazolo[1,5

- 6 049302

c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(2-methoxy-6-methylpyridin-4-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-(pyridin-2-ylamino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile;

3-(5-amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((6-methylpyridin-2-yl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile;

3-(5-amino-2-((pyridin-2-yloxy)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile;

2-((5-amino-7-(3-cyanophenyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-2yl)methoxy)nicotinonitrile;

2-((5-amino-7-(3-cyanophenyl)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile;

2-((5-amino-7-(3-cyanophenyl)-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile;

2-((5-amino-7-(3-cyanophenyl)-8-(2,6-dimethylpyridin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-2yl)methoxy)nicotinonitrile;

2-((5-amino-7-(3-cyanophenyl)-8-(2-methoxy-6-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile;

3-(5-amino-2-((5-(pyridin-2-yl)-2H-tetrazol-2-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(pyrimidin-2-yl)-III-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(pyrimidin-4-yl)-2-((5-(pyrimidin-4-yl)-III-tetrazol-1-yl)methyl)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(pyridin-3-yl)-III-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((5-(pyridin-4-yl)-III-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((6-methylpyridin-2-yl)methoxy)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-((6-methylpyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-((6-methylpyridin-2-yl)methoxy)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-((6-methylpyridin-2-yl)methoxy)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-(((6-methylpyridin-2-yl)methyl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(((6-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-2-(((6-methylpyridin-2-yl)methyl)amino)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile;

3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-(((6-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; and

3-(5-amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-(((6-methylpyridin-2-yl)methyl)amino)-

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile;

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is an (S)-enantiomer of one of the preceding compounds or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is an (R)-enantiomer of one of the preceding compounds or a pharmaceutically acceptable salt thereof.

It should also be understood that some features of the invention that are described in the context of separate embodiments for clarity may also be provided in combination in a single embodiment. Conversely, various features of the invention that are described in the context of a single embodiment for brevity are also provided separately or in any suitable subcombination.

At various points in the description of this invention, divalent linking substituents are described. In particular, each divalent linking substituent is intended to include both the direct and reverse forms of said linking substituent. For example,

- 7 049302

-NR(CR'R) _n - includes both -NR(CR'R) _n - and -(CR'R) _n NR-. It is assumed that when a linking group is explicitly required in a structure, the Markush variables listed for that group are the linking groups.

The term n-membered, where n is an integer, generally describes the number of ring-forming atoms in a moiety where the number of ring-forming atoms is n. For example, piperidinyl is an example of a 6-membered heterocycloalkyl ring, pyrazolyl is an example of a 5-membered heteroaryl ring, pyridyl is an example of a 6-membered heteroaryl ring, and 1,2,3,4-tetrahydronaphthalene is an example of a 10-membered cycloalkyl group.

As used herein, the phrase optionally substituted means unsubstituted or substituted. The substituents are independently selected and the substitution may be at any chemically accessible position. As used herein, the term substituted means that a hydrogen atom is removed and replaced by a substituent. One divalent substituent, such as oxo, may replace two hydrogen atoms. It should be understood that substitution at a given atom is limited by valence.

In the context of this document, the phrase each variable is independently selected from means essentially the same as in each case the variable is selected from.

In all definitions, the term C _n _m denotes a range that includes the endpoints where n and m are integers and indicate the number of carbon atoms. Examples include C1_ ₃ , C1_ ₄ , C1-6 and similar compounds.

As used herein, the term C _nm alkyl, used alone or in combination with other terms, refers to a saturated hydrocarbon group, which may be straight or branched, containing from n to m carbon atoms. Examples of alkyl moieties include, but are not limited to, chemical groups such as methyl (Me), ethyl (Et), n-propyl (n-Pr), isopropyl (iso-Pr), n-butyl, tert-butyl, isobutyl, sec-butyl; higher homologues such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, 1,2,2-trimethylpropyl and the like. In some embodiments, the alkyl group contains from 1 to 6 carbon atoms, from 1 to 4 carbon atoms, from 1 to 3 carbon atoms, or from 1 to 2 carbon atoms.

As used herein, the term C _nm alkoxy, used alone or in combination with other terms, refers to a group of the formula -O-alkyl, wherein the alkyl group contains from n to m carbon atoms. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), butoxy (e.g., n-butoxy and tert-butoxy), and the like. In some embodiments, the alkyl group contains from 1 to 6, from 1 to 4, or from 1 to 3 carbon atoms.

In the context of this document, the term amino means a group of the formula -NH2.

As used herein, the term halo or halogen refers to F, Cl, Br, or I. In some embodiments, the halogen is F, Cl, or Br. In some embodiments, the halogen is F or Cl. In some embodiments, the halogen is F. In some embodiments, the halogen is Cl.

As used herein, the term heteroaryl refers to a monocyclic aromatic heterocycle having at least one ring heteroatom selected from N, O, S, and B. In some embodiments, the heteroaryl ring has 1, 2, 3, or 4 ring heteroatoms independently selected from N, O, S, and B. In some embodiments, any ring-forming N in the heteroaryl moiety can be Noxide. In some embodiments, the heteroaryl is a five-membered heteroaryl ring. In some embodiments, the heteroaryl is a six-membered heteroaryl ring. In some embodiments, the heteroaryl group has 1 to 4 ring-forming heteroatoms, 1 to 3 ring-forming heteroatoms, 1 to 2 ring-forming heteroatoms, or 1 ring-forming heteroatom. When the heteroaryl group has more than one ring heteroatom, the heteroatoms can be the same or different. Examples of heteroaryl groups include, but are not limited to, pyridine, pyrimidine, pyrazine, pyridazine, dihydropyridine, dihydropyridazine, pyrrole, pyrazole, azolyl, oxazole, isoxazole, thiazole, isothiazole, imidazole, furan, thiophene, triazole, tetrazole, thiadiazole, triazine.

In some places, definitions or embodiments of the invention refer to specific rings (e.g., azetidine ring, pyridine ring, etc.). Unless otherwise specified, these rings may be attached at any atom in the ring, provided that the valency of the atom is not exceeded. For example, an azetidine ring may be attached at any position in the ring, with the pyridin-3-yl ring being attached at position 3.

In the context of this document, the term oxo refers to an oxygen atom (i.e., =O) as a divalent substituent that forms a carbonyl group when attached to a carbon atom (e.g., C=O or C(O)), or when attached to a nitrogen or sulfur heteroatom to form a nitroso, sulfinyl, or sulfonyl group.

- 8 049302

In the context of this document, the term independently selected from means that each occurrence of a variable or substituent is independently selected in each instance from the applicable list.

The compounds described herein may be asymmetric (e.g., having one or more stereocenters). All stereoisomers, e.g., enantiomers and diastereomers, are intended unless otherwise noted. Compounds of this invention that contain asymmetrically substituted carbon atoms may be isolated in optically active or racemic forms. Methods for preparing optically active forms from optically inactive starting materials are known in the art, such as by resolution of racemic mixtures or by stereoselective synthesis. Also, various geometric isomers of olefins, C=N double bonds, and the like may be present in the compounds described herein, and all such stable isomers are contemplated by this invention. Cis- and trans-geometric isomers of the compounds of this invention are described and may be isolated as a mixture of isomers or as separate isomeric forms. In some embodiments, the compound has the s-configuration. In some embodiments, the compound has the ^-configuration. The formulas (e.g., formulas (I), (II), etc.) provided herein include stereoisomers of the compounds.

The separation of racemic mixtures of compounds can be carried out by any of a variety of methods known in the art. An example of a method includes fractional recrystallization using a chiral resolving acid, which is an optically active salt-forming organic acid. Suitable resolving agents for fractional recrystallization methods are, for example, optically active acids, such as the D and L forms of tartaric acid, diacetyl tartaric acid, dibenzoyl tartaric acid, mandelic acid, malic acid, lactic acid, or various optically active camphorsulfonic acids, such as β-camphorsulfonic acid. Other resolving agents useful in fractional recrystallization methods include stereoisomerically pure forms of αmethylbenzylamine (e.g., S and R forms, or diastereomerically pure forms), 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, and similar compounds.

Separation of racemic mixtures can also be carried out by elution on a column containing an optically active resolving agent (e.g. dinitrobenzoylphenylglycine). A suitable composition of the eluting solvent can be determined by a person skilled in the art.

The compounds provided herein also include tautomeric forms. Tautomeric forms arise from the substitution of a single bond by an adjacent double bond along with the concomitant transfer of a proton. Tautomeric forms include prototropic tautomers, which are isomeric protonation states having the same empirical formula and overall charge. Examples of prototropic tautomers include keto-enol pairs, amide-imido acid pairs, lactam-lactim pairs, enamine-imine pairs, and ring forms where a proton can occupy two or more positions of a heterocyclic system, such as 1H- and 3H-imidazole, 1H-, 2H-, and 4H-1,2,4-triazole, 1H- and 2H-isoindole, 2-hydroxypyridine and 2-pyridone, and 1H- and 2H-pyrazole. Tautomeric forms can exist in equilibrium or be sterically locked into one form by using suitable substitution.

All compounds and their pharmaceutically acceptable salts may be present with other substances, such as water and solvents (e.g. hydrates and solvates) or may be isolated.

In some embodiments of the invention, the preparation of compounds may involve the addition of acids or bases to influence, for example, the catalysis of a desired reaction or to form salt forms, for example, acid addition salts.

In some embodiments, the compounds provided herein or salts thereof are substantially isolated. By substantially isolated is meant that the compound is at least partially or substantially separated from the medium in which it was prepared or found. Partial separation may include, for example, a composition enriched in the compounds provided herein. Substantial separation may include compositions containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of the compounds provided herein or salts thereof. Methods for isolating compounds and salts thereof are routine in the art.

The term compound as used herein is intended to include all stereoisomers, geometric isomers, tautomers and isotopes of the structures shown. Compounds identified herein by name or structure as one particular tautomeric form are intended to include other tautomeric forms unless otherwise indicated.

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The phrase pharmaceutically acceptable is used herein to mean those compounds, materials, compositions and/or dosage forms that are medically suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reaction or other problem or complication, proportionate to a reasonable benefit/risk ratio.

This application also includes pharmaceutically acceptable salts of the compounds described herein. The term pharmaceutically acceptable salts refers to derivatives of the disclosed compounds, wherein the parent compound is modified by converting an existing acidic or basic moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, salts of mineral or organic acids with basic residues, for example, amines; alkali or organic salts with acidic residues, for example, carboxylic acids; and similar salts. Pharmaceutically acceptable salts of the invention include conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the invention can be synthesized from a parent compound that contains a basic or acidic moiety by conventional chemical methods. Typically, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of both; In general, non-aqueous media such as ether, ethyl acetate, alcohols (e.g., methanol, ethanol, isopropanol, or butanol), or acetonitrile (ACN) are preferred. Lists of suitable salts are provided in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, and Journal of Pharmaceutical Science, 66, 2 (1977), the contents of each of which are incorporated herein by reference in their entirety.

Synthesis.

As will be appreciated by those skilled in the art, the compounds provided herein, including their salts and stereoisomers, can be prepared using known methods of organic synthesis and can be synthesized according to any of numerous possible synthetic routes.

Scheme 1.

Compounds of formulae 1-9 can be synthesized by the synthetic route suggested in Scheme 1. Starting material 1-1 is first cross-coupled with reagent 1-2 to form compound 1-3, in which M is a boronic acid, boronic ester, or appropriately substituted metal [e.g., M is B(OR) ₂ , Sn(alkyl) ₃ , or Zn-Hal], under standard Suzuki cross-coupling conditions (e.g., in the presence of a palladium catalyst and a suitable base) or under standard Stille cross-coupling conditions (e.g., in the presence of a palladium catalyst) or under standard Negishi cross-coupling conditions (e.g., in the presence of a palladium catalyst). Nucleophilic aromatic substitution reaction (SNAr) of compound 1-3 with hydrazide 14 then affords compound 1-5, which undergoes cyclization reaction at elevated temperature in the presence of a suitable reagent, for example, N,O-bis(trimethylsilyl)acetamide, to afford bicycle 1-6. Halogenation of 1-6 with a suitable reagent, for example, N-bromosuccinimide (NBS), affords compound 1-7. The final product 1-9 can be prepared by cross-coupling reaction between compound 1-7 and a derivative of formula 1-8 using procedures similar to those described for the preparation of compound 1-3 from starting material 1-1. At various stages of this synthetic sequence, the R ² group can be further functionalized.

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Scheme 2.

Compounds of formula 2-4 can be synthesized by the synthetic route suggested in Scheme 2. Improved intermediate 2-1 (which can be prepared using the synthetic procedures as shown in Scheme 1) is first subjected to a halogenation reaction (using a suitable reagent, e.g., thionyl chloride) to form compound 2-2 (Hal is a halide, e.g., F, Cl, Br, or I). Compound 2-2 can then be subjected to a nucleophilic substitution (SN2) reaction with reagents of formula 2-3 to give compound 2-4.

Scheme 3.

Su=- M

M-,LύY «AN/__.

k ^F Y_7 NH, F3 1 3 3

Compounds of formula 3-6 can be synthesized by the synthetic route suggested in Scheme 3. Improved intermediate 3-1 (which can be prepared using the synthetic procedures as shown in Scheme 1) is first cross-coupled with reagent 3-2 to afford compound 3-3, in which M is a boronic acid, boronic ester, or appropriately substituted metal [e.g., M is B(OR) ₂ , Sn(Alkyl) ₃ , or Zn-Hal], under standard Suzuki cross-coupling conditions (e.g., in the presence of a palladium catalyst and a suitable base), or under standard Stille cross-coupling conditions (e.g., in the presence of a palladium catalyst), or under standard Negishi cross-coupling conditions (e.g., in the presence of a palladium catalyst). Bromination (NBS) reaction of 3-3 affords compound 3-4. Compound 3-4 can then be cross-coupled with reagent 3-5, using procedures similar to those described for the preparation of compound 3-3 from 3-1, to give compound 3-6.

Scheme 4.

Compounds of formula 4-10 can be synthesized by the synthetic route suggested in Scheme 4. The selective nucleophilic aromatic substitution reaction ( _SN Ar) of starting material 41 with amine 4-2 (ZG is a suitable protecting group, e.g. 4-methoxybenzyl) gives

- 11 049302 compound 4-3. Compound 4-3 can then be cyclized to intermediate 4-4 by appropriate chemical transformations, for example, a two-step sequence using O-ethyl carboisothiocyanatidate and hydroxylamine hydrochloride. Cross-coupling reaction between 4-4 and a reagent of formula 4-5 in which M is a boronic acid, boronic ester, or appropriately substituted metal [e.g., M is B(OR)2, Sn(Alkyl) ₃ , or Zn-Hal], under standard Suzuki cross-coupling conditions (e.g., in the presence of a palladium catalyst and a suitable base) or under standard Stille cross-coupling conditions (e.g., in the presence of a palladium catalyst) or under standard Negishi cross-coupling conditions (e.g., in the presence of a palladium catalyst), will provide intermediate 4-6. The amino group of 4-6 can then be functionalized using suitable chemistry, such as Buchwald-Hartwig coupling conditions in the presence of a palladium catalyst (e.g., chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino1,1'-biphenyl)]palladium(II)) and a base (e.g., sodium tert-butoxide) or reductive amination conditions (e.g., in the presence of a suitable hydride source) to provide 4-7. Halogenation of 4-7 using a suitable reagent, such as N-bromosuccinimide (NBS), provides 4-8. Cross-coupling between 4-8 and a derivative of formula 4-9 using procedures similar to those described for the preparation of 4-6 from 4-4, followed by deprotection, provides 4-10.

Scheme 5.

Compounds of formula 5-7 can be synthesized by the synthetic route suggested in Scheme 5. The amino group of compound 5-1 (which can be obtained by the synthetic procedures suggested in Scheme 4) can first be converted to a halogen by suitable chemical transformations, for example, the Sandmeyer reaction (for example, in the presence of a suitable oxidizing agent, for example, isobutyl nitrite and a suitable halogen source) to give 5-2. Then, a nucleophilic aromatic substitution reaction (S _N Ar) of compound 5-2 with alcohol 5-3 in the presence of a suitable base gives compound 5-4. Halogenation of 5-4 using a suitable reagent, for example, N-bromosuccinimide (NBS), gives compound 5-5. A cross-coupling reaction between 5-5 and a derivative of formula 5-6, followed by removal of the protecting group, gives the product 5-7.

Methods of application.

The compounds of this invention can modulate the activity of adenosine receptors, such as the A2A and A2B receptor subtypes. Accordingly, the compounds, salts, or stereoisomers described herein can be used in methods of inhibiting adenosine receptors (e.g., A2A and/or A2B receptors) by contacting the receptor with any one or more of the compounds, salts, or compositions described herein. In some embodiments, the compounds or salts can be used in methods of inhibiting adenosine receptor activity in an individual/patient in need of inhibition by administering an effective amount of a compound or salt described herein. In some embodiments, modulation is inhibition. In some embodiments, the contacting occurs in vivo. In some embodiments, the contacting occurs ex vivo or in vitro.

The compounds or salts described herein may be selective. By selective is meant that the compound binds to or inhibits an adenosine receptor with greater affinity or potency, respectively, than at least one other receptor, kinase, etc. The compounds of this invention may also be dual antagonists (i.e., inhibitors) of adenosine receptors, e.g., adenosine

- 12,049,302 A2A and A2B receptors.

Another aspect of the invention relates to methods for treating a disease or disorder associated with an adenosine receptor in an individual (e.g., a patient) by administering to the individual in need of such treatment a therapeutically effective amount or dose of one or more compounds of the invention or a pharmaceutical composition thereof. A disease or disorder associated with an adenosine receptor may include any disease, disorder, or condition that is directly or indirectly associated with the expression or activity of an adenosine receptor, including overexpression and/or abnormal levels of activity.

The compounds of the present invention are useful in the treatment of diseases associated with adenosine receptor activity, including, for example, cancer, inflammatory diseases, cardiovascular diseases, neurodegenerative diseases, immunomodulatory disorders, central nervous system diseases and diabetes.

Based on the important role of adenosine receptors, such as A2A, A2B, in many immunosuppressive mechanisms, the development of inhibitors can stimulate the immune system to suppress tumor progression. Adenosine receptor inhibitors can be used to treat, alone or in combination with other therapies, bladder cancer, lung cancer (e.g., non-small cell lung cancer (NSCLC), lung metastasis), melanoma (e.g., metastatic melanoma), breast cancer, cervical cancer, ovarian cancer, colorectal cancer, pancreatic cancer, esophageal cancer, prostate cancer, kidney cancer, skin cancer, thyroid cancer, liver cancer, uterine cancer, head and neck cancer, and renal cell carcinoma (Antonioli, L. et al., Nature Reviews Cancer, 2013, 13, 842-857). See also https://globenewswire.com/newsrelease/2017/04/04/954192/0/en/Corvus-Pharmaceuticals-Announces-Interim-Results-from-Ongoing -Phase-11b-Study-Demonstrating-Safety-and-Clinical-Actrvity-of-Lead-Checkpoint-Inhibitor-CPI-444-in-Patients-withAdva.html; Cekic C et al. J Immunol 2012, 188:198-205; Lannone, R. et al., Am. J. Cancer Res. 2014, 4:172181 (study shows that blocking both A2A and CD73 enhances the antitumor activity of anti-CTLA-4 monoclonal antibody (mAb) therapy in a B16F10 murine melanoma model); Lannone, R. et al., Neoplasia, 2013, 15:1400-1410 and Beavis PA., et al., Proc Natl Acad Sci. USA, 2013, 110:14711-14716 (study shows that blocking A2A and CD73 reduces metastasis in a CD73-high 4T1 breast tumor model). In some embodiments, the prostate cancer is metastatic castration-resistant prostate cancer (mCRPC). In some embodiments, the colorectal cancer is colorectal carcinoma (CRC).

In some embodiments, the disease or disorder is lung cancer (e.g., non-small cell lung cancer), melanoma, pancreatic cancer, breast cancer, squamous cell cancer of the head and neck, prostate cancer, liver cancer, colorectal cancer, endometrial cancer, bladder cancer, skin cancer, uterine cancer, kidney cancer, gastric cancer, or sarcoma. In some embodiments, the sarcoma is Askin's tumor, botryoid sarcoma, chondrosarcoma, Ewing's sarcoma, malignant hemangioendothelioma, malignant schwannoma, osteosarcoma, alveolar soft tissue sarcoma, angiosarcoma, cystosarcoma phyllodes, dermatofibrosarcoma protuberans, desmoid tumor, desmoplastic small round cell tumor, epithelioid sarcoma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, fibrosarcoma, gastrointestinal stromal tumor (GIST), hemangiopericytoma, hemangiosarcoma, Kaposi's sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, lymphosarcoma, malignant peripheral nerve sheath tumor (MPNST), neurofibrosarcoma, rhabdomyosarcoma, synovial sarcoendothelioma, or undifferentiated pleomorphic sarcoma.

In some embodiments, the disease or disorder is mesothelioma or adrenocarcinoma. In some embodiments, the disease or disorder is mesothelioma. In some embodiments, the disease or disorder is adenocarcinoma.

MDSCs (myeloid-derived suppressor cells) are a heterogeneous group of immune cells of the myeloid lineage (a family of cells derived from bone marrow stem cells). MDSCs proliferate in pathological situations such as chronic infections and cancer as a result of altered hematopoiesis. MDSCs differ from other types of myeloid cells in that they have pronounced immunosuppressive activity rather than immunostimulatory properties. Like other myeloid cells, MDSCs interact with other types of immune cells, including T cells, dendritic cells, macrophages, and natural killer cells, to regulate their functions. In some embodiments, the compounds and others described herein can be used in methods relating to cancerous tissue (e.g., tumors) with high MDSC infiltration, including solid tumors with high basal levels of macrophage and/or MDSC infiltration.

In some embodiments of the invention, the disease or disorder is

- 13 049302 is head and neck squamous cell carcinoma (HNSCC), non-small cell lung cancer (NSCLC), colorectal cancer, melanoma, ovarian cancer, bladder cancer, renal cell carcinoma, liver cancer, or hepatocellular carcinoma.

In some embodiments, the compounds of the invention can be used to treat pulmonary inflammation, including bleomycin-induced pulmonary fibrosis and adenosine deaminase deficiency-associated injury (Baraldi, et al., Chem. Rev., 2008, 108, 238-263).

In some embodiments, the compounds of this invention can be used to treat an inflammatory disease, such as allergic reactions (e.g., adenosine receptor-dependent allergic reactions A2B) and other adenosine receptor-dependent immune reactions. Other inflammatory diseases that can be treated with the compounds of this invention include respiratory diseases, sepsis, reperfusion injury, and thrombosis.

In some embodiments, the compounds of the invention can be used to treat cardiovascular diseases, such as ischemic heart disease (myocardial infarction, angina, heart failure), cerebrovascular disease (stroke, transient ischemic attack), peripheral arterial disease and atherosclerosis and aortic aneurysm. Atherosclerosis is a major etiologic factor in many types of cardiovascular diseases. Atherosclerosis begins in adolescence with fatty streaks that progress to plaques in adulthood and finally leads to thrombotic events causing vascular occlusion, resulting in clinically significant morbidity and mortality. Adenosine A2B receptor and adenosine A2A receptor antagonists may be useful in preventing atherosclerotic plaque formation (Eisenstein, A. et al., J. Cell Physiol., 2015, 230(12), 2891-2897).

In some embodiments, the compounds of the invention can be used to treat movement disorders; deficits caused by degeneration of the striatonigral dopamine system; and Parkinson's disease; certain motivational symptoms of depression (Collins, LE et al. Pharmacol. Biochem. Behav., 2012, 100, 498-505.).

In some embodiments, the compounds of the invention can be used to treat diabetes and related disorders, such as insulin resistance. Diabetes affects adenosine production and expression of adenosine A2B receptors (A2BRs), which stimulate IL-6 and CRP production, insulin resistance, and the association between A _2B R gene single nucleotide polymorphisms (ADORA2B SNPs) and inflammatory markers. Increased A2BR signaling in diabetes may increase insulin resistance in part by increasing levels of proinflammatory mediators. Selective A2BR blockers may be useful for the treatment of insulin resistance (Figler, RA et al. Diabetes, 2011, 60 (2), 669-679).

It is believed that the compounds provided herein, for example, the compounds of formula (I) or any variants thereof, may have a satisfactory pharmacological profile and promising biopharmaceutical properties, such as toxicological profile, metabolism and pharmacokinetic properties, solubility and permeability. It should be understood that the determination of suitable biopharmaceutical properties is within the knowledge of a person skilled in the art, for example, determining cytotoxicity in cells or inhibition of certain targets or channels to determine potential toxicity.

The terms individual or patient, used interchangeably, refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, and most preferably humans.

The phrase therapeutically effective amount refers to the amount of an active compound or pharmaceutical agent that elicits in a tissue, system, animal, individual, or human the biological or medical response sought by the researcher, veterinarian, physician, or other practitioner.

The term treatment or therapy refers to one or more of (1) inhibiting a disease; for example, inhibiting a disease, condition, or disorder in an individual who is experiencing or demonstrating the pathology or symptomatology of the disease, condition, or disorder (i.e., stopping the further development of the pathology and/or symptomatology); and (2) alleviating a disease; for example, alleviating a disease, condition, or disorder in an individual who is experiencing or demonstrating the pathology or symptomatology of the disease, condition, or disorder (i.e., regressing the pathology and/or symptomatology), for example, reducing the severity of the disease.

In some embodiments, the compounds of this invention are useful for preventing or reducing the risk of developing any of the diseases mentioned herein; for example, preventing or reducing the risk of developing a disease, condition or disorder in an individual who may be predisposed to said disease, condition or disorder but does not yet experience or exhibit the pathology or

- 14 049302 symptoms of the disease.

Types of combination therapy.

I. Immune checkpoint therapy.

In some embodiments, the dual A2A and A2B inhibitors provided herein may be used in combination with one or more immune checkpoint inhibitors to treat cancer as described herein. In one embodiment, the combination with one or more immune checkpoint inhibitors as described herein may be used to treat melanoma. The compounds of the invention may be used in combination with one or more immune checkpoint inhibitors. Exemplary immune checkpoint inhibitors include inhibitors directed against immune checkpoint molecules such as CD20, CD28, CD40, CD122, CD96, CD73, CD47, GITR, CSF1R, JAK, PI3K delta, PI3K gamma, TAM, arginase, HPK1, CD137 (also known as 4-1BB), ICOS, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, VISTA, TIGIT, PD-1, PD-L1, and PD-L2. In some embodiments, the immune checkpoint molecule is a stimulatory checkpoint molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, and CD137. In some embodiments, the immune checkpoint molecule is an inhibitory checkpoint molecule selected from A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3, TIGIT, and VISTA. In some embodiments, the compounds provided herein may be used in combination with one or more agents selected from KIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors, and TGFR-beta inhibitors.

In some embodiments, the dual A2A and A2B inhibitors provided herein may be used in combination with one or more agonists of immune checkpoint molecules, such as OX40, CD27, OX40, GITR, and CD137 (also known as 4-1BB).

In some embodiments, the immune checkpoint molecule inhibitor is an anti-PD1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

In some embodiments, the immune checkpoint molecule inhibitor is a PD-1 inhibitor, such as an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab, pembrolizumab (also known as MK-3475), durvalumab (Imfinzi®), pidilizumab, SHR-1210, PDR001, mgA012, PDR001, AB122, or AMP-224. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab or pembrolizumab. In some embodiments, the anti-PD1 antibody is pembrolizumab. In some embodiments, the anti-PD-1 monoclonal antibody is mgA012. In some embodiments, the anti-PD1 antibody is SHR-1210. Other anticancer agent(s) include antibody-based therapeutic agents, such as 4-1BB (eg, urelumab or utomilumab).

In some embodiments, the immune checkpoint molecule inhibitor is a PD-L1 inhibitor, such as an anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-L1 monoclonal antibody is BMS-935559, MEDI4736, MPDL3280A (also known as RG7446), or MSB0010718C. In some embodiments, the anti-PD-L1 monoclonal antibody is MPDL3280A or MEDI4736.

In some embodiments, the PD-1 and PD-L1 inhibitor is, for example, an anti-PD-1/PD-L1 monoclonal antibody. In some embodiments, the anti-PD-1/PD-L1 is MCLA-136.

In some embodiments, the inhibitor is INCB086550.

In some embodiments, the inhibitor is MCLA-145.

In some embodiments, the immune checkpoint molecule inhibitor is a CTLA-4 inhibitor, such as an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, AGEN1884, or CP-675,206.

In some embodiments, the immune checkpoint molecule inhibitor is a LAG3 inhibitor, such as an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016, LAG525, or INCAGN2385.

In some embodiments, the immune checkpoint molecule inhibitor is a TIM3 inhibitor, such as an anti-TIM3 antibody. In some embodiments, the anti-TIM3 antibody is INCAGN2390, MBG453, or TSR-022.

- 15 049302

In some embodiments, the immune checkpoint molecule inhibitor is a GITR inhibitor, such as an anti-GITR antibody. In some embodiments, the anti-GITR antibody is TRX518, MK-4166, INCAGN1876, MK-1248, Amg228, BMS-986156, GWN323, or MEDI1873.

In some embodiments, the immune checkpoint molecule inhibitor is an OX40 inhibitor, such as an anti-OX40 antibody or an OX40L fusion protein. In some embodiments, the anti-OX40 antibody is MEDI0562, MOXR-0916, PF-04518600, GSK3174998, or BMS-986178. In some embodiments, the OX40L fusion protein is MEDI6383.

In some embodiments, the immune checkpoint molecule inhibitor is a CD20 inhibitor, such as an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is obinutuzumab or rituximab.

The compounds of the present invention can be used in combination with bispecific antibodies. In some embodiments, one of the domains of the bispecific antibody targets PD-1, PD-L1, CTLA-4, GITR, OX40, TIM3, LAG3, CD137, ICOS, CD3, tumor-specific antigens (e.g., CD70), or the TGFβ receptor.

In some embodiments, the compounds of the invention may be used in combination with one or more metabolic enzyme inhibitors. In some embodiments, the metabolic enzyme inhibitor is an IDO1, TDO, or arginase inhibitor. Examples of IDO1 inhibitors include epacadostat, NLG919, BMS-986205, PF-06840003, IOM2983, RG-70099, and LY338196.

As indicated below, additional compounds, inhibitors, agents, etc. may be combined with the claimed compound as a single or continuous dosage form, or they may be administered simultaneously or sequentially as separate dosage forms.

II. Cancer treatment.

Cancer cell growth and survival can be influenced by multiple signaling pathways. Thus, for the treatment of such conditions, it may be advantageous to combine different enzyme/protein/receptor inhibitors that exhibit different preferences for the targets whose activity they modulate. Targeting more than one signaling pathway (or more than one biological molecule involved in a given signaling pathway) may reduce the likelihood of drug resistance in a cell population and/or reduce treatment toxicity.

The compounds of the present invention may be used in combination with one or more other enzyme/protein/receptor inhibitors or with one or more therapeutic agents for the treatment of diseases, such as cancer. Examples of diseases and indications treatable with combination therapy include those described herein.

The compounds of the present invention can be used in combination with one or more additional pharmaceutical agents, such as chemotherapeutic agents, immuno-oncological agents, metabolic enzyme inhibitors, chemokine receptor inhibitors and phosphatase inhibitors, as well as targeted therapy agents, such as Bcr-Abl, Flt-3, EGFR, HER2, JAK, c-MET, VEGFR, PDGFR, c-Kit, IGF-1R, RAF and FAK kinase inhibitors. The one or more additional pharmaceutical agents can be administered to the patient simultaneously or sequentially.

For example, the compounds disclosed herein can be combined with one or more inhibitors of the following kinases to treat cancer and other diseases or disorders described herein: Akt1, Akt2, Akt3, TGF-βR, PKA, PKG, PKC, CaM kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, INS-R, IGF-1R, IR-R, PDGFαR, PDGFβR, CSFIR, KIT, FLK-II, KDR/FLK-1, FLK-4, flt-1, FGFR1, FGFR2, FGFR3, FGFR4, c-Met, Ron, Sea, TRKA, TRKB, TRKC, FLT3, VEGFR/Flt2, Flt4, EphA1, EphA2, EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYK, FRK, JAK, ABL, ALK and B-Raf. Non-limiting examples of inhibitors that can be combined with the compounds of the invention to treat cancer and other diseases and disorders described herein include an FGFR inhibitor (FGFR1, FGFR2, FGFR3, or FGFR4, e.g., INCB54828, INCB62079, and INCB63904), a JAK inhibitor (JAK1 and/or JAK2, e.g., ruxolitinib, baricitinib, or INCB39110), an IDO inhibitor (e.g., epacadostat, NLG919, or BMS986205), an LSD1 inhibitor (e.g., INCB59872 and INCB60003), a TDO inhibitor, a PI3K-delta inhibitor (e.g., INCB50797 and INCB50465), a Pim inhibitor, a CSF1R inhibitor, a TAM receptor tyrosine kinase (Tyro-3, Axl and Mer), a histone deacetylase (HDAC) inhibitor, such as an HDAC8 inhibitor, an angiogenesis inhibitor, an interleukin receptor inhibitor, bromine and additional terminal family member inhibitors (e.g. bromodomain inhibitors or BET inhibitors, such as INCB54329 and INCB57643) and an adenosine receptor antagonist, or combinations thereof.

Examples of antibodies for use in combination therapy include, but are not limited to, trastuzumab (e.g., anti-HER2), ranibizumab (e.g., anti-VEGF-A), bevacizumab

- 16 049302 (trade name Avastin, eg, anti-VEGF, panitumumab (eg, anti-EGFR), cetuximab (eg, anti-EGFR), rituxan (anti-CD20), and antibodies directed to c-MET.

One or more of the following agents may be used in combination with the compounds of the invention and are provided as a non-limiting list: cytostatic agent, cisplatin, doxorubicin, taxotere, taxol, etoposide, irinotecan, camptostar, topotecan, paclitaxel, docetaxel, epothilones, tamoxifen, 5-fluorouracil, methoxtrexate, temozolomide, cyclophosphamide, SCH 66336, R115777, L778.123, BMS 214662, IRESSA ^TM (gefitinib), TARCEVA ™ (erlotinib), anti-EGFR antibodies, intron, ara-C, adriamycin, cytoxan, gemcitabine, uracil mustard, chlormethine, ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6thioguanine, fludarabine phosphate, oxaliplatin, leucovorin, ELOXATIN™ (oxaliplatin), pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide 17.alpha-ethinylestradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testolactone, megestroacetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide, flutamide, toremifene, goserelin, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisole, navelbene, anastrazole, letrazole, capecitabine, reloxafine, droloxafine, hexamethylmelamine, avastin, HERCEPTIN™ (trastuzumab), BEXXAR™ (tositumomab), VELCADE™ (bortezomib), ZEVALIN™ (ibritumomab tiuxetan), TRISENOX™ (arsenic trioxide), XELODA™ (capecitabine), vinorelbine, porfimer, ERBITUX™ (cetuximab), thiotepa, altretamine, melphalan, trastuzumab, lerozole, fulvestrant, exemestane, ifosfomide, rituximab, C225 (cetuximab), campath (alemtuzumab), clofarabine, cladribine, aphidicolone, rituxan, sunitinib, dasatinib, tezacitabine, Sml1, fludarabine, pentostatin, triapine, didox, trimidox, amidox, 3-AP, and MDL-101,731.

The compounds of the invention can be further used in combination with other cancer treatments, such as chemotherapy, radiation therapy, tumor-directed therapy, adjuvant therapy, immunotherapy or surgery. Examples of immunotherapy include cytokine therapy (e.g., interferons, GM-CSF, G-CSF, IL-2), CRS-207 immunotherapy, cancer vaccine, monoclonal antibody, adoptive T-cell transfer, Toll receptor agonists, STING agonists, oncolytic virotherapy and immunomodulatory small molecules including thalidomide or a JAK1/2 inhibitor and similar immunotherapy. The compounds can be administered in combination with one or more anticancer drugs, such as chemotherapeutic agents. Examples of chemotherapy drugs include any of the following: abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezombine, bortezomib, intravenous busulfan, oral busulfan, calusterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, daunorubicin, decitabine, denileukin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone propionate, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib mesylate, interferon alfa 2a, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, meclorethamine, megestrol acetate, melphalan, mercaptopurine, methotrexate, methoxalen, mitomycin C, mitotane, mitoxantrone, nandrolone phenpropionate, nelarabine, nofetumomab, olaparib, oxaliplatin, paclitaxel, pamidronate, panitumumab, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobromane, plicamycin, procarbazine, quinacrine, rasburicase, rituximab, ruxolitinib, rucaparib, streptozocin, tamoxifen, temozolomide, teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, niraparib, veliparib, talazoparib and zoledronate.

Additional examples of chemotherapeutic agents include proteasome inhibitors (eg, bortezomib), thalidomide, revlimid, and DNA damaging agents such as melphalan, doxorubicin, cyclophosphamide, vincristine, etoposide, carmustine, and similar agents.

Examples of Bcr-Abl inhibitors include imatinib mesylate (GLEEVAC™), nilotinib, dasatinib, bosutinib, and ponatinib, and pharmaceutically acceptable salts thereof. Other examples of suitable Bcr-Abl inhibitors include compounds and pharmaceutically acceptable salts thereof of the genera and species disclosed in U.S. Patent No. 5,521,184, WO 04/005281, and U.S. Ser. No. 60/578,491.

Examples of suitable Flt-3 inhibitors include midostaurin, lestaurtinib, linifanib, sunitinib, sunitinib maleate, sorafenib, quizartinib, crenolanib, pacritinib, tandutinib, PLX3397, and ASP2215, and pharmaceutically acceptable salts thereof. Other examples of suitable Flt-3 inhibitors

- 17 049302 include the compounds and their pharmaceutically acceptable salts described in WO 03/037347, WO 03/099771 and WO 04/046120.

Examples of suitable RAF inhibitors include dabrafenib, sorafenib and vemurafenib and pharmaceutically acceptable salts thereof. Other examples of suitable RAF inhibitors include the compounds and pharmaceutically acceptable salts thereof described in WO 00/09495 and WO 05/028444.

Examples of suitable FAK inhibitors include VS-4718, VS-5095, VS-6062, VS-6063, BI853520 and GSK2256098 and pharmaceutically acceptable salts thereof. Other examples of suitable FAK inhibitors include the compounds and pharmaceutically acceptable salts thereof described in WO 04/080980, WO 04/056786, WO 03/024967, WO 01/064655, WO 00/053595 and WO 01/014402.

In some embodiments, the compounds of this invention may be used in combination with one or more other kinase inhibitors, including imatinib, particularly for the treatment of patients resistant to imatinib or other kinase inhibitors.

In some embodiments, the compounds of the invention may be used in combination with a chemotherapeutic agent in the treatment of cancer and may improve the response to treatment compared to the response to the chemotherapeutic agent alone, without increasing its toxic effects. In some embodiments, the compounds of the invention may be used in combination with a chemotherapeutic agent provided herein. For example, additional pharmaceutical agents used in the treatment of multiple myeloma may include, but are not limited to, melphalan, melphalan plus prednisone [MP], doxorubicin, dexamethasone, and Velcade (bortezomib). Other additional agents used in the treatment of multiple myeloma include Bcr-Abl, Flt-3, RAF, and FAK kinase inhibitors. In some embodiments, the agent is an alkylating agent, a proteasome inhibitor, a corticosteroid, or an immunomodulatory agent. Examples of an alkylating agent include cyclophosphamide (CY), melphalan (MEL), and bendamustine. In some embodiments, the proteasome inhibitor is carfilzomib. In some embodiments, the corticosteroid is dexamethasone (DEX). In some embodiments, the immunomodulatory agent is lenalidomide (LEN) or pomalidomide (POM). Additive or synergistic effects are desirable results of combining a PI3K inhibitor of the invention with an additional agent.

In some embodiments, the compounds of the invention may be used in combination with a JAK or PI3Kδ inhibitor.

The agents may be combined with the compound in a single or continuous dosage form, or the agents may be administered simultaneously or sequentially in separate dosage forms.

The compounds of the present invention may be used in combination with one or more other inhibitors or one or more therapeutic agents to treat infections. Examples of infections include viral infections, bacterial infections, fungal infections, or parasitic infections.

In some embodiments of the invention, a corticosteroid, such as dexamethasone, is administered to a patient in combination with the compounds of this invention, wherein the dexamethasone is administered intermittently rather than continuously.

The compounds of formula (I) or any of the formulas described herein, a compound specified in any of the claims and described herein, or salts thereof can be combined with another immunogenic agent, such as cancer cells, purified tumor antigens (including recombinant proteins, peptides and carbohydrate molecules), cells and cells transfected with genes encoding immunostimulatory cytokines. Non-limiting examples of tumor vaccines that can be used include melanoma antigen peptides, such as gp100 peptides, MAGE, Trp-2, MARTI and/or tyrosinase antigens, or tumor cells transfected to express the cytokine GM-CSF.

The compounds of formula (I) or any of the formulas described herein, a compound specified in any of the claims and described herein, or salts thereof, can be used in combination with a vaccination protocol for the treatment of cancer. In some embodiments, tumor cells are transduced to express GM-CSF. In some embodiments, tumor vaccines include proteins from viruses that cause human cancer, such as human papillomavirus (HPV), hepatitis viruses (HBV and HCV), and Kaposi's sarcoma herpes virus (KHSV). In some embodiments, the compounds of this invention can be used in combination with a tumor-specific antigen, such as heat shock proteins isolated from the tumor tissue itself. In some embodiments, the compounds of formula (I) or any of the formulas described herein, a compound specified in any of the claims and described herein, or salts thereof, can be combined with immunization

- 18,049,302 dendritic cells to activate potent anti-tumor responses.

The compounds of the present invention can be used in combination with bispecific macrocyclic peptides that target effector cells expressing the Fe alpha or Fe gamma receptor in tumor cells. The compounds of the present invention can also be combined with macrocyclic peptides that activate the host immune response.

In some additional embodiments, combinations of the compounds of this invention with other therapeutic agents may be administered to a patient prior to, during, and/or after bone marrow transplantation or stem cell transplantation. The compounds of this invention may be used in combination with bone marrow transplantation to treat various tumors of hematopoietic origin.

The compounds of formula (I) or any of the formulas described herein, a compound specified in any of the claims and described herein, or salts thereof may be used in combination with vaccines to stimulate an immune response to pathogens, toxins, and autoantigens. Examples of pathogens for which this therapeutic approach may be particularly useful include pathogens for which there is currently no effective vaccine or pathogens for which conventional vaccines are less than completely effective. These include, but are not limited to, HIV, hepatitis (A, B, and C), influenza, herpes, giardia, malaria, leishmaniasis, Staphylococcus aureus, Pseudomonas aeruginosa.

Viruses that cause infections treatable by the methods of the present invention include, but are not limited to, human papillomavirus, influenza, hepatitis A, B, C or D viruses, adenovirus, poxvirus, herpes simplex viruses, human cytomegalovirus, severe acute respiratory syndrome virus, Ebola virus, measles virus, herpes virus (e.g., VZV, HSV-1, HAV-6, HSV-II and CMV, Epstein-Barr virus), flaviviruses, echovirus, rhinovirus, coxsackie virus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum virus, poliovirus, rabies virus, JC virus and arbovirus encephalitis virus.

Pathogenic bacteria that cause infections treatable by the methods of the invention include, but are not limited to, chlamydia, rickettsioses, mycobacteria, staphylococci, streptococci, pneumococci, meningococci and conococci, klebsiella, proteus, serratia, pseudomonas, legionella, diphtheria, salmonella, bacilli, cholera, tetanus, botulism, anthrax, plague, leptospirosis and Lyme disease bacteria.

Pathogenic fungi that cause infections treatable by the methods of the present invention include, but are not limited to, Candida (albicans, krusei, glabrata, tropicalis, etc.), Cryptococcus neoformans, Aspergillus (fumigatus, niger, etc.), the genus Mucorales (mucor, absidia, rhizophus), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis, and Histoplasma capsulatum. Pathogenic parasites that cause infections treatable by the methods of the present invention include, but are not limited to, Entamoeba histolytica, Balantidium coli, Naegleriafowleri, Acanthamoeba sp., Giardia lambia, Cryptosporidium sp., Pneumocystis carinri, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondi, and Nippostrongylus brasiliensis.

Methods for safely and effectively administering most of these chemotherapeutic agents are known to those skilled in the art. In addition, their administration is described in the common literature. For example, administration of many chemotherapeutic agents is described in the Physicians' Desk Reference (PDR, e.g., 1996 edition, Medical Economics Company, Montvale, NJ), the disclosure of which is incorporated herein by reference in its entirety.

Pharmaceutical compositions and dosage forms.

When used as pharmaceuticals, the compounds of the invention can be administered in the form of pharmaceutical compositions. These compositions can be prepared by methods well known in the pharmaceutical art and can be administered in a variety of ways depending on whether local or systemic treatment is indicated and the site to be treated. Administration is local (including transdermal, epidermal, ocular and mucosal, including intranasal, vaginal and rectal administration), pulmonary (e.g. by inhalation or insufflation of powders or aerosols, in particular by means of a nebulizer; intratracheal or intranasal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular administration or injection or infusion; or intracranial, for example intrathecal or intraventricular administration. Parenteral administration may be in the form of a single bolus dose or, for example, continuously using a perfusion pump. Pharmaceutical compositions and formulations for topical administration include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickening agents and similar vehicles may be necessary or desirable.

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This invention also includes pharmaceutical compositions that contain, as an active ingredient, a compound of the invention or a pharmaceutically acceptable salt thereof in combination with one or more pharmaceutically acceptable carriers (excipients). In some embodiments, said composition is suitable for topical administration. In preparing the compositions of the invention, the active ingredient is typically mixed with an excipient, diluted by an excipient, or enclosed in such a carrier in the form of, for example, a capsule, sachet, paper bag, or other container. When the filler acts as a diluent, it can be a solid, semi-solid, or liquid material that acts as a drug-carrying medium, carrier, or vehicle for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, infusions, suspensions, emulsions, solutions, syrups, aerosols (solid or in a liquid medium), ointments, containing, for example, up to 10% by weight of active compound, soft and hard gelatin capsules, suppositories, sterile injection solutions or sterile packaged powders.

In preparing the composition, before combining with other ingredients, the active compound may be milled to obtain an appropriate particle size. If the active compound is substantially insoluble, it may be milled to a particle size of less than 200 mesh. If the active compound is substantially soluble in water, the particle size may be adjusted by milling to obtain a substantially uniform distribution in the composition, for example, about 40 mesh.

The compounds of the invention may be milled using known milling techniques, such as wet milling, to produce particles of a size suitable for forming tablets and other types of preparations. Micronized (nanoparticulate) preparations of the compounds of the invention may be prepared by methods known in the art, see, for example, International Application No. WO 2002/000196.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup and methylcellulose. The formulations may additionally contain: lubricating agents, for example talc, magnesium stearate and mineral oil; wetting agents; emulsifying and dispersing agents; preservatives, for example methyl and propyl hydroxybenzoates; sweetening agents and flavoring agents. The compositions of the present invention can be designed to provide quick, sustained or delayed release of the active ingredient after administration to the patient using procedures known in the art.

The compositions may be formulated in unit dosage form, each dosage containing from about 5 to about 1000 mg (1 g), more usually from about 100 to about 500 mg, of the active ingredient. The term unit dosage forms refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient.

In some embodiments, the compositions of the invention comprise from about 5 to about 50 mg of the active ingredient. One skilled in the art will appreciate that these are compositions comprising from about 5 to about 10, from about 10 to about 15, from about 15 to about 20, from about 20 to about 25, from about 25 to about 30, from about 30 to about 35, from about 35 to about 40, from about 40 to about 45, or from about 45 to about 50 mg of the active ingredient.

In some embodiments, the compositions of the invention contain from about 50 to about 500 mg of the active ingredient. One skilled in the art will appreciate that these are compositions containing from about 50 to about 100, from about 100 to about 150, from about 150 to about 200, from about 200 to about 250, from about 250 to about 300, from about 350 to about 400, or from about 450 to about 500 mg of the active ingredient.

In some embodiments, the compositions of the invention comprise from about 500 to about 1000 mg of the active ingredient. One skilled in the art will appreciate that these are compositions comprising from about 500 to about 550, from about 550 to about 600, from about 600 to about 650, from about 650 to about 700, from about 700 to about 750, from about 750 to about 800, from about 800 to about 850, from about 850 to about 900, from about 900 to about 950, or from about 950 to about 1000 mg of the active ingredient.

Similar dosages may be used for the compounds described herein in the methods and uses of the invention.

The active compound may be effective over a wide dosage range and is generally administered in a pharmaceutically effective amount. However, it should be understood that the amount of compound actually administered will generally be determined by the physician in accordance with the relevant circumstances, including the condition being treated, the route of administration chosen, the compound actually administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, and

- 20 049302 similar circumstances.

For the preparation of solid compositions, for example tablets, the principal active ingredient is mixed with a pharmaceutical excipient to form a composition prior to giving it a dosage form containing a homogeneous mixture of the compound of this invention. In the event that these compositions are said to be homogeneous prior to giving them a dosage form, it is understood that the active ingredient is generally dispersed uniformly throughout the composition so that the composition can be easily divided into equally effective unit dosage forms, for example tablets, dragees and capsules. This solid composition is then divided prior to giving it a dosage form into unit dosage forms of the types described above containing, for example, from about 0.1 to about 1000 mg of the active ingredient of this invention.

The tablets or pills of the present invention may be coated or otherwise formulated to provide a dosage form that provides the advantage of sustained action. For example, a tablet or pill may comprise an inner and an outer dosage component, the latter being in the form of a coating on the excipient. The two components may be separated by an enteric layer that serves to resist disintegration in the stomach and to allow the inner component to pass intact into the duodenum or to be released in a delayed manner. A variety of materials may be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol and cellulose acetate.

Liquid forms into which the compounds or compositions of the invention may be incorporated for oral administration or injection include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions and flavored emulsions with edible oils, for example, cottonseed oil, sesame oil, coconut oil or peanut oil, as well as with elixirs and similar pharmaceutical carriers.

Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the compositions are administered by inhalation through the mouth or nose to achieve a local or systemic effect. The compositions can be nebulized using inert gases. Nebulized solutions can be inhaled directly from a nebulizer device, or the nebulizer device can be attached to a face mask, oxygen tent or intermittent positive pressure breathing apparatus. Solution, suspension or powder compositions can be administered orally or nasally from devices that deliver the composition accordingly.

The topical formulations may contain one or more conventional carriers. In some embodiments, ointments may contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum and similar carriers. The carrier compositions in creams may be based on water in combination with glycerin and one or more other components, for example, glycerin monostearate, PEG-glycerin monostearate and cetyl stearyl alcohol. Gels may be formulated using isopropyl alcohol and water, in appropriate combination with other components, for example, glycerin, hydroxyethyl cellulose and similar components. In some embodiments, the topical formulations contain at least about 0.1, at least about 0.25, at least about 0.5, at least about 1, at least about 2 or at least about 5% by weight of compounds of the present invention. The topical formulations may be suitably packaged in tubes, for example 100 g, optionally accompanied by instructions for treating the selected symptom, for example psoriasis or other skin condition.

The amount of the compound or composition administered to a patient will vary depending on what is being administered, the purpose of administration, such as prophylaxis or treatment, the patient's condition, the route of administration, and the like. For therapeutic purposes, the compositions may be administered to a patient already suffering from a disease in an amount sufficient to treat or at least partially alleviate the symptoms of the disease or its complications. Effective doses will depend on the condition of the disease being treated, as well as on the judgment of the attending physician, depending on factors such as the severity of the disease, age, weight, general condition of the patient, and similar factors.

The compositions administered to the patient may be in the form of pharmaceutical compositions as described above. These compositions may be sterilized by conventional sterilization techniques or subjected to filtered sterilization. Aqueous solutions may be packaged for use as is, or lyophilized, with the lyophilized preparation being lyophilized prior to administration.

- 21 049302 is combined with a sterile aqueous vehicle. The pH of the compounded preparations is typically from 3 to 11, more preferably from 5 to 9, and most preferably from 7 to 8. It should be understood that the use of some of the above excipients, carriers or stabilizers will result in the formation of pharmaceutical salts.

The therapeutic dosage of a compound of this invention may vary according to, for example, the particular purpose for which treatment is being performed, the route of administration of the compound, the medical condition of the patient, and the judgment of the attending physician. The proportion or concentration of a compound of this invention in a pharmaceutical composition may vary depending on a number of factors, including the dosage, chemical characteristics (e.g., hydrophobicity), and route of administration. For example, the compounds of this invention may be provided in an aqueous physiological buffer containing from about 0.1 to about 10% w/v of the compound for parenteral administration. Some typical dosages are in the range of from about 1 μg/kg to about 1 g/kg body weight per day. In some embodiments, the dosage is in the range of from about 0.01 mg/kg to about 100 mg/kg body weight per day. The dosage will likely depend on such variables as the type and extent of progression of the disease or disorder, the general health of the individual patient, the relative biological effectiveness of the compound selected, the composition of the excipient, and the route of administration. Effective doses can be extrapolated from dose-response curves obtained in vitro in animal model test systems.

The compositions of the present invention may further comprise one or more additional pharmaceutical agents, such as a chemotherapeutic, steroidal, anti-inflammatory compound or immunosuppressant, examples of which are listed herein.

Labeled compounds and methods of analysis.

Another aspect of the present invention relates to labeled compounds of the invention (radioactively labeled, fluorescently labeled, etc.) which would be useful not only in imaging techniques but also in assays, both in vitro and in vivo, for localizing and quantifying A2A and/or A2B receptors in tissue samples, including humans, and for identifying A2A and/or A2B antagonists by inhibiting labeled compound binding. Substitution of one or more atoms of the compounds of the invention may also be useful for creating differential ADME (absorption, distribution, metabolism and excretion). Accordingly, the present invention includes assays for adenosine receptors (e.g., A2A and/or A2B) containing such labeled or substituted compounds.

The present invention further includes isotopically labeled compounds of the invention. An isotopically or radioactively labeled compound is a compound of the invention in which one or more atoms are replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (i.e., occurring in nature). Suitable radionuclides that can be included in the compounds of this invention include, but are not limited to, 2H (also referred to as D for deuterium), ^3H (also referred to as T for tritium), ^nC , ^13C , ^14C , ^13N , 15N, 15O, 17O, ^18O , ^18F , ^35S , 36Cl, ^82Br , ^75Br , ^{76Br, 77Br , 123I} , ^124I , ^{125I ,} and ^131I . For ^example , one or more hydrogen atoms in a compound of this invention can be replaced with deuterium atoms (e.g., one or more hydrogen atoms ^of a _C1-6 alkyl group of formula (I ⁾ can be optionally replaced with deuterium atoms, e.g., _-CD3 is replaced with -CH3). In some embodiments, the alkyl groups in any of the disclosed formulas, such as formula (I), may be perdeuterated.

One or more constituent atoms of the compounds provided herein may be replaced or substituted with isotopes of atoms that are abundant or uncommon in nature. In some embodiments, the compound includes at least one deuterium atom. For example, one or more hydrogen atoms in a compound provided herein may be replaced or substituted with deuterium (e.g., one or more hydrogen atoms in a _{C1_6} alkyl group may be replaced with deuterium atoms, such as -CD3 is replaced with -CH3). In some embodiments, the compound comprises two or more deuterium atoms. In some embodiments, the compound comprises 1-2, 1-3, 1-4, 1-5, or 1-6 deuterium atoms. In some embodiments, all of the hydrogen atoms in the compound may be replaced or substituted with deuterium atoms.

In some embodiments, 1, 2, 3, 4, 5, 6, 7, or 8 hydrogen atoms are attached to the carbon atoms of any alkyl, alkenyl, alkynyl, aryl, phenyl, cycloalkyl, heterocycloalkyl, or heteroaryl substituents, or each of the _{-C1_6} alkyl-, alkylene, alkenylene, and alkynylene linking groups as described herein is optionally replaced with a deuterium atom.

Synthetic methods for incorporating isotopes into organic compounds are known in the art (Deuterium Labeling in Organic Chemistry by Alan F. Thomas (New York, N.Y., Appleton-Century Crofts, 1971; The Renaissance of H/D Exchange by Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen

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Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labelling by James R. Hanson, Royal Society of Chemistry, 2011). Isotopically labeled compounds can be used in a variety of studies, such as NMR spectroscopy, metabolism experiments, and/or assays.

Substitution with heavier isotopes, such as deuterium, may confer some therapeutic advantage as a result of higher metabolic stability, such as increased half-life in vivo or reduced dosage requirements, and may therefore be advantageous in some settings (see, for example, A. Kerekes et al. J. Med. Chem. 2011, 54, 201-210; R. Xu et al. J. Label Compd. Radiopharm. 2015, 58, 308-312). In particular, substitution at one or more sites of metabolism may provide one or more therapeutic advantages.

The radionuclide included in these radiolabeled compounds will depend on the particular application of the radiolabeled compound. For example, compounds that include ³ H, ¹⁴ C, ⁸² Br, ¹²⁵ 1, ¹³¹ I, or ³⁵ S may be used for in vitro labeling of adenosine receptors and competition assays. For radioimaging applications, ⁿ C, ¹⁸ F, ¹²⁵ I, ¹²³ I, ¹²⁴ I, ¹³¹ 1, ⁷⁵ Br, ⁷⁶ Br, or ⁷⁷ Br may be used.

It is understood that a radioactively labeled or labeled compound is a compound that includes at least one radionuclide. In some embodiments of the invention, the radionuclide is selected from the group consisting of ³ H, ¹⁴ C, ¹²⁵ 1, ³⁵ S and ⁸² Br.

The present invention may further include synthetic methods for incorporating radioisotopes into the compounds of the invention. Synthetic methods for incorporating radioisotopes into organic compounds are well known in the art, and one of ordinary skill in the art will readily recognize methods applicable to the compounds of the invention.

The labeled compound of the present invention can be used in a screening assay to identify/evaluate compounds. For example, a newly synthesized or discovered compound (i.e., a test compound) that is labeled can be assessed for its ability to bind an adenosine receptor by monitoring the change in its concentration upon contact with the adenosine receptor by monitoring the labeling. For example, a test compound (labeled) can be assessed for its ability to reduce the binding of another compound known to bind to the adenosine receptor (i.e., a standard compound). Accordingly, the ability of the test compound to compete with the standard compound for binding to the adenosine receptor directly correlates with its binding affinity. Conversely, in some other screening assays, the standard compound is labeled and the test compound is unlabeled. Therefore, the concentration of the labeled standard compound is monitored to assess the competition between the standard compound and the test compound, and thus determine the relative binding capacity of the test compound.

Sets.

The present invention also includes pharmaceutical kits useful, for example, in the treatment or prevention of diseases or disorders associated with an adenosine receptor (e.g., cancer, an inflammatory disease, a cardiovascular disease, or a neurodegenerative disease), which include one or more container units containing a pharmaceutical composition containing a therapeutically effective amount of a compound of the invention. Such kits may further include, if desired, one or more different standard components of pharmaceutical kits, for example, container units with one or more pharmaceutically acceptable carriers, additional container units, etc., as will be apparent to those skilled in the art. Instructions in the form of inserts or labels indicating the amount of components to be administered, directions for use, and/or directions for mixing the components may also be included in the kit.

The invention will be described in more detail with reference to specific examples. The examples below are for illustrative purposes and are not intended to limit the invention in any way. Those skilled in the art will readily recognize many non-critical parameters that can be changed or modified to obtain exactly the same results. The compounds of the examples were found to inhibit adenosine receptor activity (e.g., A2A and/or A2B) according to at least one assay described herein.

Examples

Preparative LC-MS purification of some of the compounds obtained was performed on Waters mass-directed fractionation systems. The setup of the main equipment, protocols, and control software for operating these systems are described in detail in the literature (see, for example, Two-Pump At Column Dilution Configuration for Preparative LC-MS, K. Blom, J. Combi. Chem., 4, 295 (2002); Optimizing Preparative LC-MS Configurations and Methods for Parallel Synthesis Purification, K. Blom, R. Sparks, J. Doughty, G. Everlof, T. Haque, A. Combs, J. Combi. Chem., 5, 670 (2003); and Preparative LC-MS Purification: Improved Compound Specific Method Optimization, K. Blom, B. Glass, R.

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Sparks, A. Combs, J. Combi. Chem., 6, 874–883 (2004)). The separated compounds were typically subjected to analytical liquid chromatography and mass spectrometry (LC-MS) for purity analysis under the following conditions: instrument; Agilent 1100 series, LC/MSD, column: Waters Sunfire ^TM C ₁₈ 5 μm, 2.1 x 50 mm, buffers: mobile phase A: 0.025% TFA in water and mobile phase B: acetonitrile; gradient from 2% to 80% B in 3 minutes at a flow rate of 2.0 mL/min.

Some of the compounds obtained were also separated on a preparative scale by reversed-phase high-performance liquid chromatography (RP-HPLC) with mass spectrometric detection or by flash chromatography (silica gel), as indicated in the examples. Typical preparative RP-HPLC column conditions are as follows:

pH=2 purification: Waters Sunfire™ C ₁₈ 5 μm, 30x100 mm or Waters XBridge™ C ₁₈ 5 μm, 30x100 mm column, eluting with mobile phase A: 0.1% TFA (trifluoroacetic acid) in water and mobile phase B: acetonitrile; flow rate was 60 ml/min, the separation gradient was optimized for each compound using a compound-specific method optimization protocol as described in the literature (see, e.g., Preparative LCMS Purification: Improved Compound Specific Method Optimization, K. Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem., 6, 874-883 (2004)).

pH=10 Purification: Waters XBridge™ C ₁₈ 5 µm, 30x100 mm column, eluting with mobile phase A: 0.1% NH ₄ OH in water and mobile phase B: acetonitrile; flow rate was 60 ml/min, the separation gradient was optimized for each compound using a compound-specific method optimization protocol as described in the literature (see, e.g., Preparative LCMS Purification: Improved Compound Specific Method Optimization, K. Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem., 6, 874–883 (2004)).

Separation of some racemic compounds into enantiopure samples was performed on a preparative scale by chiral phase high performance liquid chromatography under the following conditions: instrument: Agilent 1100 Prep HPLC; column: Phenomenex Lux Cellulose-4, 21.2 x 250 mm, 5 μm; elution with an isocratic mobile phase of 45% EtOH in hexanes at a flow rate of 20 mL/min.

Example 1. 3-(5-Amino-2-((5-(3-aminophenyl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

Step 1. 3-(2-Amino-6-chloropyrimidin-4-yl)benzonitrile.

A mixture of 4,6-dichloropyrimidin-2-amine (2.5 g, 15.2 mmol), (3-cyanophenyl)boronic acid (2.02 g, 13.7 mmol), tetrakis(triphenylphosphine)palladium(0) (1.06 g, 0.92 mmol), and sodium carbonate (3.23 g, 30.5 mmol) in 1,4-dioxane (60 mL) and water (5 mL) was degassed with nitrogen, then the resulting mixture was heated and stirred at 60 °C for two days. After cooling to room temperature (rt), the mixture was concentrated, diluted with water, and extracted with DCM (30 mL x 3). The combined organic layers were dried over MgSO ₄ , filtered, and concentrated. The resulting residue was purified by flash column chromatography on silica gel eluting with 8% EtOAc in dichloromethane to afford the desired product. LCMS calculated for C11H ₈ ClN ₄ (M+H)+: 231.0. found: 231.0.

Step 2. 3-(5-Amino-2-(hydroxymethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

2-Hydroxyacetohydrazide (2.34 g, 26.01 mmol) was added to an ethanol (35 mL) solution of 3-(2-amino-6-chloropyrimidin-4-yl)benzonitrile (4.00 g, 17.34 mmol) at rt. After heating and stirring at reflux for 2 h, the reaction mixture was cooled to rt and concentrated. The resulting residue was taken up in N,O-bis(trimethylsilyl)acetamide (20 mL) and

- 24 049302 was stirred at 120 °C for 7 h. The mixture was then cooled to rt, poured onto ice and stirred at rt for 1 h. The resulting solid was collected by filtration and taken up in 20 mL of 1 N HCl solution. The resulting mixture was stirred at rt for 1 h, filtered and the aqueous layer was neutralized by adding saturated NaHCO ₃ solution. The formed precipitate was collected by filtration and dried to give the desired product as a brown solid. LCMS calculated for C13H11N6O (M+H)+: 267.1; found 267.1.

Step 3. 3-(5-Amino-8-bromo-2-(hydroxymethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

To a mixture of 3-(5-amino-2-(hydroxymethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (1.0 g, 3.76 mmol) in DMF (12 mL) at -30 °C was added NBS (0.67 g, 3.76 mmol) portionwise. The reaction mixture was allowed to warm slowly to 0 °C to give a homogeneous solution. After stirring at 0 °C for 1 h, the reaction mixture was diluted with saturated NaHCO ₃ solution and the desired product was collected by filtration and dried. LCMS calculated for C ₁₃ H ₁₀ BrN ₆ O (M+H)+: 345.0; found 345.0.

Step 4. 3-(5-Amino-2-(hydroxymethyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile.

Tetrakis(triphenylphosphine)palladium(0) (0.067 g, 0.058 mmol) was added to a mixture of 4-(tributylstannyl)pyrimidine (0.321 g, 0.869 mmol), 3-(5-amino-8-bromo-2(hydroxymethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (0.20 g, 0.579 mmol), CsF (0.176 g, 1.159 mmol), and copper(T) iodide (0.022 g, 0.116 mmol) in 1,4-dioxane (5.0 mL). The reaction mixture was flushed with _N2 and stirred at 80°C for 7 h. The resulting mixture was cooled to rt, concentrated and purified by flash column chromatography eluting with 0% to ₁₀ % methanol in DCM to give the product. _LCMS calculated for _C17H13N8O (M+H)+: 345.1; found 345.1.

Step 5. 3-(5-Amino-2-(chloromethyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile.

To a mixture of 3-(5-amino-2-(hydroxymethyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile (0.1 g, 0.290 mmol) in acetonitrile (10 mL) was added thionyl chloride (0.212 mL, 2.90 mmol) at rt. The reaction mixture was stirred at rt for 5 h, concentrated and purified by flash chromatography eluting with 0% to 5% methanol in DCM to give the product. LCMS calculated for C ₁₇ H ₁₂ ClN ₈ (M+H)+: 363.1; found 363.1.

Step 6. 3-(5-amino-2-((5-(3-aminophenyl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

A mixture of 3-(5-amino-2-(chloromethyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (10 mg, 0.028 mmol), 3-(1H-tetrazol-5-yl)aniline (8.9 mg, 0.055 mmol) and _Cs2CO3 (20.7 mg, 0.064 mmol) in DMF (1 mL) was stirred at 100 °C for 10 _min . The reaction mixture was then cooled to rt, diluted with methanol (4 mL) and purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LC-MS calculated for C ₂₄ H ₁₈ N ₁₃ (M+H)+: 488.2; found 488.2.

Example 2. 3-(5-Amino-2-((5-(6-methylpyridin-2-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

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This compound was prepared using procedures similar to those described for Example 1, substituting 5-(m-tolyl)-1H-tetrazole for 3-(1H-tetrazol-5-yl)aniline. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LC-MS calculated for C ₂₄ H ₁₈ N ₁₃ (M+H)+: 488.2; found 488.2.

Example 3. 3-(5-Amino-2-((5-(6-methoxypyridin-2-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Example 1 substituting 2-methoxy-6-(1H-tetrazol-5-yl)pyridine for 3-(1H-tetrazol-5-yl)aniline. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LC-MS calculated for C ₂₄ H ₁₈ N ₁₃ O (M+H)+: 504.2; found 504.2.

Example 4. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(pyridin-2-yl)-1Htetrazol-1-yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

Step 1. 3-(5-Amino-2-(hydroxymethyl)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

A mixture of 3-(5-amino-8-bromo-2-(hydroxymethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (20 mg, 0.046 mmol), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one (100 mg, 0.46 mmol), XPhos Pd G2 (35 mg, 47 μmol), and Na ₂ CO ₃ (200 mg, 1.9 mmol) in 1,4-dioxane (5.0 mL) and water (1.0 mL) was purged with nitrogen and sealed. The reaction mixture was stirred at 110°C for 1 h. The resulting mixture was cooled to rt, concentrated and purified by flash column chromatography eluting with 0% to 10% methanol in DCM to give the product. LCMS calculated for C ₁₉ H ₁₆ N ₇ O ₂ (M+H)+: m/z=374.1; found 374.1.

Step 2. 3-(5-Amino-2-(chloromethyl)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

- 26 049302

This compound was prepared using procedures similar to those described for Step 5 of Example 1 substituting 3-(5-amino-2-(hydroxymethyl)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile for 3-(5-amino-2-(hydroxymethyl)-8(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile. LCMS calculated for C ₁₉ H ₁₅ ClN ₇ O (M+H)+: 392.1; found 392.

Step 3. 3-(5-Amino-2-(chloromethyl)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Step 6 of Example 1 substituting 3-(5-amino-2-(chloromethyl)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile for 3-(5-amino-2-(chloromethyl)-8-(pyrimidin-4yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile. The product was purified by preparative LCMS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LCMS calculated for C ₂₄ H ₁₈ N ₁₃ O (M+H)+: 504.2; found 504.2.

Example 5. 3-(2-((5-(1H-Pyrazol-1-yl)-1H-tetrazol-1-yl)methyl)-5-amino-8-(1-methyl-6-oxo1,6-dihydropyridin-3-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Example 4 substituting 5-(1H-pyrazol-1-yl ₎ -1H-tetrazole for 2-(1H-tetrazol-5-yl)pyridine. The product was purified by preparative LC-MS ₍ pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LC-MS calculated for _C23H18N13O (M+H)+: 492.5; found 492.4.

Example 6. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(thiazol-4-yl)-1Htetrazol-1-yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Example 4 substituting 4-(1H-tetrazol-5-yl)thiazole for 2-(1H-tetrazol-5-yl)pyridine. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LC-MS calculated for C ₂₃ H ₁₇ N ₁₂ OS (M+H)+: 509.1; found 509.2.

Example 7. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(pyrimidin-2-yl)-1Htetrazol-1-yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

- 27 049302

This compound was prepared using procedures similar to those described for Example 4, substituting 2-(1H-tetrazol-5-yl)pyrimidine for 2-(1H-tetrazol-5-yl)pyridine. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LCMS calculated for C ₂₄ H ₁₈ N ₁₃ O (M+H)+: 504.2; found 504.2.

Example 8. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(pyrazin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Example 4, substituting 2-(1H-tetrazol-5-yl)pyrazine for 2-(1H-tetrazol-5-yl)pyridine. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LCMS calculated for C ₂₄ H ₁₈ N ₁₃ O (M+H)+: 504.2; found 504.2.

Example 9. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-((5-(pyridin-2-yl)1H-tetrazol-1-yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 4 substituting 2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridazin-3(2H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one in step 1. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LC-MS calculated for C ₂₄ H ₁₈ N ₁₃ O (M+H)+: 504.2; found 504.2.

Example 10. 3-(5-Amino-2-((5-(pyridin-2-yl)-III-tetrazol-1-yl)methyl)-8-(pyridin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Example 4 substituting pyridin-4-ylboronic acid for 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)pyridin-2(1H)-one in Step 1. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LC-MS calculated for C ₂₄ H ₁₇ N ₁₂ (M+H)+: 473.2; found 473.2.

Example 11. 3-(5-Amino-8-(2-(dimethylamino)pyridin-4-yl)-2-((5-(pyridin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Example 4, using (2-(dimethylamino)pyridin-4-yl)boronic acid instead of 1-methyl-5-(4,4,5,5

- 28 049302 tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one in step 1. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LCMS calculated for C ₂₆ H ₂₂ N ₁₃ (M+H)+: 515.2; found 515.2.

Example 12. 3-(5-Amino-8-(2-aminopyridin-4-yl)-2-((5-(pyridin-2-yl)-1H-tetrazol-1-yl)methyl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

NH

This compound was prepared using procedures similar to those described for Example 4 substituting (2-aminopyridin-4-yl)boronic acid for 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)pyridin-2(1H)-one in Step 1. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LC-MS calculated for C ₂₄ H ₁₈ N ₁₃ (M+H)+: 488.2; found 488.2.

Example 13 3-(5-Amino-2-(2-fluoro-6-(pyridin-4-yl)benzyl)-8-(pyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Example 1, step 2, using 2-(2-bromo-6-fluorophenyl)acetohydrazide instead of 2-hydroxyacetohydrazide. LC-MS calculated for C ₁₉ H ₁₃ BrFN ₆ (M+H)+: 423.0; found 423.0.

Step 2. 3-(5-Amino-8-bromo-2-(2-bromo-6-fluorobenzyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile.

NH _Z

NBS (126 mg, 0.709 mmol) was added to a DMF (2.00 mL) solution of 3-(5-amino-2-(2-bromo-6-fluorobenzyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (300 mg, 0.709 mmol) at rt. After stirring at rt for 1 h, it was diluted with water and the resulting precipitate was collected by filtration. The brown solid was dissolved in DCM and purified by flash column chromatography on silica gel eluting with 0 to 50% EtOAc in DCM to give the desired product. LCMS calculated for C ₁₉ H ₁₂ Br ₂ N ₆ (M+H)+: 501.0; 501.0 found.

Step 3. 3-(5-Amino-2-(2-fluoro-6-(pyridin-4-yl)benzyl)-8-(pyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

A mixture of 3-(5-amino-8-bromo-2-(2-bromo-6-fluorobenzyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile (30 mg, 0.060 mmol) and pyridin-4-ylboronic acid (8.8 mg, 0.072 mmol), XPhos Pd G2 (4.7 mg, 6.0 μmol) and sodium carbonate (13.0 mg, 0.123 mmol) in 1,4-dioxane (2.0 mL) and water (0.20 mL) was stirred at 100 °C for 3 h. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LC-MS calculated for C ₂₉ H ₂₀ FN ₈ (M+H)+: 499.2; found 499.2.

Example 14 3-(5-Amino-8-(2-aminopyridin-4-yl)-2-(2-(2-aminopyridin-4-yl)-6-fluorobenzyl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

- 29 049302

This compound was prepared using procedures similar to those described for Example 13, substituting (2-aminopyridin-4-yl)boronic acid for pyridin-4-ylboronic acid in Step 3. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LC-MS calculated for C29H22FN10 (M+H) ⁺ : 529.2; found 529.3.

Example 15. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2yl)methyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

¢)

NI 1 _g

Step 1. 6-Chloro-N,N-bis(4-methoxybenzyl)pyrimidine-2,4-diamine.

Cl

nh ₂

To a solution of 2,6-dichloropyrimidin-4-amine (5.0 g, 31 mmol) in 2-propanol (31 mL) were added N,N-diisopropylethylamine (6.4 mL, 37 mmol) and bis(4-methoxybenzyl)amine (7.9 g, 31 mmol). The resulting solution was stirred at 100 °C for 16 h, cooled to rt, diluted with water (100 mL) and extracted with EtOAc (100 mL). The organic layer was washed with water and brine, dried over anhydrous sodium sulfate and concentrated to give the crude product, which was used in the next step without further purification. LCMS calculated for C20H22CIN4O2 (M+H) ⁺ : 385.1; found 385.1.

Step 2. 7-Chloro-N,N-bis(4-methoxybenzyl)-[1,2,4]triazolo[1,5-c]pyrimidine-2,5-diamine.

^0

O-Ethyl carbonisothiocyanatidate (3.1 mL, 26 mmol) was added to a 1,4-dioxane (5.0 mL) solution of 6-chloro-Na,Na-bis(4-methoxybenzyl)pyrimidine-2,4-diamine (1.0 g, 2.6 mmol) at rt. The reaction mixture was then stirred at 90 °C overnight, cooled to rt and concentrated. The resulting material was dissolved in methanol (12 mL) and ethanol (12 mL) and N,N-diisopropylethylamine (0.91 mL, 5.2 mmol) was added followed by hydroxylamine hydrochloride (0.54 g, 7.8 mmol). The reaction mixture was stirred at 45 °C for 2 h, cooled to rt and concentrated. The resulting material was taken up in EtOAc, washed with water, dried over anhydrous sodium sulfate, and concentrated. The crude material was then purified by silica gel chromatography eluting with 0% to 50% EtOAc in hexanes to give the product. LCMS calculated for C21H22CIN6O2 (M+H) ⁺ : 425.1; found 425.2.

Step 3. 3-(2-Amino-5-(bis(4-methoxybenzyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

-30049302

Chloro(2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2yl)palladium(II) (330 mg, 0.42 mmol) was added to a mixture of (3-cyanophenyl)boronic acid (460 mg, 3.2 mmol), 7-chloro-N ⁵ ,N ⁵ -bis-(4-methoxybenzyl)-[1,2,4]triazolo[1,5-c]pyrimidine-2,5-diamine (890 mg, 2.1 mmol), and sodium carbonate (890 mg, 8.4 mmol) in 1,4-dioxane (8.8 mL) and water (1.8 mL). The mixture was purged with N ₂ and stirred at 95 °C overnight. The reaction mixture was then cooled to rt, concentrated and purified by silica gel chromatography eluting with 0% to 50% EtOAc in DCM to give the desired product. LCMS calculated for C ₂₈ H ₂₆ N ₇ O ₂ (M+H)+: 492.2; found 492.2.

Step 4. 3-(5-(bis(4-Methoxybenzyl)amino)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

3-(2-Amino-5-(bis(4-methoxybenzyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (300 mg, 0.610 mmol), triethyl orthoformate (508 µL, 3.05 mmol), 3-methylpicolinaldehyde (148 mg, 1.221 mmol) were combined and EtOH (5 mL) was added. The suspension was heated at 120 °C overnight, then cooled to room temperature, diluted with DCM (2 mL). Sodium tetrahydroborate (46.2 mg, 1.221 mmol) was carefully added to the solution. After stirring at room temperature for 1 h, the mixture was carefully quenched with saturated aqueous NH ₄ Cl, extracted with DCM and separated. _The organic layer _was dried over _Na2SO4 , _filtered and evaporated. The residue was used in the next reaction step without further purification. LC-MS calculated for _C35H33N8O2 ( _M +H)+: m/z=597.2; found 597.2.

Step 5. 3-(5-(bis(4-Methoxybenzyl)amino)-8-bromo-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

To a solution of 3-(5-(bis(4-methoxybenzyl)amino)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (300 mg, 0.503 mmol) in DCM (1 mL) at room temperature was added NBS (89 mg, 0.503 mmol). The mixture was stirred at room temperature for 1 h and then quenched with saturated aqueous NaHCO ₃ solution and separated. The organic layer was dried over Na ₂ SO ₄ , filtered, and evaporated. The residue was purified by column chromatography (10 to 50% AcOEt in hexane) to give the desired product. LC-MS calculated for C ₃₅ H ₃₂ BrN ₈ O ₂ (M+H)+: m/z=675.2; found 675.2.

Step 6. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2yl)methyl)amino)-[1,2,4]triazolo[1,5-]pyrimidin-7-yl)benzonitrile.

- 31 049302

3-(5-(bis(4-Methoxybenzyl)amino)-8-bromo-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (30 mg, 0.044 mmol), 1-methyl-5-(4,4,5,5-tetramethyl1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one (20.88 mg, 0.089 mmol), Xphos-G2 (3.49 mg, 4.44 μmol), sodium carbonate (9.41 mg, 0.089 mmol) were combined. 1,4-dioxane (1 mL) and water (0.100 mL) were added to the mixture. The mixture was heated to 100 °C and stirred for 3 h and then evaporated. TFA (1 mL) was added to the residue and the mixture was heated at 120 °C for 20 min. After cooling to room temperature, the mixture was diluted with acetonitrile, filtered and purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to give the _desired product as the TFA salt. LC-MS calculated for _C25H22N9O (M+H)+: m/z=464.2; found 464.2.

Example 16. 3-(5-Amino-8-(1-ethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2yl)methyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 15 substituting 1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC- _{MS (} pH=2, MeCN/water with TFA) to give the desired product as the TFA salt. LC-MS calculated for _C26H24N9O (M+H)+: m/z=478.2; found 478.2.

Example 17. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(((3-methylpyridin-2yl)methyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 15 substituting 2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridazin-3(2H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS ₍ pH=2, MeCN/water with TFA) to afford the desired product _as the TFA salt. LC-MS calculated for _C24H21N10O (M+H)+: m/z=465.2; found 465.2.

Example 18. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((3-methylpyridin-2yl)methoxy)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

Step 1. 3-(5-(bis(4-Methoxybenzyl)amino)-2-iodo-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

- 32 049302

To a solution of 3-(2-amino-5-(bis(4-methoxybenzyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile in _CH2Cl2 ( _3.00 mL)/acetonitrile (3 mL) (Example 15, step 3, 250 mg, 0.509 mmol) was added HI (57% in water, 201 μL, 1.526 mmol). The mixture was heated to 60 °C and then tert-butyl nitrite (134 μL, 1.017 mmol) was added. The reaction mixture was heated at 60 °C for 20 min. After cooling to room temperature, 1 N aqueous NH4OH solution was added and the mixture was extracted three times _{with CH2Cl2} . The combined organic layer was dried over sodium sulfate, filtered and the solvent was evaporated under reduced pressure. The residue was purified by silica gel chromatography (10 to 50% AcOEt in hexane) to give the desired product. LC-MS calculated for C ₂₈ H ₂₄ IN ₆ O ₂ (M+H)+: m/z=603.1; found 603.1.

Step 2. 3-(5-(bis(4-Methoxybenzyl)amino)-2-((3-methylpyridin-2-yl)methoxy)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

3-(5-(bis(4-Methoxybenzyl)amino)-2-iodo-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (250 mg, 0.415 mmol), (3-methylpyridin-2-yl)methanol (153 mg, 1.245 mmol) were combined and 1,4-dioxane (5 mL) was added. Sodium hydride (60% in mineral oil, 41.5 mg, 1.037 mmol) was added to the solution. The suspension was heated to 105 °C and stirred for 1 h, then cooled to room temperature and quenched with aqueous NH ₄ Cl, diluted with AcOEt, separated. The aqueous layer was extracted with AcOEt and the combined organic layer was washed with brine, separated, dried over _N2SO4 , filtered and evaporated. The residue was _used in the next reaction without further purification. _LCMS calculated for _C35H32N7O3 (M+H)+: m/z=598.2; found 598.2.

Step 3. 3-(5-(bis(4-Methoxybenzyl)amino)-8-bromo-2-((3-methylpyridin-2-yl)methoxy)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

To a solution of 3-(5-(bis(4-methoxybenzyl)amino)-2-((3-methylpyridin-2-yl)methoxy)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (292 mg, 0.488 mmol) (300 mg, 0.503 mmol) in DCM (5 mL) at room temperature was added NBS (87 mg, 0.488 mmol). The mixture was stirred at room temperature for 1 h and then quenched with saturated aqueous NaHCO ₃ solution and separated. The organic layer was dried over Na ₂ SO ₄ , filtered, and evaporated. The residue was purified by column chromatography (10 to 50% AcOEt in hexane) to give the desired product. LC-MS calculated for C ₃₅ H ₃₁ BrN ₇ O ₃ (M+H)+: m/z=676.2; found 676.2.

Step 4. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((3-methylpyridin-2

- 33 049302 (yl)methoxy)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

3-(5-(bis(4-methoxybenzyl)amino)-8-bromo-2-((3-methylpyridin-2-yl)methoxy)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (20 mg, 0.030 mmol) (30 mg, 0.044 mmol), 1-methyl-5(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one (13.90 mg, 0.059 mmol), Xphos-G2 (2.326 mg, 2.96 μmol), and sodium carbonate (6.27 mg, 0.059 mmol) were combined. To this mixture were added 1,4-dioxane (1 mL) and water (0.100 mL). The mixture was heated to 100 °C and stirred for 3 h and then evaporated. TFA (1 mL) was added to the residue and the mixture was heated at 120 °C for 20 min. After cooling to room temperature, the mixture was diluted with acetonitrile, filtered and purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to give the _{desired product} as the TFA salt. LC-MS calculated for _C25H21N8O2 (M+H)+: m/z=465.2; found 465.2.

Example 19. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-((3-methylpyridin-2yl)methoxy)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 18 substituting 2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridazin-3(2H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS _{( pH} =2, MeCN/water with TFA) to give the desired product as the _TFA salt. LC-MS calculated for _C24H20N9O2 (M+H)+: m/z=466.2; found 466.2.

Example 20. 3-(5-Amino-2-((3-methylpyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 18 substituting 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl-5(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2( _1H )-one. The resulting material was purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to give the desired product as the TFA salt. LC-MS calculated for _C25H21N8O (M+H)+: m/z=449.2; found 449.2.

Example 21. 3-(5-Amino-8-(2,6-dimethylpyridin-4-yl)-2-((3-methylpyridin-2-yl)methoxy)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 18 substituting 2,6-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2( _1H )-one. The resulting material was purified by preparative LC-MS (pH=2, MeCN/water with TFA) to give the desired product as the _TFA salt. LC-MS calculated for _C26H23N8O (M+H)+: m/z=463.2; found 463.2.

Example 22. 3-(5-Amino-8-(1-ethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((3-methylpyridin-2yl)methoxy)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

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This compound was prepared using procedures similar to those described in Example 18 substituting 1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS _{( pH} =2, MeCN/water with TFA) to give the desired product as the _TFA salt. LC-MS calculated for _C26H23N8O2 (M+H)+: m/z=479.2; found 479.2.

Example 23. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2yl)methyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile.

Step 1. 3-(2-Amino-5-(bis(4-methoxybenzyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile.

Chloro(2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2yl)palladium(II) (0.185 g, 0.235 mmol) was added to a mixture of (3-cyano-2-fluorophenyl)boronic acid (0.582 g, 3.53 mmol), 7-chloro-N5,N5-bis(4-methoxybenzyl)-[1,2,4]triαzolo[1,5-c]nurimidine-2,5-diαmina (Example 15, step 2, 1 g, 2.354 mmol), sodium carbonate (0.499 g, 4.71 mmol) in 1,4-dioxane (13.08 mL) and water (2.62 mL). The reaction mixture was flushed with N ₂ and heated at 110 °C overnight. The mixture was diluted with AcOEt and water and separated. The organic layer was washed with brine, dried over Na ₂ SO ₄ , concentrated and purified by column chromatography (0 to 50% AcOEt in DCM). LCMS calculated for C ₂₈ H ₂₅ FN ₇ O ₂ (M+H)+: m/z = 510.2; found 510.2.

Step 2. 3-(5-(bis(4-Methoxybenzyl)amino)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile.

3-(2-Amino-5-(bis(4-methoxybenzyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile (300 mg, 0.589 mmol) (300 mg, 0.610 mmol), triethyl orthoformate (490 µL, 2.94 mmol), 3-methylpicolinaldehyde (143 mg, 1.178 mmol) were combined and EtOH (5 mL) was added. The resulting suspension was heated at 120 °C overnight. The mixture was then cooled to room temperature, diluted with

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DCM (2 mL). Sodium tetrahydroborate (46.2 mg, 1.221 mmol) was carefully added to the solution. After stirring at room temperature for 1 h, the mixture was carefully quenched with aqueous NH ₄ Cl, extracted with DCM and separated. The organic layer was dried over Na ₂ SO ₄ , filtered and evaporated. The residue was used in the next reaction without further purification. LCMS calculated for C ₃₅ H ₃₂ FN ₈ O ₂ (M+H)+: m/z = 615.2; found 615.2.

Step 3. 3-(5-(bis(4-Methoxybenzyl)amino)-8-bromo-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile.

To a solution of 3-(5-(bis(4-methoxybenzyl)amino)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile (300 mg, 0.488 mmol) in DCM (5 mL) at room temperature was added NBS (87 mg, 0.488 mmol). The mixture was stirred at room temperature for 1 h, then quenched with saturated aqueous NaHCO ₃ and separated. The organic layer was dried over Na2SO4, filtered and evaporated. The residue was purified by column chromatography (10 to 50% AcOEt in hexane) to give the desired product. LC-MS was calculated for C ₃₅ H ₃₁ BrFN ₈ O ₂ (M+H)+: m/z=693.2; found 693.2.

Step 4. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2yl)methyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile.

3-(5-(bis(4-methoxybenzyl)amino)-8-bromo-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile (20 mg, 0.029 mmol), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one (13.56 mg, 0.058 mmol), sodium carbonate (6.11 mg, 0.058 mmol) were combined. To this mixture were added 1,4-dioxane (1 mL) and water (0.100 mL). The mixture was heated to 100°C and stirred for 3 h and then evaporated. TFA (1 mL) was added to the residue and the mixture was heated at 120°C for 20 min. After cooling to _room temperature, the mixture was diluted with acetonitrile, filtered and purified by preparative LC-MS (pH=2, MeCN/water with TFA) to afford the desired product as the _TFA salt. LC-MS calculated for _C25H21FN9O (M+H)+: m/z=482.2; found 482.2.

Example 24. 3-(5-Amino-8-(1-ethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2yl)methyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile.

This compound was prepared using procedures similar to those described in Example 23 substituting 1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS (pH=2, MeCN/water with TFA) to give the desired product as the TFA salt. LC-MS calculated for C ₂₆ H ₂₃ FN ₉ O (M+H)+: m/z=496.2; found 496.2.

Example 25. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(((3-methylpyridin-2yl)methyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile.

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This compound was prepared using procedures similar to those described in Example 23 substituting 2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridazin-3(2H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS ₍ pH=2, MeCN/water with TFA) to afford the desired product as the _TFA salt. LC-MS calculated for _C24H20FN10O (M+H)+: m/z=483.2; found 483.2.

Example 26 3-(5-Amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile.

This compound was prepared using procedures similar to those described in Example 23 substituting 2-methoxy-6-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS (pH=2, MeCN/water with TFA) to give the desired product as the TFA salt. LC-MS calculated for C ₂₆ H ₂₃ FN ₉ O (M+H)+: m/z=496.2; found 496.2.

Example 27. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(pyridin-2-ylmethyl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

Step 1. 2-(Pyridin-2-yl)acetohydrazide.

Hydrazine (4.15 mL, 132 mmol) was added to an ethanolic (66 mL) solution of methyl 2-(pyridin-2yl)acetate (10 g, 66.2 mmol) at rt. The mixture was heated and stirred at 85 °C for 4 h and then cooled to rt. A white solid formed on standing, which was collected by filtration and used in the next step without further purification. LCMS calculated for C ₇ H ₁₀ N ₃ O (M+H)+: 152.1. Found: 152.0.

Step 2. 3-(5-Amino-2-(pyridin-2-ylmethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

_NH2

2-(Pyridin-2-yl)acetohydrazide (2.62 g, 17.34 mmol) was added to an ethanol (35 mL) solution of 3-(2-amino-6-chloropyrimidin-4-yl)benzonitrile (Example 1, step 1, 4.00 g, 17.34 mmol) at rt. After heating and stirring at reflux for 2 h, the reaction mixture was cooled to rt and concentrated. The resulting residue was taken up in N,O-bis(trimethylsilyl)acetamide (20 mL) and stirred at 120 °C for 7 h. The mixture was then cooled to rt, poured onto ice and allowed to stir at rt for 1 h. The resulting solid was collected by filtration and taken up in 20 mL of 1 N HCl solution. The resulting mixture was stirred at rt for 1 h, filtered, and the aqueous layer was neutralized by adding saturated _NaHCO3 solution. The resulting _precipitate was collected by filtration and dried to give the desired product as a brown solid. _LCMS calculated for _C18H14N7 (M+H)+: 328.1; found 328.1.

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Step 3. 3-(5-Amino-8-bromo-2-(pyridin-2-ylmethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

To a mixture of 3-(5-amino-2-(pyridin-2-ylmethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (2 g, 6.11 mmol) in DMF (12 mL) at -30 °C was added NBS (1.09 g, 6.11 mmol) portionwise. The reaction mixture was allowed to warm slowly to 0 °C, resulting in a homogeneous solution. After stirring at 0 °C for 1 h, the reaction mixture was diluted with saturated NaHCO ₃ solution and the resulting solid was collected by filtration. The solid was then purified by flash column chromatography on silica gel eluting with 0 to 10% MeOH in DCM to give the desired product. LC-MS calculated for C ₁₈ H ₁₃ BrN ₇ (M+H)+: 406.0; found 406.0.

Step 4. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(pyridin-2-ylmethyl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

3-(5-Amino-8-bromo-2-(pyridin-2-ylmethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (15 mg, 0.037 mmol), 2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridazin3(2H)-one (17.43 mg, 0.074 mmol), sodium carbonate (7.48 mg, 0.070 mmol), and Xphos-G2 (1.387 mg, 1.763 μmol) were combined. To this mixture were added 1,4-dioxane (1 mL) and water (0.100 mL). The mixture was heated to 100°C and stirred for 3 h, then cooled to room temperature, diluted with acetonitrile and TFA, filtered and purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to give the _desired product as the TFA salt. LC-MS calculated for _C23H18N9O (M+H)+: m/z=436.2; found 436.2.

Example 28. 3-(5-Amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

Step 1. 3-(5-(bis(4-Methoxybenzyl)amino)-2-((3-fluoropyridin-2-yl)methoxy)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

3-(5-(bis(4-Methoxybenzyl)amino)-2-iodo-[1,2,4]triazolo[1,5-]pyrimidin-7-yl)benzonitrile (250 mg, 0.415 mmol), (3-fluoropyridin-2-yl)methanol (158 mg, 1.245 mmol) were combined and 1,4-dioxane (5 mL) was added. Sodium hydride (60% in mineral oil, 41.5 mg, 1.037 mmol) was added to the solution. The suspension was heated to 105 °C and stirred for 1 h, then cooled to room temperature and quenched with aqueous NH ₄ Cl, diluted with AcOEt, separated. The aqueous layer was extracted with AcOEt and the combined organic layer was washed with brine, separated, dried _{over Na2SO4} , filtered _{and evaporated} . The residue was used in the next reaction without further purification. _LCMS calculated for _C34H29FN7O3 (M+H)+: m/z=602.2; found 602.2.

Step 2. 3-(5-(bis(4-Methoxybenzyl)amino)-8-bromo-2-((3-fluoropyridin-2-yl)methoxy)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

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To a solution of 3-(5-(bis(4-methoxybenzyl)amino)-2-((3-fluoropyridin-2-yl)methoxy)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile (294 mg, 0.488 mmol) (300 mg, 0.503 mmol) in DCM (5 mL) at room temperature was added NBS (87 mg, 0.488 mmol). The mixture was stirred at room temperature for 1 h and then quenched with saturated aqueous NaHCO ₃ solution and separated. The organic layer was dried over Na ₂ SO ₄ , filtered and evaporated. The residue was purified by column chromatography (10 to 50% AcOEt in hexane) to give the desired product. LC-MS was calculated for C ₃₄ H ₂₈ BrFN ₇ O ₃ (M+H)+: m/z=680.2; found 680.2.

Step 3. 3-(5-Amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

3-(5-(bis(4-methoxybenzyl)amino)-8-bromo-2-((3-fluoropyridin-2-yl)methoxy)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (30 mg, 0.044 mmol), tetrakis (10.19 mg, 8.82 μmol), 4(tributylstannyl)pyrimidine (24.41 mg, 0.066 mmol) were combined and 1,4-dioxane (1 mL) was added. The mixture was heated to 110 °C and stirred for 3 h and then evaporated. TFA (1 mL) was added to the residue and the mixture was heated at 120 °C for 20 min. After cooling to room temperature, the mixture was diluted with acetonitrile, filtered and purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to afford the product as the _TFA salt. LC-MS calculated for _C22H15FN9O (M+H)+: m/z=440.2; found 440.2.

Example 29. 3-(5-Amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

3-(5-(bis(4-methoxybenzyl)amino)-8-bromo-2-((3-fluoropyridin-2-yl)methoxy)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (12 mg, 0.018 mmol), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)pyridin-2(1H)-one (8.29 mg, 0.035 mmol), sodium carbonate (3.74 mg, 0.035 mmol), Xphos-G2 (1.387 mg, 1.763 μmol) were combined. To this mixture were added 1,4-dioxane (1 mL) and water (0.100 mL). The mixture was heated to 100 °C and stirred for 3 h and then evaporated. TFA (1 mL) was added to the residue and the mixture was heated at 120 °C for 20 min. After cooling to room temperature, the mixture was diluted with acetonitrile, filtered _and purified by preparative LC-MS (pH=2, MeCN/water with TFA) to give the desired product as _{the TFA salt. LC-MS calculated for C24H18FN8O2 (} M ₊ H)+: m/z=469.2; found 469.2.

Example 30. 3-(5-Amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridazin3-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 29, substituting 2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridazin-3(2H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS (pH 2, MeCN/water with TFA) to give the desired product as

- 39 049302 TFU salts. LC-MS calculated for C ₂₃ H ₁₇ FN ₉ O ₂ (M+H)+: m/z=470.2; found 470.2.

Example 31. 3-(5-Amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 29 substituting 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl-5(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2( _1H ) _-one . The resulting material was purified by preparative LC-MS (pH=2, MeCN/water with TFA) to afford the desired product as the TFA salt. LC-MS calculated for _C24H18FN8O (M+H)+: m/z=453.2.

Example 32. 3-(5-Amino-8-(2,6-dimethylpyridin-4-yl)-2-((3-fluoropyridin-2-yl)methoxy)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 29 substituting 2,6-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2( _1H )-one. The resulting material was purified by preparative LC-MS (pH=2, MeCN/water with TFA) to give the desired product as the _TFA salt. LC-MS calculated for _C25H20FN8O (M+H)+: m/z=467.2; found 467.2.

Example 33 3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(2-methoxy-6-methylpyridin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 29 substituting 2-methoxy-6-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS ₍ pH= ₂ , MeCN/water with TFA) to afford the desired product as the _TFA salt. LC-MS calculated for _C25H20FN8O2 (M+H)+: m/z=483.2; found 483.2.

Example 34. 3-(5-Amino-2-(pyridin-2-ylamino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile.

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Step 1. 3-(5-(bis(4-Methoxybenzyl)amino)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile.

3-(2-Amino-5-(bis(4-methoxybenzyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile (Example 15, step 3, 200 mg, 0.407 mmol), 2-bromopyridine (79 µL, 0.814 mmol), sodium 2-methylpropan-2-olate (78 mg, 0.814 mmol), XPhos-G2 (64.0 mg, 0.081 mmol) were combined and tert-BuOH (5 mL) was added. The mixture was heated at 70 °C overnight, then quenched with aqueous NH4Cl, diluted with AcOEt and separated. The organic layer was washed with brine, dried over _Na2SO4 , _filtered and evaporated. The residue was used in the next reaction without further purification. LCMS calculated for C ₃₃ H ₂₉ N ₈ O ₂ (M+H)+: m/z=569.2; found 569.2.

Step 2. 3-(5-(bis(4-Methoxybenzyl)amino)-8-bromo-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

To a solution of 3-(5-(bis(4-methoxybenzyl)amino)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile (278 mg, 0.488 mmol) (300 mg, 0.503 mmol) in DCM (5 mL) at room temperature was added NBS (87 mg, 0.488 mmol). The mixture was stirred at room temperature for 1 h, then quenched with saturated aqueous NaHCO ₃ solution and separated. The organic layer was dried over Na ₂ SO ₄ , filtered, and evaporated. The residue was purified by column chromatography (10 to 60% AcOEt in hexane) to give the desired product. LC-MS calculated for C ₃₃ H ₂₈ BrN ₈ O ₂ (M+H)+: m/z=647.2; found 647.2.

Step 3. 3-(5-Amino-2-(pyridin-2-ylamino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile.

3-(5-(bis(4-methoxybenzyl)amino)-8-bromo-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile (30 mg, 0.046 mmol), tetrakis (10.71 mg, 9.27 μmol), 4(tributylstannyl)pyrimidine (25.7 mg, 0.069 mmol) were combined and 1,4-dioxane (1 mL) was added. The mixture was heated to 110 °C and stirred for 3 h and then evaporated. TFA (1 mL) was added to the residue and the mixture was heated at 120 °C for 20 min. After cooling to room temperature, the mixture was diluted with acetonitrile, filtered and purified by preparative LC-MS (pH= ₂ , MeCN/water with TFA) to afford the product as the _TFA salt. LC-MS calculated for _C21H15N10 (M+H)+: m/z=407.2; found 407.2.

Example 35. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(pyridin-2-ylamino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

3-(5-(bis(4-methoxybenzyl)amino)-8-bromo-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile (20 mg, 0.031 mmol), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane

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2-yl)pyridin-2(III)-one (14.52 mg, 0.062 mmol), sodium carbonate (6.55 mg, 0.062 mmol), Xphos-G2 (2.430 mg, 3.09 μmol) were added to the mixture and 1,4-dioxane (1 mL) and water (0.100 mL) were added. The mixture was heated at 100 °C for 3 h and then evaporated. TFA (1 mL) was added to the residue and the mixture was heated at 120 °C for 20 min. After cooling to room temperature, the mixture was diluted with acetonitrile, filtered and purified by preparative LC-MS (pH=2, MeCN/water with TFA) to give the desired product as a TFA salt. LC-MS calculated for C ₂₃ H ₁₈ N ₉ O (M+H)+: m/z=436.2.

Example 36. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(pyridin-2-ylamino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 35 substituting 2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridazin-3(2H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to afford the desired product as the _TFA salt. LC-MS calculated for _C22H17N10O (M+H)+: m/z=437.2.

Example 37. 3-(5-Amino-8-(3-methylpyridin-4-yl)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 35 substituting 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl-5(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2( _1H )-one. The resulting material was purified by preparative LC-MS (pH= ₂ , MeCN/water with TFA) to afford the desired product as the TFA salt. LC-MS calculated for _C23H18N9 (M+H)+: m/z=420.2.

Example 38. 3-(5-Amino-8-(2,6-dimethylpyridin-4-yl)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 35 substituting 2,6-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2( _1H )-one. The resulting material was purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to afford the desired product as the TFA salt. LC-MS calculated for _C24H20N9 (M+H)+: m/z=434.2.

Example 39. 3-(5-Amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 35, with

- 42 049302 using 2-methoxy-6-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine instead of 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS (pH=2, MeCN/water with TFA) _to afford the desired product as the _TFA salt. LC-MS calculated for _C24H20N9O (M+H)+: m/z=450.2.

Example 40. 3-(5-Amino-2-((6-methylpyridin-2-yl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

N

_NH2

This compound was prepared using procedures similar to those described in Example 34, substituting 2-bromo-6-methylpyridine for 2-bromopyridine. The resulting material was purified by preparative LC-MS (pH=2, MeCN/water with TFA) to afford the _desired product as the TFA salt. LC-MS calculated for _C22H17N10 (M+H)+: m/z=421.2; found 421.2.

Example 41. 3-(5-Amino-2-((pyridin-2-yloxy)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

N

_NH2

To a 1,4-dioxane (1 mL) solution of 3-(5-amino-2-(hydroxymethyl)-8-(pyrimidin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (Example 1, step 4, 15 mg, 0.044 mmol) and 2-fluoropyridine (0.011 mL, 0.131 mmol) was added sodium hydride (60% in mineral oil, 3.48 mg, 0.087 mmol). The mixture was heated at 60 °C for 3 h and then cooled to room temperature, diluted with acetonitrile and TFA, filtered and purified by preparative LC-MS (pH 2, MeCN/water with TFA) to give the desired product as the TFA salt. LC-MS calculated for C ₂₂ H ₁₆ N ₉ O (M+H)+: m/z=422.2; found 422.2.

Example 42. 2-((5-Amino-7-(3-cyanophenyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-2yl)methoxy)nicotinonitrile.

Step 1. 2-((5-Amino-7-(3-cyanophenyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)methoxy)nicotinonitrile.

To a solution of 3-(5-amino-2-(hydroxymethyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile in 1,4-dioxane (5 mL) (Example 1, step 2, 100 mg, 0.376 mmol) and 2-fluoronicotinonitrile (138 mg, 1.127 mmol) was added sodium hydride (60% in mineral oil, 30.0 mg, 0.751 mmol). The mixture was heated at 60 °C for 3 h, then cooled to room temperature, quenched with aqueous NH4Cl, _{diluted with DCM, and separated. The organic layer was dried over Na2SO4 ,} filtered, and evaporated. The residue was used in the next reaction without further purification. LC-MS calculated for C ₁₉ H ₁₃ N ₈ O (M+H)+: m/z=369.2; found 369.2.

Step 2. 2-((5-Amino-8-bromo-7-(3-cyanophenyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2yl)methoxy)nicotinonitrile.

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To a suspension of 2-((5-amino-7-(3-cyanophenyl)-[1,2,4]triazolo[1,5-c]pyrimidin-2yl)methoxy)nicotinonitrile (185 mg, 0.503 mmol) in DCM (1 mL) was added NBS (89 mg, 0.503 mmol) at room temperature. The mixture was stirred at room temperature for 1 h. After cooling to room temperature, the mixture was quenched with saturated aqueous NaHCO ₃ , diluted with DCM and separated. The organic layer was dried over Na ₂ SO ₄ , filtered and evaporated. The residue was purified by column chromatography (0 to 10% MeOH in DCM) to give the desired product. LC-MS calculated for C ₁₉ H ₁₂ BrN ₈ O (M+H)+: m/z = 447.0; 447.0 found.

Step 3. 2-((5-Amino-7-(3-cyanophenyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-2yl)methoxy)nicotinonitrile.

2-((5-Amino-8-bromo-7-(3-cyanophenyl)-[1,2,4]triazolo[1,5-c]pyrimidin2-yl)methoxy)nicotinonitrile (15 mg, 0.034 mmol), tetrakis (7.75 mg, 6.71 μmol), 4(tributylstannyl)pyrimidine (18.57 mg, 0.050 mmol) were combined and 1,4-dioxane (1 mL) was added. The mixture was heated at 110 °C for 3 h, then cooled to room temperature, quenched and diluted with TFA (0.5 mL), filtered and purified by preparative LC-MS (pH = 2, MeCN/water with TFA) to afford the desired product as the TFA salt. LC-MS calculated for C ₂₃ H ₁₅ N ₁₀ O (M+H)+: m/z=447.2; found 447.2.

Example 43. 2-((5-Amino-7-(3-cyanophenyl)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3 yl)-[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)methoxy)nicotinonitrile.

2-((5-(bis(4-methoxybenzyl)amino)-8-bromo-7-(3-cyanophenyl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile (20 mg, 0.029 mmol), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)pyridin-2(1H)-one (13.68 mg, 0.058 mmol), sodium carbonate (6.17 mg, 0.058 mmol), Xphos-G2 (2.289 mg, 2.91 μmol) were combined. 1,4-Dioxane (1 mL) and water (0.100 mL) were added and the mixture was heated to 100 °C and stirred for 3 h, then evaporated. TFA (1 mL) was added to the residue and the mixture was heated at 120 °C for 20 min. After cooling to room temperature, the mixture was diluted with acetonitrile, filtered and purified by preparative LC-MS _{( pH} =2, MeCN/water with TFA) to give the desired product as the _TFA salt. LC-MS calculated for _C25H18N9O2 (M+H)+: m/z=476.2; found 476.2.

Example 44. 2-((5-Amino-7-(3-cyanophenyl)-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)[1,2,4]triazolo[1,5-c]pyrimidin-2-yl)methoxy)nicotinonitrile.

This compound was prepared using procedures similar to those described in Example 43 substituting 2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridazin-3(2H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by _preparative LC-MS (pH=2, MeCN/water with TFA) to afford the desired product as the _TFA salt. LC-MS calculated for _C24H17N10O2 (M ₊ H)+: m/z=477.2; found 477.2.

Example 45. 2-((5-Amino-7-(3-cyanophenyl)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile.

- 44 049302

This compound was prepared using procedures similar to those described in Example 43 substituting 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl-5(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2( _1H ) _-one . The resulting material was purified by preparative LC-MS (pH=2, MeCN/water with TFA) to afford the desired product as the TFA salt. LC-MS calculated for _C25H18N9O (M+H)+: m/z=460.2.

Example 46. 2-((5-Amino-7-(3-cyanophenyl)-8-(2,6-dimethylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile.

This compound was prepared using procedures similar to those described in Example 43 substituting 2,6-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2( _1H )-one. The resulting material was purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to give the desired product as the TFA salt. LC-MS calculated for _C26H20N9O (M+H)+: m/z=474.2; found 474.2.

Example 47. 2-((5-Amino-7-(3-cyanophenyl)-8-(2-methoxy-6-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile.

This compound was prepared using procedures similar to those described in Example 43 substituting 2-methoxy-6-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS ₍ pH= ₂ , MeCN/water with TFA) to give the desired product as the _TFA salt. LC-MS calculated for _C26H20N9O2 (M+H)+: m/z=490.2; found 490.2.

Example 48. 3-(5-Amino-2-((1-(pyridin-2-yl)ethyl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

Step 1. 3-(2-Amino-5-(bis(4-methoxybenzyl)amino)-8-bromo-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile.

- 45 049302

To a solution of 3-(2-amino-5-(bis(4-methoxybenzyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile (Example 15, step 3, 330 mg, 0.66 mmol) in DMF (1.4 mL) was slowly added NBS (120 mg, 0.66 mmol) at 0 °C. The reaction mixture was then stirred at rt for 30 min, after which water (10 mL) was added. The resulting solid was collected by filtration and dried to give the desired product. LCMS calculated for C ₂₈ H ₂₅ BrN ₇ O ₂ (M+H)+: m/z = 570.1.

Step 2. 3-(2-Amino-5-(bis(4-methoxybenzyl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

A mixture of 3-(2-amino-5-(bis(4-methoxybenzyl)amino)-8-bromo-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile (350 mg, 0.61 mmol), 4-(tributylstannyl)pyrimidine (0.21 mL, 0.67 mmol), tetrakis(triphenylphosphine)palladium(0) (70 mg, 0.060 mmol), copper(T) iodide (23 mg, 0.12 mmol), and cesium fluoride (180 mg, 1.2 mmol) in dioxane (4.7 mL) was heated and stirred at 140 °C for 30 min in a microwave reactor. The reaction mixture was then cooled to room temperature, filtered through a pad of celite (washing with DCM), and concentrated. The resulting material was purified by column chromatography on silica gel eluting with 0-20% MeOH/DCM to give the desired product. LC-MS calculated for C ₃₂ H ₂₈ N ₉ O ₂ (M+H)+: m/z=570.2.

Step 3. 3-(5-(Bis(4-methoxybenzyl)amino)-2-bromo-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

To a solution of copper(II) bromide (91 mg, 0.407 mmol) and tert-butyl nitrite (0.054 mL, 0.407 mmol) in acetonitrile (3 mL) under nitrogen atmosphere at 50 °C was added dropwise 3-(2-amino-5-(bis(4methoxybenzyl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (100 mg, 0.203 mmol) in acetonitrile (3 mL). The mixture was stirred at 50 °C for 2 h. After cooling to room temperature, 1 N aqueous NH ₄ OH (20 mL) was added and the mixture was extracted three times with CH ₂ Cl ₂ (20 mL). The combined organic layers were dried over sodium sulfate, filtered and the solvent was evaporated under reduced pressure. The residue was purified by silica gel _column chromatography eluting with 50-100% ethyl _acetate /hexane _to give the desired product. LC-MS calculated for _C32H26BrN8O2 (M+H)+: m/z=633.2.

Step 4. 3-(5-Amino-2-((1-(pyridin-2-yl)ethyl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5

- 46 049302

c]pyrimidin-7-yl)benzonitrile.

3-(5-(bis(4-methoxybenzyl)amino)-2-bromo-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile (750 mg, 1.184 mmol), 1-(pyridin-2-yl)ethan-1-amine (289 mg, 2.368 mmol), sodium tert-butoxide (228 mg, 2.368 mmol), and (tert-Bu)PhCPhosPd G3 (92 mg, 0.118 mmol) were combined and 1,4-dioxane (10 mL) was added. The mixture was heated at 60 °C for 3 h, then cooled to room temperature and evaporated. TFA (5 mL) was added to the residue and the mixture was heated at 120°C for 20 min. After cooling to room temperature, the mixture was diluted with acetonitrile, filtered and purified by preparative LC-MS (pH=2, MeCN/ _water with TFA) to afford the desired product as the TFA salt. LC-MS calculated for _C23H19N10 (M ₊ H)+: m/z=435.2.

Example 49. 3-(5-Amino-2-((2-(pyridin-2-yl)propan-2-yl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 48, substituting 2-(pyridin-2-yl)propan-2-amine for 1-(pyridin-2-yl)ethan-1-amine. The resulting material was purified by _preparative LC-MS (pH=2, MeCN/water with TFA) to afford the desired product as the TFA _salt . LC-MS calculated for _C24H21N10 (M+H)+: m/z=449.2.

Example 50. 3-(5-Amino-2-((5-(pyridin-2-yl)-2H-tetrazol-2-yl)methyl)-8-(pyrimidin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Example 1, with 2-(1H-tetrazol-5-yl)pyridine replacing 3-(1H-tetrazol-5-yl)aniline. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LCMS calculated for C ₂₃ H ₁₆ N ₁₃ (M+H)+: 474.2; found 474.2.

Example 51. 3-(5-amino-2-((5-(pyrimidin-2-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Example 1, with 2-(1H-tetrazol-5-yl)pyrimidine replacing 3-(1H-tetrazol-5-yl)aniline. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LCMS calculated for C ₂₂ H ₁₅ N ₁₄ (M+H)+: 475.2; found 475.2.

Example 52. 3-(5-amino-8-(pyrimidin-4-yl)-2-((5-(pyrimidin-4-yl)-1H-tetrazol-1-yl)methyl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Example 1, with 4(1H-tetrazol-5-yl)pyrimidine replacing 3-(1H-tetrazol-5-yl)aniline. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LC

- 47 049302

MS was calculated for C ₂₂ H ₁₅ N ₁₄ (M+H)+: 475.2; detected 475.2.

Example 53. 3-(5-Amino-2-((5-(pyridin-3-yl)-III-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Example 1, with 3(1H-tetrazol-5-yl)pyridine replacing 3-(1H-tetrazol-5-yl)aniline. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LCMS calculated for C ₂₃ H ₁₆ N ₁₃ (M+H)+: 474.2; found 474.2.

Example 54. 3-(5-Amino-2-((5-(pyridin-4-yl)-III-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described for Example 1, with 4(1H-tetrazol-5-yl)pyridine replacing 3-(1H-tetrazol-5-yl)aniline. The product was purified by preparative LC-MS (pH 2, acetonitrile/water with TFA) to afford the product as the TFA salt. LCMS calculated for C ₂₃ H ₁₆ N ₁₃ (M+H)+: 474.2; found 474.2.

Example 55. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((6-methylpyridin-2yl)methoxy)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

Step 1. 3-(5-(bis(4-Methoxybenzyl)amino)-8-bromo-2-((6-methylpyridin-2-yl)methoxy)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 28 substituting (6-methylpyridin-2-yl)methanol for (3-fluoropyridin-2-yl)methanol in Step 1. The product was purified by column chromatography (10 to 60% AcOEt in hexanes) to give the desired product. LCMS calculated for C ₃₅ H ₃₁ BrN7O ₃ (M+H)+: m/z=676.2; found 676.2.

Step 2. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((6-methylpyridin2-yl)methoxy)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

Combined 3-(5-(bis(4-methoxybenzyl)amino)-8-bromo-2-((6-methylpyridin-2-yl)methoxy)

- 48 049302

[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (20 mg, 0.030 mmol), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2dioxaborolan-2-yl)pyridin-2(1H)-one (6.95 mg, 0.030 mmol), sodium carbonate (6.27 mg, 0.059 mmol), XphosG2 (2.326 mg, 2.96 μmol). To the above mixture were added 1,4-dioxane (1 mL) and water (0.100 mL). The mixture was heated to 100 °C and stirred for 3 h and then evaporated. To the residue was added TFA (1 mL), and the mixture was heated at 120 °C for 20 min. It was then diluted with acetonitrile, filtered and purified by preparative LC-MS (pH=2, MeCN/water with TFA) to give the products as the TFA salt. LCMS calculated for C^H^N^ (M+H)+: m/z=465.2; found 465.2.

Example 56. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-((6-methylpyridin-2yl)methoxy)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 55 substituting 2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridazin-3(2H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one in Step 2. The resulting material was purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to afford the desired product as the TFA salt. LC-MS calculated for _C24H20N9O2 (M+H)+: m/z=466.2.

Example 57 3-(5-Amino-2-((6-methylpyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 55 substituting 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl-5(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one in Step 2. The resulting material was purified by preparative LC-MS (pH=2, MeCN/water with TFA) to give the desired product as the TFA salt. LC-MS calculated for C2 ₅ H ₂₁ N ₈ O (M+H)+: m/z=449.2; found 449.2.

Example 58. 3-(5-Amino-8-(2,6-dimethylpyridin-4-yl)-2-((6-methylpyridin-2-yl)methoxy)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 55 substituting 2,6-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2( _1H )-one in Step 2. The resulting material was purified by preparative LC-MS (pH=2, MeCN/water with TFA) to give the desired product as the TFA salt. LC-MS calculated for _C26H23N8O (M+H)+: m/z=463.2; found 463.2.

Example 59

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This compound was prepared using procedures similar to those described in Example 55 substituting 2-methoxy-6-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one in Step 2. The resulting material was purified by _preparative LC- _MS (pH=2, MeCN/water with TFA) to give the desired product as the _TFA salt. LC-MS calculated for _C26H23N8O2 (M+H)+: m/z=479.2; found 479.2.

Example 60. 3-(5-Amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

Step 1. 3-(5-(bis(4-Methoxybenzyl)amino)-8-bromo-2-((3,6-dimethylpyridin-2-yl)methoxy)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 28, substituting (3,6-dimethylpyridin-2-yl)methanol for (3-fluoropyridin-2-yl)methanol in Step 1. The product was purified by column chromatography (10 to 60% AcOEt in hexanes) to give the desired product. LCMS calculated for C ₃₆ H ₃₃ BrN ₇ O ₃ (M+H)+: m/z=690.2.

Step 2. 3-(5-Amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

3-(5-(bis(4-methoxybenzyl)amino)-8-bromo-2-((6-methylpyridin-2-yl)methoxy)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (20 mg, 0.030 mmol), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one (6.95 mg, 0.030 mmol), sodium carbonate (6.27 mg, 0.059 mmol), Xphos-G2 (2.326 mg, 2.96 μmol) were combined. To the mixture were added 1,4-dioxane (1 mL) and water (0.100 mL). The mixture was heated to 100°C and stirred for 3 h and then evaporated. TFA (1 mL) was added to the residue and the mixture was heated at 120°C for 20 min. The mixture was then diluted with acetonitrile, filtered and purified _{by preparative} LC-MS (pH=2, _MeCN /water with TFA) to afford the products as the TFA salt. LC-MS calculated for _C26H23N8O2 (M+H)+: m/z=479.2.

Example 61. 3-(5-Amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

- 50 049302

This compound was prepared using procedures similar to those described in Example 60 substituting 2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridazin-3(2H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one in Step 2. The resulting material _was purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to afford the desired product as the TFA salt. LC-MS calculated for _C25H22N9O2 (M ₊ H)+: m/z=480.2.

Example 62. 3-(5-Amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 60 substituting 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl-5(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one in Step 2. The resulting material was purified by preparative LC-MS ( _pH = ₂ , MeCN/water with TFA) to afford the desired product as the TFA salt. LC-MS calculated for _C26H23N8O (M+H)+: m/z=463.2.

Example 63. 3-(5-Amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(2,6-dimethylpyridin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 60 substituting 2,6-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2( _1H )-one in Step _2. The resulting material was purified by preparative LC-MS (pH=2, MeCN/water with TFA) to afford the desired product as the TFA salt. LC-MS calculated for _C27H25N8O (M+H)+: m/z=477.2.

Example 64. 3-(5-amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(2-methoxy-6-methylpyridin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 60, substituting 2-methoxy-6-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one in Step 2. The resulting material was purified by preparative LC-MS (pH=2, MeCN/water with TFA) to give the desired product.

- 51 049302 as TFA salt. LC-MS calculated for C ₂₇ H ₂₅ N ₈ O ₂ (M+H)+: m/z=493.2.

Example 65. 3-(5-Amino-2-(((6-methylpyridin-2-yl)methyl)amino)-8-(pyrimidin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

Step 1. 3-(5-(bis(4-Methoxybenzyl)amino)-8-bromo-2-(((6-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 15, substituting 6-methylpicolinaldehyde for 3-methylpicolinaldehyde in Step _4. The product _was purified by column chromatography (10 to 60% _AcOEt in hexanes) to give the desired product. LCMS calculated for _C35H32BrN8O2 (M+H)+: m/z=675.2; found 675.2.

Step 2. 3-(5-Amino-2-(((6-methylpyridin-2-yl)methyl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile.

3-(5-(bis(4-methoxybenzyl)amino)-8-bromo-2-(((6-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5- c]pyrimidin-7-yl)benzonitrile (20 mg, 0.030 mmol), tetrakis (6.84 mg, 5.92 μmol), 4-(tributylstannyl)pyrimidine (16.39 mg, 0.044 mmol) were combined. 1,4-dioxane (1 mL) was added to the mixture. The mixture was heated at 110°C for 3 h and evaporated. TFA (1 mL) was added to the residue and the mixture was heated at 120°C for 20 min, then diluted with acetonitrile, filtered and purified by preparative LC-MS (pH 2, MeCN/water with TFA) to afford the product as the TFA salt. LC-MS calculated for C ₂₃ H ₁₉ N ₁₀ (M+H)+: m/z = 435.2; found 435.2.

Example 66. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((6-methylpyridin-2yl)methyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

3-(5-(bis(4-methoxybenzyl)amino)-8-bromo-2-(((6-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile (20 mg, 0.030 mmol), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one (6.96 mg, 0.030 mmol), sodium carbonate (6.28 mg, 0.059 mmol), Xphos-G2 (2.329 mg, 2.96 μmol) were combined. To the mixture were added 1,4-dioxane (1 mL) and water (0.100 mL). The mixture was heated at 100°C for 3 h and then evaporated. TFA (1 mL) was added to the residue and the mixture was heated at 120°C for 20 min and then diluted with acetonitrile, filtered and purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to give the products as the _TFA salt. LC-MS calculated for _C25H22N9O (M+H)+: m/z=464.2.

Example 67. 3-(5-Amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(((6-methylpyridin2-yl)methyl)amino)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

- 52 049302

This compound was prepared using procedures similar to those described in Example 66 substituting 2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridazin-3(2H)-one for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to give the desired product _as the TFA salt. LC-MS calculated for _C24H21N10O (M+H)+: m/z=465.2; found 465.2.

Example 68. 3-(5-Amino-2-(((6-methylpyridin-2-yl)methyl)amino)-8-(3-methylpyridin-4-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 66 substituting 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl-5(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2( _1H )-one. The resulting material was purified by preparative LC-MS (pH= ₂ , MeCN/water with TFA) to give the desired product as the TFA salt. LC-MS calculated for _C25H22N9 (M+H)+: m/z=448.2; found 448.2.

Example 69. 3-(5-Amino-8-(2,6-dimethylpyridin-4-yl)-2-(((6-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile.

This compound was prepared using procedures similar to those described in Example 66 substituting 2,6-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1-methyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2( _1H )-one. The resulting material was purified by preparative LC-MS (pH=2, _MeCN /water with TFA) to give the desired product as the TFA salt. LC-MS calculated for _C26H24N9 (M+H)+: m/z=462.2; found 462.2.

Example 70

This compound was prepared using procedures similar to those described in Example 66, substituting 2-methoxy-6-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine for 1methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one. The resulting material was purified by preparative LC-MS (pH 2, MeCN/water with TFA) to afford the desired product as

- 53 049302 TFU salts. LC-MS calculated for C ₂₆ H ₂₄ N ₉ O (M+H)+: m/z=478.2; found 478.2.

Example A. Cyclic AMP GS assay of the A2A adenosine receptor.

Stably transfected HEK-293 cells expressing human adenosine receptor A2A (Perkin Elmer) were maintained in MEM culture medium with 10% FBS and 400 μg/ml geneticin (Life Technologies). Geneticin was removed from the culture 18-24 h before the assay. The cisbio cAMP-GS Dynamic kit using FRET (fluorescence resonance energy transfer) technology was used to measure cAMP accumulation in the cells. The compounds of the invention at the appropriate concentration were mixed with 10,000 cells/well in white 96-well half-area plates (Perkin Elmer) for 30 min at room temperature (RT) with gentle shaking. The agonist CGS21680 (R&D Technologies) at a concentration of 4 nM was added to each well for 60 min at RT with gentle shaking. The detection reagents, d2-labeled cAMP (acceptor) and anti-cAMP cryptate (donor) were added to each well for 60 min at RT with gentle shaking. The plates were read on a Pherastar (BMG Labtech), the 665/620 fluorescence ratio was calculated and the EC ₅₀ was determined by curve fitting the percent control to the logarithm of the compound concentration using GraphPad Prism.

Example B. Cyclic AMP GS assay of the A2B adenosine receptor.

Stably transfected HEK-293 cells expressing human adenosine receptor A2B (Perkin Elmer) were maintained in MEM culture medium with 10% FBS and 100 μg/ml geneticin (Life Technologies). Geneticin was removed from the culture 18-24 h before the assay. The cisbio cAMP-GS Dynamic kit using FRET (fluorescence resonance energy transfer) technology was used to measure cAMP accumulation in the cells. The compounds of the invention at the appropriate concentration were mixed with 10,000 cells/well in white 96-well half-area plates (Perkin Elmer) for 30 min at RT with gentle shaking. The agonist NECA (R&D Technologies) at a concentration of 12 nM was added to each well for 60 min at RT with gentle shaking. Detection reagents, d2-labeled cAMP (acceptor) and anti-cAMP cryptate (donor) were added to each well for 60 min at RT with gentle shaking. Plates were read on a Pherastar (BMG Labtech), the 665/620 fluorescence ratio was calculated, and the EC ₅₀ was determined by fitting the percent control curve to the logarithm of the compound concentration using GraphPad Prism. The EC ₅₀ data obtained using this method are shown in Table 1.

Example C. A2A Tag-lite® HTRF Analysis.

Assays were performed in black 384-well low-volume polystyrene plates (Greiner 784076-25) in a final volume of 10 μl. Test compounds were first serially diluted in DMSO and 100 nl were added to the wells of the plate before addition of other reaction components. The final DMSO concentration was 1%. Cells labeled with adenosine Tag-lite® A2A (CisBio C1TT1A2A) were diluted 1:5 in Tag-lite buffer (CisBio LABMED) and centrifuged at 1200 g for 5 min. The pellet was resuspended in 10.4 times the original cell suspension volume in Tag-lite buffer and the fluorescent ligand, adenosine receptor antagonist A2A Red (CisBio L0058RED), was added at a final concentration of 12.5 nM. 10 μl of the cell-ligand mixture was added to the assay wells and incubated at room temperature for 45 min before reading on a PHERAstar FS plate reader (BMG Labtech) with an HTRF 337/620/665 optical module. The percentage of fluorescent ligand binding was calculated; where 100 nM of the control A2A antagonist ZM 241385 (Tocris 1036) displaces 100% of the ligand and 1% DMSO displaces 0%. The % binding versus log inhibitor concentration data were fitted to a single-site competitive binding model (GraphPad Prism version 7.02) where ligand constant = 12.5 nM and ligand K _d = 1.85 nM. The Ki data obtained using this method are shown in Table 1.

Example D. A2B Filter Binding Analysis.

Assays were performed in deep-well polypropylene plates (Greiner 786201) in a final volume of 550 μl. Test compounds were first serially diluted in DMSO and 5.5 μl were added to the wells of the plate before adding other reaction components. The final DMSO concentration was 3%. HEK293 cell membranes overexpressing the human adenosine A2B receptor (Perkin Elmer ES-113-M400UA) were diluted to 40 μgμl/ml in 50 mM HEPES, pH 7.0, 5 mM _MgCl2 , 1 mM EDTA (assay buffer). [3H]8-Cyclopentyl-1,3-dipropylxanthine (Perkin Elmer NET974001MC) was diluted with assay buffer+22% DMSO to 24.2 nM and then further diluted to 1 nM by adding to the diluted membranes. 545 μl of the membrane-ligand mixture was added to the assay wells and incubated on a shaker at room temperature for 1 h. The membrane mixture was then filtered through a UniFilter GF/C filter plate (Perkin Elmer 6005174) pre-soaked with 50 mM HEPES, pH 6.5, 5 mM _MgCl2 , 1 mM EDTA, 0.5% BSA, and then washed with 5 ml of ice-cold 50 mM HEPES. pH 6.5, 5 mM _MgCl2 , 1 mM EDTA, 0.2% BSA. 50 μL MicroScint™ Cocktail (Perkin Elmer 6013621) was added and the plates were read on a Topcount NXT FS (Perkin Elmer). The percentage of [3H]-ligand binding was calculated, where 1000 nM control LUF 5834 (Tocris 4603) displaces 100% of the ligand and 3% DMSO displaces 0%. The % binding data are compared to

- 54 049302 logarithm of inhibitor concentration correspond to the competitive binding model with one site (GraphPad Prism version 7.02), where ligand constant = 2 nM and ligand Kd = 13 nM.

Example E. SPA A1 and A3 binding assays.

Both assays were performed in white 384-well polystyrene plates (Greiner 781075) in a final volume of 50 μl. Inhibitors were first serially diluted in DMSO and 100 nl were added to the wells of the plate before adding other reaction components. The final DMSO concentration was 2%.

Yttrium silicate SPA beads coated with wheat germ agglutinin (Perkin Elmer RPNQ0023) and CHO-K1 cell membranes overexpressing each human adenosin receptor were incubated in 50 mM HEPES pH 7.0, 5 mM _MgCl2 , 1 mM EDTA (assay buffer) on a rotary shaker for 2 h at 4°C. Beads were pelleted by centrifugation at 6000 g for 1 min, after which the supernatant with unbound membrane was discarded. Beads were resuspended to the original volume in assay buffer. Each radioligand was diluted in assay buffer+22% DMSO to 12.2-fold the final concentration and then added to the SPA bead suspension. 50 μl of the SPA bead reaction mixture was added to the assay wells and the plates were shaken at 600 rpm for 1 h at room temperature. The beads were then allowed to settle for 1 h before being counted on a Topcount NXT FS instrument (Perkin Elmer). The percent binding of the radiolabeled ligand was calculated, where the control at >100X Ki displaces the ligand by 100% and 2% DMSO has 0% displacement. The % binding versus log inhibitor concentration data are consistent with a single-site competitive binding model (GraphPad Prism version 7.02). The assay conditions are listed in the table below.___________________________

Analysis component A1 A3 SPA balls in Hepes buffer 3 mg/ml 1.25 mg/ml Membrane 60 mcg/ml Perkin Elmer ES-010 20 mcg/ml Perkin Elmer ES-012 Radioligand 1 nM [3H] DP-CPX (Perkin Elmer NET974) K _D =1 nM 0.1 nM [125I] MECA (Perkin Elmer NEX312) Kd=0.8 nM Control 1 μM DPCPX (Tocris 0439) 0.1 µM IB-MECA (Tocris 1066)

Table 1

The data of A _2A _Ki (Example C) and the data of A _2B _cAMP_EC ₅₀ (Example B) are presented below

Example No. A2A_Ki (nM) A2B_cAMP_EC ₅₀ (nM) 1 t t 2 t t 3 t tt 4 t tt 5 t tt 6 t tt 7 t tt 8 t tt 9 t tt 10 t t 11 t t 12 t t 13 t t 14 t t 15 t t 16 t t 17 t t 18 t t 19 t t 20 t t 21 t tt 22 t tt 23 t tt 24 t tt 25 t t 26 t tt 27 t t

- 55 049302

28 t t 29 t t 30 t t 31 n/a n/a 32 t tt 33 t t 34 t t 35 n/a n/a 36 n/a n/a 37 n/a n/a 38 n/a n/a 39 n/a n/a 40 t t 41 t t 42 t t 43 t t 44 t tt 45 n/a n/a 46 t ttt 47 t tt 48 n/a n/a 49 n/a n/a 50 t t 51 t t 52 t t 53 t t 54 t t 55 t tt 56 n/a n/a 57 t tt 58 t ttt 59 t tt 60 n/a n/a 61 n/a n/a 62 n/a n/a 63 n/a n/a 64 n/a n/a 65 t tt 66 n/a n/a 67 t tt 68 t t 69 t tttt 70 t ttt

t indicates A _2A _Ki or A _2B _cAMP_EC ₅₀ < 10 nM, tt indicates A _2A _K or A _2B _cAMP_EC ₅₀ > 10 nM but < 100 nM, ttt indicates A _2A _K; or A _2B _cAMP_EC ₅₀ > 100 nM but < 1 μM, tttt indicates A _2A _K; or A _2B _cAMP_EC ₅₀ > 1 μM, and n/a indicates no data.

From the foregoing description, various modifications of the invention in addition to those described herein will be apparent to those skilled in the art. Such modifications are also intended to be within the scope of the appended claims. Each reference, including all patents, patent applications, and publications cited in this application, is incorporated herein by reference in their entirety.

Claims (14)

1. A compound selected from: 3-(5-amino-2-((5-(3-aminophenyl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((5-(6-methylpyridin-2-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((5-(6-methoxypyridin-2-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(pyridin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(2-((5-(1H-pyrazol-1-yl)-1H-tetrazol-1-yl)methyl)-5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(thiazol-4-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(pyrimidin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((5-(pyrazin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-((5-(pyridin-2-yl)-1H-tetrazol-1yl)methyl)-[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((5-(pyridin-2-yl)-III-tetrazol-1-yl)methyl)-8-(pyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(2-(dimethylamino)pyridin-4-yl)-2-((5-(pyridin-2-yl)-III-tetrazol-1-yl)methyl)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(2-aminopyridin-4-yl)-2-((5-(pyridin-2-yl)-III-tetrazol-1-yl)methyl)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-(2-fluoro-6-(pyridin-4-yl)benzyl)-8-(pyridin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile; 3-(5-amino-8-(2-aminopyridin-4-yl)-2-(2-(2-aminopyridin-4-yl)-6-fluorobenzyl)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-ethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((3-methylpyridin-2-yl)methoxy)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-((3-methylpyridin-2-yl)methoxy)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((3-methylpyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-((3-methylpyridin-2-yl)methoxy)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-ethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((3-methylpyridin-2-yl)methoxy)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile; 3-(5-amino-8-(1-ethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile; 3-(5-amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-(((3-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)-2-fluorobenzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(pyridin-2-ylmethyl)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile; 3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; - 57 049302 3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-((3-fluoropyridin-2-yl)methoxy)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((3-fluoropyridin-2-yl)methoxy)-8-(2-methoxy-6-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-(pyridin-2-ylamino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(pyridin-2-ylamino)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(3-methylpyridin-4-yl)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile; 3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile; 3-(5-amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-(pyridin-2-ylamino)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((6-methylpyridin-2-yl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile; 3-(5-amino-2-((pyridin-2-yloxy)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-7yl)benzonitrile; 2-((5-amino-7-(3-cyanophenyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-2yl)methoxy)nicotinonitrile; 2-((5-amino-7-(3-cyanophenyl)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile; 2-((5-amino-7-(3-cyanophenyl)-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile; 2-((5-amino-7-(3-cyanophenyl)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-2yl)methoxy)nicotinonitrile; 2-((5-amino-7-(3-cyanophenyl)-8-(2,6-dimethylpyridin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin-2yl)methoxy)nicotinonitrile; 2-((5-amino-7-(3-cyanophenyl)-8-(2-methoxy-6-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-2-yl)methoxy)nicotinonitrile; 3-(5-amino-2-((1-(pyridin-2-yl)ethyl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5-c]pyrimidin7-yl)benzonitrile; 3-(5-amino-2-((2-(pyridin-2-yl)propan-2-yl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((5-(pyrimidin-2-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(pyrimidin-4-yl)-2-((5-(pyrimidin-4-yl)-1H-tetrazol-1-yl)methyl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((5-(pyridin-3-yl)-III-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((5-(pyridin-4-yl)-1H-tetrazol-1-yl)methyl)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-((6-methylpyridin-2-yl)methoxy)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-((6-methylpyridin-2-yl)methoxy)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((6-methylpyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-((6-methylpyridin-2-yl)methoxy)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-((6-methylpyridin-2-yl)methoxy)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; - 58 049302 3-(5-amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(2,6-dimethylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-((3,6-dimethylpyridin-2-yl)methoxy)-8-(2-methoxy-6-methylpyridin-4-yl)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-(((6-methylpyridin-2-yl)methyl)amino)-8-(pyrimidin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(((6-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-2-(((6-methylpyridin-2-yl)methyl)amino)[1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-2-(((6-methylpyridin-2-yl)methyl)amino)-8-(3-methylpyridin-4-yl)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; 3-(5-amino-8-(2,6-dimethylpyridin-4-yl)-2-(((6-methylpyridin-2-yl)methyl)amino)-[1,2,4]triazolo[1,5c]pyrimidin-7-yl)benzonitrile; and 3-(5-amino-8-(2-methoxy-6-methylpyridin-4-yl)-2-(((6-methylpyridin-2-yl)methyl)amino)- [1,2,4]triazolo[1,5-c]pyrimidin-7-yl)benzonitrile; or a pharmaceutically acceptable salt thereof. 2. A pharmaceutical composition comprising a compound according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier. 3. A method of treating a disease or disorder in a patient, wherein the disease or disorder is bladder cancer, lung cancer, breast cancer, ovarian cancer, colorectal cancer, pancreatic cancer, prostate cancer, or head and neck cancer, comprising administering to said patient a therapeutically effective amount of a compound according to π. 1 or a pharmaceutically acceptable salt thereof. 4. The method according to claim 3, characterized in that the disease or disorder is head and neck cancer. 5. The method according to i.4, characterized in that the head and neck cancer is head and neck squamous cell carcinoma (HNSCC). 6. The method according to claim 3, characterized in that the disease or disorder is lung cancer. 7. The method according to item 6, characterized in that the lung cancer is non-small cell lung cancer (NSCLC). 8. The method according to claim 3, characterized in that the disease or disorder is colorectal cancer. 9. The method according to claim 3, characterized in that the disease or disorder is ovarian cancer. 10. The method according to claim 3, characterized in that the disease or disorder is prostate cancer. 11. The method according to claim 10, characterized in that the prostate cancer is metastatic castration-resistant prostate cancer. 12. The method according to claim 3, characterized in that the disease or disorder is bladder cancer. 13. The method according to claim 3, characterized in that the disease or disorder is breast cancer. 14. The method according to claim 3, characterized in that the disease or disorder is pancreatic cancer.