Azaspires for the treatment of cancer-related diseases

Abstract

Described herein are compounds that are inhibitors of at least one of the A 2A and A 2B receptors, as well as compositions containing the compounds and methods for synthesizing the compounds. The compounds and compositions are useful for treating a variety of diseases, disorders, and conditions including cancer and immune-related disorders that are mediated at least in part by the adenosine A 2A receptors and / or the adenosine A 2B receptors.

Description

[0001] This application is a divisional application of Chinese invention patent application (filed on January 19, 2018, application number 201880007563.X (PCT application number: PCT / US2018 / 014352), entitled "Zyzopyrimidine for the treatment of cancer-related diseases").

[0002] Cross-references to related applications

[0003] This application, filed March 30, 2017, claims priority to U.S. Provisional Application No. 62 / 448,608, filed January 20, 2017, and U.S. Provisional Application No. 62 / 479,005, filed March 30, 2017, the contents of which are incorporated herein by reference for all purposes.

[0004] Declaration of Invention Rights for Federally Sponsored Research and Development

[0005] not applicable.

[0006] Refer to the "Sequence List," table, or computer program list appendix submitted on the CD.

[0007] not applicable. Background of the Invention

[0008] Adenosine is a purine nucleoside compound comprising a complex of adenine and a ribose molecule (furanose). Adenosine is naturally occurring in mammals and plays important roles in several biochemical processes, including energy transfer (as adenosine triphosphate and adenosine monophosphate) and signal transduction (as cyclic adenosine monophosphate). Adenosine also functions in processes related to vasodilation, including cardiac vasodilation, and acts as a neuromodulator (e.g., it is thought to be involved in promoting sleep). In addition to its involvement in these biochemical processes, adenosine is used as a therapeutic antiarrhythmic agent to treat conditions such as supraventricular tachycardia. As discussed further in this paper, tumors evade host responses by suppressing immune function and promoting tolerance, and adenosine has been shown to play an important role in mediating tumor evasion of the immune system. Adenosine has been identified through expression on various immune cell subsets and endothelial cells. 2A Rs and A 2B Adenosine signaling in Rs plays an important role in protecting tissues during inflammatory responses. Therefore, under certain conditions, adenosine protects tumors from immune destruction (see, for example, Fishman, P et al. (2009) Handbook of Experimental Pharmacology 193:399-441).

[0009] Adenosine receptors are a class of purine G protein-coupled receptors that use adenosine as an endogenous ligand. The four types of adenosine receptors in the human body are called A1, A2, A3, A4, A5, A6, A7, A8, A9, A1, A2, A1, A2, A2, A3, A4, A5, A6, A7, A8, A9, A1, A2, A2, A2, 2A A 2B And A3. Regulation of A1 has been suggested for the management and treatment of conditions such as neurological disorders, asthma, and heart and kidney failure; A 2A Antagonists have been suggested for the management and treatment of conditions such as Parkinson's disease; A 2B Regulation of A3 has been suggested for the management and treatment of chronic lung diseases, including asthma; regulation of A3 has been suggested for the management and treatment of diseases such as asthma and chronic obstructive pulmonary disease, glaucoma, cancer, and stroke.

[0010] Historically, modulators of adenosine receptors have been non-selective. This was acceptable in certain indications, such as the parenteral administration of adenosine, an endogenous agonist that acts on all four adenosine receptors in cardiac tissue, to treat severe tachycardia. However, the use of subtype-selective adenosine receptor agonists and antagonists offers the potential to achieve the desired outcome while minimizing or eliminating adverse effects.

[0011] Therefore, there is a need in the art for subtype-selective adenosine receptor agonists. This invention addresses this need and also provides related advantages. Summary of the Invention

[0012] This invention relates to the regulation of adenosine A 2A Receptor (A) 2A R) and / or adenosine A 2B Receptor (A) 2B Compounds of R and compositions comprising said compounds (such as pharmaceutical compositions). These compounds, including their methods of synthesis and compositions, are described in detail below.

[0013] This invention also relates to these compounds and compositions for the treatment and / or prevention of various diseases wholly or partially caused by adenosine A. 2A Receptor (A) 2A R) and / or adenosine A 2B Receptor (A) 2B Use in treating diseases, symptoms, and conditions mediated by R. These diseases, symptoms, and conditions are described in detail elsewhere herein. Unless otherwise stated, when using the compounds of the invention described herein, it should be understood that these compounds may be in the form of compositions (e.g., pharmaceutical compositions).

[0014] As will be described later, although the compounds of the present invention are believed to work by inhibiting adenosine A 2A Receptor (A) 2A R) and / or adenosine A 2B Receptor (A) 2BR) to achieve their activities, but practicing this invention does not require a precise understanding of the compounds' mechanisms of action. It is conceivable that the compounds may alternately achieve their activities by directly or indirectly inhibiting adenylate cyclase. It is also conceivable that the compounds may also achieve their activities by inhibiting A 2A Receptor (A) 2A R) and adenosine A 2B Receptor (A) 2B R) and adenylate cyclase are used to achieve their activities. Although the compounds of the present invention generally refer to adenosine A herein. 2A Receptor (A) 2A R) and / or adenosine A 2B Receptor (A) 2B R) inhibitors should be understood as, the term "A" 2A R / A 2B "R inhibitors" contain substances that inhibit A 2A R, A 2B Compounds that act alone or by inhibiting A or adenylate cyclase, and / or by inhibiting A. 2A R, A 2B Compounds that function with R and adenylate cyclase.

[0015] Discover A 2A and A 2B Cell surface adenosine receptors are upregulated in various tumor cells. Therefore, A 2A and / or A 2B Adenosine receptor antagonists represent a new and promising class of oncology therapeutics.

[0016] A 2A Activation of adenosine receptors suppresses the immune response against tumors by inhibiting T-regulatory cell function, suppressing the cytotoxicity of natural killer cells, and inhibiting tumor-specific CD4+ / CD8+ activity. Therefore, inhibiting this receptor subtype with specific antagonists can enhance immunotherapy in cancer treatment. 2B Activation of adenosine receptors plays a role in tumor development by upregulating the expression levels of angiogenic factors within microvascular endothelial cells. [See, for example, P. Fishman et al., Handbook of Experimental Pharmacology (2009); 193:399-441]. Furthermore, adenosine receptor 2A blockade has been shown to increase anti-PD-1 potency through enhanced anti-tumor T-cell responses (P. Beavis et al., Cancer Immunol Res. DOI: 10.1158 / 2326-6066. CIR-14-0211, published February 11, 2015). The effects of A are listed below. 2A Rs and A 2B A more comprehensive discussion of the role of Rs.

[0017] Adenosine 2A receptor (A 2A R)

[0018] A 2A R (also known as ADORA2A) is a G protein-coupled receptor (GPCR), and its family members have seven transmembrane α-helices. Based on its crystal structure, A 2A R contains a ligand-binding pocket that distinguishes it from other structurally defined GPCRs (e.g., β-2 adrenergic receptors).

[0019] As discussed elsewhere in this article, adenosine is involved in mediating tumor evasion by the immune system. 2A R plays a crucial, non-redundant role in mediating adenosine-induced anti-inflammatory responses. A 2A R negatively regulates the immune response, and therefore pharmacologically inhibits A. 2A Activation of R has proven to be a viable approach to enhance immunotherapy.

[0020] As mentioned earlier, A 2A Activation of R affects the adaptive immune response; for example, A 2A R protects the host from excessive tissue damage not only by acutely suppressing T cell function, but also by promoting the development of regulatory T cells. Because A 2A R activation is an effective inhibitor of adaptive immune response, so tumor-derived adenosine is associated with blocking anti-tumor immunity.

[0021] In addition to its other functions, A 2A R also involves selectively enhancing anti-inflammatory cytokines, promoting the upregulation of PD-1 and CTLA-4, promoting the generation of LAG-3 and Foxp3+ regulatory T cells, and mediating the suppression of regulatory T cells. PD-1, CTLA-4, and other immune checkpoints are discussed further in this article. Since all these immunosuppressive properties have been identified as mechanisms by which tumors evade host responses, A... 2A Cancer immunotherapy regimens with R antagonists can lead to enhanced tumor immunotherapy. [See overall, Naganuma, M et al. (2006) Journal of Immunology (J Immunol) 177:2765-769].

[0022] A 2A R-antagonists may play an important role in chemotherapy and radiotherapy. Mechanically, the concomitant administration of A-antagonists has been recommended during chemotherapy or radiotherapy. 2A R antagonists induce the expansion of tumor-specific T cells while simultaneously preventing the induction of tumor-specific regulatory T cells. Furthermore, given their different mechanisms of action, A... 2A The combination of R antagonists and tumor vaccines is believed to provide at least one additive effect. Finally, A 2AR-blockers are most effectively used in combination with tumor vaccines and other checkpoint inhibitors. For example, they block PD-1 involvement and inhibit A... 2A R may reduce the ability of tumors to shut down tumor-specific effector T cells (see, for example, Fishman, P et al., (2009) Handbook of Experimental Pharmacology 193:399-441). Furthermore, it has been found that A... 2A Adenosine signaling of the R receptor is a promising negative feedback loop, and preclinical studies have confirmed that blocking A... 2A R activation can significantly enhance anti-tumor immunity (Sitkovsky, MV et al., (2014) Cancer Immunology Research 2:598-605).

[0023] Adenosine 2B receptor (A 2B R)

[0024] A 2b R (also known as ADORA2B) is a GPCR found in many different cell types. It differs from other adenosine receptor subtypes (e.g., A1R, A...). 2A Compared to A3R, it requires a higher concentration of adenosine for activation (Fredholm BB et al., (2001) Biochem Pharmacol 61:443-448). This condition has been observed in tumors, for example, where hypoxia is commonly observed. Unlike other adenosine receptor subtypes, A... 2B R may play an important role in the pathophysiological conditions associated with massive adenosine release. Therefore, selective blocking or stimulation of this adenosine receptor subtype may not interfere with many important physiological functions of adenosine mediated by other adenosine receptor subtypes. However, it can lead to A 2B The pathways of R-mediated inhibition are not fully understood.

[0025] Angiogenesis represents a key mechanism of tumor growth. The angiogenesis process is highly regulated by a series of angiogenic factors and is triggered by adenosine under specific hypoxia-related conditions. 2B R is expressed in human microvascular endothelial cells, where it plays an important role in regulating the expression of angiogenic factors such as vascular endothelial growth factor (VEGF). In certain tumor types, hypoxia-induced A... 2B The upward adjustment of Rs indicates that A 2B Rs play a crucial role in mediating the effects of adenosine on angiogenesis. Therefore, A 2B Blocking Rs can limit tumor growth by restricting oxygen supply to tumor cells. Furthermore, experiments involving adenylate cyclase activation showed that A 2B Rs is the only adenosine receptor subtype in some tumor cells, which indicates that A 2BR antagonists may show efficacy against specific tumor types (see, for example, Feoktistov, I et al. (2003) Circulation Research (Circ Res) 92:485-492).

[0026] Recent data makes A 2B Understanding the exact role of R regulators has become more complex. As discussed earlier, data confirm that A 2B Rs play a crucial role in mediating the effects of adenosine on tumor growth and progression. In fact, inhibition of angiogenesis and ERK1 / 2 phosphorylation represent key mechanisms for mediating A-based tumor growth and progression. 2B R is one of the most interesting potential targets for anticancer therapy. However, although inhibiting angiogenesis requires the use of A 2B R antagonist, but by using A 2B Treatment with R agonists can inhibit growth signaling via other clinically relevant pathways, such as the MAP kinase pathway (see, for example, Graham, S et al., (2001) European Journal of Pharmacology 420:19-26). Further experimental results may suggest that both agonists and antagonists, if used at different stages of the disease and its treatment, could provide useful options in combination with other therapies.

[0027] In one particular aspect, the present invention provides compounds having formula (I):

[0028]

[0029] Or its pharmaceutically acceptable salts, hydrates or solvates, wherein,

[0030] G 1 For N or CR 3a ;

[0031] G 2 For N or CR 3b ;

[0032] G 3 For N or CR 3c ;

[0033] R 3a R 3b and R 3c Each independently can be H, deuterium, or C. 1-3 alkyl;

[0034] R 1a and R 1b Each person independently selects from the following groups:

[0035] i)H or deuterium,

[0036] ii) Optionally placed by 1-3 R 5 C replaced by substituent 1-8 alkyl,

[0037] iii) Optionally placed by 1-3 R 5 -X replaced by substituent 1 -OC 1-8 alkyl,

[0038] iv)-C(O)-R 6 ,

[0039] v) Optionally selected by 1-3 R 7 Y replaced by the substituent, and

[0040] vi) Optionally assigned to 1-3 R 7 -X replaced by substituent 1 -Y; or

[0041] vii)R 1a and R 1b Together with the nitrogen attached to them, they are optionally formed by 1-3 R 8 The 5-6 membered heterocyclic alkyl ring substituted by the substituent, wherein the heterocyclic alkyl ring has 0-2 heteroatom ring vertices selected from the group consisting of O, N and S;

[0042] Each Y is C 3-8 Cycloalkyl or 4- to 6-membered heterocyclic alkyl groups having 1-3 heteroatom ring vertices selected from the O, N, and S grouping groups;

[0043] R 2 and R 4 Each independently can be H, deuterium, or C. 1-3 alkyl;

[0044] Ar 1 It is a phenyl or a 5- to 6-membered heteroaryl group, wherein each is optionally surrounded by 1-3 R groups. 9 Replaced;

[0045] Ar 2 It is a phenyl or a 5- to 6-membered heteroaryl group, wherein each is optionally surrounded by 1-3 R groups. 10 Replaced;

[0046] Among them, Ar 1 and Ar 2 Each of the 5- to 6-membered heteroaryl groups independently possesses 1-3 selectable O, N, N + -O - The vertices of the heteroatom rings formed by S;

[0047] Each X 1 C 1-6 Alkylene;

[0048] Each R 5 Independently selected from the following group: hydroxyl, C 3-8 Cycloalkyl, phenyl, -O-phenyl, -C(O)OR a and oxygenation;

[0049] Each R 6 It is C 1-8 Alkyl or Y, wherein each is optionally substituted by 1 to 3 substituents selected from the group consisting of: hydroxyl, -O-phenyl, phenyl, and -OC. 1-8 alkyl;

[0050] Each R 7 Select independently from the following group: C 1-8 Alkyl, hydroxyl, -OC 1-8 Alkyl, oxo, and C(O)OR a ;

[0051] Each R 8 Select independently from the following group: C 1-8 Alkyl, hydroxyl, and oxo;

[0052] Each R 9 Select independently from the following group: C 1-8 Alkyl, C 1-8 Deuterated alkyl, -OC 1-8 Alkyl, -OC 1-8 Deuterated alkyl groups, -X 1 -OC 1-8 Alkyl, -OX 1 -OC 1-8 Alkyl, -X 1 -OX 1 -OC 1-8 Alkyl, -C(O)OR a Halogen, cyano, -NR b R c Y, -X 1 -C 3-8 cycloalkyl and -X 2 -Z, where X 2 Selected from the following group: C 1-6 Alkylene, -C 1-6 Alkylenes -O-, -C(O)-, and –S(O)2-, Z being 4- to 6-membered heterocyclic alkyl groups having 1-3 heteroatom ring vertices selected from the group consisting of O, N, and S, and wherein each of the R 9 The substituents are optionally replaced by 1-3 Rs 11 Replaced;

[0053] Each R 10 Select independently from the following group: C1-8 Alkyl, C 1-8 Deuterated alkyl, halogen, cyano, -OC 1-8 Alkyl, -OC 1-8 Deuterated alkyl groups, -X 1 -OC 1-8 Alkyl, -OX 1 -OC 1-8 Alkyl, -S(O)2-C 1-6 Alkyl, -C(O)NR d R e and 4-6 membered heteroaryl groups having 1-3 heteroatom ring vertices selected from the group consisting of O, N and S, wherein each of the R 10 Substituents are independently replaced by 1-3 R 12 Replaced, or in Ar 2 Two R's at adjacent ring vertices 10 Optionally combined to form a 5-membered heterocycle optionally replaced by 1-2 halogens;

[0054] Each R 11 Independently selected from the following group: hydroxyl, halogen, cyano, -NR d R e -C(O)OR a , phenyl, C 3-8 cycloalkyl, and optionally C(O)OR a The C that was replaced 1-4 alkyl;

[0055] Each R 12 Independently selected from the group consisting of: halogen, cyano, hydroxyl, -C(O)OR; and

[0056] Each R a H, deuterium, or C 1-6 alkyl;

[0057] Each R b and R c Independently selected from the following groups: H, deuterium, C 1-8 Alkyl, -S(O)2-C 1-6 Alkyl, -C(O)OR a and -X 1 -C(O)OR a ;

[0058] Each R d and R e Independently selected from the following groups: H, deuterium, C 1-8 Alkyl, -S(O)2-C 1-6 Alkyl groups; and

[0059] The condition is when G 1 and G2 Each is N, G 3 For CH, R 2 CH3, and R 1a and R 1b When each is H or deuterium, then Ar 2 It is not 2-thienyl, phenyl, 2-,3- or 4-methoxyphenyl, 3- or 4-halophenyl, 2,4-dimethoxyphenyl, 2,4-dichlorophenyl or 2- or 4-methylphenyl.

[0060] In some embodiments, the present invention contemplates a compound having the structure shown in the following formula:

[0061]

[0062] Or its pharmaceutically acceptable salt, hydrate, or solvation. As described below, compound I is an effective A 2A R and A 2B R antagonists, whose potency against both receptors is less than 10 nM.

[0063] In some embodiments, the present invention contemplates a method for treating or preventing cancer in a subject (e.g., a person), comprising administering to said subject a therapeutically effective amount of at least one of the A described herein. 2A R / A 2B R inhibitors. In some embodiments, the invention includes methods to effectively reverse or prevent A. 2A The progression of R-mediated immunosuppression involves administering at least one compound described herein to a subject for the treatment or prevention of cancer in the subject. In some embodiments, A 2A R-mediated immunosuppression is mediated by antigen-presenting cells (APCs).

[0064] Examples of cancers that can be treated with the compounds and compositions described herein include, but are not limited to: prostate cancer, colorectal cancer, pancreatic cancer, cervical cancer, gastric cancer, endometrial cancer, brain cancer, liver cancer, bladder cancer, ovarian cancer, testicular cancer, head cancer, neck cancer, skin cancer (including melanoma and basal carcinoma), mesothelial lining cancer, leukocyte cancer (including lymphoma and leukemia), esophageal cancer, breast cancer, muscle cancer, connective tissue cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), adrenal cancer, thyroid cancer, kidney cancer, or bone cancer; glioblastoma, mesothelioma, renal cell carcinoma, gastric cancer, sarcoma, choriocarcinoma, basal cell carcinoma of the skin, and seminoma of the testis. In some embodiments of the invention, the cancer is melanoma, colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, leukemia, brain tumor, lymphoma, sarcoma, ovarian cancer, head and neck cancer, cervical cancer, or Kaposi's sarcoma. Candidate cancers to be treated with the compounds and compositions of the present invention are discussed further below.

[0065] This invention relates to a method for treating subjects who have received bone marrow transplantation or peripheral blood stem cell transplantation, which involves administering a therapeutically effective dose of A sufficient to increase delayed-type hypersensitivity to tumor antigens, delay the recurrence time of post-transplant malignancies, increase recurrence-free survival time after transplantation, and / or increase long-term survival after transplantation. 2A R / A 2B R inhibitors.

[0066] In some embodiments, the invention contemplates a method for treating or preventing an infectious condition (e.g., viral infection) in a subject (e.g., a person), comprising administering to the subject a therapeutically effective amount of at least one A 2A R / A 2B R inhibitors (e.g., novel inhibitors of the present invention). In some embodiments, the infectious condition is a viral infection (e.g., a chronic viral infection), a bacterial infection, a fungal infection, or a parasitic infection. In some embodiments, the viral infection is human immunodeficiency virus or cytomegalovirus.

[0067] In other embodiments, the invention contemplates methods for treating or preventing immune-related diseases, symptoms, or conditions in a subject (e.g., a person), comprising administering to the subject a therapeutically effective amount of at least one of the A described herein. 2A R / A 2B R inhibitors. Examples of immune-related diseases, symptoms, and conditions are described below.

[0068] Other adjustments can be made via A. 2A R / A 2B The use of R activity to treat or prevent diseases, symptoms, and conditions is the A of this invention. 2A R / A 2BCandidate indications for R inhibitor compounds.

[0069] The present invention further considers the A described herein. 2A R / A 2B The use of R inhibitors in combination with one or more other agents. These one or more other agents may contain some adenosine A. 2A Receptor and / or adenosine A 2B Receptor-modulating activity; or they may act through distinct mechanisms of action. In some embodiments, such agents include radiation (e.g., local or total radiotherapy) and / or other non-pharmacological modalities of treatment. When using combination therapy, the compounds described herein and one or more additional agents may be in the form of a single composition or multiple compositions, and the modalities of treatment may be administered simultaneously, sequentially, or through some other regimen. For example, the present invention contemplates a treatment regimen in which a radiation phase is followed by a chemotherapy phase. Combination therapies may have additive or synergistic effects. Other benefits of combination therapy are described below.

[0070] In specific embodiments, the present invention considers the A described herein. 2A R / A 2B Use of R inhibitors in combination with one or more immune checkpoint inhibitors. Blockade of immune checkpoints that amplify antigen-specific T-cell responses has proven to be a promising approach in the treatment of human cancers. Examples of immune checkpoints (ligands and receptors), some of which are selectively upregulated in various types of tumor cells, are candidates for blockade, including PD1 (programmed cell death protein 1); PDL1 (PD1 ligand); BTLA (B and T lymphocyte attenuator); CTLA4 (cytotoxic T lymphocyte-associated antigen 4); TIM3 (T cell membrane protein 3); LAG3 (lymphocyte activation gene 3); TIGIT (T cell immune receptor with Ig and ITIM domains); and cytotoxic inhibitory receptors. Immune checkpoint inhibitors and their combination therapies are discussed in detail elsewhere in this article.

[0071] In other embodiments, the present invention provides a method for treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of at least one A 2A R / A 2BR inhibitors and at least one chemotherapeutic agent, including but not limited to, alkylating agents (e.g., nitrogen mustard, such as chlorambucil, cyclophosphamide, isofamide, nitrogen mustard, melphalan, and uracil mustard; azacyclopropane, such as thiotepa; methanesulfonates, such as busulfan; nucleoside analogs (e.g., gemcitabine); nitrosoureas, such as carmustine, lomustine, and streptozotocin; topoisomerase 1 inhibitors (e.g., irinotecan); platinum complexes, such as cisplatin, carboplatin, and oxaliplatin; bioreductive alkylating agents, such as mitomycin, procarbazine, dacarbazine, and altretamine); anthracycline agents (e.g., doxorubicin). Bicin, daunorubicin, epirubicin, and idarubicin; DNA strand breaking agents (such as bleomycin); topoisomerase II inhibitors (such as acridine, daunorubicin, idarubicin, mitoxantrone, doxorubicin, etoposide, and teniposide); DNA minor groove binding agents (such as plicamydin); antimetabolites (e.g., folic acid antagonists such as methotrexate and trimethoprim; pyrimidine antagonists such as fluorouracil, fluorodeoxyuridine, CB3717, azacitidine, etc.). Cytidine and fluorouridine; purine antagonists, such as mercaptopurine, 6-thioguanine, fludarabine, pentostatin; asparginase; and ribonucleotide reductase inhibitors, such as hydroxyurea; microtubule interactors (e.g., vincristine, estramustine, vinblastine, docetaxel, epormycin derivatives, and paclitaxel); hormones (e.g., estrogens; conjugated estrogens; ethinylestradiol; diethylstilbestrol). ol); chlortrianisen; idenestrol; progestins, such as hydroxyprogesterone caproate, medroxyprogesterone acetate, and megestrol acetate; and androgens, such as testosterone, testosterone propionate, fluoromethesterone, and methyltestosterone; corticosteroids (e.g., prednisone, dexamethasone, methylprednisolone, and prednisolone); luteinizing hormone-releasing hormone (LH-REM) or gonadotropin-releasing hormone (GnRH) antagonists (e.g., leuprorelin acetate and goserelin acetate); and anti-hormone antigens (e.g., tamoxifen, anti-androgen agents such as flutamide; and anti-adrenergic agents such as mitotane and aminoglutethimide). The present invention also contemplates A 2A R / A 2B Use of R inhibitors in combination with other agents known in the art (e.g., arsenic trioxide) and other chemotherapeutic agents that may be developed in the future.

[0072] In some embodiments of methods relating to cancer treatment, a therapeutically effective amount of the A described herein is administered in combination with at least one chemotherapy agent. 2A R / A 2B R inhibitors result in cancer survival rates greater than those observed by administration of either agent alone. In other embodiments of methods relating to cancer treatment, the therapeutically effective amount of A described herein is administered in combination with at least one chemotherapeutic agent. 2A R / A 2B R inhibitors resulted in a greater reduction in tumor size or a slowdown in tumor growth than the reduction in tumor size or growth observed when either agent was administered alone.

[0073] In other embodiments, the invention contemplates a method for treating or preventing cancer in a subject, comprising administering to the subject a therapeutically effective amount of at least one of the A methods described herein. 2A R / A 2B R inhibitors and at least one signal transduction inhibitor (STI). In a specific embodiment, the at least one STI is selected from the group consisting of bcr / abl kinase inhibitors, epidermal growth factor (EGF) receptor inhibitors, HER-2 / neu receptor inhibitors, and farnesyltransferase inhibitors (FTIs). Other candidate STIs are listed elsewhere herein.

[0074] The present invention also considers a method for enhancing tumor cell rejection in subject subjects, which includes A 2A R / A 2B R inhibitors are administered in combination with at least one chemotherapy agent and / or radiotherapy, where the resulting tumor cell rejection is greater than that achieved by administering A alone. 2A R / A 2B Tumor rejection resulting from R inhibitors, chemotherapy agents, or radiation therapy.

[0075] In other embodiments, the present invention provides a method for treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of at least one A 2A R / A 2B R inhibitors and A-removal drugs 2A R / A 2BAt least one immunomodulatory agent other than R inhibitors. In a specific embodiment, the at least one immunomodulatory agent is selected from the group consisting of: CD4OL, B7, B7RP1, anti-CD40, anti-CD38, anti-ICOS, 4-IBB ligand, dendritic cell cancer vaccines, IL2, IL12, ELC / CCL19, SLC / CCL21, MCP-1, IL-4, IL-18, TNF, IL-15, MDC, IFN-α / -13, M-CSF, IL-3, GM-CSF, IL-13, anti-IL-10, and indoleamine 2,3-dioxygenase 1 (IDO1) inhibitors. Other candidate immunomodulatory agents are listed elsewhere in this document.

[0076] The present invention contemplates embodiments including methods for treating or preventing infectious conditions (e.g., viral infections) in a subject (e.g., a person), comprising administering a therapeutically effective amount of at least one of the A described herein. 2A R / A 2B R inhibitors and therapeutically effective doses of anti-infective agents.

[0077] In some embodiments of the invention, the additional therapeutic agents are cytokines, including, for example, granulocyte-macrophage colony-stimulating factor (GM-CSF) or flt3 ligand. The invention also contemplates methods for treating or preventing viral infections (such as chronic viral infections), such as, but not limited to, hepatitis C virus (HCV), human papillomavirus (HPV), cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella-zoster virus, Coxsackie virus, and human immunodeficiency virus (HIV). The use of the compounds described herein for treating (alone or as part of a combination therapy) infections is further discussed below.

[0078] In another embodiment, the vaccine is administered with a therapeutically effective amount of the present invention A. 2A R / A 2B Co-administration of a combination of R inhibitors achieves treatment of infectious diseases. In some embodiments, the vaccine is an antiviral vaccine, including, for example, an anti-HIV vaccine. In other embodiments, the vaccine is effective against tuberculosis or malaria. In other embodiments, the vaccine is an oncology vaccine (e.g., a vaccine effective against melanoma); the oncology vaccine may comprise genetically modified tumor cells or genetically modified cell lines, including genetically modified tumor cells or genetically modified cell lines that have been transfected to express granulocyte-macrophage stimulating factor (GM-CSF). In a specific embodiment, the vaccine comprises one or more immunogenic peptides and / or dendritic cells.

[0079] In some embodiments, the present invention contemplates methods of using the compounds described herein in combination with one or more antimicrobial agents.

[0080] Involving the application of A 2A R / A 2B In some embodiments of treating infection with an R inhibitor and at least one other therapeutic agent, A is administered... 2A R / A 2B The symptoms of infection observed after administration of R inhibitors and additional therapeutic agents were improved compared to the same symptoms observed after administration alone. In some embodiments, the observed symptoms of infection may be a reduction in viral load and CD4 count. + Increased T-cell count, reduced opportunistic infections, increased survival time, eradication of chronic infections, or a combination thereof. Brief description of the attached figures

[0081] not applicable. Invention Details

[0082] Before further describing the invention, it should be understood that the invention is not limited to the specific embodiments described herein, and that the terminology used herein is for the purpose of describing specific embodiments only and is not restrictive.

[0083] When providing ranges of values, it should be understood that, unless the context explicitly specifies otherwise, intermediate values ​​(to one-tenth of the lower limit unit) between the upper and lower limits of the range, as well as any other specified or intermediate values ​​within the stated range, are included in this invention. The upper and lower limits of these smaller ranges may be independently included within the smaller ranges and also within this invention, subject to any specific exclusions within the range. Where the range includes one or two limits, the range excluding any one or two of those included limits is also included in this invention. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0084] As used herein, unless the context clearly specifies otherwise, the singular forms “a,” “an,” and “the” include the plural forms. It should also be noted that claims may be drafted to exclude any optional elements. Therefore, this statement is intended as an exclusionary term used in connection with the detailed description of the claim elements, such as “solely,” “only,” etc., or as a precondition for a “negative” limitation.

[0085] The publications discussed herein are only those that were published prior to the filing date of this application. Furthermore, the publication dates provided may differ from the actual publication dates and may require separate verification.

[0086] General

[0087] The example provided in this article is for inhibiting adenosine A. 2AReceptor (A) 2A R) and / or adenosine A 2B Receptor (A) 2B Compounds and compositions thereof (R), and pharmaceutical compositions comprising them. This document also provides, for example, methods for treating or preventing diseases using adenosine A. 2A Receptor (A) 2A R) and / or adenosine A 2B Receptor (A) 2B Methods for suppressing diseases, conditions or illnesses or their symptoms mediated by R.

[0088] definition

[0089] Unless otherwise stated, the following terms are intended to have the meanings described below. Other terms are defined elsewhere throughout the specification.

[0090] Unless otherwise stated, the term "alkyl", either on its own or as part of another substituent, refers to an alkyl group having a specified number of carbon atoms (i.e., C40, C50, C60, C70, C6 ... 1-8 Alkyl groups refer to straight-chain or branched hydrocarbon groups (1 to 8 carbons). Alkyl groups can include any number of carbons, for example, C10, C20, C30, C40, C50, C60, C70, C80, C9 ... 1-2 C 1-3 C 1-4 C 1-5 C 1-6 C 1-7 C 1-8 C 1-9 C 1-10 C 2-3 C 2-4 C 2-5 C 2-6 C 3-4 C 3-5 C 3-6 C 4-5 C 4-6 and C 5-6 Examples of alkyl groups include: methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc.

[0091] The term "alkylene" refers to a straight-chain or branched saturated aliphatic group having a specified number of carbon atoms and being attached to at least two other groups, i.e., divalent hydrocarbon groups. The two parts attached to the alkylene group can be connected to the same atoms or different atoms of the alkylene group. For example, a straight-chain alkylene group can be a divalent group -(CH2). n - where n is 1, 2, 3, 4, 5, or 6. Representative alkylene compounds include, but are not limited to: methylene, ethylene, propylene, isopropylene, butylene, isobutylene, secondary-butylene, pentylene, and hexylene. Alkylenes (generally referred to as X in this invention) 1 or X 2The group can be substituted or unsubstituted. When X is included... 1 or X 2 When the group is optionally substituted, it should be understood that the optional substituent may be at the alkylene site of that moiety.

[0092] The term "cycloalkyl" refers to a hydrocarbon ring having a specified number of ring atoms (e.g., C3-6 cycloalkyl) and being fully saturated or having no more than one double bond between the ring vertices. "Cycloalkyl" can also mean bicyclic and polycyclic hydrocarbons, such as bicyclic [2.2.1]heptane, bicyclic [2.2.2]octane, etc. In some embodiments, the cycloalkyl group of the present invention is a monocyclic C3-6 cycloalkyl group. 3-6 Cycloalkyl moiety.

[0093] The term "heterocycloalkyl" refers to a cycloalkyl ring having a specified number of ring vertices (or members) and having one to five heteroatoms selected from N, O, and S replacing one to five carbon vertices, wherein nitrogen and sulfur atoms are optionally oxidized and nitrogen atoms are optionally quaternized. The cycloheteroalkyl can be a monocyclic, bicyclic, or polycyclic system. Non-limiting examples of cycloheteroalkyl groups include: pyrrolidine, imidazoline, pyrazolidine, butyrolactam, valproic acid, imidazolinone, hydantoin, dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrrolidine, thiaran, pyranone, tetrahydrofuran, tetrahydrothiophene, quinine ring, and similar groups. Cycloheteroalkyl groups can be linked to other parts of the molecule via cyclic carbons or heteroatoms.

[0094] As used herein, wavy lines intersecting with single, double, or triple bonds in any chemical structure described herein. This indicates the point of connection between a single, double, or triple bond and the rest of the molecule. Furthermore, a bond extending to the center of the ring (e.g., a benzene ring) signifies a connection at any available ring vertices. Those skilled in the art will understand that multiple substituents shown attached to the ring will occupy ring vertices, providing a stable compound and additionally being spatially compatible. For divalent components, this designation means including either direction (forward or reverse). For example, the group “–C(O)NH-” signifies a bond including either direction: -C(O)NH- or –NHC(O)-, and similarly, “-O-CH2CH2-” is intended to include both -O-CH2CH2- and -CH2CH2-O-.

[0095] Unless otherwise stated, the terms "halogenated" or "halogen," either on their own or as part of another substituent, refer to a fluorine, chlorine, bromine, or iodine atom. Additionally, terms such as "halogenated alkyl" are intended to include both monohalogenated and polyhalogenated alkyl groups. For example, the term "C"...1-4 "Halogenated alkyl" is intended to include: trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and similar groups.

[0096] Unless otherwise stated, the term "aryl" refers to a polyunsaturated, typically aromatic, hydrocarbon group, which can be monocyclic or fused together or covalently linked polycyclic (up to three rings). Non-limiting examples of aryl groups include phenyl, naphthyl, and biphenyl.

[0097] The term "heteroaryl" refers to an aryl group (or ring) containing one to five heteroatoms selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom is optionally quaternized. Heteroaryl groups can be linked to other parts of the molecule via heteroatoms. Non-limiting examples of heteroaryl groups include: pyridinyl, pyrazinyl, pyrazinyl, pyrimidinyl, triazinyl, quinolinyl, quinoxolinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl, purineyl, benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzoisoxazoleyl, isobenzofuranyl, isoindolyl, indolyl, benzotriazinyl, thienopyridinyl, thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridinyl, benzothiaxolyl, benzofuranyl, benzothienopyridinyl, indolyl, quinolinyl, isoquinolinyl, isothiazolyl, pyrazolyl, inzonoyl, pteridinyl, imidazoleyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrroleyl, thiazolyl, furanyl, and similar groups. The substituents of the heteroaryl ring can be selected from the acceptable substituents described below.

[0098] In some embodiments, the terms above (e.g., "alkyl", "aryl", and "heteroaryl") may be optionally substituted. Substituents for various types of groups are provided below.

[0099] The optional substituents of alkyl groups (including those commonly referred to as alkylene, alkenyl, ynyl, and cycloalkyl) can be zero to (2m'+1) groups selected from the following: halogen, -OR', -NR'R”, -SR', -SiR'R”R”', -OC(O)R', -C(O)R', -CO2R', -CONR'R”, -OC(O)NR'R”, -NR”C(O)R', -NR'-C(O)NR”R”', - NR”C(O)2R', -NH-C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)=NR', -S(O)R', -S(O)2R', -S(O)2NR'R”, -NR'S(O)2R”, -CN (cyano), -NO2, aryl, aryloxy, oxo, cycloalkyl, and heterocyclic alkyl, where m' is the total number of carbon atoms in the group. R', R”, and R”’ each independently refer to hydrogen, unsubstituted C 1-8 Alkyl, unsubstituted aryl, aryl substituted with 1-3 halogens, C 1-8 Alkoxy or C 1-8 thioalkoxy groups, or unsubstituted aryl-C 1-4 Alkyl groups. When R' and R” are attached to the same nitrogen atom, they can combine with the nitrogen atom to form 3, 4, 5, 6, or 7-membered rings. For example, -NR'R” means including 1-pyrrolidinyl and 4-morpholinyl.

[0100] The optional substituents of cycloalkyl and heterocycloalkyl groups can be selected from the following groups: optionally substituted with C(O)OR', halogen, -OR', -NR'R”, -SR', -SiR'R”R”', -OC(O)R', -C(O)R', -CO2R', -CONR'R”, -OC(O)NR'R”, -NR”C(O)R', -NR'-C(O)NR”R”', -NR”C(O)2R', -NH-C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)=NR', -S(O)R', -S(O)2R', -S(O)2NR'R”, -NR'S(O)2R”, -CN (cyano), -NO2, aryl, aryloxy, and oxo alkyl groups. R', R”, and R”' each independently refer to hydrogen, unsubstituted C 1-8 Alkyl, unsubstituted aryl, aryl substituted with 1-3 halogens, C 1-8 Alkoxy or C 1-8 Thioalkoxy, or unsubstituted aryl-C 1-4 alkyl.

[0101] Similarly, the substituents that can be selected for aryl and heteroaryl groups are varied and are generally selected from: -halogen, -OR', -OC(O)R', -NR'R”, -SR', -R', -CN, -NO2, -CO2R', -CONR'R”, -C(O)R', -OC(O)NR'R”, -NR”C(O)R', -NR”C(O)2R', -NR'-C(O)NR”R”', -NH-C( NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)=NR', -S(O)R', -S(O)2R', -S(O)2NR'R”, -NR'S(O)2R”, -N3, perfluoro(C1-C4)alkoxy, and perfluoro(C1-C4)alkyl, in the range from zero to the total number of open valences on the aromatic ring system; and R', R” and R”' are independently selected from: hydrogen, C 1-8 Alkyl, C 1-8 Haloalkyl, C 3-6 cycloalkyl, C 2-8 Alkenyl and ynyl groups. Other suitable substituents include the aforementioned aryl substituents connected to the ring atoms via an alkylene group of 1-6 carbon atoms.

[0102] Two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be of the formula -TC(O)-(CH2). q The substituent is replaced by a -U- substituent, where T and U are independently -NH-, -O-, -CH2-, or a single bond, and q is an integer from 0 to 2. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced by a -A-(CR)-substituent. f R g ) r The -B- substituent replaces A and B, where A and B are independently -CH2-, -O-, -NH-, -S-, -S(O)-, -S(O)2-, -S(O)2NR'- or a single bond, r is an integer from 1 to 3, and R f and R g Each can be independently H or a halogen. One of the single bonds in the newly formed ring can optionally be replaced by a double bond. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring can optionally be replaced by the formula -(CH2). s -X-(CH2) t The substituent is replaced by a substituent, where s and t are independently integers from 0 to 3, and X is -O-, -NR'-, -S-, -S(O)-, -S(O)2-, or -S(O)2NR'-. The substituent R' in NR'- and -S(O)2NR'- is selected from hydrogen or unsubstituted C. 1-6 alkyl.

[0103] As used herein, the term “heteroatoms” is intended to include oxygen (O), nitrogen (N), sulfur (S), and silicon (Si).

[0104] The term "pharmaceutically acceptable salt" refers to salts of active compounds prepared with relatively non-toxic acids or bases, depending on the specific substituents present on the compounds described herein. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of these compounds with a sufficient amount of the desired base in a solvent-free or suitable inert solvent. Examples of salts derived from pharmaceutically acceptable inorganic bases include aluminum, ammonium, calcium, copper, iron, ferrous, lithium, magnesium, trivalent manganese, divalent manganese, potassium, sodium, zinc, and the like. Salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary, and tertiary amines, including substituted amines, cyclic amines, and naturally occurring amines, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and similar groups. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of these compounds with a sufficient amount of the desired acid, which may be solvent-free or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrocarbonic acid, phosphoric acid, monohydrophosphoric acid, dihydrophosphoric acid, sulfuric acid, monohydrosulfuric acid, hydroiodic acid, or phosphorous acid, as well as salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. Salts of amino acids, such as arginine, and salts of organic acids, such as glucuronic acid or galacturonic acid, are also included (see, for example, Berge, SM, et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present invention contain both basic and acidic functional groups, such that the compounds can be converted into basic or acid addition salts.

[0105] The neutral form of the compound can be regenerated by contacting the salt with a base or acid and separating the parent compound in a conventional manner. The parent form of the compound differs from its various salt forms in certain physical properties, such as solubility in polar solvents, but in other respects the salt is equivalent to the parent form of the compound for the purposes of this invention. In addition to the salt form, this invention provides compounds existing in prodrug form. The prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of this invention. Furthermore, prodrugs can be converted into the compounds of this invention in an in vitro environment by chemical or biochemical methods. For example, when a prodrug is placed in a transdermal patch reservoir containing suitable enzymes or chemical reagents, the prodrug is slowly converted into the compounds of this invention. Prodrugs are described in more detail elsewhere herein.

[0106] In addition to the salt form, the present invention provides compounds existing in the form of prodrugs. The prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present invention. Furthermore, prodrugs can be converted into the compounds of the present invention in an in vitro environment by chemical or biochemical methods. For example, when a prodrug is placed in a transdermal patch reservoir containing suitable enzymes or chemical reagents, the prodrug is slowly converted into the compounds of the present invention.

[0107] Some compounds of the present invention may exist in both solvated and hydrated forms, including hydrated forms. Generally, the hydrated form is equivalent to the hydrated form and is intended to be included within the scope of the present invention. Some compounds of the present invention may exist in polycrystalline or amorphous forms. Generally, all physical forms are equivalent to the uses contemplated by the present invention and are intended to be within the scope of the present invention.

[0108] Certain compounds of the present invention have asymmetric carbon atoms (optical centers) or double bonds; racemates, diastereomers, geometric isomers, regio isomers, and single isomers (e.g., individual enantiomers) are all intended to be included within the scope of the present invention. When a stereochemical description is shown, it means a compound in which one isomer is present and which is substantially free of the other isomer; "substantially free of" the other isomer means a ratio of at least 80 / 20 between the two isomers, more preferably 90 / 10 or 95 / 5 or more. In some embodiments, the content of one isomer is at least 99%.

[0109] The compounds of the present invention may also contain, on one or more atoms constituting these compounds, atomic isotopes in non-natural proportions. Non-natural proportions of isotopes can be defined as amounts ranging from those found in nature to those consisting of 100% of the atoms in question. For example, the compounds may be doped with radioactive isotopes, such as tritium (…). 3 H), Iodine-125 125 I) or carbon-14 (14 C) or non-radioactive isotopes, such as deuterium ( 2 H) or carbon-13 ( 13 C). Such isotopic variations can provide additional utility for those described elsewhere in this application. For example, isotopic variants of the compounds of the present invention may be found to have other uses, including but not limited to, as diagnostic and / or imaging agents, or as therapeutic agents for cytotoxicity / radiotoxicity. Additionally, isotopic variants of the compounds of the present invention may have altered pharmacokinetic and pharmacodynamic characteristics, which may contribute to improved safety, tolerability, or efficacy during treatment. All isotopic variants of the compounds of the present invention, whether radioactive or not, are intended to be included within the scope of this invention.

[0110] The terms “patient” and “object” can be used interchangeably or to refer to human or non-human animals (e.g., mammals).

[0111] When applied to, for example, an object, cell, tissue, organ, or biological fluid, the terms "application," "administration," etc., refer to, for example, administering A... 2A R / A 2B R inhibitors, pharmaceutical compositions or diagnostic reagents containing them, are brought into contact with a subject, cell, tissue, organ, or biological fluid. In the case of cells, administration includes contacting the reagent with cells (e.g., in vitro or ex vivo) and contacting the reagent with a fluid, wherein the fluid contacts the cells.

[0112] The terms "treatment," "therapeutic," and "treatment method" refer to a course of action initiated after a disease, condition, or illness or its symptoms have been diagnosed or observed (e.g., administering A). 2A R / A 2B R inhibitors (or pharmaceutical compositions containing them) are used to temporarily or permanently eliminate, reduce, suppress, alleviate, or improve at least one underlying cause of a disease, symptom, or condition affecting a subject, or at least one symptom associated with the subject's disease, symptom, or condition. Therefore, treatment includes inhibiting (e.g., preventing the development or further development of a disease, symptom, or condition or associated clinical symptoms) the active disease.

[0113] As used in this article, “requiring treatment” refers to a judgment made by a physician or other caregiver that an individual needs or will benefit from treatment, based on various factors within the physician’s or caregiver’s professional field.

[0114] The terms “prevention,” “preventive,” and “preventive method” refer to a course of action (such as administering A) initiated in a certain way (e.g., before the onset of a disease, condition, or illness, or its symptoms). 2A R / A 2BR inhibitors (or pharmaceutical compositions containing them) are used to temporarily or permanently prevent, suppress, inhibit, or reduce the risk of a subject developing a disease, condition, or illness (as determined by the absence of clinical symptoms) or delay its onset, typically when the subject is susceptible to a particular disease, condition, or illness. In some cases, the term also refers to slowing the progression of a disease, condition, or illness or inhibiting its development into a harmful or other undesirable condition.

[0115] As used in this article, “in need of prevention” refers to a judgment made by a physician or other nursing staff that an individual needs or will benefit from preventive care, based on a variety of factors within the physician’s or nurse’s professional field.

[0116] The phrase "therapeutic effective dose" refers to an amount of medicine, administered alone or as part of a pharmaceutical composition and in a single dose or as part of a series of doses, that, when administered to the subject, has any detectable positive effect on any symptom, aspect, or characteristic of a disease, condition, or illness. Therapeutic effective doses can be determined by measuring the relevant physiological effects and can be adjusted based on the dosing regimen and diagnostic analysis of the subject's condition. For example, measuring A at a specific time after administration... 2A R / A 2B Serum levels of R inhibitors (or, for example, their metabolites) can indicate whether a therapeutically effective dose has been used.

[0117] The phrase "a quantity sufficient to achieve change" refers to a detectable difference between the level of an indicator measured before (e.g., baseline level) and after the administration of a particular therapy. Indicators include any objective parameter (e.g., serum concentration) or subjective parameter (e.g., the subject's well-being).

[0118] The term "small molecule" refers to chemical compounds with a molecular weight of less than about 10 kDa, less than about 2 kDa, or less than about 1 kDa. Small molecules include, but are not limited to, inorganic molecules, organic molecules, organic molecules containing inorganic components, molecules containing radioactive atoms, and synthetic molecules. In therapeutic applications, small molecules may be more easily absorbed by cells, less susceptible to degradation, and larger molecules are less likely to trigger an immune response.

[0119] The term "ligand" refers to, for example, a peptide, polypeptide, membrane-associated or membrane-bound molecule, or a complex thereof, that can act as a receptor agonist or antagonist. Ligands include natural and synthetic ligands, such as cytokines, cytokine variants, analogs, mutant proteins, and binding components derived from antibodies, as well as small molecules. The term also includes agents that are neither agonists nor antagonists but can bind to receptors without significantly affecting their biological properties, such as signal transduction or adhesion. Furthermore, the term includes membrane-bound ligands that have been modified, for example, by chemical or recombinant methods, into a soluble form of the membrane-bound ligand. A ligand or receptor may be entirely intracellular; that is, it may be located in the cytoplasm, the nucleus, or some other intracellular compartment. A complex of a ligand and a receptor is called a "ligand-receptor complex."

[0120] The terms “inhibitor” and “antagonist,” or “activator” and “agonist,” refer to molecules that inhibit or activate, for example, ligands, receptors, cofactors, genes, cells, tissues, or organs. An inhibitor is a molecule that reduces, blocks, prevents, delays, inactivates, desensitizes, or downregulates, for example, genes, proteins, ligands, receptors, or cells. An activator is a molecule that increases, activates, promotes, enhances activation, sensitizes, or upregulates, for example, genes, proteins, ligands, receptors, or cells. An inhibitor can also be defined as a molecule that reduces, blocks, or inactivates constitutive activity. An “agonist” is a molecule that interacts with a target to cause or promote an increase in target activation. An “antagonist” is a molecule that acts oppositely to an agonist. Antagonists prevent, reduce, inhibit, or neutralize the activity of agonists, and antagonists can also prevent, inhibit, or reduce the constitutive activity of a target, such as a target receptor, even in the absence of a definite agonist.

[0121] The terms "regulation" and "regulatory method" refer to the direct or indirect increase or decrease of A by molecules (such as activators or inhibitors). 2A R / A 2B The ability of R to function or activate. Modulators can act alone or with the help of cofactors, such as proteins, metal ions, or small molecules. Examples of modulators include small molecule compounds and other bioorganic molecules. Many libraries of small molecule compounds (e.g., combinatorial libraries) are commercially available and can serve as a starting point for identifying modulators. Those skilled in the art can develop one or more analytical methods (e.g., biochemical or cell-based analyses) in which these compound libraries can be screened to identify one or more compounds with the desired properties; thereafter, a skilled medicinal chemist can optimize such one or more compounds by, for example, synthesizing and evaluating their analogues and derivatives. Synthetic and / or molecular modeling studies can also be used to identify activators.

[0122] The "activity" of a molecule can describe or refer to the binding of a molecule to a ligand or receptor; catalytic activity; the ability to stimulate gene expression or cell signaling, differentiation, or maturation; antigenic activity; and the regulation of the activity of other molecules, etc. The term "proliferative activity" includes activities that promote normal cell division, as well as those required for or specifically associated with cancer, tumors, developmental abnormalities, cell transformation, metastasis, and angiogenesis.

[0123] As used herein, terms such as “comparable,” “comparable activity,” “comparable activity to,” “comparable effect,” and “comparable effect to” are relative terms that can be observed quantitatively and / or qualitatively. The meaning of these terms generally depends on their context. For example, from a qualitative point of view, two drugs that both activate receptors may be considered to have comparable effects, but from a quantitative point of view, if the activity of one drug, as determined by analyses accepted in the art (e.g., dose-response analysis) or in animal models accepted in the art, is only 20% that of the activity of the other drug, the two drugs may be considered to lack comparable effects. When comparing one result to another (e.g., comparing one result to a reference standard), “comparable” often (though not always) means that a result deviates from the reference standard by less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 7%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%. In a particular embodiment, a result is considered comparable to a reference standard if it deviates from the reference standard by less than 15%, less than 10%, or less than 5%. As an example rather than a limitation, activity or effect can refer to potency, stability, solubility, or immunogenicity.

[0124] "Substantially pure" means that the component constitutes more than about 50% of the total composition, and typically more than about 60% of the total polypeptide content. More typically, "substantially pure" means that at least 75%, at least 85%, at least 90% or more of all components in the composition are the components of interest. In some cases, the polypeptide will constitute more than about 90% or more than about 95% of the total composition.

[0125] When referring to ligand / receptor, antibody / antigen, or other binding pairs, the terms "specific binding" or "selective binding" indicate a binding reaction that determines the presence of a protein in a heterogeneous population of proteins and other biological agents. Thus, under specified conditions, a specified ligand binds to a specific receptor and does not significantly bind to other proteins present in the sample. The antibody or antigen-binding composition derived from an antibody in the considered method has an affinity for its antigen or its variants or mutant proteins that is at least two, at least ten, at least 20, or at least 100 times greater than that for any other antibody or binding composition derived from them. In specific embodiments, the antibody will have an affinity greater than about 10, as determined by, for example, Scatchard analysis (Munsen et al., 1980 Analyt. Biochem. 107: 220-239). 9 Affinity per liter / molar.

[0126] The term "response," such as the response of a cell, tissue, organ, or organism, includes changes in biochemical or physiological behavior, such as concentrations, densities, adhesions, or migrations within biological compartments, changes in gene expression rates, or differentiation states, where the changes are related to activation, stimulation, or treatment, or to internal mechanisms such as gene programming. In some cases, the terms "activation," "stimulation," etc., refer to cell activation regulated by internal mechanisms and external or environmental factors; while the terms "inhibition," "downregulation," etc., refer to the opposite effect.

[0127] The terms “polypeptide,” “peptide,” and “protein,” used interchangeably herein, refer to a polymer of amino acids of any length, which may include genetically encoded and non-genetically encoded amino acids, chemically or biochemically modified or derived amino acids, and polypeptides having a modified polypeptide backbone. The term includes fusion proteins, including but not limited to fusion proteins having heterologous amino acid sequences, fusion proteins with or without N-terminal methionine residues and heterologous and homologous leader sequences; immunolabeled proteins, etc.

[0128] As used herein, the terms “variant” and “homologue” are used interchangeably to refer to amino acid or DNA sequences that are similar to a reference amino acid or nucleic acid sequence, respectively. The term includes both naturally occurring variants and non-naturally occurring variants. Naturally occurring variants include homologues (amino acid or nucleotide sequences that differ from one species to another, each distinct polypeptide and nucleic acid) and allelic variants (amino acid or nucleotide sequences that differ within a species from one individual to another, each distinct polypeptide and nucleic acid). Thus, variants and homologues include naturally occurring DNA sequences and the proteins they encode, as well as their isotypes, and splicing variants of proteins or genes. The term also covers nucleic acid sequences that differ from one or more bases of a naturally occurring DNA sequence, but which, due to the degeneracy of the genetic code, are still translated into the amino acid sequence corresponding to a naturally occurring protein. Non-naturally occurring variants and homologues include polypeptides and nucleic acids that respectively contain changes in amino acid or nucleotide sequences, where the sequence changes (e.g., mutant proteins) are artificially introduced; for example, the changes are produced in a laboratory by human intervention (“manual”). Therefore, non-natural variants and homologues can also refer to those that differ from naturally occurring sequences through one or more conserved substitutions and / or labels and / or conjugates.

[0129] As used in this article, the term "mutant protein" broadly refers to a mutated recombinant protein. These proteins typically carry one or more amino acid substitutions and are usually derived from cloned genes that have undergone site-directed or random mutagenesis, or from fully synthetic genes.

[0130] The terms “DNA,” “nucleic acid,” “nucleic acid molecule,” and “polynucleotide” are used interchangeably in this document to refer to a polymer of nucleotides of any length, whether deoxyribonucleotides, ribonucleotides, or analogs thereof. Non-limiting examples of polynucleotides include linear and circular nucleic acids, messenger RNA (mRNA), complementary DNA (cDNA), recombinant polynucleotides, vectors, probes, primers, etc.

[0131] Adenosine A 2A Receptor and adenosine A 2B Receptors and their inhibition

[0132] As stated above, a precise understanding of the potential mechanisms of action of the compounds affecting the activity of the present invention is not necessary for carrying out the present invention, and the compounds (or subsets thereof) are believed to inhibit adenosine A. 2A Receptor (A) 2A R) and / or adenosine A 2B Receptor (A) 2B Alternatively, the compound (or a subset thereof) may inhibit adenylate cyclase function. The compound (or a subset thereof) may also have activity against A. 2A Receptor (A) 2AR), adenosine A 2B Receptor (A) 2B R) and adenylate cyclase inhibitory activity. Although the compounds of the present invention generally refer to adenosine A herein. 2A Receptor (A) 2A R) and / or adenosine A 2B Receptor (A) 2B R) inhibitors should be understood as, the term "A" 2A R / A 2B "R inhibitors" contain substances that inhibit A 2A R, A 2B Compounds that act alone or by inhibiting A or adenylate cyclase, and / or by inhibiting A. 2A R, A 2B Compounds that function with R and adenylate cyclase.

[0133] Identification of adenosine A 2A Receptor and adenosine A 2B Receptor inhibitors possess the desired properties

[0134] In part, the present invention relates to the identification of at least one adenosine A having therapeutically relevant properties or characteristics. 2A Receptor and adenosine A 2B Receptor inhibitors. Candidate inhibitors can be identified using, for example, field-acceptable assays or models, such as those described herein.

[0135] Following identification, candidate inhibitors can be further evaluated using techniques that provide data on inhibitor characteristics, such as pharmacokinetic parameters, methods for determining solubility and stability. Comparison of candidate inhibitors with reference standards (which may be the "best" among existing inhibitors) demonstrates the potential feasibility of these candidates.

[0136] Compounds of the present invention

[0137] This article provides compounds of formula (I):

[0138]

[0139] Or its pharmaceutically acceptable salts, hydrates or solvates, wherein,

[0140] G 1 For N or CR 3a ;

[0141] G 2 For N or CR 3b ;

[0142] G 3 For N or CR 3c ;

[0143] R3a R 3b and R 3c Each independently can be H, deuterium, or C. 1-3 alkyl;

[0144] R 1a and R 1b Each person independently selects from the following groups:

[0145] viii)H or deuterium,

[0146] ix) can be arbitrarily assigned to 1-3 Rs 5 C replaced by substituent 1-8 alkyl,

[0147] x) Optionally assigned to 1-3 R 5 -X replaced by substituent 1 -OC 1-8 alkyl,

[0148] xi)-C(O)-R 6 ,

[0149] xii) Optionally assigned to 1-3 R 7 Y replaced by the substituent, and

[0150] xiii) Optionally assigned to 1-3 1-3R 7 -X replaced by substituent 1 -Y; or

[0151] xiv)R 1a and R 1b Together with the nitrogen attached to them, they are optionally formed by 1-3 R 8 The 5-6 membered heterocyclic alkyl ring substituted by the substituent, wherein the heterocyclic alkyl ring has 0-2 heteroatom ring vertices selected from the group consisting of O, N and S;

[0152] Each Y is C 3-8 Cycloalkyl or 4- to 6-membered heterocyclic alkyl groups having 1-3 heteroatom rings selected from the O, N, and S groups;

[0153] R 2 and R 4 Each independently can be H, deuterium, or C. 1-3 alkyl;

[0154] Ar 1 It is a phenyl or a 5- to 6-membered heteroaryl group, wherein each is optionally surrounded by 1-3 R groups. 9 Replaced;

[0155] Ar 2 It is a phenyl or a 5- to 6-membered heteroaryl group, wherein each is optionally surrounded by 1-3 R groups. 10Replaced;

[0156] Among them, Ar 1 and Ar 2 Each of the 5- to 6-membered heteroaryl groups independently possesses 1-3 selectable O, N, N + -O - The heteroatom ring of the group composed of S is located at the site.

[0157] Each X 1 C 1-6 Alkylene;

[0158] Each R 5 Independently selected from the following group: hydroxyl, C 3-8 Cycloalkyl, phenyl, -O-phenyl, -C(O)OR a and oxygenation;

[0159] Each R 6 It is C 1-8 Alkyl or Y, wherein each is optionally substituted by 1 to 3 substituents selected from the group consisting of: hydroxyl, -O-phenyl, phenyl, and -OC. 1-8 alkyl;

[0160] Each R 7 Select independently from the following group: C 1-8 Alkyl, hydroxyl, -OC 1-8 Alkyl, oxo, and C(O)OR a ;

[0161] Each R 8 Select independently from the following group: C 1-8 Alkyl, hydroxyl, and oxo;

[0162] Each R 9 Select independently from the following group: C 1-8 Alkyl, C 1-8 Deuterated alkyl, -OC 1-8 Alkyl, -OC 1-8 Deuterated alkyl groups, -X 1 -OC 1-8 Alkyl, -OX 1 -OC 1-8 Alkyl, -X 1 -OX 1 -OC 1-8 Alkyl, -C(O)OR a Halogen, cyano, -NR b R c Y, -X 1 -C 3-8 cycloalkyl and -X 2 -Z, where X 2 Selected from the following group: C1-6 Alkylene, -C 1-6 Alkylenes -O-, -C(O)-, and –S(O)2-, Z being 4- to 6-membered heterocyclic alkyl groups having 1-3 heteroatom ring vertices selected from the group consisting of O, N, and S, and wherein each of the R 9 The substituents are optionally replaced by 1-3 Rs 11 Replaced;

[0163] Each R 10 Select independently from the following group: C 1-8 Alkyl, C 1-8 Deuterated alkyl, halogen, cyano, -OC 1-8 Alkyl, -OC 1-8 Deuterated alkyl groups, -X 1 -OC 1-8 Alkyl, -OX 1 -OC 1-8 Alkyl, -S(O)2-C 1-6 Alkyl, -C(O)NR d R e and 4-6 membered heteroaryl groups having 1-3 heteroatom ring vertices selected from the group consisting of O, N and S, wherein each of the R 10 Substituents are independently replaced by 1-3 R 12 Replaced, or in Ar 2 Two R's at adjacent ring vertices 10 Optionally combined to form a 5-membered heterocycle optionally replaced by 1-2 halogens;

[0164] Each R 11 Independently selected from the following group: hydroxyl, halogen, cyano, -NR d R e -C(O)OR a , phenyl, C 3-8 cycloalkyl, and optionally C(O)OR a The C that was replaced 1-4 alkyl;

[0165] Each R 12 Independently selected from the group consisting of: halogen, cyano, hydroxyl, -C(O)OR; and

[0166] Each R a H, deuterium, or C 1-6 alkyl;

[0167] Each R b and R c Independently selected from the following groups: H, deuterium, C 1-8 Alkyl, -S(O)2-C 1-6 Alkyl, -C(O)OR a and -X1 -C(O)OR a ;

[0168] Each R d and R e Independently selected from the following groups: H, deuterium, C 1-8 Alkyl, -S(O)2-C 1-6 Alkyl groups; and

[0169] The condition is when G 1 and G 2 Each is N, G 3 For CH, R 2 CH3, and R 1a and R 1b When each is H or deuterium, then Ar 2 It is not 2-thienyl, phenyl, 2-,3- or 4-methoxyphenyl, 3- or 4-halophenyl, 2,4-dimethoxyphenyl, 2,4-dichlorophenyl or 2- or 4-methylphenyl.

[0170] In one preferred embodiment, a compound of formula (I) is provided, wherein Ar 1 To be arbitrarily assigned to 1-3 R 9 The 5 to 6 heteroaryl groups that are replaced.

[0171] In another alternative embodiment, a compound of formula (I) is provided, wherein Ar 1 Select from the following group: 1-3 R's of any land cover 9 The substituted pyridyl, pyridyl N-oxide, imidazole, pyrazolyl, and thiazolyl groups. In some selected embodiments, Ar 1 It is a pyridinyl or pyridinyl N-oxide, and optionally surrounded by 1-3 R groups. 9 What it replaced.

[0172] In some selected embodiments, compounds of formula (I) are provided, wherein G 3 For CR 3c .

[0173] In some selected embodiments, the compound of formula (I) is represented by formula (Ia).

[0174]

[0175] Where n is an integer from 0 to 2.

[0176] In some selected embodiments, the compound of formula (I) is represented by formula (Ib).

[0177]

[0178] In some selected embodiments, compounds of formulas (I), (Ia), and (Ib) are provided, wherein Ar 2 Optional land is covered by 1-3 R 10 Replaced. In some embodiments, at least one R 10 It is a cyano group.

[0179] In some selected embodiments, the compound of formula (I) is represented by formula (Ic).

[0180]

[0181] Where m is an integer from 0 to 2.

[0182] In some selected embodiments, the compound of formula (I) is represented by formula (Id).

[0183]

[0184] In some selected embodiments, compounds of formulas (I), (Ia), (Ib), (Ic), and (Id) are provided, wherein each R 9 Select independently from the following group: C 1-8 Alkyl, C 1-8 Deuterated alkyl, -OC 1-8 Alkyl, -OC 1-8 Deuterated alkyl groups, -X 1 -OC 1-8 Alkyl, -OX 1 -OC 1-8 Alkyl, -X 1 -OX 1 -OC 1-8 Alkyl groups, wherein each of the R groups 9 The substituents are optionally replaced by 1-3 Rs 11 What it replaced.

[0185] In some selected embodiments, compounds of formulas (I), (Ia), (Ib), (Ic), and (Id) are provided, wherein each R 9 Independently selected from the following group: -C(O)OR a -NR b R c Y, -X 1 -C 3-8 cycloalkyl 、 and -X 2 -Z, where X 2 Selected from the following group: C 1-6 Alkylene, -C 1-6Alkylenes -O-, -C(O)-, and –S(O)2-, Z being 4- to 6-membered heterocyclic alkyl groups having 1-3 heteroatom ring vertices selected from the group consisting of O, N, and S, and wherein each of the R 9 The substituents are optionally replaced by 1-3 Rs 11 What it replaced.

[0186] In some selected embodiments, the compound of formula (I) is represented by formula (Ie).

[0187]

[0188] In some selected embodiments, compounds of formulas (I), (Ia), (Ib), (Ic), (Id), and (Ie) are provided, wherein G 2 Let N be the number of elements in the array.

[0189] In some selected embodiments, compounds of formulas (I), (Ia), (Ib), (Ic), (Id), and (Ie) are provided, wherein G 1 Let N be the number of elements in the array.

[0190] In some selected embodiments, compounds of formulas (I), (Ia), (Ib), (Ic), (Id), and (Ie) are provided, wherein G 1 For CR 3a .

[0191] In some selected embodiments, compounds of formulas (I), (Ia), (Ib), (Ic), (Id), and (Ie) are provided, wherein R 2 It can be H or deuterium.

[0192] In some selected embodiments, compounds of formulas (I), (Ia), (Ib), (Ic), (Id), and (Ie) are provided, wherein R 4 It can be H or deuterium.

[0193] In some selected embodiments, compounds of formulas (I), (Ia), (Ib), (Ic), (Id), and (Ie) are provided, wherein R 1b It is H or deuterium. In some selected embodiments, compounds of formulas (I), (Ia), (Ib), (Ic), (Id), and (Ie) are provided, wherein R is H or deuterium. 1b Selected from the following group:

[0194] i)H or deuterium,

[0195] ii) Optionally placed by 1-3 R 5 C replaced by substituent 1-8 Alkyl groups, and

[0196] iii) Optionally placed by 1-3 R 5-X replaced by substituent 1 -OC 1-8 alkyl.

[0197] In some selected embodiments, compounds of formulas (I), (Ia), (Ib), (Ic), (Id), and (Ie) are provided, wherein R 1b Selected from the following group:

[0198] i)H or deuterium,

[0199] iv)-C(O)-R 6 ,

[0200] v) Optionally selected by 1-3 R 7 Y replaced by the substituent, and

[0201] vi) Optionally assigned to 1-3 1-3Rs 7 -X replaced by substituent 1 -Y.

[0202] In some selected embodiments, compounds of formulas (I), (Ia), (Ib), (Ic), (Id), and (Ie) are provided, wherein each R 10 Select independently from the following group: C 1-8 Alkyl, halogen, cyano, -OC 1-8 Alkyl, -X 1 -OC 1-8 Alkyl, -OX 1 -OC 1-8 Alkyl groups, wherein each of the R groups 10 The substituents are optionally replaced by 1-3 Rs 12 What it replaced.

[0203] In some embodiments, compounds of formulas (I), (Ia), (Ib), (Ic), (Id), and (Ie) are provided, wherein each R 10 Select independently from the following group: C 1-8 Alkyl, halogen, cyano, -OC 1-8 alkyl.

[0204] In some selected embodiments, compounds of formula (Ic) are provided, wherein m is at least 1 and at least one R. 10 It is cyano. In some selected embodiments, compounds of formulas (Id) and (Ie) are provided, wherein at least one R 10 It is a cyano group.

[0205] In some selected embodiments, any of the compounds in Table 1 are provided.

[0206] In some selected embodiments, any one of the following compounds is provided:

[0207]

[0208] In some selected embodiments, compound I is provided.

[0209]

[0210] In some preferred embodiments, deuterated forms of compounds of formulas (I), (Ia), (Ib), (Ic), (Id), and (Ie) are provided. Deuterium can independently replace hydrogen at any position where hydrogen might be present.

[0211] Synthesis method

[0212] Typically, the compounds provided herein can be prepared using the conventional methods described in the examples below.

[0213] Prodrugs and other methods of drug delivery and / or half-life prolongation

[0214] In some aspects of the invention, the compounds described herein are administered as prodrugs.

[0215] To prolong therapeutic activity, drug molecules can be designed for delivery using carriers. Such carriers may be used in a non-covalent manner, with the drug partially physicochemically formulated as a solvent-carrier mixture, or via a carrier reagent permanently covalently linked to one of the functional groups of the drug (see WO 20150202317 for general reference).

[0216] Several non-covalent methods are favored. For example, but not limited to, in some embodiments, long-acting formulations comprising a non-covalent drug encapsulated within a polymer carrier are employed. In such formulations, drug molecules are bound to and processed with the carrier material such that the drug molecules are distributed within a large-volume carrier. Examples include particulate polymer-drug aggregates (e.g., Microspheres (Phosphorex, Inc.) are used as injectable suspensions; polymer-drug molecule aggregates formulated into gels (e.g., (Lupron (AbbVie Inc.) , which is used as a single-dose bolus injection; and liposome formulations (e.g., (Pacira Pharmaceuticals) In these formulations, the carrier can be a polymer or non-polymer entity capable of dissolving the drug. In these formulations, drug molecules may be released when the carrier swells or physically breaks down. In other cases, chemical degradation allows the drug to diffuse into the biological environment; this chemical degradation process can be autohydrolysis or enzymatically catalyzed. Among other limitations, non-covalent drug encapsulation requires the prevention of uncontrolled drug release, and the dependence of the drug release mechanism on biodegradation can lead to inter-patient variability.

[0217] In specific embodiments, drug molecules, including both small and large molecules, are coupled to a carrier via permanent covalent bonds. Certain small-molecule therapeutics exhibiting low solubility in aqueous fluids can be dissolved by coupling with a hydrophilic polymer, examples of which are described elsewhere herein. Regarding large protein molecules, half-life extension can be achieved, for example, through permanent covalent modification with the palmitoyl moiety, and through coupling with another protein that itself has an extended half-life (e.g., ...). This is achieved through permanent covalent modification of the drug carrier. Typically, when a drug carrier is covalently bound to a drug, the drug molecule exhibits reduced biological activity.

[0218] In some cases, the limitations associated with drug molecules containing non-covalent polymer mixtures or permanent covalent linkages can be successfully overcome by employing a prodrug approach to chemically couple a drug to a polymer carrier. In this case, an inactive or less active therapeutic agent than the drug moiety itself will predictably be converted into an active molecular entity. The reduced bioactivity of the prodrug is advantageous when slow or controlled drug release is required, compared to the released drug. In this case, drug release occurs over time, reducing the need for repeated and frequent drug administration. The prodrug approach may also be advantageous when the drug moiety itself is not absorbed in the gastrointestinal tract or has below-optimal absorption; in these cases, the prodrug promotes the absorption of the drug moiety, which is then cleaved at a later time (e.g., via first-pass metabolism). Bioactive drug molecules are typically linked to the polymer carrier moiety via temporary bonds formed between the carrier moiety and the hydroxyl, amino, or carboxyl groups of the drug molecule.

[0219] The above methods are subject to several limitations. Prodrug activation can occur via enzymatic or non-enzymatic cleavage of the temporary bond between the carrier and the drug molecule, or a combination of both in sequence (e.g., an enzymatic step followed by a non-enzymatic modification). In an enzyme-free in vitro environment (e.g., an aqueous buffer), temporary bonds such as esters or amides may undergo hydrolysis, but the corresponding hydrolysis rates may keep them outside the therapeutically useful range. In contrast, in the in vivo environment, esterases or amidases are commonly present, and esterases and amidases can cause significant catalytic accelerations of hydrolysis kinetics ranging from two to several orders of magnitude (see, for example, Greenwald et al., (1999) Journal of Medicinal Chemistry (J Med Chem) 42(18): 3857-67).

[0220] As described herein, prodrugs can be classified into i) biological precursors and ii) carrier-linked prodrugs. Biological precursors do not contain a carrier group and are activated through the metabolism of functional groups. In contrast, in carrier-linked prodrugs, the active substance is complexed with the carrier moiety via a temporary link at a functional group of the biological active entity. Preferred functional groups are hydroxyl or amino. The linking chemistry and hydrolysis conditions depend on the type of functional group used. The carrier can be biologically inert (e.g., PEG) or can have targeting properties (e.g., antibodies). Cleavage of the carrier moiety of a carrier-linked prodrug yields the biological active entity of interest, and the deprotected nature of the functional groups of the biological active entity generally contributes to its biological activity.

[0221] Patents and scientific literature describe many macromolecular prodrugs in which temporary linkages are unstable ester bonds. In these cases, the functional group of the bioactive entity is a hydroxyl or carboxylic acid (see, for example, Cheng et al. (2003), Bioconjugate Chemistry, 14: 1007-17). Furthermore, it is generally advantageous for biomacromolecules and certain small molecule drugs to link the carrier to an amino group of the bioactive entity (e.g., the N-terminus or lysine amino group of a protein). During prodrug preparation, the amino group can be resolved more chemoselectively because it has higher nucleophilicity compared to hydroxyl or phenolic groups. This is particularly relevant for proteins and peptides containing a variety of different reactive functional groups, where non-selective binding reactions result in undesirable mixtures of products requiring extensive characterization or purification, thus reducing reaction yields and the therapeutic efficiency of the active moiety.

[0222] Generally, amide bonds are more stable to hydrolysis than ester bonds, and the cleavage rate of amide bonds may be too slow for the therapeutic efficacy of a carrier-linked prodrug. Therefore, it may be advantageous to add structural chemical components to control the cleavability of the prodrug amide bond. These additional cleavage-controlling chemical components, provided by neither the carrier entity nor the drug itself, are commonly referred to as “linking groups.” Prodrug linking groups can have a major influence on the hydrolysis rate of the temporary bond, and changes in the chemistry of the linking group often result in specific properties. Activation of the amine-containing bioactive moiety of the prodrug by a specific enzyme for targeted release requires the linking group to exhibit a structural motif that is recognized as a substrate by the corresponding endogenous enzyme. In these cases, the cleavage of the temporary bond occurs in a one-step enzymatic process. For example, the enzymatic release of cytarabine is influenced by the protease plasminogen activator, which is present in relatively high concentrations in various tumor masses.

[0223] Patient-to-patient variability is a major drawback of dominant enzymatic cleavage. Enzyme levels can vary significantly between subjects, leading to biological differences in how enzymatic cleavage induces prodrug activation. Enzyme levels can also vary depending on the administration site (e.g., for subcutaneous injection, certain areas of the body produce more predictable therapeutic effects than others). Furthermore, it is difficult to establish in vivo-in vitro correlations of the pharmacokinetic properties of enzyme-dependent carrier-linked prodrugs.

[0224] The use of other carrier prodrugs temporarily linked to an amino group in the drug moiety is based on a cascade mechanism. Cascade cleavage is achieved through a linker compound consisting of a structural combination of a masking group and an activating group. The masking group is linked to the activating group via a first temporary link, such as an ester or carbamate. The activating group is linked to the amino group of the drug molecule via a second temporary link (e.g., a carbamate). The stability of the second temporary link, or its sensitivity to hydrolysis, depends on the presence or absence of the masking group. In the presence of the masking group, the second temporary link is highly stable and it is impossible to release the drug molecule with therapeutically useful kinetics, while in the absence of the masking group, this link becomes highly unstable, leading to rapid cleavage and release of the drug moiety.

[0225] The cleavage of the first temporary link is the rate-limiting step in the cascade mechanism. The first step can induce a molecular rearrangement of the activating group (e.g., 1,6-elimination as described by Greenwald et al. (1999), Journal of Medicinal Chemistry (J Med Chem) 42: 3657-67), and this rearrangement makes the second temporary link more unstable, thus inducing its cleavage. Ideally, the cleavage rate of the first temporary link is the same as the desired release rate of the drug molecule in a given therapeutic regimen. Furthermore, it is ideal that the cleavage of the second temporary link is substantially instantaneous after the instability induced by the cleavage of the first temporary bond.

[0226] Another implementation includes a polyamino-containing prodrug based on trimethyl lock lactonization (see, for example, Greenwald et al. (2000), Journal of Medicinal Chemistry, 43(3): 457-87). In this prodrug system, a substituted o-hydroxyphenyl-dimethylpropionic acid is linked to a PEG via an ester, carbonate, or carbamate group as a first temporary link, and to an amino group of the drug molecule via an amide bond as a second temporary link. The rate-determining step in drug release is the enzymatic cleavage of the first link, followed by rapid amide cleavage via lactonization, releasing aromatic lactone byproducts. The main disadvantages of the prodrug system described by Greenwald et al. are the highly reactive release and the potentially toxic aromatic small molecule byproducts such as quinone methylates or aromatic lactones following the cleavage of the temporary links. The potentially toxic entities are released in a 1:1 stoichiometric ratio with the drug and can exhibit high in vivo concentrations.

[0227] In some embodiments of cascade prodrugs comprising an aromatic activating group based on 1,6-elimination, the masking group is structurally separate from the support. This can be achieved by employing a stable bond between the polymer support and the activating group, wherein the stable bond does not participate in the cascade fragmentation mechanism. If the support is not used as a masking group and the activating group is linked to the support via a stable bond, the release of potentially toxic byproducts (e.g., the activating group) is avoided. The stable connection between the polymer and the activating group also inhibits the release of drug-linking intermediates with indeterminate pharmacology.

[0228] The first example of the method described in the preceding paragraphs includes a polymeric prodrug system based on a mandelic acid activating group (see, for example, Shabat et al., (2004) Chem Eur J 10: 2626-34). In this method, a masking group is linked to an activating group via a carbamate bond. The activating group is permanently coupled to a polyacrylamide polymer via an amide bond. After the masking group is enzymatically activated by a catalytic antibody, the masking group is cleaved by cyclization to release the drug; after drug release, the activating group remains linked to the polyacrylamide polymer. A similar prodrug system is based on a mandelic acid activating group and an enzymatically cleavable ester-linked masking group (see, for example, Lee et al., (2004) Angew Chem 116: 1707-10).

[0229] When using the aforementioned linker groups, the 1,6-elimination step still produces highly reactive aromatic intermediates. Even if the aromatic moiety remains permanently attached to the polymer support, side reactions with potentially toxic byproducts or immunogenic effects may occur. Therefore, it is advantageous to use aliphatic prodrug linker groups to generate linker groups for forming polymeric prodrugs containing amine-active reagents, wherein the aliphatic prodrug linker groups are not enzyme-dependent and do not produce reactive aromatic intermediates during lysis. One such example is the use of PEG5000-maleic anhydride for the reversible modification of amino groups in tissue plasminogen activator and urokinase (see, for example, (1987) Garman et al., FEBS Letters 223(2): 361-65). The functional enzyme regeneration from the PEG-uPA complex, incubated in pH 7.4 buffer and by cleavage of maleamic acid-linked complexes, follows first-order kinetics and has a half-life of about 6 hours. A disadvantage of maleamic acid linking is the lack of stability of the complex at lower pH values.

[0230] Another approach includes PEG-cascade prodrug systems based on the N,N-bis-(2-hydroxyethyl)glycine amide (N-diaglycine) linker group (see, for example, Journal of Medicinal Chemistry (2004) 47:726-34). In this system, two PEG carrier molecules are temporarily linked to an N-diaglycine molecule coupled to the amino group of a drug molecule. The first step of prodrug activation involves the enzymatic cleavage of the first temporary link between the two PEG carrier molecules and the hydroxyl group of the N-diaglycine activating group. Different links between PEG and N-diaglycine lead to different prodrug activation kinetics. The second step of prodrug activation involves the cleavage of the second temporary link between the N-diaglycine activating group and the amino group of the drug molecule. A disadvantage of this system is the slow hydrolysis rate of the second temporary N-diaglycine amide linker, which may result in the release of N-diaglycine-modified prodrug intermediates exhibiting different pharmacokinetic, immunogenic, toxic, and pharmacodynamic properties compared to the native parent drug molecule.

[0231] In specific implementations, dipeptides are used for prodrug development involving targeted or targeted transport because they are substrates for enzymes or biological transport systems. The ability of non-enzymatic pathways for dipeptide prodrug formation—i.e., intramolecular cyclization to form the corresponding diketopiperazine (DKP) and release the active drug—has not been established.

[0232] In some embodiments, the dipeptide is linked to the drug moiety via an ester bond, as described for the dipeptide ester of the drug paracetamol (Gomes et al., (2005) Bio & MedChem Lett). In this case, the cyclization reaction proceeds from a nucleophilic attack of the N-terminal amine of the peptide on the carbon atom of the ester to form a tetrahedral intermediate, after which a proton is transferred from the amine to the leaving group oxoanion, simultaneously forming a peptide bond, yielding the cyclic DKP product and the free drug. This method is applicable to hydroxyl-containing drugs in vitro, but it has been found to compete with the enzymatic hydrolysis of ester bonds in vivo, as the rate of release of acetaminophen from the corresponding dipeptide ester is much faster than in buffer (Gomes et al., (Molecules) 12 (2007) 2484-2506). Susceptibility of dipeptide-based prodrugs to peptidases can be addressed by incorporating at least one non-natural amino acid into the dipeptide motif. However, endogenous enzymes capable of cleaving ester bonds are not limited to peptidases, and this enzyme-dependent prodrug cleavage still leads to unpredictable in vivo performance.

[0233] In some embodiments, the enzyme-dependent formulation is intentionally modified into a DKP prodrug, for example, in which a dipeptide ester prodrug is formylated at the amino terminus of the dipeptide, and enzymatic deformylation is used to initiate the formation of diketopiperazine and subsequent cleavage of the ester-dipeptide bond, followed by the release of the drug molecule (see, for example, USP 7,163,923). By a further example, the octapeptide is linked to the 4-hydroxyl group of vinblastine via an ester linker, and the ester bond is cleaved by the formation of DKP following specific enzymatic removal of the N-terminal hexapeptide (see Brady et al., (2002) Journal of Medicinal Chemistry 45: 4706-15).

[0234] The scope of DKP formation reactions has also been extended to amide prodrugs. For example, USP 5,952,294 describes the use of diketopiperazine to form a prodrug activation for a dipeptide amide prodrug of cytarabine. In this case, a temporary link is formed between the carbonyl group of the dipeptide and the aromatic amino group of cytarabine. However, sustained-release effects are unlikely to be achieved for such complexes because there is no carrier or other half-life-extending moieties or functional groups present.

[0235] Dipeptide prodrugs containing bioactive peptides such as GLP-1 are also described, which release the peptide through the formation of a dipeptide-elongated diketopiperazine (see, for example, WO2009 / 099763). The bioactive peptide moiety may contain an additional PEG chain on one of its amino acid side chain residues to achieve elongated cycling of the bioactive peptide. However, this approach is associated with several significant drawbacks. First, the PEG chain must be linked to the peptide without impairing its bioactivity, which may be difficult to achieve for many peptide-based bioactive agents. Second, since the pegylated peptide itself is bioactive, the dipeptide precursor influences the peptide's bioactivity and may negatively impact its receptor-binding properties.

[0236] Specific exemplary technologies that can be used with the compounds of this invention include those developed by ProLynx (San Francisco, CA) and Ascendis Pharma (Palo Alto, CA). The ProLynx technology platform utilizes multiple sets of novel linkers pre-programmed to cleave at different rates, thereby allowing controlled, predictable, and sustained release of small molecules and peptides from circulating semi-solid macromolecular complexes. This technology allows for maintaining the steady-state serum levels required for therapeutic agents for weeks to months.

[0237] The Ascendis technology platform combines the advantages of prodrug and sustained-release technologies to enhance the performance of small molecules and peptides. During circulation, the proprietary prodrug releases the unmodified active parent therapeutic agent at a predetermined rate controlled by physiological pH and temperature conditions. Because the therapeutic agent is released in its unmodified form, it retains its original mechanism of action.

[0238] Modifications to enhance inhibitory characteristics

[0239] Improving one or more physical properties of the therapeutic modalities disclosed herein and / or their mode of administration is often beneficial and sometimes necessary. Improvements to physical properties include, for example, methods to increase water solubility, bioavailability, serum half-life and / or therapeutic half-life; and / or modulate biological activity.

[0240] Modifications known in the art include polyethylene glycol (PEG) fusion, Fc-fusion, and albumin fusion. While typically associated with macromolecular agents such as peptides, such modifications have recently been evaluated with specific small molecules. For example, Chiang, M et al. (J. Am. Chem. Soc., 2014, 136(9):3370-73) described a small molecule agonist of the adenosine 2a receptor coupled to the Fc domain of immunoglobulin. The small molecule-Fc complex retained efficient Fc and adenosine 2a receptor interactions and exhibited superior performance compared to the uncomplexed small molecule. Covalent linkage of PEG molecules to small molecule therapeutics has also been described (Li, W. et al., Progress in Polymer Science, 2013).

[0241] 38:421-44).

[0242] Other known modifications include deuteration to improve pharmacokinetic, pharmacodynamic, and toxicological characteristics. Due to the larger atomic mass of deuterium, breaking the carbon-deuterium bond requires more energy than breaking the carbon-hydrogen bond. Because these stronger bonds are more difficult to break, the drug is metabolized more slowly than in its non-deuterated form, which allows for lower frequency dosing and may further reduce toxicity. (Charles Schmidt, Nature Biotechnology, 2017, 35(6):493-494; Harbeson, S. and Tung, R., Medchem News, 2014(2):8-22).

[0243] Therapeutic and preventive uses

[0244] This invention considers the A described herein. 2A R / A 2B Use of R inhibitors in the treatment or prevention of a wide range of diseases, conditions and / or illnesses, and / or their symptoms. Although specific uses are described in detail below, it should be understood that the invention is not limited thereto. Furthermore, while general categories of specific diseases, conditions and illnesses are listed below, some diseases, conditions and illnesses may be members of more than one category, and others may not be members of any of the disclosed categories.

[0245] In some embodiments, the diseases, conditions, and / or ailments described herein are at least in part caused by adenosine A. 2A Receptor (A) 2A Adenosine A (A) mediates. In some embodiments, diseases, conditions, and / or ailments as described herein are at least partially mediated by adenosine A. 2B Receptor (A) 2BR) mediated. In some embodiments, the disease, condition, and / or ailment as described herein is at least partially mediated by A. 2A R and A 2B R-mediated.

[0246] In some embodiments, such as A as described herein 2A R / A 2B R inhibitors effectively reverse or stop A 2A Dosage administration of R-mediated immunosuppression.

[0247] Oncology-related diseases According to the present invention, A 2A R / A 2B R inhibitors can be used to treat or prevent proliferative diseases or conditions, including cancers such as uterine cancer, cervical cancer, breast cancer, prostate cancer, testicular cancer, gastrointestinal cancers (e.g., esophageal cancer, oropharyngeal cancer, stomach cancer, small bowel cancer, large bowel cancer, colon cancer, or rectal cancer), kidney cancer, renal cell carcinoma, bladder cancer, bone cancer, bone marrow cancer, skin cancer, head or neck cancer, liver cancer, gallbladder cancer, heart cancer, lung cancer, pancreatic cancer, salivary gland cancer, adrenal cancer, thyroid cancer, brain cancer (e.g., glioma), ganglion cancer, cancers of the central nervous system (CNS) and peripheral nervous system (PNS), as well as cancers of the hematopoietic system and the immune system (e.g., spleen or thymus). This invention also provides methods for treating or preventing, for example, immunogenic tumors, non-immunogenic tumors, dormant tumors, virus-induced cancers (e.g., epithelial carcinoma, endothelial carcinoma, squamous cell carcinoma, and papillomavirus), adenocarcinoma, lymphoma, carcinoma, melanoma, leukemia, myeloma, sarcoma, teratoma, chemically induced cancer, metastasis, and other cancer-related diseases, symptoms, or conditions involving angiogenesis. This invention contemplates reducing tolerance to tumor cell or cancer cell antigens, for example, by modulating the activity of regulatory T cells and / or CD8+ T cells (see, for example, Ramirez-Montagut et al. (2003) Oncogene 22:3180-87; and Sawaya et al. (2003) New Engl. J. Med. 349:1501-09). In specific embodiments, the tumor or cancer is colon cancer, ovarian cancer, breast cancer, melanoma, lung cancer, glioblastoma, or leukemia. The term “cancer-related diseases, symptoms and conditions” is used broadly to refer to conditions that are directly or indirectly related to cancer, and includes, for example, angiogenesis and precancerous conditions such as dysplasia.

[0248] In some embodiments, the cancer is metastatic or at risk of metastasis, or may be present in diffuse tissues, including blood cancers or bone marrow cancers (e.g., leukemia). In some further embodiments, the compounds of the present invention can be used to overcome T-cell tolerance.

[0249] In some embodiments, the present invention provides the use of A 2A R / A 2B Examples of methods for treating proliferative conditions, cancers, tumors, or precancerous conditions with R inhibitors and at least one other therapeutic or diagnostic agent are described elsewhere in this article.

[0250] Immune or inflammatory conditions. As used herein, terms such as “immune disease,” “immune condition,” “immune disorder,” “inflammatory disease,” “inflammatory condition,” and “inflammatory disorder” are intended to broadly encompass any A that can be described herein. 2A R / A 2B R-inhibitor therapy provides therapeutic benefits for immune-related conditions (such as autoimmune diseases) or diseases with inflammatory components. These conditions are often inextricably linked to other diseases, symptoms, and illnesses. For example, an "immunocondition" can refer to proliferative conditions such as cancer, tumors, and angiogenesis; or to infections (acute and chronic), tumors, and cancers that resist eradication by the immune system.

[0251] A of the present invention 2A R / A 2B R inhibitors can be used to increase or enhance immune responses; to improve immunity, including enhancing vaccine efficacy; and to increase inflammation. The compounds disclosed herein can be used to treat immunodeficiency diseases, immunosuppressive medical treatments, acute and / or chronic infections, and age-related immunodeficiency. The A... 2A R / A 2B R inhibitors can also be used to stimulate the immune system of patients with iatrogenic immunosuppression, including those who have undergone bone marrow transplantation, chemotherapy, or radiation therapy.

[0252] In a specific embodiment of this disclosure, by providing adjuvant activity, the A 2A R / A 2B R inhibitors are used to increase or enhance the immune response to antigens. In specific embodiments, at least one antigen or vaccine is combined with at least one A of the present invention. 2A R / A 2B R inhibitors are administered in combination to subjects to prolong the immune response to antigens or vaccines. Therapeutic compositions are also provided, comprising at least one of the A components of the present invention. 2A R / A 2B The R inhibitor is combined with at least one antigenic reagent or vaccine component, said antigenic reagent or vaccine component including but not limited to viruses, bacteria and fungi or parts thereof, proteins, peptides, tumor-specific antigens, and nucleic acid vaccines.

[0253] A non-limiting list of immune and inflammatory-related diseases, conditions, and illnesses that can be treated or prevented by the compounds and compositions of the present invention includes: arthritis (such as rheumatoid arthritis), kidney failure, lupus, asthma, psoriasis, colitis, pancreatitis, allergies, fibrosis, surgical complications (such as sites where inflammatory cytokines prevent healing), anemia, and fibromyalgia. Other diseases and illnesses that may be associated with chronic inflammation include: Alzheimer's disease, congestive heart failure, stroke, aortic stenosis, arteriosclerosis, osteoporosis, Parkinson's disease, infections, inflammatory bowel diseases (such as Crohn's disease and ulcerative colitis), allergic contact dermatitis and other eczema, systemic sclerosis, transplantation, and multiple sclerosis.

[0254] In other immune-related conditions, inhibition of A can be expected. 2A R / A 2B R function can also play a role in immune tolerance and prevention of intrauterine fetal rejection.

[0255] In some implementations, as described herein, A 2A R / A 2B R inhibitors can be combined with immunosuppressants to reduce immune effector cells.

[0256] The following text describes some of the aforementioned diseases, symptoms, and conditions in more detail, including their A aspects. 2A R / A 2B R inhibitors may be particularly effective (due to, for example, the limitations of current therapies).

[0257] Rheumatoid arthritis (RA), typically characterized by chronic inflammation in the synovial membrane of joints, affects approximately 1% of the US population (-2.1 million people). A better understanding of the roles of cytokines, including TNF-α and IL-1, in the inflammatory process has enabled the development and introduction of a new class of disease-modifying antirheumatic drugs (DMARDs). These agents (some of which overlap with existing treatments for RA) include ENBREL (etanercept), REMICADE (infliximab), HUMIRA (adalimumab), and KINERET (anakinra). While some of these agents alleviate symptoms, inhibit the progression of structural damage, and improve function in certain patient populations, there is still a need for alternative agents with improved potency, complementary mechanisms of action, and fewer / less severe side effects.

[0258] Psoriasis, a common group of immune-mediated chronic skin diseases, affects more than 4.5 million people in the United States, of whom 1.5 million are considered to have moderate to severe forms of the disease. Furthermore, more than 10% of people with psoriasis develop psoriatic arthritis, which damages the bone and connective tissue surrounding the joints. A better understanding of the underlying physiology of psoriasis has led to the introduction of agents, such as those targeting T lymphocytes and the activity of cytokines responsible for the inflammatory nature of the disease. These agents include TNF-α inhibitors (also used to treat rheumatoid arthritis (RA)), including ENBREL (etanercept), REMICADE (infliximab), HUMIRA (adalimumab), and T-cell inhibitors such as AMEVIVE (alefacept) and RAPTIVA (efalizumab). While several of these agents are effective to some extent in certain patient populations, none have shown efficacy in treating all patients.

[0259] Microbial-related diseases The present invention envisions the A described herein. 2A R / A 2B The use of R inhibitors in the treatment and / or prevention of any viral, bacterial, fungal, parasitic, or other infectious diseases, conditions, or illnesses, with the use of A 2A R / A 2B R inhibitors may be beneficial in treating these diseases, symptoms, or conditions.

[0260] Examples of viral diseases, symptoms, and conditions to be considered include, but are not limited to, hepatitis B virus (HBV), hepatitis C virus (HCV), human papillomavirus (HPV), HIV, AIDS (including its clinical manifestations such as cachexia, dementia, and diarrhea), herpes simplex virus (HSV), Epstein-Barr virus (EBV), varicella-zoster virus, Coxsackie virus, and cytomegalovirus (CMV).

[0261] Other examples of such diseases and conditions include: staphylococcal and streptococcal infections (e.g., *Staphylococcus aureus* and *Streptococcus sanguinis*, respectively), *Leishmania*, *Toxoplasma gondii*, *Trichomonas vaginalis*, *Giardia lamblia*, *Candida albicans*, *Bacillus anthracis*, and *Pseudomonas aeruginosa*. In some embodiments, diseases and conditions include mycobacterial infections (e.g., *Mycobacterium leprae* or *Mycobacterium tuberculosis*) or infections caused by *Listeria monocytogenes* or *Toxoplasma gondii*. The compounds of the present invention can be used to treat sepsis, reduce or inhibit bacterial growth, and reduce or inhibit inflammatory cytokines.

[0262] Other implementations consider treating parasitic infections, including but not limited to Leishmania donovani, Leishmania tropicalis, Leishmania macrophylla, Leishmania esculenta, Leishmania mexicana, Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale, or Plasmodium malariae. Antiparasitic therapy is often administered prophylactically (e.g., before the subject travels to an area with a high frequency of parasitic infections).

[0263] CNS-related and neurological disorders Inhibit A 2A R / A 2B R can also be an important treatment strategy for patients with neurological, neuropsychiatric, neurodegenerative, or other diseases, conditions, and illnesses related to the central nervous system, including those associated with cognitive and motor impairments. Examples include Parkinson's disease, extrapyramidal syndrome (EPS), dystonia, akathisia, tardive dyskinesia, restless legs syndrome (RLS), epilepsy, periodic limb movement disorder (PLMS), attention deficit disorder, depression, anxiety disorder, dementia, Alzheimer's disease, Huntington's disease, multiple sclerosis, cerebral ischemia, hemorrhagic stroke, subarachnoid hemorrhage, and traumatic brain injury.

[0264] Subjects with multiple sclerosis (MS) (a severely debilitating autoimmune disease involving multiple inflammatory areas and scarring of myelin in the brain and spinal cord) may be particularly helpful with the A described in this article. 2A R / A 2B R inhibitors are currently used because existing treatments only alleviate symptoms or delay the progression of disability.

[0265] Similarly, A 2A R / A 2B R inhibitors may be particularly beneficial for patients with neurodegenerative diseases such as Alzheimer's disease (AD), brain conditions that severely impair a patient's thinking, memory, and language processes, and Parkinson's disease (PD), which are progressive CNS diseases characterized by, for example, abnormal movement, rigidity, and tremor. These diseases are progressive and debilitating, and there are no effective drugs available.

[0266] Other symptoms Embodiments of the present invention consider applying the A described herein to an object. 2A R / A 2B R inhibitors, used for treatment or prevention, can be administered by at least A 2A R / A 2BAny other condition that benefits from a certain level of inhibition of R. These diseases, conditions, and illnesses include, for example, cardiovascular diseases (e.g., ischemic heart disease), gastrointestinal diseases (e.g., Crohn's disease), metabolic diseases (e.g., diabetes), liver diseases (e.g., liver fibrosis, NASH, and NAFLD), lung diseases (e.g., COPD and asthma), ophthalmological diseases (e.g., diabetic retinopathy), and kidney diseases (e.g., kidney failure).

[0267] Pharmaceutical Composition

[0268] A of the present invention 2A R / A 2B R inhibitors can be in the form of compositions suitable for administration to a target. Typically, such compositions contain A. 2A R / A 2B A "pharmaceutical composition" comprising an R inhibitor and one or more pharmaceutically or physiologically acceptable diluents, carriers, or excipients. In some embodiments, A 2A R / A 2B R inhibitors are present in therapeutically acceptable amounts. Pharmaceutical compositions can be used in the methods of the present invention; thus, for example, the pharmaceutical compositions can be administered to a subject ex vivo or in vivo to perform the treatment and prevention methods and uses described herein.

[0269] The pharmaceutical compositions of the present invention can be formulated to be compatible with the intended method or route of administration; exemplary routes of administration are set forth herein. Furthermore, the pharmaceutical compositions can be used in combination with other therapeutically active agents or compounds described herein to treat or prevent the diseases, symptoms, and conditions considered in this invention.

[0270] Contains active ingredients (e.g., A) 2A R / A 2B Pharmaceutical compositions of R-function inhibitors can be in forms suitable for oral administration, such as tablets, capsules, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups, solutions, microbeads, or elixirs. Pharmaceutical compositions for oral administration can be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions can contain one or more agents, such as sweeteners, flavoring agents, coloring agents, and preservatives, to provide a pharmaceutically elegant and palatable formulation. Tablets, capsules, etc., contain an active ingredient mixed with non-toxic, pharmaceutically acceptable excipients suitable for manufacturing tablets. These excipients can be, for example, diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrants such as corn starch or alginate; binders such as starch, gelatin, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc.

[0271] Tablets, capsules, etc., suitable for oral administration can be uncoated or coated using known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide sustained action. For example, delay-release materials such as glyceryl monostearate or glyceryl distearate can be used. They can also be coated using techniques known in the art to form osmotic therapeutic tablets for controlled release. Other agents include biodegradable or biocompatible particles or polymers such as polyesters, polyamines, hydrogels, polyvinylpyrrolidone, polyanhydride, polyglycolic acid, ethylene-vinyl acetate, methylcellulose, carboxymethylcellulose, protamine sulfate, or lactide / glycolic acid copolymers, polylactide / glycolic acid copolymers, or ethylene-vinyl acetate copolymers to control the delivery of the administered composition. For example, oral dosage forms can be encapsulated in microcapsules prepared by coagulation techniques or by interfacial polymerization, using hydroxymethylcellulose or gelatin microcapsules or poly(methyl methacrylate) microcapsules or colloidal drug delivery systems, respectively. Colloidal dispersion systems include macromolecular complexes, nanocapsules, microspheres, microbeads, and lipid-based systems, including oil-in-water emulsions, micelles, mixed micelles, and liposomes. Methods for preparing the above formulations will be readily apparent to those skilled in the art.

[0272] Preparations for oral use may also be hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate, kaolin, or microcrystalline cellulose, or soft gelatin capsules in which the active ingredient is mixed with an aqueous or oily medium such as peanut oil, liquid paraffin, or olive oil.

[0273] Aqueous suspensions contain active materials mixed with excipients suitable for their manufacture. Such excipients can be suspending agents, such as sodium carboxymethyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and gum arabic; dispersants or wetting agents, such as naturally occurring phospholipids (e.g., lecithin) or condensation products of ethylene oxides and fatty acids (e.g., polyoxyethylene stearate), or condensation products of ethylene oxides and long-chain fatty alcohols (e.g., heptadecanyloxycetyl alcohol), or condensation products of ethylene oxides and esters derived from fatty acids and hexitols (e.g., polyoxyethylene sorbitan monooleate), or condensation products of ethylene oxides and esters derived from fatty acids and hexitol anhydrides (e.g., polyvinyl dehydrated sorbitan monooleate). Aqueous suspensions may also contain one or more preservatives.

[0274] Oil suspensions can be formulated by suspending the active ingredient in vegetable oils (such as peanut oil, olive oil, sesame oil, or coconut oil) or mineral oils (such as liquid paraffin). Oil suspensions may contain thickeners such as beeswax, hard paraffin, or hexadecyl alcohol. Sweeteners and flavorings, as mentioned above, can be added to provide a palatable oral formulation.

[0275] Dispersible powders and granules suitable for preparing aqueous suspensions by adding water provide active ingredients that can be mixed with dispersants or wetting agents, suspending agents, and one or more preservatives. Suitable dispersants or wetting agents and suspending agents are illustrated herein.

[0276] The pharmaceutical compositions of the present invention may also be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, such as olive oil or peanut oil, or a mineral oil, such as liquid paraffin, or a mixture thereof. Suitable emulsifiers may be naturally occurring gums, such as gum arabic or tragacanth; naturally occurring phospholipids, such as soybean or lecithin, and esters or metaesters derived from fatty acids; hexitan anhydrides, such as sorbitan monooleate; and condensation products of partial esters with vinyl oxides, such as polyoxyethylene dehydrated sorbitan monooleate.

[0277] The pharmaceutical composition typically contains a therapeutically effective amount of A as described in this invention. 2A R / A 2B R inhibitors and one or more pharmaceutically and physiologically acceptable formulation agents. Suitable pharmaceutically or physiologically acceptable diluents, carriers, or excipients, including but not limited to antioxidants (e.g., ascorbic acid and sodium bisulfate), preservatives (e.g., benzyl alcohol, methylparaben, ethylparaben, or n-propylparaben), emulsifiers, suspending agents, dispersants, solvents, fillers, fillers, detergents, buffers, carriers, diluents, and / or adjuvants. For example, a suitable carrier may be a physiological saline solution or a citrate-buffered saline solution, possibly supplemented with other substances common in pharmaceutical compositions for parenteral administration. Neutral buffered saline solutions or saline solutions mixed with serum albumin are additional exemplary carriers. Those skilled in the art will readily recognize the various buffers that may be used in the pharmaceutical compositions and dosage forms considered herein. Typical buffers include, but are not limited to, pharmaceutically acceptable weak acids, weak bases, or mixtures thereof. As an example, the buffer component may be a water-soluble substance such as phosphoric acid, tartaric acid, lactic acid, succinic acid, citric acid, acetic acid, ascorbic acid, aspartic acid, glutamic acid, and their salts. Acceptable buffers include, for example, tromethamine buffer (Tris buffer), N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES), 2-(N-morpholine)ethanesulfonic acid (MES), sodium 2-(N-morpholine)ethanesulfonate (MES), 3-(N-morpholine)propanesulfonic acid (MOPS), and N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid (TAPS).

[0278] After formulation, the pharmaceutical composition can be stored in sterile vials as a solution, suspension, gel, emulsion, solid, or dehydrated or lyophilized powder. This formulation can be stored in ready-to-use form, lyophilized form requiring reconstitution before use, liquid form requiring dilution before use, or other acceptable forms. In some embodiments, the pharmaceutical composition is in a single-use container (e.g., a single-use vial, ampoule, syringe, or auto-injector (similar to, for example...)). The original text provides a reference to a specific implementation, while other implementations provide a reusable container (e.g., a reusable vial).

[0279] The formulation may also include a carrier to protect the composition from rapid degradation or elimination from the body, such as controlled-release formulations, which include liposomes, hydrogels, prodrugs, and microencapsulated delivery systems. For example, delayed-release materials, such as glyceryl monostearate or glyceryl stearate alone in combination with a wax, can be used to deliver A. 2A R / A 2B R inhibitors, including implants (e.g., implantable pumps) and catheter systems, slow-infusion pumps and devices, are all well known to those skilled in the art.

[0280] Depot injections, typically administered subcutaneously or intramuscularly, can also be used to release the A disclosed herein within a limited timeframe. 2A R / A 2B R inhibitors. Accumulated injections are typically solid- or oil-based and usually contain at least one formulation component described herein. Those skilled in the art are familiar with the possible formulations and uses of accumulated injections.

[0281] The pharmaceutical composition may be in the form of a sterile injectable aqueous solution or an oily suspension. The suspension may be formulated using suitable dispersants or wetting agents and suspending agents mentioned herein, according to known techniques. The sterile injectable formulation may also be a sterile injectable solution or suspension in a non-toxic, parenteral-acceptable diluent or solvent (e.g., a solution in 1,3-butanediol). Acceptable diluents, solvents, and dispersion media include water, Ringer's solution, isotonic sodium chloride solution, Cremophor EL... TM (BASF, Parsipani, New Jersey) or phosphate-buffered saline (PBS), ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. Additionally, sterile, fixed oils are often used as solvents or suspension media. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. Furthermore, fatty acids such as oleic acid can be used to prepare injectable formulations. Prolonged absorption of specific injectable formulations can be achieved by including agents that delay absorption (e.g., aluminum monostearate or gelatin).

[0282] This invention considers administering A in the form of a suppository for rectal use. 2A R / A 2B Inhibitors. Suppositories can be prepared by mixing a drug with a suitable, non-irritating excipient that is solid at room temperature but liquid at rectal temperature, thus melting in the rectum to release the drug. Such materials include, but are not limited to, cocoa butter and polyethylene glycol.

[0283] The present invention considers A 2A R / A 2B R inhibitors can be in the form of any other suitable pharmaceutical composition currently known or to be developed in the future (e.g., a spray for nasal or inhalation use).

[0284] Application method

[0285] This invention contemplates the application of A in any suitable manner. 2A R / A 2B R inhibitors and combinations thereof. Suitable routes of administration include oral, parenteral (e.g., intramuscular, intravenous, subcutaneous (e.g., injection or implantation), intraperitoneal, intracisional, intra-articular, intraperitoneal, intracerebral (within brain parenchyma), and intraventricular), nasal, vaginal, sublingual, intraocular, rectal, local (e.g., transdermal), oral, and inhalation. Depot injection, usually administered subcutaneously or intramuscularly, can also be used to release the A2AR / A2BR inhibitors disclosed herein over a defined time period.

[0286] The specific embodiments of the present invention cover oral administration.

[0287] Combination therapy

[0288] This invention considers A 2A R / A 2B R inhibitors can be used alone or in combination with one or more active therapeutic agents. Additional active therapeutic agents can be small chemical molecules; macromolecules such as proteins, antibodies, peptides, DNA, RNA, or fragments of these macromolecules; or cell or gene therapies. In such combination therapies, the various active agents typically have different complementary mechanisms of action. This combination therapy can be particularly beneficial by allowing for dose reductions of one or more agents, thus reducing or eliminating side effects associated with one or more agents. Furthermore, this combination therapy may have a synergistic therapeutic or preventative effect on underlying diseases, symptoms, or conditions.

[0289] As used herein, “combination” means including therapies that can be administered separately, such as those formulated separately for individual administration (e.g., as may be provided in a kit) and therapies that can be administered together in a single formulation (i.e., “co-formulated”).

[0290] In some implementations, A 2A R / A 2B R inhibitors are administered or applied sequentially, for example, one agent is administered before one or more other agents. In other embodiments, A is administered simultaneously. 2A R / A 2B R inhibitors, for example, two or more drugs administered at or approximately the same time; two or more agents may exist in two or more separate formulations or be combined into a single formulation (i.e., co-formulation). Whether the two or more agents are administered sequentially or simultaneously, they are considered to be administered in combination for the purposes of this invention.

[0291] In this case, the A of the present invention 2A R / A 2B R inhibitors can be used in combination with at least one other (active) agent in any suitable manner. In one embodiment, the use of at least one active agent and at least one of the A agents of the present invention is maintained for a period of time. 2A R / A 2B R inhibitors are used for treatment. In another embodiment, when using the A of the present invention... 2A R / A 2B When treatment with an R inhibitor maintains a constant dosing regimen, treatment with at least one active agent is reduced or discontinued (e.g., when the subject is stable). In another embodiment, when using A of the present invention... 2A R / A 2B When treatment with an R inhibitor is reduced (e.g., lower doses, less frequent dosing, or shorter treatment regimens), treatment with at least one active agent is reduced or discontinued (e.g., when the subject is stable). In yet another embodiment, treatment with at least one active agent is reduced or discontinued (e.g., when the subject is stable), and the use of A of the present invention is increased. 2A R / A 2B Treatment with R inhibitors (e.g., higher doses, more frequent doses, or longer treatment regimens). In yet another embodiment, treatment with at least one active agent is maintained and treatment with A of the present invention is reduced or discontinued. 2A R / A 2B R inhibitor therapy (e.g., lower doses, less frequent dosing, or shorter treatment regimens). In yet another embodiment, treatment with at least one active agent and with the A of the present invention is reduced or discontinued. 2A R / A 2B R inhibitor therapy (e.g., lower doses, less frequent dosing, or shorter treatment regimens).

[0292] Oncology-related diseases This invention provides the use of A 2A R / A 2BA method of treating and / or preventing proliferative disorders, cancers, tumors, or precancerous conditions, diseases, or illnesses using an R inhibitor and at least one additional therapeutic or diagnostic agent. In some embodiments, the additional therapeutic or diagnostic agent is a radiation agent, an immunomodulator or chemotherapeutic agent, or a diagnostic agent. Suitable immunomodulators that may be used in this invention include: CD4OL, B7, and B7RP1; activated monoclonal antibodies (mAbs) against stimulating receptors, such as anti-CD40, anti-CD38, anti-ICOS, and 4-IBB ligands; dendritic cell antigen loads (in vitro or in vivo); anticancer vaccines such as dendritic cell cancer vaccines; cytokines / chemokines such as ILL, IL2, IL12, IL18, ELC / CCL19, SLC / CCL21, MCP-1, IL-4, IL-18, TNF, IL-15, MDC, IFNa / b, M-CSF, IL-3, GM-CSF, IL-13, and anti-IL-10; bacterial lipopolysaccharides (LPS); indoleamine 2,3 dioxygenase 1 (IDO1) inhibitors and immune-activating oligonucleotides.

[0293] In some embodiments, the present invention provides a method for tumor suppression of tumor growth, comprising administering A as described herein. 2A R / A 2B R inhibitors are administered in combination with signal transduction inhibitors (STIs) to achieve additive or synergistic inhibition of tumor growth. As used herein, the term "signal transduction inhibitor" refers to an agent that selectively inhibits one or more steps in a signal transduction pathway. The signal transduction inhibitors (STIs) of this invention include: (i) bcr / abl kinase inhibitors (e.g., GLEEVEC); (ii) epidermal growth factor (EGF) receptor inhibitors, including kinase inhibitors and antibodies; (iii) HER-2 / neu receptor inhibitors (e.g., Herceptin); (iv) Akt family kinase or Akt pathway inhibitors (e.g., rapamycin); (v) cell cycle kinase inhibitors (e.g., flavopiridol); and (vi) phosphatidylinositol kinase inhibitors. Agents involved in immune regulation may also be used in combination with the ATIs described herein. 2A R / A 2B R inhibitor combinations are used to inhibit tumor growth in cancer patients.

[0294] Examples of chemotherapeutic agents include, but are not limited to, alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, indomethacin, and piperazine; aziridines such as benzoxadipa, caloquinone, mitdopa, and uredopa; ethyleneimine and methylmelamine, including hexamethylmelamine, triethylenemelamine, triethylenephosphamide, triethylenethiophosphamide, and tris(hydroxymethylmelamine); nitrogen mustards such as chiorambucil, naphthiamethoxam, chlorophosphamide, estradiol, ifosfamide, dichloromethyldiethylamine, dichloromethyldiethylamine oxide hydrochloride, melphalan, novombhichin, phenesterine, prednisolone, trelophosphamide, and uracil mustard; nitrosamines. Ureas, such as carmustine, chlorurea, formustine, lomustine, nimustine, lamustine; antibiotics, such as aclacinomysin, actinomycin, autramycin, diazoserine, bleomycin, actinomycin C, galicariin, carabicin, caminomycin, carzinophilin, chromomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-leucine, doxorubicin, epirubicin, arubicin, idarubicin, ephedrine, mitosis. Mycotoxin, mycophenolic acid, nogamycin, oligomycin, pepromycin, prednisone, puromycin, quelamycin, rodorubicin, streptomycin, strepzotocin, tuberculin, ubenimex, fenestrated statin, zorubicin; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs, such as folate, methotrexate, pterin, trimethoprim; purine analogs, such as fludarabine, 6-mercaptopurine, thiamine, thioguanine; pyrimidine analogs, such as ancitabine, azacitidine, 6-azouridine, carmoflurane, cytarabine, dideoxyuridine, deoxyfluorouridine, enoxabin, fluorouridine, 5-FU; androgen Hormones, such as capprogesterone, drotaldone propionate, cyclothionol, meandranone, and testosterone; antiadrenergics, such as aminoglutethimide, mitotane, and trilosterone; folic acid supplements, such as folinic acid; acetoglucan lactone; aldehyde phosphoramide glycoside; aminolevulinic acid; acridine; bestrabucil; bifenthrin; edatraxate; defofamine; dimethicone; diaziquone; elformithine; elifonitrile; etoglucopyranoside; gallium nitrate; hydroxyurea; lentinan; chlordamine; mitoguazone; mitoxantrone; mopidamol;Nitracrine; Pentostatin; Phenamet; Pirarubicin; Podophyllin; 2-Ethylhydrazide; Procarbazine; Razoxane; Sizofiran; Spirogermanium; Tenuazonic acid acid); triaziquone; 2,2',2”-trichlorotriethylamine; urethan; vindesine; dacarbazine; mannitol; dibromomannitol; mitolactone; piperobromane; gacytosine; Ara-C arabinoside; cyclophosphamide; thiotepa; paclitaxel-like compounds, such as paclitaxel and docetaxel; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum and platinum coordination compounds, such as cisplatin and carboplatin. And oxaliplatin; vincristine; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; donomycin; aminopterin; capecitabine; ibandronate; CPT11; topoisomerase inhibitors; difluoroacetyl-ornithine (DMFO); retinoic acid; esperamicin; capecitabine; anthracyclines; and pharmaceutically acceptable salts, acids, or derivatives of any of the above.

[0295] Chemotherapy agents also include anti-hormonal agents used to modulate or inhibit the effects of tumor hormones, such as anti-estrogens including, for example, tamoxifen, raloxifene, aromatase inhibitor 4(5)-imidazole, 4-hydroxytamoxifen, trioxifene, keoxifene, onapristone, and toremifene; and anti-androgens such as flutamide, nilutamide, bicalutamide, leuprorelin, and goserelin; and pharmaceutically acceptable salts, acids, or derivatives of any of the foregoing. In some embodiments, combination therapy includes a chemotherapy regimen comprising one or more chemotherapeutic agents. In some embodiments, combination therapy includes administration of hormones or related hormonal agents.

[0296] Can be with A 2A R / A 2B Other treatment modalities that use R inhibitors in combination include radiotherapy, monoclonal antibodies against tumor antigens, complexes of monoclonal antibodies and toxins, T-cell adjuvants, bone marrow grafts, or antigen-presenting cells (e.g., dendritic cell therapy) that include TLR agonists to stimulate such antigen-presenting cells.

[0297] In some embodiments, the invention contemplates the use of the compounds described herein in combination with adoptive cell therapy, a novel and promising form of personalized immunotherapy in which immune cells with anti-tumor activity are administered to a cancer patient. Adoptive cell therapy is being investigated using tumor-infiltrating lymphocytes (TILs) and T cells engineered to express, for example, chimeric antigen receptors (CARs) or T-cell receptors (TCRs). Adoptive cell therapy typically involves collecting T cells from an individual, genetically modifying them to target specific antigens or enhance their anti-tumor effects, expanding them to a sufficient number, and infusing the genetically modified T cells into a cancer patient. T cells can be collected from a patient and the expanded cells (e.g., autologous) can be re-infused into that patient, or T cells can be collected from a donor patient (e.g., allogeneic).

[0298] In some embodiments, the invention contemplates the use of the compounds described herein in combination with RNA interference-based therapies for silencing gene expression. RNAi begins by cleaving a long double-stranded RNA into small interfering RNAs (siRNAs). One strand of the siRNA is incorporated into a ribonucleoprotein complex called an RNA-induced silencing complex (RISC), which is then used to identify mRNA molecules that are at least partially complementary to the incorporated siRNA strand. The RISC can bind to or cleave the mRNA, both of which inhibit translation.

[0299] Immune checkpoint inhibitors. This invention considers the A described herein. 2A R / A 2B The use of R-function inhibitors in combination with immune checkpoint inhibitors.

[0300] A multitude of genetic and epigenetic alterations characteristic of all cancers provide multiple sets of antigens that the immune system can use to distinguish tumor cells from their normal counterparts. In the case of T cells, the final magnitude (e.g., the level of cytokine production or proliferation) and quality (e.g., the type of immune response produced, such as the pattern of cytokine production) of the response initiated via antigen recognition via T cell receptors (TCRs) are regulated by a balance between co-stimulatory and inhibitory signals (immune checkpoints). Under normal physiological conditions, immune checkpoints are essential for the prevention of autoimmunity (i.e., maintaining self-tolerance) and also for protecting tissues from damage when the immune system responds to pathogenic infections. The expression of immune checkpoint proteins can be dysregulated by tumors as an important mechanism of immune resistance.

[0301] T cells have been a major focus of therapeutic control of endogenous antitumor immunity because: i) their ability to selectively recognize protein-derived peptides in all cellular compartments; ii) their ability to directly recognize and kill antigen-expressing cells (via CD8+ effector T cells; also known as cytotoxic T lymphocytes (CTLs)); and iii) their ability to coordinate multiple immune responses via CD4+ helper T cells, which integrate adaptive and innate effector mechanisms.

[0302] In the clinical setting, blocking immune checkpoints—which amplifies antigen-specific T-cell responses—has proven to be a promising approach for the treatment of human cancers.

[0303] T-cell-mediated immunity involves multiple sequential steps, each regulated by counterbalancing stimulatory and inhibitory signals to optimize the response. While almost all inhibitory signals in the immune response ultimately regulate intracellular signaling pathways, many are initiated via membrane receptors whose ligands are membrane-bound or soluble (cytokines). Although costimulatory and inhibitory receptors and ligands regulating T-cell activation are generally not overexpressed in cancer relative to normal tissues, inhibitory ligands and receptors regulating T-cell effector function in tissues are often overexpressed on tumor cells or on non-transformed cells associated with the tumor microenvironment. The function of soluble and membrane-bound receptor-ligand immune checkpoints can be modulated using agonist antibodies (for costimulatory pathways) or antagonist antibodies (for inhibitory pathways). Therefore, unlike most antibodies currently approved for cancer therapy, antibodies that block immune checkpoints do not directly target tumor cells but instead target lymphocyte receptors or their ligands to enhance endogenous antitumor activity. [See Pardoll, (April 2012) Nature Rev. Cancer 12: 252-64].

[0304] Examples of immune checkpoints (ligands and receptors), some of which are selectively upregulated in various types of tumor cells, are candidates for blockade, including PD1 (programmed cell death protein 1); PDL1 (PD1 ligand); BTLA (B and T lymphocyte attenuator); CTLA4 (cytotoxic T lymphocyte-associated antigen 4); TIM3 (T cell membrane protein 3); LAG3 (lymphocyte activation gene 3); TIGIT (T cell immune receptor with Ig and ITIM domains); and cytotoxic cell inhibitory receptors, which can be divided into two classes based on their structural features: i) cytotoxic cell immunoglobulin-like receptors (KIRs) and ii) type C lectin receptors (members of the type II transmembrane receptor family). Other less well-defined immune checkpoints have been described in the literature, including receptors (e.g., the 2B4 (also known as CD244) receptor) and ligands (e.g., certain B7 family inhibitory ligands, such as B7-H3 (also known as CD276) and B7-H4 (also known as B7-S1, B7x, and VCTN1)). [See Pardoll, (April 2012) Nature Rev. Cancer 12: 252-64].

[0305] This invention considers the A described herein. 2A R / A 2B Use of R-function inhibitors in combination with inhibitors of the aforementioned immune checkpoint receptors and ligands, and those described below. Some modulators of immune checkpoints are currently available, while others are in late-stage development. For example, when the fully humanized CTLA4 monoclonal antibody ipilimumab (YERVOY; Bristol-Myers Squibb) was approved in 2011 for the treatment of melanoma, it became the first immune checkpoint inhibitor to receive regulatory approval in the United States. Fusion proteins containing CTLA4 and an antibody (CTLA4-Ig; abatcept (ORENCIA; Bristol-Myers Squibb)) have been used to treat rheumatoid arthritis, and other fusion proteins have shown efficacy in kidney transplant patients sensitive to Epstein-Barr virus. PD-1 antibodies are under development (e.g., nivolumab (Bristol-Myers Squibb) and lambrolizumab (Merck)), and anti-PD-L1 antibodies (e.g., MPDL3280A (Roche)) are also being evaluated. Nivolumab has shown promise in patients with melanoma, lung cancer, and kidney cancer.

[0306] In one aspect of the invention, the claimed A 2A R / A 2BR inhibitors are used in combination with immuno-oncology agents, which are either agonists of (i) stimulating (including co-stimulating) receptors or antagonists of inhibitory (including co-inhibitory) signals on T cells, both of which lead to an amplified antigen-specific T cell response. Some of the stimulating and inhibitory molecules are members of the immunoglobulin superfamily (IgSF). An important family of membrane-bound ligands that bind to co-stimulating or co-inhibitory receptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1), B7-DC (PD-L2), B7-H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA), and B7-H6. Other families of membrane-bound ligands that bind to co-stimulatory or co-inhibitory receptors are TNF family molecules that bind to members of the homologous TNF receptor family, including CD40 and CD4OL, OX-40, OX-40L, CD70, CD27L, CD30, CD3OL, 4-1BBL, CD137 (4-1BB), TRAIL / Apo2-L, TRAILR1 / DR4, TRAILR2 / DR5, TRAILR3, TRAILR4, OPG, RANK, RANKL, TWEAKR / Fn14, TWEAK, BAFFR, EDAR, XEDAR, TACI, APRIL, BCMA, LT13R, LIGHT, DcR3, HVEM, VEGI / TL1A, TRAMP / DR3, EDAR, EDA1, XEDAR, EDA2, TNFR1, lymphotoxin a / TNF13, TNFR2, TNFa, LT13R, and lymphotoxin a. 1132, FAS, FASL, RELT, DR6, TROY, NGFR.

[0307] In other respects, immuno-oncology agents are cytokines that inhibit T cell activation (e.g., IL-6, IL-10, TGF-β, VEGF, and other immunosuppressive cytokines) or stimulate T cell activation, which are used to promote immune responses.

[0308] In one respect, T cell responses can be achieved by disclosing A... 2A R / A 2BR inhibitors and antagonists of one or more (i) proteins that inhibit T cell activation (e.g., immune checkpoint inhibitors), such as CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, TIM-3, galectin 9, CEACAM-1, BTLA, CD69, galectin-1, TIGIT, CD113, GPR56, VISTA, 2B4, CD48, GARP, PD1H, LAIR1, TIM-1, and TIM-4, and / or protein agonists that stimulate T cell activation, such as B7-1, B7-2, CD28, 4-1BB (CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX4OL, GITR, GITRL, CD70, CD27, CD40, DR3, and CD2. These can be used in conjunction with the A of the present invention. 2A R / A 2B Other agents used in combination with KIR inhibitors for cancer treatment include antagonists of inhibitory receptors on NK cells or agonists of activating receptors on NK cells. For example, the compounds described herein can be combined with KIR antagonists such as lirilumab.

[0309] It is also used in combination therapy with other agents, including agents that inhibit or deplete macrophages or monocytes, including but not limited to: CSF-1R antagonists, such as CSF-1R antagonist antibodies including RG7155 (W011 / 70024, W011 / 107553, W011 / 131407, W013 / 87699, W013 / 119716, W013 / 132044) or FPA-008 (W011 / 140249; W013169264; W014 / 036357).

[0310] On the other hand, the disclosed A 2A R / A 2B R inhibitors may be combined with one or more agonists that bind to positive co-stimulatory receptors, blockers that attenuate signal transduction through inhibitory receptors, antagonists, and one or more agents that systemically increase the frequency of anti-tumor T cells, agents that overcome different immunosuppressive pathways in the tumor microenvironment (e.g., blocking inhibitory receptor involvement (e.g., PD-L1 / PD-1 interaction), depleting or inhibiting Tregs (e.g., using anti-CD25 monoclonal antibodies (such as dacrolimus) or depleting them through ex vivo anti-CD25 beads), or reversing / preventing T cell dysfunction or depletion), and agents that trigger innate immune activation and / or inflammation at the tumor site.

[0311] In one aspect, the immuno-oncology agent is a CTLA-4 antagonist, such as an antagonistic CTLA-4 antibody. Suitable CTLA-4 antibodies include, for example, ipilimumab (YERVOY) or tremelimumab.

[0312] On the other hand, the immuno-oncology agent is a PD-1 antagonist, such as an antagonistic PD-1 antibody. Suitable PD-1 antibodies include, for example, OPDIVO (nivolumab), KEYTRUDA (pembrolizumab), or MEDI-0680 (AMP-514; WO2012 / 145493). The immuno-oncology agent may also include Pidilizumab (CT-011), although its specificity for PD-1 binding is questionable. Another approach to targeting the PD-1 receptor is a recombinant protein, called AMP-224, composed of the extracellular domain of PD-L2 (B7-DC) fused to the Fc portion of IgG1.

[0313] On the other hand, the immuno-oncology agent is a PD-L1 antagonist, such as an antagonistic PD-L1 antibody. Suitable PD-L1 antibodies include, for example, MPDL3280A (RG7446; WO2010 / 077634), durvalumab (MEDI4736), BMS-936559 (WO2007 / 005874), and MSB0010718C (WO2013 / 79174).

[0314] On the other hand, the immuno-oncology agent is a LAG-3 antagonist, such as an antagonistic LAG-3 antibody. Suitable LAG3 antibodies include, for example, BMS-986016 (W010 / 19570, W014 / 08218), or IMP-731 or IMP-321 (W008 / 132601, W009 / 44273).

[0315] On the other hand, the immuno-oncology agent is a CD137 (4-1BB) agonist, such as an agonistic CD137 antibody. Suitable CD137 antibodies include, for example, urelumab and PF-05082566 (W012 / 32433).

[0316] On the other hand, the immuno-oncology agent is a GITR agonist, such as an agonistic GITR antibody. Suitable GITR antibodies include, for example, BMS-986153, BMS-986156, TRX-518 (W006 / 105021, W009 / 009116) and MK-4166 (W011 / 028683).

[0317] On the other hand, the immuno-oncology agent is an OX40 agonist, such as an agonistic OX40 antibody. Suitable OX40 antibodies include, for example, MEDI-6383 or MEDI-6469.

[0318] On the other hand, the immuno-oncology agent is an OX4OL antagonist, such as an antagonistic OX4OL antibody. Suitable OX4OL antibodies include, for example, RG-7888 (W006 / 029879).

[0319] In another embodiment, the immuno-oncology agent is a CD40 agonist, such as an agonist CD40 antibody. In yet another embodiment, the immuno-oncology agent is a CD40 antagonist, such as an antagonist CD40 antibody. Suitable CD40 antibodies include, for example, lucarumumab or dacetuzumab.

[0320] On the other hand, the immuno-oncology agent is a CD27 agonist, such as an agonistic CD27 antibody. Suitable CD27 antibodies include, for example, varlilumab.

[0321] On the other hand, the immuno-oncology reagent is MGA271 (against B7H3) (W011 / 109400).

[0322] This invention includes any of the pharmaceutically acceptable salts, acids, or derivatives described above.

[0323] Metabolic and cardiovascular diseases This invention provides the use of A 2A R / A 2B Methods of treating and / or preventing certain cardiovascular and / or metabolic diseases, conditions and ailments, and related symptoms, using R inhibitors and at least one other therapeutic or diagnostic agent.

[0324] Examples of therapeutic agents that can be used to treat hypercholesterolemia (and atherosclerosis) include statins that inhibit the enzymatic synthesis of cholesterol (e.g., CRESTOR, LESCOL, LIPITOR, MEVACOR, PRAVACOL, and ZOCOR); bile acid resins (e.g., COLESTID, LO-CHOLEST, PREVALIITE, QUESTRAN, and WELCHOL) that chelate cholesterol and prevent its absorption; ezetimibe (ZETIA) that blocks cholesterol absorption; fibrinolytic acids (e.g., TRICOR) that reduce triglycerides and moderately increase HDL; niacin (e.g., NIACOR) that moderately lowers LDL cholesterol and triglycerides; and / or combinations of the foregoing (e.g., VYTORIN (ezetimibe with simvastatin)). These can be combined with the A described herein. 2A R / A 2B Alternative cholesterol therapies for use in combination with R inhibitors include a variety of supplements and herbs (such as garlic, eicosanol, and guggul).

[0325] This invention includes any of the pharmaceutically acceptable salts, acids, or derivatives described above.

[0326] Immune and inflammatory related diseases This invention provides the use of A 2A R / A 2B R inhibitors and at least one other therapeutic or diagnostic agent for the treatment and / or prevention of immune-related diseases, conditions and illnesses; and methods for treating diseases, conditions and illnesses with inflammatory components.

[0327] Examples of chemotherapeutic agents useful in combination therapy include, but are not limited to: nonsteroidal anti-inflammatory drugs (NSAIDs) such as aspirin, ibuprofen, and other propionic acid derivatives (amenoprofen, benzoxalofen, bucloxic acid, carboprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin, pirprofen, fenoprofen), suprafen, tiaprofenic acid, etc. Acetic acid derivatives (indomethacin, acemetacin, alclofenac, clidanac, diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac, isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin, and zomepirac), fenamic acid derivatives (flufenamic acid, meclofenamic acid, mefenamic acid, niflumic acid), and tranexamic acid. Combinations include biphenyl acid derivatives (diflunisal and flufenisal), oxicams (isoxicam, piroxicam, sudoxicam, and tenoxican), salicylates (acetylsalicylic acid, sulfasalazine), and pyrazolone derivatives (apazone, bezpiperylon, feprazone, murprozin, hydroxyphenylbutazone, and phenylbutazone). Other combinations include cyclooxygenase-2 (COX-2) inhibitors.

[0328] Other active pharmaceutical agents used in combination include steroids such as dehydrocortisone, prednisone, methylprednisolone, betamethasone, dexamethasone, or hydrocortisone. Such combinations can be particularly advantageous because one or more side effects of the steroid can be reduced or even eliminated by gradually decreasing the required dose.

[0329] Other examples of active agents that can be used in combination to treat conditions such as rheumatoid arthritis include: cytokine-suppressing anti-inflammatory drugs (CSAIDs); antibodies or antagonists of other human cytokines or growth factors, such as TNF, LT, IL-10, IL-2, IL-6, IL-7, IL-8, IL-15, IL-16, IL-18, EMAP-II, GM-CSF, FGF, or PDGF.

[0330] Specific combinations of active agents may interfere with different sites in the autoimmune and subsequent inflammatory cascade, and include: TNF antagonists, such as chimeric, humanized, or human TNF antibodies, remicades, anti-TNF antibody fragments (e.g., CDP870), and soluble p55 or p75 TNF receptors, their derivatives, p75 TNF RIgG (ENBREL) or p55 TNF R1gG (LENERCEPT), soluble IL-13 receptors (sIL-13), and TNFα-converting enzyme (TACE) inhibitors; similarly, IL-1 inhibitors (e.g., interleukin-1 converting enzyme inhibitors) may be effective. Other combinations include interleukin-11, anti-P7, and p-selectin glycoprotein ligands (PSGL). (The last sentence appears to be incomplete and possibly refers to a different topic.) 2A R / A 2B Other examples of useful agents in R inhibitor combinations include: interferon 131a (AVONEX); interferon 13lb (BETASERON); glatiramer acetate (copaxone); hyperbaric oxygen; intravenous immunoglobulin; clabribine; and antibodies or antagonists of other human cytokines or growth factors (such as antibodies to CD40 ligands and CD80).

[0331] Microbial diseases This invention provides the use of A 2A R / A 2B Methods of treating and / or preventing viral, bacterial, fungal, and parasitic diseases, conditions, and illnesses, and related symptoms, using R inhibitors and at least one other therapeutic or diagnostic agent (e.g., one or more other antiviral agents and / or one or more agents unrelated to viral therapy).

[0332] This combination therapy includes antiviral agents that target various stages of the viral life cycle and have different mechanisms of action, including but not limited to: viral uncoating inhibitors (e.g., amantadine and liamtidine); reverse transcriptase inhibitors (e.g., acyclovir, zidovudine, and lamivudine); drugs that target integrase; drugs that block the binding of transcription factors to viral DNA; drugs that affect translation (e.g., antisense molecules) (e.g., fomivirsen); drugs that modulate translation / ribonuclease function; protease inhibitors; viral assembly modulators (e.g., rifampin); antiretroviral drugs, such as nucleoside analog reverse transcriptase inhibitors (e.g., azidothymidine (AZT), dd1, ddC, 3TC, d4T); non-nucleoside reverse transcriptase inhibitors (e.g., efavirenz, nevirapine); nucleotide analog reverse transcriptase inhibitors; and drugs that prevent the release of viral particles (e.g., zanamivir and oseltamivir). Treatment and / or prevention of certain viral infections (e.g., HIV) often require a combination of antiviral agents (“cocktail”).

[0333] With A 2A R / A 2BOther antiviral agents used in combination with R inhibitors include, but are not limited to, the following: abacavir, adefovir, amantadine, ampradil, arbidol, atazanavir, lipitor, boceprevirertet, cidofovir, combivir, darunavir, delavudine, didanoxin, docosanol, edoxudine, emtricitabine, enfuvirtide, entecavir, famciclovir, fosanavir, fosfonet, ganciclovir, ibatabin, imunovir, idoxuridine, imiquimod, and indinavir. Inosine, various interferons (e.g., pegylated interferon alpha-2a), lopinavir, loferidine, maraviro, moroxydine, methisazone, nelfinavir, nexavir, penciclovir, peramivir, proconali, podophyllotoxin, raltegravir, ribavirin, ritonavir, pyramidine, saquinavir, stavudine, telaprevir, tenofovir, titranavir, trifluridine, trizivir, triamantadine, truvada, valacyclovir, valganciclovir, vicriviroc, vidarabine, viramidine, and zalcitabine.

[0334] This invention considers the A described herein. 2A R / A 2B Use of R-function inhibitors in combination with antiparasitic agents. These agents include, but are not limited to, thiabendazole, pyrantel pamoate, mebendazole, praziquantel, niclosamide, thiamethoxam, ivermectin, albendazole, efornithine, melarisol, pentamidine, benznidazole, nifurimoxazole, and nitroimidazole. Other agents known to those skilled in the art for the treatment of parasitic diseases are also available.

[0335] Embodiments of the present invention consider the A described herein. 2A R / A 2BR inhibitors are used in combination with agents useful in the treatment or prevention of bacterial diseases. Antimicrobial agents can be classified in various ways, including based on mechanism of action, chemical structure, and activity spectrum. Examples of antibacterial agents include those that target the bacterial cell wall (e.g., cephalosporins and penicillins) or cell membrane (e.g., polymyxins) or interfere with essential bacterial enzymes (e.g., sulfonamides, rifamycins, and quinolines). Most antimicrobial agents that target protein synthesis (e.g., tetracyclines and macrolides) are bacteriostatic agents, while agents such as aminoglycosides are bactericidal agents. Another way to classify antimicrobial agents is based on their targeting specificity; “narrow-spectrum” agents target specific types of bacteria (e.g., Gram-positive bacteria such as streptococci), while “broad-spectrum” agents have activity targeting a wider range of bacteria. Those skilled in the art know the types of antimicrobial agents suitable for specific bacterial infections.

[0336] The embodiments of the present invention take into account the A described herein. 2A R / A 2B R inhibitors are used in combination with agents useful for the treatment or prevention of fungal conditions. Antifungal agents include polyenes (e.g., amphotericin, nystatin, and pimmicin); azoles (e.g., fluconazole, itraconazole, and ketoconazole); allylamines (e.g., naftifine and terbinafine) and morpholines (e.g., amorolfine); and antimetabolites (e.g., 5-fluorocytosine).

[0337] This invention includes pharmaceutically acceptable salts, acids, or derivatives of the aforementioned pharmaceutical agents (and members of the pharmaceutical class).

[0338] dose

[0339] A of the present invention 2A R / A 2B R inhibitors can be administered to subjects in amounts that depend on factors such as, for example, the target of administration (e.g., the degree of resolution desired); the age, weight, sex, health, and physical condition of the subject to which the formulation is administered; the route of administration; and the nature of the disease, condition, illness, or its symptoms. The dosing regimen may also take into account the presence, nature, and extent of any adverse effects associated with the administered drug. Effective doses and dosing regimens can be readily determined from, for example, safety and dose escalation studies, in vivo studies (e.g., animal models), and other methods known to those skilled in the art.

[0340] Typically, dosing parameters specify a dose less than the amount that may have irreversible toxicity to the subject (maximum tolerated dose (MTD)) and not less than the amount required to produce a measurable effect on the subject. Taking into account the route of administration and other factors, these amounts are determined by pharmacokinetic and pharmacodynamic parameters, such as those related to ADME.

[0341] The effective dose (ED) is the amount of a drug that produces a therapeutic response or desired effect in a subset of the population in which it is administered. The “median effective dose” or ED50 of a drug is the amount of a drug that produces a therapeutic response or desired effect in 50% of the population in which it is administered. Although ED50 is generally used to measure a reasonable expectation of a drug’s effect, a clinician may consider an appropriate dose, after taking all relevant factors into account, not necessarily an appropriate dose. Therefore, in some cases, the effective dose is greater than the calculated ED50; in others, the effective dose is less than the calculated ED50; and in still others, the effective dose is the same as the calculated ED50.

[0342] In addition, the A of the present invention 2A R / A 2B An effective dose of an R inhibitor can be the amount that produces the desired result relative to a healthy subject when administered one or more doses. For example, for a subject experiencing a specific condition, an effective dose can be the amount that improves the diagnostic parameters, measurements, markers, etc., of that condition by at least about 5%, at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or more than 90%, where 100% is defined as the diagnostic parameters, measurements, markers, etc., exhibited by a normal subject.

[0343] In some embodiments, the A contemplated by the present invention 2A R / A 2B R inhibitors can be administered once or more daily at dose levels of about 0.01 mg / kg to about 50 mg / kg, or about 1 mg / kg to about 25 mg / kg, based on the subject's daily body weight (e.g., orally) to achieve the desired therapeutic effect.

[0344] For oral administration, the composition may be provided in the form of tablets, capsules, etc., containing 1.0-1000 mg of active ingredient, specifically 1.0, 3.0, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 mg of active ingredient.

[0345] In some implementations, the required A 2A R / A 2B The dosage of R inhibitors is included in the "unit dosage form." The phrase "unit dosage form" refers to physically discontinuous units, each containing a predetermined amount of A, alone or in combination with one or more other reagents, sufficient to produce the desired effect. 2A R / A 2BR inhibitors. It should be understood that the parameters of the unit dosage form will depend on the specific drug and the desired effect.

[0346] Reagent test kit

[0347] The present invention also considers the inclusion of A 2A R / A 2B Kits and pharmaceutical compositions for R inhibitors. As described below, the kits are typically in the form of a physical structure containing various components and can be used, for example, to carry out the methods described above.

[0348] The kit may include one or more compounds disclosed herein (e.g., provided in a sterile container) in the form of a pharmaceutical composition suitable for administration to a subject. Compounds as described herein may be provided in a ready-to-use form (e.g., tablets or capsules) or in a form requiring, for example, reconstitution or dilution prior to administration (e.g., powder). When the compounds as described herein are in a form requiring reconstitution or dilution by the user, the kit may also include diluents (e.g., sterile water), buffers, pharmaceutically acceptable excipients, etc., packaged together with or separately from the compounds as described herein. When considering combination therapy, the kit may contain several reagents individually, or they may have already been combined in the kit. Each component of the kit may be contained in a separate container, and all the various containers may be in a single package. The kits of the present invention may be designed to maintain the components contained therein under the conditions required (e.g., refrigeration or freezing).

[0349] The kit may include a label or packaging insert containing identification information for its components and instructions for use (e.g., dosage parameters, clinical pharmacology of the active ingredient, including mechanism of action, pharmacokinetics and pharmacodynamics, adverse effects, contraindications, etc.). The label or insert may include manufacturer information such as batch number and expiration date. The label or packaging insert may be integrated into the physical structure containing the components, contained separately within the physical structure, or attached to the components of the kit (e.g., ampoules, tubes, or vials).

[0350] Labels or inserts may be additionally included or incorporated into computer-readable media such as disks (e.g., hard disks, cards, storage discs), optical discs, such as CDs or DVD-ROM / RAM, DVDs, MP3s, magnetic tapes, or combinations thereof, electrical storage media such as RAM and ROM, or media such as magnetic / optical storage media, FLASH media, or memory cards. In some embodiments, actual instructions are not included in the kit, but means for obtaining instructions from a remote source, such as via the Internet, are provided.

[0351] experiment

[0352] The following embodiments are provided to provide a complete disclosure and description of how the invention can be made and used to those skilled in the art, and are not intended to limit the scope of what the inventors consider their invention, nor do they imply that the experiments described below have been completed or that they are all possible experiments. It should be understood that the exemplary description written in the present tense is not necessarily performed, but rather can be performed to generate data, etc., of the properties described therein. Efforts have been made to ensure the accuracy of the figures used (e.g., quantities, temperatures, etc.), but some experimental errors and biases should be taken into account.

[0353] Unless otherwise stated, parts are by weight, molecular weight is by weight-average molecular weight, temperature is in degrees Celsius (°C), and pressure is at or near atmospheric pressure. Standard abbreviations were used, including the following: Wt = wild type; bp = base pair; kb = kilobase; nt = nucleotide; as = amino acid; s or sec = second; min = minute; h or hr = hour; ng = nanogram; [tg = microgram; mg = milligram; g = gram; kg = kilogram; dl or dL = deciliter; pl or 1AL = microliter; ml or mL = milliliter; l or L = liter; [iM = micromolar; mM = millimole; M = mole; kDa = kilodalton; im = intramuscular; ip = intraperitoneal; SC or SQ = subcutaneous; QD = daily; BID = twice daily; QW = weekly; QM = monthly; HPLC = high performance liquid chromatography; BW = body weight; U = unit; ns = no statistical significance; PBS = phosphate buffered saline; IHC = immunohistochemistry; DMEM = Dulbeco's Modification of Eagle's Medium); EDTA = ethylenediaminetetraacetic acid.

[0354] Materials and methods

[0355] The following general materials and methods are used where indicated or in the examples below:

[0356] Standard methods in molecular biology are described in the scientific literature (see, for example, Sambrook and Russell (2001) Molecular Cloning, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; and Ausubel et al. (2001) Laboratory Guide to Molecular Biology, Volumes 1–4, John Wiley & Sons, NY, which describes cloning and DNA mutagenesis in bacterial cells (Vol. 1), cloning in mammalian cells and yeast (Vol. 2), glycoconjugates and protein expression (Vol. 3) and bioinformatics (Vol. 4)).

[0357] Scientific literature describes methods for protein purification, including immunoprecipitation, chromatography, electrophoresis, centrifugation and crystallization, as well as chemical analysis, chemical modification, post-translational modification, fusion protein generation and protein glycosylation (see, for example, Coligan et al. (2000), Current Protocols in Protein Science, Volumes 1-2, John Wiley & Sons, New York).

[0358] Software packages and databases are available for identifying, for example, antigen fragments, leader sequences, protein folds, functional domains, glycosylation sites, and sequence alignments (see, for example, GCG Wisconsin Package (Accelrys, San Diego, CA); and DeCypher™ (TimeLogic, Crystal Bay, Nevada).

[0359] The literature contains a wealth of analytical and other experimental techniques that can be used as a basis for evaluating the compounds described herein. For example, mass spectrometry-based ligand binding analysis (see, for instance, Massink, A et al., Purinergic Signaling (2015) 11:581. https: / / doi.org / 10.1007 / s11302-015-9477-0; Dionisotti S et al., Journal of Pharmacology and Experimental Therapeutics (1996) 298:726-732) can be used to determine various properties of the compounds of the present invention.

[0360] Functional analysis can also be used to evaluate the compounds of this invention. The cAMP analysis detailed below was used to evaluate some of the compounds described herein.

[0361] Yuan et al. describe an optional exemplary functional analysis that assesses IFN-γ secretion (International Journal of Medicinal Chemistry (Int J Med Chem); Vol. 2017 (2017), Article ID 4852537);

[0362] (https: / / doi.org / 10.1155 / 2017 / 4852537). In short, during CD3 ligand-activated T-cell receptor (TCR) activation, incubating C57BL / 6 mouse spleen T cells with a receptor agonist inhibited IFN-γ secretion induced by receptor A2A—immunosuppression induced by intracellular cAMP. Effective receptor antagonists block receptor activation signals, thereby restoring cytokine secretion to enhance and prolong the immune response.

[0363] Use A 2A R / TREx CHO cAMP functional analysis was used to measure the adenosine receptor activity of compound I.

[0364] The dose-response of NECA (5-ethylformamide adenosine) (a non-selective adenosine receptor agonist) was analyzed daily to determine the EC50 of NECA used in cAMP functional assays. 80 Stable expression of A at 1000-2500 cells / well 2A R TRex CHO cells were seeded onto 384-well Opti plates (Perkin Elmer) and incubated at 37°C for 30 min with different concentrations (ranging from 10 μM to 0 μM) of NECA. After 30 min of incubation, 5 μL of L Light-anti-cAMP (diluted 1:150 with the conjugate and lysis buffer provided by Perkin Elmer) and 5 μL of Eu-cAMP tracer (diluted 1:50 with the conjugate and lysis buffer provided by Perkin Elmer) were added to the cell stimulation, and the cells were incubated for 1 h. FRET signals were detected using an Envision multilabel plate analyzer (Perkin Elmer) when the Eu-cAMP tracer was excited at 615 nm and emitted at 665 nm. Data analysis was performed using a GraphPad Prism to determine the EC50 of NECA. 80 .

[0365] In A 2A cAMP antagonist function analysis was performed on R TRex CHO stable cell lines (Perkin Elmer). 1x10 6 Cells were seeded into T75 flasks and grown overnight at 37°C and 5% CO2. 2A R TRex CHO was used to induce confluency with ~70% to 80% tetracycline at 1 μg / mL for at least 16 hours. Stable A expression was achieved using 1000-2500 cells / well. 2A R TREx CHO cells were seeded onto white 384-well Opti plates and then incubated at 37°C for 30 min with different concentrations (ranging from 10 μM to 0 mM) of compound 1. NECA (Sigma Aldrich) EC was added to the cell stimulation mixture. 80 The cells were incubated at 37°C for 30 min. After 30 min of incubation, 5 μL of Ulight-anti-cAMP and 5 μL of Eu-cAMP tracer were added to the cell stimulation and the cells were incubated for 1 h. FRET signals were detected when the Eu-cAMP tracer was excited at 615 nm and emitted at 665 nm.

[0366] Data analysis was performed on GraphPad Prism to measure the K of compound I. B (less than 10 nM). Example

[0367] General method:

[0368] Those skilled in the art will recognize that various methods are available for preparing the molecules as described in the claims. Typically, a useful method for synthesizing the compounds described in the claims consists of four parts, which can be performed in any order: connecting fragments a and b (or forming the abc moiety via b cyclization), connecting fragments b and c (or forming the abc moiety via b cyclization), connecting fragments c and d, and modifying the functional groups in all fragments. The retrosynthesis of the compounds of the present invention, involving the breaking down of fragments a and b, is useful for constructing the compounds shown below:

[0369]

[0370] Several methods for preparing the claimed compound are exemplary (eq. 1-7). Reaction 1 illustrates one method of forming a bond between fragments c and d via the Suzuki reaction. In the case of eq. 1, Z can be selected from suitable groups such as Cl, Br, I, OTf, etc., and B(OR)2 is a boric acid or borate ester, and the coupling is mediated by a transition metal (preferably palladium with a suitable ligand).

[0371]

[0372] This coupling can be facilitated by the use of organic or inorganic bases, and a wide variety of conditions known in the art contribute to Suzuki coupling. The functionalization of the coupling partner can also be reversed as illustrated in eq.2. Those skilled in the art will recognize that other combinations may also produce the desired product.

[0373]

[0374] Reaction 3 illustrates another method for forming the CD fragment. In eq. 3, a suitable arylacrylic acid is condensed with Meldrum's acid and a suitable promoting agent (such as EDCI and DMAP, although other agents will also provide the desired product) to give the corresponding arylketoester. The ketone ester is then condensed with guanidine to form the corresponding 2-amino-3-hydroxy-arylpyrimidine, which can be converted to the corresponding chloride by treatment with POCl3 or other suitable agents.

[0375]

[0376] The formation of the bond between fragments c and b can occur before or after the formation of the connection between fragments c and d, and can be further modified with groups before or after the connection of fragments c and b. Reaction 4 illustrates one method of connecting fragments c and b via Suzuki coupling.

[0377]

[0378] In eq. 4, Z can be selected from suitable groups such as Cl, Br, I, OTf, etc., and B(OR)2 is a boric acid or borate ester, and the coupling is mediated by a transition metal (preferably palladium with a suitable ligand). This coupling can be facilitated by the use of organic or inorganic bases. A wide variety of conditions known in the art will contribute to Suzuki coupling. The functionalization of the coupling coupler can also be reversed as illustrated in eq. 5. Those skilled in the art will recognize that other combinations may also produce the desired product.

[0379]

[0380] Alternatively, fragment b can be formed via a cycloaddition between fragments a and c through an azide-alkyne Huisgen1 1,3-dipolar cycloaddition (reaction 6). In eq. 6, appropriately functionalized fragments a and c can be combined in a cycloaddition reaction between an azide and an alkyne. The reaction can be promoted by using a copper catalyst or other catalysts.

[0381]

[0382] When fragment b is a triazole, the ring can also be synthesized via palladium-mediated sodium azide to alkenyl halide addition (Barluenga et al., Angewandte Chemie International Edition, 2006, 45, 6893-6896), Amberlyst-15-catalyzed azide to nitroalkene addition (Zhang et al., Synthesis, 2016, 48, 131-135), I2 / TBPB-mediated N-tosylhydrozones to aniline oxidative cycloaddition (Cai et al., Organic Communications, 2014, 16, 5108-5111), and many other methods (see “Synthesis of 1,2,3-triazoles” at www.organic-chemistry.org / synthesis / heterocycles / 1,2,3-triazoles.shtm). The synthesis was carried out using 1,2,3-triazoles. Those skilled in the art will understand that various methods exist for achieving this transformation.

[0383] Reaction 7 illustrates one method of forming a bond between fragments a and b via alkylation. In the case of eq. 7, Z is a suitable electrophile such as Cl, Br, I, OTf, etc., and the coupling can be mediated by the use of an organic or inorganic base. Those skilled in the art will recognize that the timing and sequence of fragment linking and the modification of the functionality present in any fragment can be varied in any given preparation in order to most efficiently prepare any particular compound of the invention.

[0384]

[0385] The various methods described above have been used to prepare the compounds of the present invention, some of which are illustrated in the examples. The deuterated forms of the following examples can be synthesized using suitable deuterated intermediates.

[0386] Example 1: Synthesis of 3-[2-amino-6-(1-{[6-(2-hydroxypropane-2-yl)pyridin-2-yl]methyl}-1H-1,2,3-triazol-4-yl)pyrimidin-4-yl]-2-methylbenzyl nitrile

[0387]

[0388] Step 1: In a 250 mL round-bottom flask equipped with a magnetic stirrer, boronic ester (3.89 g, 16 mmol) and 2-amino-4,6-dichloropyrimidine (3.67 g, 22.4 mmol) were successively added. Anhydrous ethanol (100 mL) was added, followed by a solution of KHCO3 (4.81 g, 48 mmol) in deionized water (19 mL). The resulting suspension was degassed with nitrogen for 5 minutes. PdCl2(PPh3)2 (112 mg, 1 mol%) was then added, and the mixture was heated to 78 °C under a nitrogen atmosphere for 3 hours. The ethanol was evaporated under reduced pressure, and deionized water (150 mL) was added. The suspension was filtered, and the solid was washed with additional water (100 mL). The solid was then dissolved in acetone (220 mL) and collected in a 500 mL round-bottom flask. A mixture of silica and prionate salt (1:1, 150 g) was added, and the solvent was removed under reduced pressure. The obtained starting material was purified by silica gel rapid chromatography (gradient 0% to 15% dichloromethane / ethyl acetate). The desired product (1.91 g, 49%) was obtained as a white solid. LCMS: Method A, retention time = 2.93 min, ESI MS [M+H] + For C 12 H9ClN4, calculated value 245.7, measured value 245.2.

[0389] Step 2: In a round-bottom flask, 5.1 g (20.8 mmol) of chloropyrimidine was suspended in 42 mL of degassed THF. 8.68 mL (62.4 mmol) of Et3N and 5.95 mL (25.0 mmol) of TIPS-acetylene were added to the suspension. The reaction mixture was stirred for 5 min, followed by the addition of 219 mg (0.312 mmol) of PdCl2(PPh3)2 and 119 mg (0.624 mmol) of CuI. The reaction mixture was stirred in N2 at 50 °C for 5 h. After cooling the reaction to room temperature, the solvent was removed, and the starting material was suspended in 100 mL of EtOAc, from which insoluble solids were filtered off. The filtrate was washed with (1:1) NH4Cl / NH4OH (2 x 100 mL) and 10% Na2S2O4 (1 x 100 mL). The organic layer was dried over Na2SO4, concentrated, and used for the next step without further purification.

[0390] Step 3: In a round-bottom flask, dissolve the crude TIPS product from the previous step in 42 mL of dry THF and cool to 0°C. Add 25 mL (25.0 mmol) of TBAF (1.0 M in THF). Stir the reaction at 0°C for 15 min. Quench the reaction with saturated NH4Cl (100 mL). Extract the organic matter from the aqueous layer with EtOAc (2 x 100 mL). Wash the combined organic layers with (1:1) NH4Cl / NH4OH (2 x 100 mL) and 10% Na2S2O4 (1 x 100 mL). Dry the organic layers with Na2SO4, concentrate, and grind with 40% CH2Cl2 / hexane to obtain a light brown solid, pure product 5. Yield: 3.71 g (76%, step 2).

[0391] Step 4: In N2 at 0°C, for 30 minutes, a solution of methyl 2-(hydroxymethyl)pyridine-2-carboxylic acid (5.0 g, 29.9 mmol) in THF (70 mL, 0.4 M) was added to a solution of methyl magnesium bromide (3 M in Et2O, 40 mL, 120 mmol, 4.0 equivalence). The resulting mixture was allowed to be heated to room temperature and stirred for 3 h. The reaction mixture was quenched with an aqueous solution of NH4Cl (55 mL) and EtOAc (50 mL) was added. The organic phase was separated, and the aqueous phase was extracted with EtOAc (3 x 40 mL). The combined organic extracts were washed with a saturated aqueous solution of sodium bisulfite (7 x 20 mL), dried (Na2SO4), filtered, and concentrated under vacuum to give the title compound as a pale yellow liquid (3.45 g, 69% yield; 96% purity, as determined by LCMS). LCMS: Method A, retention times = 0.722 and 1.06 min, ESI MS [M+H] + For C9H 13 NO2, calculated value 167.09, measured value 167.2

[0392] Step 5: In N2 at 0°C, diphenylphosphoryl azide (7.73 mL, 35.9 mmol, 1.2 equivalents) was added to a solution of 2-hydroxymethyl-6-(1-hydroxy-1-methylethyl)pyridine (5 g, 29.9 mmol, 1.0 equivalents) in PhMe (33 mL, 0.9 M), followed by 1,8-diazabicyclo[5.4.0]undec-7-ene (5.37 mL, 35.9 mmol, 1.2 equivalents). The resulting mixture was heated to room temperature and stirred for 14 h. After completion, the mixture was diluted with ethyl acetate, washed with water, dried (Na2SO4), filtered, and the organic layer was concentrated. The residue was dissolved in 1N HCl aqueous solution (2 eq, 60 mmol) and extracted with MTBE in hexane solution (3:7, 100 mL). The organic layer was washed with water (50 mL) and neutralized with 2N NaOH aqueous solution. The combined aqueous layer was extracted with ethyl acetate (3 × 75 mL). The organic layer (Na₂SO₄) was dried, filtered through a cotton plug, and the filtrate was concentrated to provide a pure compound (3.75 g, 75%) as a pale yellow liquid. LCMS: Method A, retention time = 2.67 min, ESI MS [M+H] + C9H 12 N4O, calculated value 193.1, measured value 193.2.

[0393] Step 6: A mixture of azide (3.34 g, 17.4 mmol), alkyne (3.71 g, 15.8 mmol), copper(II) sulfate (39 mg, 0.158 mmol), and sodium ascorbate (156 mg, 0.790 mmol) in a 2:1 t-BuOH / H2O (158 mL) solution was heated at 60 °C for 13 h. The solvent was removed under vacuum, and the dried residue was loaded onto silica gel and purified by silica gel chromatography (0-100% EtOAc in hexane solution) to provide the desired product (6.08 g, 90%) as a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.69(s,1H),7.90(d,J=7.8Hz,1H),7.80(t,J=7.8Hz,1H),7.76(d,J=7.8Hz,1H),7.61(d,J=8.0Hz,1H), 7.51(t,J=7.8Hz,1H),7.28(s,1H),7.10(d,J=7.6Hz,2H),6.90(s,2H),5.81(s,2H),5.23(s,1H),2.55(s,3H),1.38(s,6H).ESI MS[M+H] + C 23 H 23N8O, calculated value 427.2, measured value 427.3.

[0394] Example 2: Synthesis of 3-[2-amino-6-(1-{[6-(2-hydroxypropane-2-yl)pyridin-2-yl]methyl}-1H-1,2,3-triazol-4-yl)pyrimidin-4-yl]-2-methoxybenzyl nitrile

[0395] process

[0396]

[0397] Step 1: In a round-bottom flask, 26 g (157.5 mmol) of 3-cyano-2-fluorobenzoic acid was suspended in 315 mL (0.5 M) of dry MeOH. 144 mL (630 mmol) of NaOMe (25 wt% MeOH solution) was added to the suspension. The resulting reaction mixture was refluxed in N2 for 2 h. After cooling to room temperature, excess MeOH was evaporated under reduced pressure to obtain a concentrated slurry. 158 mL (473 mmol) of 3 M HCl aqueous solution was added to this slurry. The product precipitated as a white solid and was separated by filtration. Residual water was removed by azeotropic distillation with toluene to obtain 26.2 g (94%) of pure product.

[0398] Step 2: 43 g (297 mmol) of Michaelis-Menten acid and 35 g (198 mmol) of 3-cyano,2-methoxybenzoic acid were suspended in 660 mL (0.3 M) CH₂Cl₂. 57 g (297 mmol) of N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC) and 24 g (198 mmol) of 4-dimethylaminopyridine (DMAP) were added to the suspension. The reaction mixture was stirred in N₂ at room temperature for 2 h. The reaction mixture became homogeneous at this point. The reaction mixture was then transferred to a separatory funnel and 200 mL of CH₂Cl₂ was added. The organic layer was washed with 1 M HCl (2 x 300 mL) and saturated NaCl (300 mL). The organic layer was dried over MgSO₄, concentrated, and the starting material was used in the next step without further purification. The crude Michaelis-Menten acid adduct from the previous step was suspended in 400 mL of anhydrous EtOH and refluxed for 1.5 h. After cooling to room temperature, the reaction was concentrated to 1 / 4 (~100 mL) of the initial volume under reduced pressure. The β-keto ester product in EtOH was used directly in the next step without further purification.

[0399] Step 3: In a round-bottom flask, dissolve 19 g (198 mmol) guanidine hydrochloride in 300 mL (0.7 M) EtOH. Add 74 mL (198 mmol) NaOEt (21 wt% EtOH solution). Stir the resulting turbid solution at room temperature for 10 min, then add 4 g (from the previous step) to 100 mL of EtOH. Reflux the reaction mixture in N2 for 72 h. After cooling the reaction to room temperature, add 300 mL of hexane. The precipitated product is obtained by filtration and used directly in the next step without further purification.

[0400] Step 4: Suspend the crude product 5 from the previous step in 200 mL of dioxane. Add POCl3 (186 mL, 2000 mmol). Heat the reaction mixture at 70 °C for 1.5 h. After cooling to room temperature, pour the reaction mixture into crushed ice (~1000 g) and stir. (Warning: Raise the temperature slowly to room temperature so that excess POCl3 is quenched by the melting of the ice, thus avoiding a violent reaction). After quenching with POCl3, add small portions of solid K2CO3 (691 g, 5000 mmol) to quench the obtained HCl and H3PO4. Extract the aqueous layer with CH2Cl2 (3 x 500 mL). Wash the combined organic layers with saturated NaCl (500 mL) and dry with MgSO4. Remove the solvent under reduced pressure to obtain a brown solid. Grind the crude product with 10% CH2Cl2 / hexane to obtain a pure product (28 g, 54% after 4 steps).

[0401] Step 5: In a round-bottom flask, 7.7 g (29.3 mmol) of chloro-pyrimidine 6 was suspended in 60 mL (1:1 dioxane / Et3N). TMS-acetylene (20.3 mL, 146 mmol) was added to the suspension, followed by PdCl2(PPh3)2 (2.6 g, 2.93 mmol) and CuI (558 mg, 2.93 mmol). The reaction mixture was stirred at 80 °C for 1 h in N2. After cooling the reaction to room temperature, silica gel (~100 g) was added, and the solvent was removed under reduced pressure. The starting material adsorbed on the silica gel was purified by chromatography with 80% (EtOAc / hexane). The yield of chloro-pyrimidine 6 was found to be 5.5 g (58%).

[0402] Step 6: In a round-bottom flask, dissolve 5.1 g (15.7 mmol) of 7 in 30 mL of dry THF. Add 16.5 mL (16.5 mmol) of TBAF (1.0 M THF solution). Stir the reaction mixture at room temperature for 30 min. Add silica gel (~100 g) to the reaction mixture and evaporate the solvent under reduced pressure. The starting material adsorbed on the silica gel is purified by chromatography using 50% (1:1 hexane:CH2Cl2 / EtOAc). Yield: 3.2 g (80%).

[0403] Step 7: CuSO4 (7.2 mg, 0.029 mmol, 1.0 mol%) and sodium ascorbate (60.0 mg, 0.305 mmol, 20 mol%) were added to a solution of azide (Example 1, Step 5, 294 mg, 1.53 mmol, 1.0 mol%) and alkyne (382 mg, 1.53 mmol, 1.0 mol%) in 2:1 t-BuOH / H2O (5 mL, 0.3 M). The resulting mixture was stirred at 55 °C for 0.5 h. After completion, the reaction mixture was cooled to room temperature and diluted with CH2Cl2 (10 mL). The organic phase was separated and the aqueous phase was extracted again with CH2Cl2 (10 mL). The combined extracts were concentrated and purified by column chromatography (CH2Cl2 → 95:5 CH2Cl2:MeOH) to obtain the title compound (604 mg, 89% yield) as a pale beige solid. 1 ¹H NMR (400MHz, CDCl₃) δ 8.30 (d, J = 1.0Hz, 1H), 8.04–7.98 (m, 1H), 7.92 (d, J = 0.8Hz, 1H), 7.78–7.64 (m, 2H), 7.37 (d, J = 7.9Hz, 1H), 7.28 (td, J = 7.8Hz, 0.8Hz, 1H), 7.14 (d, J = 7.6Hz, 1H), 5.75 (brs, 2H), 5.15 (brs, 2H), 4.74 (s, 1H), 3.94 (d, J = 0.8Hz, 3H), 1.54 (d, J = 0.8Hz, 6H). ESI calculated value C 23 H 23 N8O2[M+H]: 443.19, measured value: 443.2.

[0404] LCMS retention time: 2.8 minutes, Method A

[0405] Example 3: 3-[2-amino-6-(1-{[6-(2-hydroxypropane-2-yl)pyridine-2-]

[0406] Synthesis of [methyl]-1H-1,2,3-triazol-4-yl)pyrimidin-4-yl]-2-fluorobenzyl nitrile

[0407]

[0408] Step 1: At 0°C, iPrMgCl₂ was added dropwise to a stirred solution of 26 g (130 mmol) of 3-bromo-2-fluorobenzyl nitrile (26 g, 130 mmol) for 20 min. LiCl solution (100 mL, 130 mmol, 1.3 M in THF) was added. The resulting solution was stirred at 0°C for 50 min, and ZnCl₂ (17.72 g, 130 mmol) was added at 0°C. The reaction mixture was warmed to room temperature and stirred at the same temperature for 25 min. Then, compound 1 (16.4 g, 100 mmol) was added and stirred for 10 min. Pd(PPh₃)₄ (2.32 g, 2 mmol) was then added and stirred at room temperature for 12 h. The reaction mixture was quenched with saturated NH₄Cl aqueous solution (500 mL), extracted with EtOAc (3 x 300 mL), and dried over Na₂SO₄. The combined organic layers were evaporated to give 28 g of crude product 2, which was used in the next step without further purification.

[0409] Step 2: At room temperature, PdCl2(PPh3)2 (3.38 g, 4.82 mmol), CuI (1.84 g, 9.65 mol), and THF / Et3N (1:1, 482 mL) were added to a stirred solution of compound 3 (24 g, 96.52 mmol) and degassed with N2 for 30 min. Then, triisopropylacetylene (130 mL, 579.15 mmol) was added dropwise over 15 min (the reaction mixture turned slightly red), and the reaction mixture was refluxed for 90 min. LCMS and TLC showed complete consumption of compound 3. The solvent was evaporated by rotary evaporation. Excess Et3N was removed using toluene (2 x 200 mL) azeotropes. The crude reaction mixture was mixed with silica gel and directly loaded onto a fast column. The solvent gradient changed from 10% in hexane to 20% to 30% to 40% to 50% EA. At 40% EA in hexane, pure solid product 3 (15.13 g, 46% from 2 steps) was given.

[0410] Step 3: TBAF (37.97 mL, 1 M THF solution) was added dropwise to a stirred solution of compound 4 (15 g, 37.97 mmol) at 0 °C for 15 min, and the mixture was stirred at 0 °C for <30 min. TLC showed no SM (due to nBuN). + Positive ion LC MS No letterThe reaction mixture was quenched at 0°C with a saturated aqueous solution of NH4Cl (200 mL), extracted with EtOAc (3 x 250 mL), dried with Na2SO4, and evaporated to give crude product 4. 200 mL of a 10% hexane solution of EtOAc was added to the crude product, followed by sonication. The upper liquid fraction was separated, and EtOAc / CH2Cl2 (200 mL, 1:1) was added to the solid residue. Hexane (600 mL) was added to the resulting slurry to precipitate the precipitate, and the mixture was sonicated for 5 min. The precipitate was filtered and dried under high vacuum to give 4 (7.2 g) in 80% yield.

[0411] Step 4: Performed in the same manner as in Example 1

[0412] 1 ¹H NMR (400 MHz, acetone-d6) δ 8.62 (s, 1H), 8.45–8.38 (m, 1H), 8.02–7.95 (m, 1H), 7.88–7.81 (m, 2H), 7.65 (dd, J = 8.0 Hz, 1H), 7.62–7.56 (m, 1H), 7.24 (d, J = 7.8 Hz, 1H), 6.30 (brs, 2H), 5.86 (s, 2H), 4.62 (s, 1H), 1.48 (s, 6H). ESI MS [M+H] + For C 22 H 19 FN8O, calculated value 431.4, measured value 431.2.

[0413] Example 4: 3-[2-amino-6-(1-{[6-(2-hydroxypropane-2-yl)pyridin-2-yl]methyl}-1H-1,2,3-triazol-4-yl)pyrimidin-4-yl]benzylnitrile

[0414]

[0415] The title compound was prepared from 3-cyanobenzoic acid, similar to Example 2. 1 H NMR(400MHz, DMSO-d6)8.71(d,J=1.2Hz,1H),8.59(q,J=1.5Hz,1H),8.47(dq,J=8.2,1.4Hz,1H),8.00(dq,J=7.7,1.4Hz,1H),7.87–7.69(m ,3H),7.61(dt,J=8.0,1.2Hz,1H),7.11(dt,J=7.7,1.1Hz,1H),6.92(s,2H),5.83(s,2H),5.23(d,J=1.2Hz,1H),1.38(d,J=1.2Hz,6H).ESI MS[M+H] +For C 22 H 20 N8O, calculated value 413.2, measured value 413.3.

[0416] Example 5: 3-[2-amino-6-(1-{[6-(2-hydroxypropane-2-yl)pyridin-2-yl]methyl}-1H-1,2,3-triazol-4-yl)pyrimidin-4-yl]-2-chlorobenzylnitrile

[0417]

[0418] The title compound was prepared from 2-chloro-3-cyanoboronic acid, similar to Example 1. 1 H NMR (400MHz, acetone-d6) δ8.67(s,1H),8.02–7.93(m,2H),7.84(t,J=7.8Hz,1H),7.70(t,J=7.8Hz,1H),7.66(d,J=7.6Hz 1H),7.62(s,1H),7.25(d,J=7.7Hz,1H),6.37(brs,2H),5.87(s,2H),4.63(s,1H),1.48(s,6H).ESI MS[M+H] + For C 22 H 19 ClN8O, calculated value 447.9, measured value 447.2.

[0419] Example 6: 2-[6-({4-[2-amino-6-(2,3-dichlorophenyl)pyrimidin-4-yl]-1H-1,2,3-triazol-1-yl}methyl)pyridin-2-yl]prop-2-ol

[0420]

[0421] The title compound was prepared from 2,3-dichloroboronic acid and the starting material, similar to Example 1. 1 ¹H NMR (400 MHz, acetone-d6) δ 8.62 (s, 1H), 7.84 (t, J = 7.8 Hz, 1H), 7.74–7.62 (m, 2H), 7.61–7.45 (m, 3H), 7.23 (d, J = 7.5 Hz, 1H), 6.21 (s, 1H), 5.85 (s, 2H), 1.48 (m, 9H); LC-MS retention time 2.96 min, Method B, ESI MS [M+H] + For C 21 H 19 Cl2N7O, calculated value 456.1, measured value 456.2.

[0422] Example 7: 3-[2-amino-6-(1-{[6-(2-hydroxypropane-2-yl)pyridin-2-yl]methyl}-1H-pyrazol-4-yl)pyrimidin-4-yl]-2-methoxybenzylnitrile

[0423]

[0424] Step 1: In N2, benzoyl peroxide (538 mg, 2.2 mmol, 0.1 equivalent) was added to a solution of sprayed 2-acetyl-6-methylpyridine (3.0 g, 22.2 mmol, 1.0 equivalent) in MeCN (100 mL, 0.2 M), followed by N-bromosuccinimide (4.7 g, 26.6 mmol, 1.2 equivalent). The flask was fitted with a reflux condenser, and the mixture was heated to 85 °C and stirred for 28 h. After this, saturated aqueous solution of Na2S2O3 (50 mL) was added, and the two-phase mixture was stirred for 10 min. The mixture was transferred to a separatory funnel containing EtOAc (100 mL) and a 1:1 water:saturated Na2S2O3 (100 mL) mixture. The organic phase was collected, and the aqueous phase was extracted with 2 x 50 mL EtOAc. The combined organic extracts were dried over MgSO4 and concentrated under vacuum. The resulting residue was purified by column chromatography (hexane → 9:1 hexane:EtOAc) to give the title compound (2.65 g, 56% yield) as a light orange oil.

[0425] Step 2: 1-(6-(bromomethyl)pyridin-2-yl)ethyl-1-one (1.0 g, 4.7 mmol, 1.0 equivalent) and pinacol ester of 4-pyrazoloborate (997 mg, 5.1 mmol, 1.1 equivalent) were contained in MeCN (23 mL, 0.2 M) and Cs₂CO₃ (1.7 g, 5.1 mmol, 1.1 equivalent) was added. The resulting mixture was stirred at room temperature for 4 h. After completion, the mixture was diluted with CH₂Cl₂ (20 mL) and filtered through a filter funnel. The filtrate was concentrated under vacuum to provide the title compound, which was used in subsequent reactions without further purification.

[0426] Step 3: A solution of 1-(6-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)methyl)pyridin-2-yl)ethyl-1-one (301 mg, 0.92 mmol, 1.2 equivalents) and 3-(2-amino-6-chloropyrimidin-4-yl)-2-methoxybenzyl nitrile (Example 2, Step 4, 200 mg, 0.77 mmol, 1.0 equivalents) in DMF (33 mL, 0.9 M) and 2.0 M K2CO3 aqueous solution (0.8 mL, 2.0 equivalents) was injected with N2 jet for 10 min. Subsequently, Pd(dppf)Cl2 (55.6 mg, 0.04 mmol, 0.1 equivalents) was added, and the reaction mixture was heated to 100 °C for 16 h. After completion, the reaction mixture was prepared with CH2Cl2 (10 mL) and H2O (10 mL). The biphase mixture was transferred to a separatory funnel and the organic phase was collected. The aqueous phase was extracted with 2 x 10 mL CH2Cl2, and the combined organic extracts were dried over MgSO4 and concentrated under vacuum. The brown residue was purified by column chromatography (7:3 hexane:EtOAc→EtOAc) to give the title compound (190 mg, 58% yield) as a yellow oil.

[0427] Step 4: In N2 at 0°C, MeMgBr (0.8 mL, 1.1 mmol, 2.5 equivalents, 1.4 M in 3:1 THF:toluene) was added to a solution of 3-(6-(1-((6-acetylpyridin-2-yl)methyl)-1H-pyrazol-4-yl)-2-aminopyrimidin-4-yl)-2-methoxybenzyl nitrile (190 mg, 0.45 mmol, 1.0 equivalent) in THF (8.2 mL, 0.05 M). The resulting mixture was warmed to room temperature and stirred for 21 h. After completion, the reaction was quenched by adding 10 mL of saturated NH4Cl aqueous solution. The biphase mixture was transferred to a separatory funnel and extracted with 3 x 10 mL EtOAc. The combined organic extracts were washed with 10 mL of brine, dried over MgSO4, and concentrated under vacuum. The residue was purified by reversed-phase HPLC (19:1→1:19H2O:MeCN with 0.1%CF3CO2H) to obtain the title compound as a white solid (10 mg, 5% yield). 1¹H NMR (400 MHz, acetone-d6) δ 8.80 (d, J = 14.1 Hz, 1H), 8.35 (d, J = 6.3 Hz, 1H), 8.15–8.07 (m, 1H), 7.97–7.77 (m, 3H), 7.65–7.54 (m, 1H), 7.17–7.09 (m, 1H), 5.58 (s, 2H), 4.00 (s, 3H), 2.65 (s, 3H), 1.48 (s, 9H); LC-MS retention time 2.52 min. LC-MS, Method B, ESI MS [M+H] + For C 24 H 23 N7O2, calculated value 441.2, measured value 441.3

[0428] Example 8: 2-[6-({4-[2-amino-6-(3-fluoro-2-methoxyphenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]-2-propanol

[0429]

[0430] The title compound was prepared in a manner similar to that of Example 1. 1 ¹H NMR (400MHz, chloroform-d) δ 8.30–8.25 (m, 1H), 7.94–7.89 (m, 1H), 7.73 (t, J = 7.9Hz, 1H), 7.56 (d, J = 7.7Hz, 1H), 7.37 (d, J = 7.9Hz, 1H), 7.23–7.05 (m, 3H), 5.75 (s, 2H), 5.07 (s, 2H), 4.74 (s, 1H), 3.94 (s, 3H), 1.55 (s, 6H); LC-MS retention time 2.89 min. LC-MS, Method A, ESI MS [M+H] + C 22 H 23 FN7O2, calculated value 436.2, measured value 436.3.

[0431] Example 9: 2-[6-({4-[2-amino-6-(3-chloro-2-methoxyphenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]-2-propanol

[0432]

[0433] The title compound was prepared in a manner similar to that of Example 1. 1¹H NMR (400MHz, chloroform-d) δ 8.27 (d, J = 1.2Hz, 1H), 7.95 (d, J = 1.3Hz, 1H), 7.77–7.64 (m, 2H), 7.48 (dt, J = 8.0, 1.5Hz, 1H), 7.37 (d, J = 8.0Hz, 1H), 7.19–7.08 (m, 2H), 5.75 (s, 2H), 5.11 (s, 2H), 4.73 (s, 1H), 3.77 (s, 3H), 1.55 (s, 6H); LC-MS retention time 3.04 min. LC-MS, Method A, ESI MS [M+H] + For C 22 H 23 ClN7O2, calculated value 452.2, measured value 452.3.

[0434] Example 10: m-(2-amino-6-{1-[(2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)benzylnitrile

[0435]

[0436] The title compound was prepared in a manner similar to that of Example 4. 1 ¹H NMR (400MHz, chloroform-d) δ 8.65 (s, 1H), 8.46 (t, J = 1.6Hz, 1H), 8.41–8.34 (m, 1H), 8.31 (dt, J = 8.0, 1.4Hz, 1H), 7.91 (s, 1H), 7.80–7.68 (m, 2H), 7.60 (t, J = 7.8Hz, 1H), 7.34–7.28 (m, 1H), 5.76 (s, 3H), 5.22 (s, 2H); ESI MS [M+H] + For C 19 H 14 N8, calculated value 355.1, measured value 355.2.

[0437] Example 11: m-(2-amino-6-{1-[(3-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)benzylnitrile.

[0438]

[0439] The title compound was prepared in a manner similar to Example 4, thereby providing 59 mg of brown (tan) solid. 1H NMR (400MHz, DMSO-d6) δ8.73(d,J=1.1Hz,1H),8.68(dd,J=2.2,1.0Hz,1H),8.57(dq,J=3.1,1.4Hz,2H),8.46(ddd,J=8.0,1.9,1.1H z,1H),7.99(dq,J=7.8,1.3Hz,1H),7.82–7.78(m,2H),7.74(td,J=7.8,1.0Hz,1H),7.48–7.40(m,1H),6.89(s,2H),5.78(s,2H).ESI MS[M+H] + For C 19 H 14 N8, calculated value 355.1, measured value 355.3.

[0440] Example 12: m-(2-amino-6-{1-[(4-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)benzylnitrile.

[0441]

[0442] The title compound was prepared in a manner similar to Example 4, thereby providing 66 mg of brown solid. 1 H NMR(400MHz, DMSO-d6)δ8.76(d,J=1.2Hz,1H),8.63–8.54(m,3H),8.47(ddd,J=8.0,1.8,1.1Hz,1H),8.03 –7.96(m,1H),7.81(d,J=1.3Hz,1H),7.78–7.68(m,1H),7.31–7.23(m,2H),6.90(s,2H),5.81(s,2H).ESI MS[M+H] + For C 19 H 14 N8, calculated value 355.1, measured value 355.3.

[0443] Example 13: m-(2-amino-6-{1-[(6-methyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)benzylnitrile.

[0444]

[0445] The title compound was prepared in a manner similar to Example 4, thereby providing 14 mg of brown solid. 1H NMR (400MHz, DMSO-d6)8.68(d,J=2.0Hz,1H),8.59(s,1H),8.47(d,J=8.1Hz,1H),8.03–7.96(m,1H),7.81(d,J=1.9Hz,1 H),7.78–7.69(m,2H),7.24(d,J=7.8Hz,1H),7.11(d,J=7.7Hz,1H),6.92(s,2H),5.78(d,J=2.0Hz,2H),2.45(s,3H).ESI MS[M+H] + For C 20 H 16 N8, calculated value 369.2, measured value 369.3.

[0446] Example 14: 3-(2-amino-6-{1-[(6-methyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)-2-fluorobenzylnitrile.

[0447]

[0448] The title compound was prepared in a manner similar to Example 3, thereby providing 13 mg of brown solid. 1 H NMR(400MHz, DMSO-d6)8.67(s,1H),8.35–8.27(m,1H),8.26–8.18(m,1H),8.09(dd,J=7.7,6.1Hz,2H),7.7 2(t,J=7.7Hz,1H),7.62(d,J=2.4Hz,1H),7.23(d,J=7.8Hz,1H),6.98(s,2H),5.78(s,2H),2.45(s,3H).ESI MS[M+H] + For C 20 H 15 FN8, calculated value 387.1, measured value 387.3.

[0449] Example 15: 6-(2-amino-6-{1-[(6-methyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)-2-methylbenzonitrile

[0450]

[0451] The title compound was prepared in a manner similar to Example 1, thereby providing 75 mg of brown solid. 1H NMR(400MHz, DMSO-d6)8.66(d,J=1.3Hz,1H),7.95–7.86(m,1H),7.80–7.67(m,2H),7.52(t,J=7.8Hz,1H),7.27(d, J=1.4Hz,1H),7.23(d,J=7.8Hz,1H),7.09(d,J=7.7Hz,1H),6.90(s,2H),5.77(s,2H),2.55(s,3H),2.45(s,3H).ESI MS[M+H] + For C 21 H 18 N8, calculated value 383.2, measured value 383.3.

[0452] Example 16: 3-(2-amino-6-{1-[(6-methyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)-2-methoxybenzonitrile

[0453]

[0454] The title compound was prepared in a manner similar to Example 2, thereby providing 84 mg of brown solid. 1 H NMR(400MHz, DMSO-d6)8.65(d,J=1.1Hz,1H),8.07(ddd,J=7.8,1.8,1.0Hz,1H),7.99–7.90(m,1H),7.72(t,J=7.7Hz,1H),7.63(d,J=1.0Hz,1 H),7.49–7.39(m,1H),7.23(d,J=7.8Hz,1H),7.11(d,J=7.7Hz,1H),6.90(s,2H),5.77(s,2H),3.84(d,J=1.7Hz,3H),2.45(s,3H).ESIMS[M+H] + For C 21 H 18 N8O, calculated value 399.2, measured value 399.3.

[0455] Example 17: m-(2-amino-6-{1-[(3-methyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)benzylnitrile

[0456]

[0457] The title compound was prepared in a similar manner to that in Example 4. 1H NMR (400MHz, DMSO-d6) δ8.69(d,J=2.1Hz,1H),8.60–8.57(m,1H),8.50–8.44(m,1H),8.37(d,J=4.6Hz,1H),8.03–7.98(m,1H),7.8 5(d,J=2.3Hz,1H),7.75(td,J=7.7,2.4Hz,2H),7.35(dd,J=7.0,4.3Hz,1H),5.91(d,J=2.3Hz,2H),4.70(bs,2H),2.41(s,3H); ESI MS[M+H] + For C 20 H 16 N8O3, calculated value 369.2, measured value 369.2.

[0458] Example 18: Synthesis of m-[2-amino-6-(1-{[6-(trifluoromethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0459]

[0460] The title compound was prepared in a manner similar to Example 4, thereby providing 74 mg of brown solid. 1 H NMR (400MHz, DMSO-d6) δ8.77(d,J=1.1Hz,1H),8.59(d,J=1.4Hz,1H),8.47(ddt,J=8.0,1.9, 1.2Hz,1H),8.16(t,J=7.9Hz,1H),8.00(dq,J=7.7,1.3Hz,1H),7.91(d,J=7.8Hz,1H),7.85–

[0461] 7.80(m,1H),7.75(t,J=7.9Hz,1H),7.61(d,J=7.9Hz,1H),6.93(s,2H),5.99(s,2H).ESI MS[M+H] + For C 20 H 13 F3N8, calculated value 423.1, measured value 423.2.

[0462] Example 19: m-[2-amino-6-(1-{[6-(hydroxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0463]

[0464] Step 1: Cool the mixture of diol (696 mg, 5 mmol) and DBU (0.9 mL, 6 mmol) in dichloromethane (15 mL) to 0 °C. Add DPPA (1.3 mL, 6 mmol) dropwise, and stir the resulting mixture at 0 °C for 15 minutes and then at room temperature overnight. Add 5 g of cinnamic salt (Celite), and evaporate the mixture to dryness and purify by silica gel chromatography (hexane / EtOAc 90:10 to 60:40) to provide the desired azide (83 mg, 10%).

[0465] Step 2: The title compound was synthesized using an azide derivative and m-(2-amino-6-ethynyl-4-pyrimidinyl)benzyl nitrile (from Example 4) in a manner similar to step 6 of Example 1. 1 H NMR (400MHz, DMSO-d6) δ8.69 (s, 1H), 8.59 (dd, J = 1.8, 1.8Hz, 1H), 8.47 (ddd, J=8.0,1.8,1.1Hz,1H),8.00(ddd,J=7.7,1.7,1.2Hz,1H),7.85(dd,J=7.7,7. 7Hz,1H),7.82(s,1H),7.75(dd,J=7.7,7.7Hz,1H),7.46(d,J=7.8Hz,1H),7. 18(d,J=7.8Hz,1H),6.93(s,2H),5.81(s,2H),4.54(d,J=5.8Hz,3H).MS[M+H] + For C 20 H 16 N8O, calculated value 385.2, measured value 385.2.

[0466] Example 20: m-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0467]

[0468] Step 1: At room temperature under a nitrogen atmosphere, 12 g of sodium hydride (300 mmol, 60% dispersed in mineral oil, 1 e.g.) in dioxane (100 mL) was added to a suspension of 2,6-pyridinediethanol (41.8 g, 300 mmol) in dioxane (600 mL). The suspension was stirred for 15 minutes. Iodomethane (42.6 g, 300 mmol) was added and the resulting mixture was heated to 50 °C for 2 hours. TLC analysis showed that ~50% of the starting material was converted. The reaction was quenched with water, followed by extraction with ethyl acetate (500 mL x 3). The ethyl acetate layer was washed with water (200 mL) and brine. The ethyl acetate solution was dried over sodium sulfate for 1 hour, filtered, and concentrated. The resulting oily residue was purified by silica gel column chromatography, eluting with dichloromethane / methanol (from 2% to 5% methanol) to give 6.6 g of compound 1 as a pink oil in 15% yield.

[0469] Step 2: The product of Step 1 (3.4 g, 17.0 mmol) in SOCl2 (30 mL) was stirred overnight at 40 °C. The mixture was concentrated to give a white solid product (3.6 g, 100%).

[0470] Step 3: The product from Step 2 (1.16 g, 6.8 mmol, 1.0 eq) and NaN3 (1.3 g, 20.3 mmol, 3.0 eq) in EtOH were heated to reflux overnight. The mixture was concentrated to obtain a crude product, which was purified on an FCC (PE / EA = 5 / 1) to give a white solid product (0.9 g, 74%).

[0471] Step 4: Using the general steps of Example 1, synthesize the title compound to provide 64 mg of yellowish-brown solid. 1 HNMR(400MHz, DMSO-d6)δ8.70(s,1H),8.59(td,J=1.8,0.6Hz,1H),8.47(ddd,J=8.0,1.8,1.1Hz,1H),8.04–7.96(m,1H),7.86(t,J=7.8 Hz,1H),7.81(s,1H),7.77–7.69(m,1H),7.40–7.36(m,1H),7.25–7.19(m,1H),6.92(bs,2H),5.83(s,2H),4.46(s,2H),3.35(s,3H).ESI MS[M+H] + For C 21 H 18 N8O, calculated value 399.2, measured value 399.3.

[0472] The title compounds described above can also be obtained as shown below.

[0473]

[0474] Example 21: 6-(3-fluoro-2-methoxyphenyl)-4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0475]

[0476] The title compound was prepared in a similar manner to that in Example 20. 1 H NMR (400MHz, DMSO-d6) δ8.64(s,1H),7.85(t,J=7.8Hz,1H),7.64(s,1H),7.60(dt,J=7.9,1.5Hz,1H),7. 45–7.36(m,2H),7.28–7.18(m,2H),6.81(s,2H),5.81(s,2H),4.46(s,H),3.84(s,3H),3.35(s,3H).ESI MS[M+H] + For C 21 H 21 FN7O2, calculated value 422.2, measured value 422.3.

[0477] Example 22:3-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-2-methoxybenzonitrile

[0478]

[0479] The title compound was prepared by starting with the corresponding azide and alkyne, similar to that in Example 20. 1 H NMR (400MHz, DMSO-d6) δ8.82(s,1H),8.08(dd,J=7.9,1.7Hz,1H),7.99(dd,J=7.7,1.7Hz,1H),7.88(t,J=7.7Hz,1H),7.69(s,1 H),7.47(t,J=7.8Hz,1H),7.41(d,J=7.9Hz,1H),7.28(d,J=8.0Hz,1H),5.86(s,2H),4.47(s,2H),3.88(s,3H),3.35(s,3H).ESI MS[M+H] + For C 22 H 21 N8O2, calculated value 429.2, measured value 429.3.

[0480] Example 23: 6-(3-chloro-2-methoxyphenyl)-4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0481]

[0482] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR(400MHz, CDCl3)δ8.28(d,J=0.9Hz,1H),7.94–7.88(m,1H),7.73–7.62(m,2H),7.50–7.43(m,1H),7.40(d ,J=7.8Hz,1H),7.19–7.07(m,2H),5.71(s,2H),5.24(s,2H),4.58(s,2H),3.88–3.61(s,3H),3.48(s,3H).ESI MS[M+H] + For C 21 H 20 ClN7O2, calculated value 438.1, measured value 438.3.

[0483] Example 24: 5-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-3-methylbenzonitrile

[0484]

[0485] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR (400MHz, DMSO-d6) δ8.70(s,1H),8.42–8.36(m,1H),8.32(d,J=0.8Hz,1H),7.86(t,J=7.8Hz,1H),7.83(s,1H),7.81(s ,1H),7.39(dd,J=7.8,0.9Hz,1H),7.26–7.20(m,1H),6.92(s,2H),5.83(s,2H),4.47(s,2H),3.35(s,3H),2.46(s,3H).ESI MS[M+H] + For C 22 H 20 N8O, calculated value 413.2, measured value 413.3.

[0486] Example 25: 3-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-5-chlorobenzylnitrile

[0487]

[0488] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR (400MHz, DMSO-d6) δ8.72(s,1H),8.62(t,J=1.5Hz,1H),8.54(dd,J=2.1,1.6Hz,1H),8.22(dd,J=2.1,1.4Hz,1H ),7.90–7.82(m,2H),7.42–7.36(m,1H),7.26–7.19(m,1H),7.00(s,2H),5.83(s,2H),4.47(s,2H),3.35(s,3H).ESI MS[M+H] + For C 21 H 17 ClN8O, calculated value 433.1, measured value 433.2.

[0489] Example 26: m-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-5-methyl-4-pyrimidinyl]benzylnitrile

[0490]

[0491] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR (400MHz, DMSO-d6) δ8.63(s,1H),8.02(s,1H),7.97–7.92(m,1H),7.88(m,2H),7.71(t,J=7.8Hz,1H),7.39 (d,J=7.7Hz,1H),7.21(d,J=7.7Hz,1H),6.61(s,2H),5.83(s,2H),4.47(s,2H),3.35(s,3H),2.37(s,3H).ESI MS[M+H] + For C 22 H 20 N8O, calculated value 413.2, measured value 413.2.

[0492] Example 27: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(o-methoxyphenyl)-2-pyrimidinylamine

[0493]

[0494] The title compound was synthesized using 2-(azidomethyl)-6-(methoxymethyl)pyridine and 4-ethynyl-6-(o-methoxyphenyl)-2-pyrimidinylamine (which was prepared in a manner similar to steps 1-3 of Example 1) and in a manner similar to step 6 of Example 1. 1 H NMR (400MHz, CDCl3) δ8.27(s,1H),7.99(s,1H),7.83(dd,J=7.7,1.8Hz,1H),7.69(dd,J=7.8,7.8Hz,1H),7 .44-7.36(m,2H),7.11–6.96(m,3H),5.71(s,2H),5.12(s,2H),4.58(s,2H),3.90(s,3H),3.49(s,3H).ESI MS[M+H] + For C 21 H 21 N7O2, calculated value 404.2, measured value 404.2.

[0495] Example 28: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(o-methylphenyl)-2-pyrimidinylamine

[0496]

[0497] The title compound was synthesized using 2-(azidomethyl)-6-(methoxymethyl)pyridine and 4-ethynyl-6-(o-methylphenyl)-2-pyrimidinylamine (which was prepared in a manner similar to steps 1-3 of Example 1) and in a manner similar to step 6 of Example 1. 1 H NMR(400MHz, CDCl3)δ8.29(s,1H),7.70(dd,J=7.8,7.8Hz,1H),7.56(s,1H),7.50–7.38(m,2H),7.37–7 .27(m,3H),7.09(d,J=8.0Hz,1H),5.72(s,2H),5.19(s,2H),4.59(s,2H),3.49(s,3H),2.44(s,3H).ESI MS[M+H] + For C 21 H 21 N7O, calculated value 388.2, measured value 388.3.

[0498] Example 29: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(p-fluorophenyl)-2-pyrimidinylamine

[0499]

[0500] The title compound was synthesized using 2-(azidomethyl)-6-(methoxymethyl)pyridine and 4-ethynyl-6-(p-fluorophenyl)-2-pyrimidinylamine (which was prepared in a manner similar to steps 1-3 of Example 1) and in a manner similar to step 6 of Example 1. 1 H NMR (400MHz, CDCl3) δ8.29(s,1H),8.16–8.07(m,2H),7.88(s,1H),7.71(d,J=7.8,7.8Hz,1H),7.40(d,J=7.8H z,1H),7.21–7.13(m,2H),7.10(d,J=7.8Hz,1H),5.73(s,2H),5.08(s,2H),4.59(s,2H),3.50(s,3H).MS[M+H] + For C 20 H 18 FN7O, calculated value 392.2, measured value 392.2.

[0501] Example 30: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(o-fluorophenyl)-2-pyrimidinylamine

[0502]

[0503] The title compound was synthesized using 2-(azidomethyl)-6-(methoxymethyl)pyridine and 4-ethynyl-6-(o-fluorophenyl)-2-pyrimidinylamine (which was prepared in a manner similar to steps 1-3 of Example 1) and in a manner similar to step 6 of Example 1. 1 H NMR (400MHz, CDCl3) δ8.29(s,1H),8.00(ddd,J=7.8,7.8,1.9Hz,1H),7.92(d,J=2.2Hz,1H),7.71(dd,J=7.8,7.8Hz,1H),7.48–7.37(m,2H),7.27(d dd,J=7.8,7.8,1.2Hz,1H),7.17(ddd,J=11.3,8.3,1.1Hz,1H),7.0.9(d,J =8.3Hz,1H),5.72(s,2H),5.16(s,2H),4.59(s,2H),3.49(s,3H).MS[M+H]+ For C 20 H 18 FN7O, calculated value 392.2, measured value 392.3.

[0504] Example 31: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(o-chlorophenyl)-2-pyrimidinylamine

[0505]

[0506] The title compound was synthesized using 2-(azidomethyl)-6-(methoxymethyl)pyridine and 4-ethynyl-6-(o-chlorophenyl)-2-pyrimidinylamine (similar to the preparation in steps 1-3 of Example 1) and in a manner similar to step 6 of Example 1. 1 H NMR(400MHz, CDCl3)δ8.30(s,1H),7.74(s,1H),7.69(dd,J=7.8,7.8Hz,1H),7.66–7.54(m,1H),7.54–7.45(m,1H),7.40 (d,J=8.0Hz,1H),7.40–7.34(m,2H),7.10(d,J=8.0Hz,1H),5.72(s,2H),5.16(s,2H),4.59(s,2H),3.50(s,3H).MS[M+H] + For C 20 H 18 ClN7O, calculated value 408.1, measured value 408.3.

[0507] Example 32: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(o-trifluoromethoxyphenyl)-2-pyrimidinylamine

[0508]

[0509] The title compound was synthesized using 2-(azidomethyl)-6-(methoxymethyl)pyridine and 4-ethynyl-6-(o-trifluoromethoxyphenyl)-2-pyrimidinylamine (which was prepared in a manner similar to steps 1-3 of Example 1) and in a manner similar to step 6 of Example 1. 1H NMR(400MHz, CDCl3)δ8.29(s,1H),7.83–7.80(dd,J=7.8,1.6Hz,1H),7.77(s,1H),7.71(dd,J=7.8,7.8Hz, 1H),7.52–7.36(m,4H),7.11(d,J=7.8Hz,1H),5.72(s,2H),5.15(s,2H),4.59(s,2H),3.49(s,3H).MS[M+H] + For C 21 H 18 F3N7O2, calculated value 458.1, measured value 458.2.

[0510] Example 33: 4-[o-(methoxymethyl)phenyl]-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0511]

[0512] The title compound was synthesized using 2-(azidomethyl)-6-(methoxymethyl)pyridine and 6-ethynyl-4-(o-(methoxymethyl)phenyl)-2-pyrimidinylamine (which was prepared in a manner similar to steps 1-3 of Example 1) and in a manner similar to step 6 of Example 1. 1 H NMR (400MHz, CDCl3) δ8.29(s,1H),7.70(dd,J=7.7,7.7Hz,1H),7.65(s,1H),7.60–7.52(m,2H),7.51–7.34(m,3H),7.10(d,J=7.7Hz 1H),5.72(s,2H),5.12(s,2H),4.64(s,2H),4.59(s,2H),3.50(s,3H),3.35(s,3H).MS[M+H] + For C 22 H 23 N7O2, calculated value 418.2, measured value.

[0513] Example 34: 4-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-2-methylbenzonitrile

[0514]

[0515] The title compound was synthesized using 2-(azidomethyl)-6-(methoxymethyl)pyridine and 4-(2-amino-6-ethynyl-4-pyrimidinyl)-2-methylbenzonitrile (which was prepared in a manner similar to steps 1-3 of Example 1) and in a manner similar to step 6 of Example 1. 1 H NMR (400MHz, CDCl3) δ8.41(s,1H),8.32(s,1H),8.19(d,J=8.0Hz,1H),7.88(s,1H),7.72(dd,J=8.0,8.0Hz,1H),7.43( dd,J=8.0,8.0Hz,2H),7.12(d,J=7.6Hz,1H),5.73(s,2H),5.17(s,2H),4.59(s,2H),3.50(s,3H),2.62(s,3H).MS[M+H] + For C 22 H 20 N8O, calculated value 413.2, measured value 413.3.

[0516] Example 35: 6-(3,5-difluorophenyl)-4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0517]

[0518] The title compound was synthesized using 2-(azidomethyl)-6-(methoxymethyl)pyridine and 4-(3,5-difluorophenyl)-6-ethynyl-2-pyrimidinylamine (which was prepared in a manner similar to steps 1-3 of Example 1) and in a manner similar to step 6 of Example 1. 1 H NMR (400MHz, DMSO-d6) δ8.81(s,1H),7.92–7.83(m,5H),7.51–7.42(m,1H),7.40(d,J=7.6H z,1H),7.26(d,J=7.6Hz,1H),6.95(bs,1H),5.86(s,1H),4.47(s,3H),3.35(s,3H).MS[M+H] + For C 20 H 17 F2N7O, calculated value 410.1, measured value 410.2.

[0519] Example 36: 6-(3,5-dimethoxyphenyl)-4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0520]

[0521] The title compound was synthesized using 2-(azidomethyl)-6-(methoxymethyl)pyridine and 4-(3,5-dimethoxyphenyl)-6-ethynyl-2-pyrimidinylamine (which was prepared in a manner similar to steps 1-3 of Example 1) and in a manner similar to step 6 of Example 1. 1 H NMR (400MHz, DMSO-d6) δ8.75(brs,1H),7.86(dd,J=7.7Hz,1H),7.70(s,1H),7.39(d,J=7.7Hz,1H),7.28(d,J=2.3Hz, 2H),7.23(d,J=7.7Hz,1H),6.82(brs,2H),6.67(s,1H),5.84(s,2H),4.47(s,2H),3.84(s,6H),3.35(s,3H).MS[M+H] + For C 22 H 23 N7O3, calculated value 434.2, measured value 434.3.

[0522] Example 37: p-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0523]

[0524] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR(400MHz,DMSO-d6)δ8.68(s,1H),8.34–8.26(m,2H),8.03–7.90(m,1H),7.88–7.79(m,1H),7.76(s,1H),7.37 (dd,J=7.9,7.9Hz,1H),7.21(dd,J=7.9,7.9Hz,1H),6.90(s,2H),5.81(s,2H),4.45(s,2H),3.35(s,3H).MS[M+H] + For C 21 H 18 N8O, calculated value 399.2, measured value 399.3.

[0525] Example 38: o-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0526]

[0527] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR (400MHz, CDCl3) δ8.34(s,1H),7.88(d,J=7.8Hz,1H),7.82(d,J=7.8Hz,1H),7.80(s,1H),7.74–7.63(m,2H),7.55(d,J= 7.8Hz,1H),7.40(d,J=7.8Hz,1H),7.11(d,J=7.8Hz,1H),5.72(s,2H),5.31(s,2H),4.58(s,2H),3.51–3.43(s,3H).MS[M+H] + For C 21 H 18 N8O, calculated value 399.2, measured value 399.3.

[0528] Example 39: 2-{m-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]phenyl}-2-propanol

[0529]

[0530] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR (400MHz, CDCl3) δ8.30(s,1H),8.24(s,1H),7.96(d,J=8.0Hz,1H),7.92(s,1H),7.70(dd,J=8.0Hz,1H),7.61(d,J=8.0Hz,1H),7.45(dd,J=8 .0Hz,1H),7.40(d,J=8.0Hz,1H),7.10(d,J=8.0Hz,1H),5.72(s,2H),5. 30(s,1H),5.16(s,2H),4.58(s,1H),3.48(s,3H),1.64(s,6H).MS[M+H] + For C 25 H 25 N7O2, calculated value 432.2, measured value 432.2.

[0531] Example 40: 6-(m-Cumenyl))-4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0532]

[0533] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 ¹H NMR (400 MHz, chloroform-d) δ 8.30 (s, ¹H), 7.96 (s, ¹H), 7.94–7.87 (m, ²H), 7.71 (dd, J = 7.7, 7.7 Hz, ¹H), 7.49–

[0534] 7.33(m,3H),7.10(d,J=7.7Hz,1H),5.73(s,2H),5.10(brs,2H),4.59(s,2H),3.50(s,3H),3.00(h,J=6.9Hz,1H),1.31(d,J=6.9Hz,6H).MS[M+H] + For C 23 H 25 N7O, calculated value 416.2, measured value 416.4.

[0535] Example 41: Ethyl 3-{m-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]phenyl}propionate

[0536]

[0537] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR(400MHz, chloroform-d)δ8.29(s,1H),7.97(s,1H),7.96–7.91(m,1H),7.90(s,1H),7 .71(dd,J=7.8Hz,1H),7.46–7.36(m,2H),7.33(d,J=7.8Hz,1H),7.10(d,J=7.8H z,1H),5.73(s,2H),5.10(brs,2H),4.59(s,2H),4.14(q,J=7.2Hz,2H),3.49(s, 3H),3.04(t,J=7.9Hz,3H),2.68(t,J=7.9Hz,3H),1.24(t,J=7.2Hz,3H).MS[M+H] + For C 25 H 27 N7O3, calculated value 474.2, measured value 474.3.

[0538] Example 42: 4-[m-(2-methoxyethoxy)phenyl]-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0539]

[0540] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 ¹H NMR (400MHz, chloroform-d) δ 8.29 (s, 1H), 7.89 (s, 1H), 7.75–7.64 (m, 3H), 7.50–7.29 (m, 2H), 7.11 (d, J = 7.6Hz, 1H), 7.09–7.03 (m, 1H), 5.72 (s, 2H), 5.08 (brs, 2H), 4.59 (s, 2H), 4.32–4.18 (m, 2H), 3.83–3.75 (m, 2H), 3.50 (s, 3H), 3.47 (s, 3H). MS [M+H] + For C 23 H 25 N7O3, calculated value 448.2, measured value 448.3.

[0541] Example 43: 3-{m-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]phenyl}propionic acid

[0542]

[0543] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR (400MHz, DMSO-d6) δ12.15(brs,1H),8.63(s,1H),7.98(s,1H),7.93(d,J=7.6Hz,1H),7.84(dd,J=7.6Hz,1H),7.68(s,1H),7.47–7.30 (m,3H),7.20(d,J=7.6Hz,1H),6.74(s,1H),5.80(s,2H),4.45(s,2H),3.34(s,3H),2.90(t,J=7.6Hz,2H),2.58(t,J=7.6Hz,2H).MS[M+H] + For C 23 H 23 N7O3, calculated value 446.2, measured value 446.3.

[0544] Example 44: 3-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-4-fluorobenzyl nitrile

[0545]

[0546] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR(400MHz,DMSO-d6)δ8.70(s,1H),8.50–8.43(m,1H),8.14–8.03(m,1H),7.84(dd,J=8.0,8.0Hz,1H),7.7 0–7.57(m,2H),7.38(d,J=7.7Hz,1H),7.22(d,J=8.0Hz,1H),5.82(s,2H),4.45(s,2H),3.53(s,3H).MS[M+H] + For C 21 H 17 FN8O, calculated value 417.2, measured value 417.3.

[0547] Example 45: 3-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-2-fluorobenzyl nitrile

[0548]

[0549] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR (400MHz, DMSO-d6) δ8.66(s,1H),8.29(dd,J=7.9,7.9Hz,1H),8.07(ddd,J=7.6,7.6Hz,1H),7.84(dd,J=7.9,7.9Hz 1H),7.61(s,1H),7.56(dd,J=7.9,7.9Hz,1H),7.37(d,J=7.6Hz,1H),7.21(d,J=7.6Hz,1H),6.94(s,2H),5.79(s,2H),4.44(s,2H).MS[M+H] + For C 21 H 17 FN8O, calculated value 417.2, measured value 417.3.

[0550] Example 46: 6-(2,3-difluorophenyl)-4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0551]

[0552] 1H NMR(400MHz,DMSO-d6)δ8.69(s,1H),7.89–7.72(m,2H),7.63–7.49(m,2H),7.41–7.29(m, 2H),7.21(d,J=7.8Hz,1H),6.99(brs,2H),5.81(s,2H),4.45(s,2H),3.55(s,3H).MS[M+H] + For C 20 H 17 F2N7O2, calculated value 410.1, measured value.

[0553] Example 47: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(m-tolyl)-2-pyrimidinylamine

[0554]

[0555] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 9 mg of yellow-brown solid. 1 H NMR (400MHz, DMSO-d6) δ8.25(s,1H),7.89–7.85(m,1H),7.83(d,J=7.2Hz,1H),7.65(t,J=7.7Hz,1H),7.37–7.33(m,1H),7.31(d,J=7.6 Hz,1H),7.26–7.22(m,1H),7.21(s,1H),7.07–7.03(m,1H),5.67(s,2H),5.13(s,2H),4.53(s,2H),3.44(s,3H),2.40–2.37(m,3H).ESI MS[M+H] + For C 21 H 21 N7O, calculated value 388.1, measured value 388.3.

[0556] Example 48: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(m-methoxyphenyl)-2-pyrimidinylamine

[0557]

[0558] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 37 mg of yellow-brown solid. 1H NMR(400MHz, DMSO-d6)δ8.67(s,1H),7.85(t,J=7.8Hz,1H),7.71–7.63(m,3H),7.48–7.34(m,2H),7.24–7.17(m ,1H),7.09(ddd,J=8.2,2.7,0.9Hz,1H),6.80(bs,2H),5.81(s,2H),4.46(s,2H),3.84(s,3H),3.34(s,3H).ESI MS[M+H] + For C 21 H 21 N7O2, calculated value 404.2, measured value 404.2.

[0559] Example 49: 6-(m-fluorophenyl)-4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0560]

[0561] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20, to provide 32 mg of yellow solid. 1 H NMR (400MHz, DMSO-d6) δ8.69(s,1H),8.01–7.98(m,1H),7.94(ddd,J=10.6,2.7,1.5Hz,1H),7.86(t,J=7.7Hz,1H),7.73(s,1H ),7.58(td,J=8.0,6.0Hz,1H),7.42–7.34(m,2H),7.26–7.20(m,1H),6.87(bs,2H),5.83(s,2H),4.47(s,2H),3.35(s,3H).ESI MS[M+H] + For C 20 H 18 FN7O, calculated value 392.2, measured value 392.2.

[0562] Example 50: 6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-[m-(trifluoromethyl)phenyl]-2-pyrimidinylamine

[0563]

[0564] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20, thereby providing an orange-yellow solid. 1H NMR (400MHz, DMSO-d6) δ8.70(s,1H),8.48(d,J=2.1Hz,1H),8.45(d,J=7.9Hz,1H),7.95–7.83(m,2H),7.79(d,J=11.5Hz ,2H),7.67–7.52(m,2H),7.42–7.36(m,1H),7.25–7.18(m,1H),6.94(s,2H),5.83(s,2H),4.47(s,2H),3.35(s,3H).ESI MS[M+H] + For C 21 H 18 F3N7O, calculated value 442.2, measured value 442.2.

[0565] Example 51: 6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-[m-(methanesulfonyl)phenyl]-2-pyrimidinylamine

[0566]

[0567] The title compound was prepared from the corresponding azide and alkyne in a manner similar to Example 20, thereby providing a yellowish-brown solid. 1 H NMR(400MHz, DMSO-d6)δ8.77–8.59(m,2H),8.49(ddd,J=7.9,1.8,1.1Hz,1H),8.08(ddd,J=7.8,1.9,1.1Hz,1H),7.90–7 .76(m,3H),7.45–7.36(m,1H),7.31–7.17(m,1H),6.96(s,2H),5.83(s,2H),4.46(s,2H),3.35(s,3H),3.30(s,3H).ESI MS[M+H] + For C 21 H 21 N7O3S, calculated value 452.2, measured value 452.2.

[0568] Example 52: 6-(m-chlorophenyl)-4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0569]

[0570] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 71 mg of brown solid. 1H NMR (400MHz, DMSO-d6) δ8.69(s,1H),8.25–8.15(m,1H),8.10(dt,J=7.4,1.6Hz,1H),7.86(t,J=7.7Hz,1H),7.73(s,1 H),7.66–7.50(m,2H),7.44–7.34(m,1H),7.30–7.16(m,1H),6.88(s,2H),5.82(s,2H),4.46(s,2H),3.35(s,3H).ESI MS[M+H] + For C 20 H 18 ClN7O, calculated value 408.1, measured value 408.2.

[0571] Example 53: 3-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-5-fluorobenzylnitrile

[0572]

[0573] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 3 mg of white solid. 1 H NMR(400MHz, CDCl3)δ8.33(s,1H),8.25(t,J=1.2Hz,1H),8.15–8.05(m,1H),7.88(s,1H),7.73–7.71(m, 1H),7.44–7.27(m,2H),7.13(d,J=8.4Hz,1H),5.74(s,2H),5.16(bs,2H),4.60(s,2H),3.51(s,3H).ESI MS[M+H] + For C 21 H 17 FN8O, calculated value 417.2, measured value 417.3.

[0574] Example 54: 3-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-5-methoxybenzonitrile

[0575]

[0576] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 56 mg of white solid. 1H NMR (400MHz, DMSO-d6) δ8.72(s,1H),8.18(d,J=1.2Hz,1H),7.99(d,J=1.6Hz,1H),7.87–7.85(m,1H),7.81(s,1H),7.60– 7.56(m,2H),7.39(d,J=8Hz,1H),7.22(J=8Hz,1H),6.92(bs,1H),5.83(s,2H),4.47(s,2H),3.92(s,3H),3.35(s,3H).ESI MS[M+H] + For C 22 H 20 N8O2, calculated value 429.2, measured value 429.3.

[0577] Example 55: 6-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-2-methylbenzonitrile

[0578]

[0579] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 78 mg of brown solid. 1 H NMR(400MHz, DMSO-d6)8.67(d,J=0.8Hz,1H),7.94–7.81(m,2H),7.75(dd,J=7.8,1.3Hz,1H),7.52(t,J=7.8Hz,1H),7.39(d,J= 7.8Hz,1H),7.27(d,J=0.9Hz,1H),7.21(d,J=7.7Hz,1H),6.88(s,2H),5.81(s,2H),4.46(s,2H),3.35(s,3H),2.55(s,3H).ESI MS[M+H] + For C 22 H 20 N8O, calculated value 413.2, measured value 413.3.

[0580] Example 56: 6-(2,2-difluoro-2H-1,3-benzodioxolane-5-yl)-4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0581]

[0582] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 30 mg of yellow solid.1 H NMR(400MHz, DMSO-d6)δ8.67(s,1H),8.15(dd,J=1.8,0.4Hz,1H),8.07(dd,J=8.5, 1.8Hz,1H),7.86(t,J=7.8Hz,1H),7.73(s,1H),7.56(dd,J=8.5,0.4Hz,1H),7.43–

[0583] 7.35(m,1H),7.22(dd,J=7.7,0.9Hz,1H),6.84(s,2H),5.82(s,2H),4.46(s,2H),3.35(s,3H).ESI MS[M+H] + For C 21 H 17 F2N7O3, calculated value 454.1, measured value 454.3.

[0584] Example 57: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(m-trifluoromethoxyphenyl)-2-pyrimidinylamine

[0585]

[0586] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 100 mg of white solid. 1 H NMR (400MHz, DMSO-d6) δ8.70(s,1H),8.20–8.17(m,1H),8.12(s,1H),8.12(bs,1H),7.86(t,J=8Hz,1H),7.76(s,1H),7.66(t,J=8 Hz,1H),7.56–7.54(m,1H),7.39(d,J=3.2Hz,1H),7.22(d,J=3.2Hz,1H),6.90(bs,2H),5.83(s,2H),4.47(s,2H),3.35(s,3H).ESI MS[M+H] + For C 21 H 18 F3N7O2, calculated value 458.2, measured value 458.3.

[0587] Example 58: {m-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]phenyl}(dimethylamino)formaldehyde

[0588]

[0589] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 48 mg of brown solid. 1 H NMR(400MHz,DMSO-d6)δ8.68(dd,J=3.7,1.1Hz,1H),8.23–8.12(m,2H),7.86(td,J =7.8,4.1Hz,1H),7.74(dd,J=3.6,1.1Hz,1H),7.63–7.51(m,2H),7.39(dd,J=7.8,3 .5Hz,1H),7.22(dd,J=7.7,3.5Hz,1H),6.82(d,J=3.5Hz,2H),5.82(d,J=3.6Hz,2H ),4.47(d,J=3.8Hz,2H),3.35(dd,J=3.7,1.1Hz,3H),3.02(s,3H),2.94(s,3H).ESI MS[M+H] + For C 23 H 24 N8O2, calculated value 445.2, measured value 445.3.

[0590] Example 59: {m-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]phenylaminohydroxysulfeno}methane

[0591]

[0592] The title compound was prepared from 3-(methanesulfonylamino)phenylboronic acid, similar to Example 1. ¹H NMR (400 MHz, D₂O) δ 8.65 (m, 1H), 8.16–7.94 (m, 1H), 7.68–7.04 (m, 8H), 5.79 (m, 2H), 3.31 (m, 3H), 3.19 (m, 1H), 2.95 (m, 3H); LC-MS retention time 2.28 min. LC-MS, Method A, ESI MS [M+H] + For C 21 H 23 N8O3S, calculated value 467.2, measured value 467.2

[0593] Example 60: 6-(m-ethylphenyl)-4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0594]

[0595] The title compound was prepared from 3-ethylphenylboronic acid, similar to Example 1. 1 ¹H NMR (400MHz, DMSO-d⁶) δ 8.97 (s, 1H), 8.04 (s, 1H), 7.99 (d, J = 7.1Hz, 1H), 7.92–7.81 (m, 2H), 7.53–7.43 (m, 2H), 7.39 (d, J = 7.8Hz, 1H), 7.28 (d, J = 7.7Hz, 1H), 5.86 (s, 2H), 4.45 (s, 2H), 3.33 (s, 2H), 2.71 (q, J = 7.6Hz, 2H), 1.23 (td, J = 7.6, 1.0Hz, 3H); LC-MS retention time 2.66 min. LC-MS, Method A, ESIMS [M+H] + For C 22 H 24 N7O, calculated value 402.2, measured value 402.3.

[0596] Example 61: {m-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]phenyl}acetonitrile

[0597]

[0598] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 103 mg of brown solid. 1 H NMR (400MHz, DMSO-d6) δ8.66(d,J=1.1Hz,1H),8.14(s,1H),8.09(d,J=7.7Hz,1H),7.90–7.79(m,1H),7.72(d,J=1.1Hz,1H),7.59–7.4 7(m,2H),7.39(d,J=7.8Hz,1H),7.22(d,J=7.8Hz,1H),6.82(s,2H),5.82(s,2H),4.47(s,2H),4.16(s,2H),3.35(d,J=1.2Hz,3H).ESI MS[M+H] + For C 22 H 20 N8O, calculated value 413.2, measured value 413.3.

[0599] Example 62: 6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-[m-(1,3-oxazol-2-yl)phenyl]-2-pyrimidinylamine

[0600]

[0601] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 110 mg of brown solid. 1 H NMR(400MHz, DMSO-d6)δ8.79(d,J=1.8Hz,1H),8.73–8.67(m,1H),8.32–8.24(m,2 H),8.13(d,J=7.7Hz,1H),7.91–7.82(m,1H),7.78(d,J=2.0Hz,1H),7.71(dt,J=8. 6,4.3Hz,1H),7.47–7.43(m,1H),7.39(d,J=7.8Hz,1H),7.23(d,J=7.8Hz,1H),6. 91(s,2H),5.83(d,J=2.0Hz,2H),4.47(d,J=2.0Hz,2H),3.35(q,J=1.4Hz,3H).ESI MS[M+H] + For C 23 H 20 N8O2, calculated value 441.2, measured value 441.3.

[0602] Example 63: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(3-pyridyl)-2-pyrimidinylamine

[0603]

[0604] The title compound was synthesized using 2-(azidomethyl)-6-(methoxymethyl)pyridine and 6-ethynyl-4-(3-pyridyl)-2-pyrimidinylamine (which was prepared in a manner similar to steps 1-3 of Example 1) and in a manner similar to step 6 of Example 1. 1 ¹H NMR (400MHz, chloroform-d) δ 9.32 (d, J = 2.4 Hz, 1H), 8.71 (dd, J = 4.8, 1.7 Hz, 1H), 8.37 (ddd, J = 8.0, 2.3, 1.7 Hz, 1H), 8.31 (s, 1H), 7.92 (s, 1H), 7.71 (dd, J = 7.8, 7.8 Hz, 1H), 7.46–7.39 (m, 2H), 7.11 (d, J = 7.8 Hz, 1H), 5.73 (s, 2H), 5.15 (s, 2H), 4.59 (s, 2H), 3.50 (s, 3H). MS [M+H] + For C 19 H 18N8O, calculated value 375.2, measured value 375.3.

[0605] Example 64: 6-(2-furanyl)-4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0606]

[0607] The title compound was synthesized using 2-(azidomethyl)-6-(methoxymethyl)pyridine and 6-ethynyl-4-(2-furanyl)-2-pyrimidinylamine in a manner similar to step 6 of Example 1. 1 H NMR (400MHz, chloroform-d) δ8.27(s,1H),7.80(s,1H),7.71(t,J=7.8Hz,1H),7.61(dd,J=1.8,0.8Hz,1H),7.43–7.39(m,1H),7.20(d ESI MS[M+H] + For C 18 H 17 N7O2, calculated value 364.1, measured value 364.2.

[0608] Example 65: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(1,3-thiazolyl)-2-pyrimidinylamine

[0609]

[0610] Step 1: At 0°C, CDI (9.72 g, 60.0 mmol) was added in a single batch to a solution of thiazolic acid derivative (6.46 g, 50.0 mmol) and THF (100 mL). The mixture was stirred at room temperature for 4 hours. In separate flasks, a mixture of potassium monoethyl malonate (25.5 g, 150 mmol), MgCl2 (14.3 g, 150 mmol), and THF (100 mL) was stirred at 75°C for 4 hours. After both reactions were complete, the derivative containing the activated acid was added to the other flasks at room temperature. The combined reaction mixtures were stirred at 50°C for 16 hours. The mixture was cooled to room temperature and 2 M HCl was added. (aq)(100 mL). The mixture was extracted with ethyl acetate (2 x 150 mL), washed with saturated NaHCO3 brine and filtered through a silica gel stopper to provide the desired brown oily β-ketoester product (9.11 g; 91%).

[0611] Step 2: Sodium ethoxide (1.36 g, 20.0 mmol) was added to a solution of guanidine hydrochloride (1.91 g, 20.0 mmol) and ethanol (40 mL). The mixture was stirred at room temperature for 10 minutes, at which point the above-mentioned β-keto ester (3.98 g, 20.0 mmol) was added. The mixture was stirred at 100 °C for 16 hours. After cooling to room temperature, hexane (100 mL) was added. The precipitated solid was collected by filtration, thus providing the desired product (2.88 g, 74%) as a yellow solid.

[0612] Step 3: The mixture of the product from Step 2 (2.88 g, 14.8 mmol) and POCl3 (13.8 mL, 148 mmol) in dioxane (59 mL) was stirred at 70 °C for two hours. The mixture was then cooled, poured onto ice (75 g), neutralized with saturated NaHCO3, extracted with ethyl acetate (2 x 150 mL), and dried over Na2SO4. The crude product was purified by silica gel chromatography (0 to 5% MeOH in CH2Cl2 solution) to provide the desired product, a brown solid chloropyrimidine derivative (1.29 g; 41%).

[0613] Steps 4 and 5: A mixture of the product from step 3 (1.29 g; 6.07 mmol), trimethylsilylacetylene (2.59 mL, 18.2 mmol), bis(triphenylphosphine)palladium chloride (428 mg; 0.61 mmol), copper iodide (I) (116 mg, 0.61 mmol), triethylamine (3 mL), and DMF (3 mL) was stirred at 80 °C for 12 hours. Volatiles were removed, and the crude product was purified by silica gel chromatography (0 to 100% EtOAc in hexane solution) to provide the alkyne derivative. This was dissolved in MeOH (30 mL), ammonia (4.3 mL, 7 M MeOH solution) was added, and the mixture was stirred at room temperature for 30 minutes. Volatiles were removed, and the crude product was purified by silica gel chromatography (0 to 50% EtOAc in CH2Cl2 / hexane (1:1) solution) to provide the desired product (254 mg; 21%) as an orange-yellow solid.

[0614] Step 6: The product was synthesized in a manner similar to that in Step 6 of Example 1: a grayish-white solid (34 mg, 30%). 1HNMR(400MHz,DMSO-d6)δ8.85(s,1H),8.13(s,1H),8.05(s,1H),7.97–7.85(m,2H),7. 43(d,J=8.4Hz,1H),7.30(d,J=7.6Hz,1H),5.87(s,2H),4.49(s,2H),3.36(s,3H).ESI MS[M+H] + For C 17 H 17 N8OS, calculated value 381.1, measured value 381.2.

[0615] Example 66: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(4-methyl-1,3-thiazolyl-2-yl)-2-pyrimidinylamine

[0616]

[0617] The title compound was synthesized in a manner similar to that described in Example 65 above. 1 ¹H NMR (400MHz, chloroform-d) δ 8.30 (s, 1H), 8.13 (s, 1H), 7.71 (t, J = 7.8 Hz, 1H), 7.44–7.36 (m, 1H), 7.14–7.09 (m, 1H), 7.07 (t, J = 0.9 Hz, 1H), 5.72 (s, 2H), 5.32 (s, 2H), 4.59 (s, 2H), 3.49 (s, 3H), 2.54 (s, 3H). ESI MS [M+H] + For C 18 H 18 N8OS, calculated value 395.1, measured value 395.2.

[0618] Example 67: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(3-methyl-2-pyrazinyl)-2-pyrimidinylamine

[0619]

[0620] Steps 1-4: Synthesize TMS alkyne derivatives in a manner similar to that of Example 65: brown solid (193 mg, 1.3% (4 steps)).

[0621] Step 5: At 0°C, TBAF (3.4 mL, 0.750 mmol, 1 M in THF) was added dropwise to a solution of TMS alkyne derivative (193 mg, 0.682 mmol) in THF (3.4 mL). The mixture was stirred at 0°C for 15 minutes. The mixture was concentrated and purified by silica gel chromatography (0 to 5% MeOH in CH2Cl2 solution) to provide the desired product (93 mg; 65%) as a brown solid.

[0622] Step 6: The product was synthesized in a manner similar to that in Step 6 of Example 1: a grayish-white solid (7 mg, 6%). 1 HNMR(400MHz,DMSO-d6)δ8.81(s,1H),8.70–8.62(m,2H),7.93–7.83(m,1H),7.76(s,1H),7.40( d,J=7.9Hz,1H),7.26(d,J=7.8Hz,1H),5.85(s,2H),4.47(s,2H),3.34(s,3H),2.80(s,3H).ESI MS[M+H] + For C 19 H 20 N9O, calculated value 390.2, measured value 390.2.

[0623] Example 68: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(2H-pyrazol-3-yl)-2-pyrimidinylamine

[0624]

[0625] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 92 mg of yellowish-brown solid. 1 H NMR(400MHz,DMSO-d6)δ8.60(s,1H),7.90–7.81(m,2H),7.77(s,1H),7.38(d,J=7.7Hz,1H),7.21 (d,J=7.7Hz,1H),6.82(t,J=2.1Hz,1H),6.68(bs,2H),5.80(s,2H),4.46(s,2H),3.35(s,3H).ESI MS[M+H] + For C 17 H 17 N9O, calculated value 364.2, measured value 364.3.

[0626] Example 69: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(1H-pyrazol-4-yl)-2-pyrimidinylamine

[0627]

[0628] Similar to Example 20, the title compound was prepared from the corresponding azide and alkyne, thereby providing 2.4 mg of yellowish-brown solid. 1 H NMR (400MHz, DMSO-d6) δ8.57(s,1H),8.45(s,1H),8.09(s,1H),7.85(t,J=7.8Hz,1H),7.49(s,1H),7.3 8(dd,J=7.9,0.9Hz,1H),7.20(d,J=7.7Hz,1H),6.57(s,2H),5.81(s,2H),4.46(s,2H),3.35(s,3H).ESI MS[M+H] + For C 17 H 17 N9O, calculated value 364.2, measured value 364.2.

[0629] Example 70: 6-(1H-indol-6-yl)-4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0630]

[0631] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR(400MHz,DMSO-d6)δ9.23(s,1H),8.50(s,1H),8.22(s,1H),8.08(s,1H),8.01–7.85(m,3H) ,7.43(d,J=7.9Hz,1H),7.36(d,J=7.9Hz,1H),5.93(s,2H),4.48(s,2H),3.33(s,3H).MS[M+H] + For C 21 H 19 N9O, calculated value 414.2, measured value 414.3.

[0632] Example 71: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(7-quinolinyl)-2-pyrimidinylamine

[0633]

[0634] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 61 mg of brown solid. 1 H NMR (400MHz, DMSO-d6) δ8.99(dd,J=4.3,1.9Hz,1H),8.79(s,1H),8.73(d,J=1. 5Hz,1H),8.45(d,J=8.2Hz,1H),8.38–8.31(m,1H),8.13(d,J=8.2Hz,1H),7.94–

[0635] 7.80(m,2H),7.66–7.54(m,1H),7.40(d,J=7.7Hz,1H),7.24(d,J=7.6Hz,1 H),6.90(s,2H),5.84(s,2H),4.52–4.41(m,2H),3.36(d,J=1.3Hz,3H).ESI MS[M+H] + For C 23 H 20 N8O, calculated value 425.2, measured value 425.3.

[0636] Example 72: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(8-quinolinyl)-2-pyrimidinylamine

[0637]

[0638] The title compound was prepared from the corresponding azide and alkyne in a similar manner to Example 20, thereby providing 31 mg of brown solid. 1 H NMR(400MHz, DMSO-d6)δ8.98(dt,J=4.2,1.5Hz,1H),8.65(d,J=1.2Hz,1H),8.49(dd,J=8 .4,1.8Hz,1H),8.21–8.09(m,2H),7.93(d,J=1.2Hz,1H),7.89–7.79(m,1H),7.75(ddd,J =8.2,7.2,1.1Hz,1H),7.62(ddd,J=8.3,4.2,1.1Hz,1H),7.39(d,J=7.8Hz,1H),7.21(d, J=7.8Hz,1H),6.73(s,2H),5.82(s,2H),4.47(s,2H),3.36(d,J=1.1Hz,3H).ESIMS[M+H] + For C 23 H 20N8O, calculated value 425.2, measured value 425.3.

[0639] Example 73: 4-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(1H-pyrazol-1-yl)-2-pyrimidinylamine

[0640]

[0641] Step 1: NaH (60% dispersed in mineral oil, 840 mg, 21 mmol, 1.05 equivalent) was suspended in DMF (80 mL), and the suspension was cooled in an ice / water bath. Pyrazole (1.43 g, 21 mmol, 1.05 equivalent) was added. After 45 minutes, solid dichloropyrimidine (3.26 g, 20 mmol, 1 equivalent) was added, and the ice bath was removed. After 2 hours, water was added, and the reaction mixture was filtered to provide 2.99 g of the title compound as a pale yellow solid.

[0642] Steps 2 and 3: Similar to Example 1, obtain 30 mg of yellow solid. 1 H NMR (400MHz, DMSO-d6) δ8.64(s,1H),8.52(dd,J=2.7,0.7Hz,1H),7.90(dd,J=1.7,0.7Hz,1H),7.85(t,J=7.8Hz,1H),7.68(s,1H), 7.38(d,J=7.7Hz,1H),7.22(d,J=7.7Hz,1H),7.01(s,2H),6.62(dd,J=2.7,1.6Hz,1H),5.81(s,2H),4.46(s,2H),3.35(s,3H).ESI MS[M+H] + For C 17 H 17 N9O, calculated value 364.2, measured value 364.2.

[0643] Example 74: m-[2-amino-6-(1-{[6-(methoxymethyl)-2-pyridyl]methyl}-5-methyl-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0644]

[0645] Step 1. Prepare the alkyne by using propyne as a reagent in a manner similar to Step 2 of Example 1, thereby providing the product.

[0646] Step 2: In a sealed tube, a mixture of the azide derivative (18 mg, 0.1 mmol) and the alkyne (23 mg, 0.1 mmol) in toluene (1 mL) was heated to 120 °C for 20 hours. The mixture was cooled to room temperature, evaporated to dryness, and purified by silica gel chromatography (hexane / EtOAc 70:30 to 0:100) to provide the desired product (4 mg, 10%) along with its regioisomer (2 mg, 5%). 1 ¹H NMR (400MHz, chloroform-d) δ 8.47 (ddd, J = 1.7, 1.7, 0.6Hz, 1H), 8.32 (ddd, J = 8.0, 1.9, 1.2Hz, 1H), 7.99 (s, 1H), 7.75 (ddd, J = 7.7, 1.7, 1.2Hz, 1H), 7.67 (dd, J = 7.8, 7.8Hz, 1H), 7.60 (ddd, J = 7.8, 7.8, 0.6Hz, 1H), 7.37 (d, J = 7.8Hz, 1H), 6.89 (d, J = 7.8Hz, 1H), 5.69 (s, 2H), 5.09 (s, 2H), 4.57 (s, 2H), 3.49 (s, 3H), 2.71 (s, 3H). MS [M+H] + For C 22 H 20 N8O, calculated value 413.2, measured value 413.3.

[0647] Example 75: m-[2-amino-6-(1-{[6-(ethoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0648]

[0649] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR (400MHz, DMSO-d6) δ8.90(s,1H),8.63(s,1H),8.50(d,J=7.9Hz,1H),8.06(d,J=7.8Hz,1H),7.95(s,1H),7.90(t,J=7.8Hz,1H),7.79(t,J =7.7Hz,1H),7.43(d,J=7.9Hz,1H),7.28(d,J=7.3Hz,1H),5.88(s,2H),4.51(s,2H),3.54(q,J=7.0Hz,2H),1.16(t,J=7.0Hz,3H).ESIMS[M+H] + For C 22 H 21N8O, calculated value 413.2, measured value 413.3.

[0650] Example 76: m-[2-amino-6-(1-{[6-(isopropoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0651]

[0652] The title compound was prepared from the corresponding azide and alkyne, similar to Example 20. 1 H NMR (400MHz, DMSO-d6) δ8.93(s,1H),8.64(s,1H),8.51(d,J=8.0Hz,1H),8.06(d,J=7.7Hz,1H),7.97(d,J=3.4Hz,1H),7.93–7.86(m,1H) ,7.82–7.76(m,1H),7.44(d,J=7.8Hz,1H),7.29(d,J=7.3Hz,1H),5.88(s,2H),4.52(s,2H),3.74–3.62(m,1H),1.14(d,J=6.1Hz,6H).ESI MS[M+H] + For C 23 H 23 N8O, calculated value 427.2, measured value 427.3.

[0653] Example 77: m-[2-amino-6-(1-{[6-(1-methoxyethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0654]

[0655] Step 1: The diol (700 mg, 4.6 mmol) was dissolved in CH₂Cl₂ (10 mL). Imidazole (640 mg, 9.4 mmol) and TBSCl (754 mg, 5 mmol) were added and the mixture was stirred until the starting diol was completely converted. The crude mixture was placed directly on a silica gel column (Hex / EtOAc 95:5) to provide a monoprotected alcohol (794 mg, 65%). The TBS-protected alcohol (794 mg, 3 mmol) from the above step was dissolved in THF (6 mL) and NaH (60% in mineral oil, 144 mg, 3.6 mmol) was added. The mixture was stirred for 10 min and iodomethane (374 μL, 6 mmol) was added. After complete conversion of the starting alcohol, the mixture was quenched with saturated NH₄Cl and purified by silica gel chromatography (Hex / EtOAc 95:5) of the residue after routine treatment to provide the desired ether (800 mg, 96%).

[0656] The above-mentioned TBS derivative (800 mg, 2.8 mmol) was dissolved in THF (5 mL), and the solution was cooled to 0 °C. At this point, a solution of TBAF (1 M in THF, 3 mL) was added dropwise. After the reaction was complete, the mixture was quenched with saturated NH4Cl, and after routine treatment, the residue was purified by silica gel chromatography (Hex / EtOAc 90:10 to 0:40) to provide the desired primary alcohol (475 mg, quantified).

[0657] Step 2: Dissolve the alcohol (475 mg, 2.8 mmol) obtained in Step 1 in CH₂Cl₂ (3 mL) and add SOCl₂ (397 μL, 2 equivalents, 5.6 mmol). Stir the resulting solution until the starting alcohol is completely converted, then evaporate the mixture to dryness. Use the resulting residue directly without further purification. Dissolve the resulting feedstock in DMF (5 mL) and add sodium azide (273 mg, 4.2 mmol). Stir the resulting mixture at 50 °C for 8 hours, then cool to room temperature. Partition the crude product between water and dichloromethane. Evaporate the organic layer to dryness and purify the residue by silica gel chromatography (Hex / EtOAc 90:10) to provide the desired azide (200 mg, 37% from two steps).

[0658] Step 3: The title compound was synthesized using an azide derivative and m-(2-amino-6-ethynyl-4-pyrimidinyl)benzyl nitrile (from Example 4) in a manner similar to Step 6 of Example 1. 1H NMR (400MHz, chloroform-d) δ8.46(s,1H),8.39–8.24(m,2H),7.92(s,1H),7.82–7.66(m,2H),7.61(dd,J=7.8,7.8Hz1H),7.40(d,J= 7.8Hz,1H),7.09(d,J=7.8Hz,1H),5.74(s,2H),5.14(s,2H),4.49–4.36(m,1H),3.33(s,3H),1.47(d,J=6.9Hz,3H).MS[M+H] + For C 22 H 20 N8O, calculated value 413.2, measured value 413.3.

[0659] Example 78: 4-(1-{[6-(1-methoxyethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-6-(1,3-oxazol-2-yl)-2-pyrimidinylamine

[0660]

[0661] Steps 1 and 2: A mixture of pyrimidine derivative (3.64 g, 10.0 mmol), 2-(tri-n-butylstannyl)oxazole (2.10 mL, 10.0 mmol), and Pd(PPh3)4 (1.16 g; 1.00 mmol) in DMF (20 mL) was stirred for 5 hours at 100 °C. The mixture was cooled to room temperature and ethyl acetate (200 mL) was added. The organic matter was washed with brine (4 x 200 mL) and dried over MgSO4. The crude product was purified by silica gel chromatography (0 to 30% EtOAc in hexane) to provide the desired product. TFA (1 mL) and CH2Cl2 (5 mL) were added and the mixture was stirred at room temperature for 10 minutes. The mixture was neutralized with saturated NaHCO3, diluted with ethyl acetate, and dried over Na2SO4 to provide the desired product (322 mg; 8%) as a yellow solid.

[0662] Steps 3 and 4: Synthesize the final alkyne in a manner similar to Example 65: brown solid (63 mg, 21%, 2 steps).

[0663] Step 5: Synthesize the product in a manner similar to that in Example 1, Step 6: yellow solid (13 mg, 10%). 1H NMR (400MHz, DMSO-d6) δ8.75(s,1H),8.37(s,1H),7.87(t,J=7.8Hz,1H),7.82(s,1H),7.54(s,1H),7.39(d,J= 7.8Hz,1H),7.19(d,J=7.7Hz,1H),5.85(s,2H),4.35(q,J=6.4Hz,1H),3.19(s,3H),1.32(d,J=6.6Hz,3H).ESI MS[M+H] + For C 18 H 19 N8O2, calculated value 379.2, measured value 379.3.

[0664] Example 79: m-[2-amino-6-(1-{[6-(1-methoxypropyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0665]

[0666] Step 1: Cool the solution of aldehyde A (1.0 g, 4 mmol) in THF (20 mL) in a dry ice / acetone bath. Add EtMgBr (3 M in THF, 2 mL, 6 mmol, 1.5 equivalence) along the side of the flask. After 1.5 hours, quench the reaction with NH4Cl and extract with EtOAc. Concentrate the organic layer to... The alcohol B was purified on SiO2 by rapid chromatography to provide a white solid (537 mg).

[0667] Step 2: Add NaH (60% dispersed in mineral oil, 99 mg, 2.5 mmol, 1.3 equivalents) to a solution of alcohol B (537 mg, 1.9 mmol) in THF (8 mL). After 30 minutes, add MeI (0.18 mL, 2.9 mmol, 1.5 equivalents). Stir the reaction mixture overnight and quench with H2O. Extract with MTBE, dry, and concentrate to provide ether C (559 mg) as a yellow oil.

[0668] Step 3: Add TBAF (1M in THF, 2mL) to a solution of ether C (1.9mmol) in THF (2mL). After 1.5 hours, concentrate the reaction mixture and purify the crude residue by rapid chromatography on SiO2 to provide colorless oily alcohol D (299mg).

[0669] Steps 4 and 5: Using the process of Example 1, the title compound was synthesized, thereby providing 81 mg of brown solid. 1HNMR(400MHz,DMSO-d6)δ8.71(dd,J=5.4,1.9Hz,1H),8.58(q,J=3.1,1.7Hz,1 H),8.54–8.40(m,1H),8.06–7.94(m,1H),7.91–7.69(m,3H),7.35(t,J=7.1Hz, 1H),7.17(t,J=6.6Hz,1H),6.90(s,2H),5.84(d,J=6.0Hz,2H),4.13(t,J=6.3 Hz,1H),3.19(dd,J=5.4,1.8Hz,3H),1.75–1.62(m,2H),0.86–0.70(m,3H).ESI MS[M+H] + For C 23 H 22 N8O, calculated value 427.2, measured value 427.3.

[0670] Example 80: m-[2-amino-6-(1-{[6-(1-methoxy-2-methylpropyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0671]

[0672] The title compound was prepared in a manner similar to Example 79, thereby providing 86 mg of brown solid. 1 H NMR (400MHz, DMSO-d6) δ8.71(d,J=2.0Hz,1H),8.58(t,J=2.0Hz,1H),8.47(dd,J=8.1,1.7Hz,1H) ,8.04–7.96(m,1H),7.90–7.79(m,2H),7.74(td,J=7.9,2.0Hz,1H),7.30(d,J=7.8Hz,1H),7.18(d d,J=7.5,1.8Hz,1H),6.89(s,2H),5.83(d,J=1.7Hz,2H),3.92(dd,J=6.2,2.0Hz,1H),3.20–3.14( m,3H),1.95(dt,J=12.2,7.3Hz,1H),0.80(dd,J=6.8,2.0Hz,3H),0.71(dd,J=6.8,2.0Hz,3H).ESI MS[M+H] + For C 24 H 24 N8O, calculated value 441.2, measured value 441.5.

[0673] Example 81: m-[2-amino-6-(1-{[6-(cyclopropylmethoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0674]

[0675] The title compound was prepared in a manner similar to Example 79, thereby providing 87 mg of brown solid. 1 H NMR (400MHz, DMSO-d6) δ8.73–8.69(m,1H),8.58(q,J=1.8Hz,1H),8.50–8.44(m,1H),8. 02–7.96(m,1H),7.89–7.79(m,2H),7.79–7.71(m,1H),7.38(d,J=7.5Hz,1H),7.20(d,J =7.3Hz,1H),6.90(s,2H),5.84(d,J=3.0Hz,2H),3.67(dd,J=7.9,3.1Hz,1H),3.22–3.1 5(m,3H),1.11–1.02(m,1H),0.55–0.36(m,2H),0.25(ddd,J=37.8,9.0,4.6Hz,2H).ESI MS[M+H] + For C 24 H 22 N8O, calculated value 439.2, measured value 439.3.

[0676] Example 82: m-[2-amino-6-(1-{[6-(cyclopentylmethoxymethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0677]

[0678] The title compound was prepared in a manner similar to Example 79, thereby providing 81 mg of brown solid. 1H NMR(400MHz,DMSO-d6)δ8.71(d,J=1.1Hz,1H),8.61–8.58(m,1H),8.51–8.42(m ,1H),8.01–7.97(m,1H),7.89–7.79(m,2H),7.74(t,J=7.8Hz,1H),7.33(d,J=7. 8Hz,1H),7.19(d,J=7.7Hz,1H),6.89(s,2H),5.84(s,2H),4.01–3.93(m,1H),3 .13(s,3H),2.16(q,J=7.9Hz,1H),1.60(d,J=8.8Hz,1H),1.52–1.09(m,7H).ESI MS[M+H] + For C 26 H 26 N8O, calculated value 467.2, measured value 467.3.

[0679] Example 83: m-[2-amino-6-(1-{[6-(methoxyphenylmethyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0680]

[0681] The title compound was prepared in a manner similar to Example 79, thereby providing 90 mg of brown solid. 1 H NMR(400MHz, DMSO-d6)δ8.72–8.65(m,1H),8.60(t,J=2.0Hz,1H),8.52–8.45(m,1H),8 .00(ddd,J=7.8,2.7,1.5Hz,1H),7.88–7.80(m,2H),7.75(td,J=7.9,2.0Hz,1H),7.49 (d,J=7.8Hz,1H),7.34(dd,J=7.2,1.8Hz,2H),7.26(tt,J=7.4,1.4Hz,2H),7.23–7.12 (m,2H),6.91(s,2H),5.85–5.76(m,2H),5.32(d,J=1.9Hz,1H),3.33–3.31(m,3H).ESI MS[M+H] + For C 27 H 22 N8O, calculated value 475.2, measured value 475.3.

[0682] Example 84: m-{6-[1-({6-[(R)-1-methoxyethyl]-2-pyridyl}methyl)-1H-1,2,3-triazol-4-yl]-2-amino-4-pyrimidinyl}benzylnitrile

[0683]

[0684] Step 1: Formic acid (8.0 mL) was added dropwise to triethylamine (13.6 mL) at 0 °C. The mixture was degassed before adding copper (2.68 g, 10.0 mmol), RuCl(p-cymene)-[(R,R)-Ts-DPEN] (129 mg, 0.200 mmol), and CH2Cl2 (2.6 mL). The mixture was stirred at room temperature for 14 hours and then irradiated with saturated NaHCO3. 3(aq) The product was quenched, diluted with EtOAc (200 mL), washed with brine, and dried over Na2SO4. The crude product was purified by silica gel chromatography (0 to 5% MeOH in CH2Cl2 solution) to provide the desired product as a brown oil (896 mg; 33%).

[0685] Step 2: NaH (204 mg, 5.11 mmol, 60% in oil) was added in a single addition to a solution of the product from Step 2 (1.14 g, 4.26 mmol) in THF (21 mL) at 0 °C. The mixture was stirred at room temperature for 15 minutes, cooled to 0 °C, and iodomethane (265 μL, 4.26 mmol) was added dropwise. The mixture was stirred at room temperature for 2 hours and concentrated onto silica gel. The crude product was purified by silica gel chromatography (0 to 30% EtOAc in hexane solution) to provide the desired product (803 mg; 67%) as a colorless oil.

[0686] Steps 3-4: Azide was synthesized in a manner similar to that of Example 79, and a colorless oily product (373 mg, 68% (2 steps)) was obtained.

[0687] Step 5: The product was synthesized in a manner similar to Step 6 of Example 1: a grayish-white solid (97 mg, 79%). The compound was synthesized in a manner similar to Step 6 of Example 1. 1H NMR(400MHz, DMSO-d6)δ8.76(s,1H),8.62–8.56(m,1H),8.52–8.43(m,1H),8.04–7.97(m,1H),7.90–7.82(m,2H),7.75(t,J=7.8 Hz,1H),7.39(d,J=8.2Hz,1H),7.18(d,J=6.7Hz,1H),5.85(s,2H),4.34(q,J=6.5Hz,1H),3.19(s,3H),1.32(d,J=6.5Hz,3H).ESI MS[M+H] + For C 22 H 21 N8O, calculated value 413.2, measured value 413.3.

[0688] Example 85: 3-{6-[1-({6-[(R)-1-methoxyethyl]-2-pyridyl}methyl)-1H-1,2,3-triazol-4-yl]-2-amino-4-pyrimidinyl}-2-fluorobenzylnitrile

[0689]

[0690] The compound was synthesized from the corresponding alkyne in a manner similar to that of Example 84. 1 H NMR(400MHz,DMSO-d6)δ8.74(s,1H),8.35–8.26(m,1H),8.14–8.05(m,1H),7.91–7.82(m,1H),7.68–7.62(m,1H),7.62–7.53 (m,1H),7.39(d,J=7.6Hz,1H),7.18(d,J=7.0Hz,1H),5.84(s,2H),4.39–4.30(m,1H),3.19(s,3H),1.32(d,J=6.6Hz,3H).ESI MS[M+H] + For C 22 H 20 FN8O, calculated value 431.2, measured value 431.3.

[0691] Example 86: m-{6-[1-({6-[(S)-1-methoxyethyl]-2-pyridyl}methyl)-1H-1,2,3-triazol-4-yl]-2-amino-4-pyrimidinyl}benzylnitrile

[0692] Synthesis: Except for the use of RuCl(p-cymene)-[(S,S)-Ts-DPEN] as a catalyst in step 2, the azides were synthesized in a manner similar to that of Example 84.

[0693] Step 6: The product was synthesized in a manner similar to Step 6 of Example 1: a grayish-white solid (96 mg, 78%). 1 HNMR(400MHz,DMSO-d6)δ8.78(s,1H),8.63–8.57(m,1H),8.53–8.44(m,1H),8.04–7.99(m,1H),7.92–7.82(m,2H),7.80–7.73 (m,1H),7.39(d,J=8.2Hz,1H),7.19(d,J=7.9Hz,1H),5.85(s,2H),4.38–4.30(m,1H),3.19(s,3H),1.32(d,J=6.5Hz,3H).ESI MS[M+H] + For C 22 H 21 N8O, calculated value 413.2, measured value 413.3.

[0694] Example 87: 3-{6-[1-({6-[(S)-1-methoxyethyl]-2-pyridyl}methyl)-1H-1,2,3-triazol-4-yl]-2-amino-4-pyrimidinyl}-2-fluorobenzylnitrile

[0695]

[0696] The compound was synthesized from the corresponding alkyne in a manner similar to that of Example 86. 1 H NMR(400MHz, DMSO-d6)δ8.76(s,1H),8.31(td,J=7.8,1.8Hz,1H),8.13–8.06(m,1H),7.87(t,J=7.8Hz,1H),7.65(d,J=2.4Hz,1H),7.58( t,J=7.8Hz,1H),7.40(d,J=7.8Hz,1H),7.19(d,J=7.3Hz,1H),5.85(s,2H),4.35(q,J=6.5Hz,1H),3.19(s,3H),1.32(d,J=6.5Hz,3H).ESI MS[M+H] + For C 22 H 20 FN8O, calculated value 431.2, measured value 431.2.

[0697] Example 88: m-[2-amino-6-(1-{1-[6-(methoxymethyl)-2-pyridyl]ethyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0698]

[0699] Step 1. Cool the solution of aldehyde (756 mg, 5.0 mmol) in THF (10 mL) to -78 °C. Add MeMgBr (3N in Et₂O, 2 mL, 1.2 equivalences) dropwise. Slowly heat the resulting mixture to 0 °C over 2 hours and then quench with a saturated NH₄Cl solution. After conventional treatment (H₂O / EtOAc), dry the organic matter with sodium sulfate, filter, and evaporate to dryness. Purify the residue by silica gel chromatography (Hex / EtOAc 90:10 to 60:40) to provide the corresponding alcohol (635 mg, 76%).

[0700] The alcohol (600 mg, 3.6 mmol) obtained in step 1 was dissolved in CH₂Cl₂ (4 mL), and SOCl₂ (525 μL, 2 equivalents, 7.4 mmol) was added. The resulting solution was stirred until the initial alcohol was completely converted, at which point the mixture was evaporated to dryness. The resulting residue was used directly without further purification.

[0701] Step 2: The feedstock obtained in Step 1 was dissolved in DMF (7 mL) and sodium azide (325 mg, 5 mmol) was added. The resulting mixture was stirred at 80 °C for 8 hours, then cooled to room temperature. The crude product was partitioned between water and dichloromethane. The organic layer was evaporated to dryness, and the residue was purified by silica gel chromatography (Hex / EtOAc 90:10) to provide the desired azide (580 mg, 84% after step 2).

[0702] Step 3: The title compound was synthesized using an azide derivative and m-(2-amino-6-ethynyl-4-pyrimidinyl)benzyl nitrile (from Example 4) in a manner similar to Step 6 of Example 1. 1 ¹H NMR (400MHz, chloroform-d) δ 8.47–8.43 (m, 1H), 8.40 (s, 1H), 8.30 (ddd, J = 8.0, 1.8, 1.2Hz, 1H), 7.89 (s, 1H), 7.75 (ddd, J = 7.7, 1.7, 1.2Hz, 1H), 7.69 (dd, J = 7.8, 7.8Hz, 1H), 7.60 (dd, J = 7.8, 7.8Hz, 1H), 7.39 (d, J = 7.8Hz, 1H), 7.12 (d, J = 7.8Hz, 1H), 5.18 (s, 2H), 4.59 (s, 2H), 3.49 (s, 3H), 2.03 (d, J = 7.2Hz, 3H). MS [M+H] + For C 22 H 20 N8O, calculated value 413.2, measured value 413.3.

[0703] Example 89: m-(6-{1-[(6-{[(S)-tetrahydrofuran-3-yloxy]methyl}-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-2-amino-4-pyrimidinyl)benzylnitrile

[0704]

[0705] Step 1. Add NaH (60%, 400 mg, 10 mmol) in five portions to a solution of 3(S)-hydroxytetrahydrofuran (440 mg, 5 mmol) in dry THF (20 mL) under stirring at 0 °C. Stir at this temperature for 30 min. A gray suspension is obtained. Add 2,6-bis(chloromethyl)pyridine hydrochloride (1.06 g, 5 mmol) to this reaction mixture in a single batch at 0 °C. Stir the reaction mixture overnight at room temperature. Cool to 0 °C, quench with a saturated aqueous solution of NH4Cl, dilute with MTBE (10 mL), separate the layers, extract the aqueous layer with MTBE, combine the organic matter, dry (Na2SO4), filter, and concentrate on rotary evaporation. Dissolve the oily residue in dichloromethane and purify by rapid column chromatography (ISCO, 40 g column, 5-60% ethyl acetate in hexane solution) to give a pure compound (480 mg, 42%) as a colorless liquid.

[0706] Step 2: The above product (480 mg, 2.1 mmol) was dissolved in dry DMSO (2 mL), NaN3 (164 mg, 2.53 mmol) was added, and the mixture was stirred at room temperature for 2 hours. The reaction was indicated by LCMS to be complete. The product was diluted with water (15 mL), extracted with MTBE (3 × 15 mL), dried (Na2SO4), filtered, and concentrated on rotary evaporation. The oily residue was dried under high vacuum to provide the product (455 mg, 92%).

[0707] Step 3: The title compound is prepared from the above azide and the corresponding alkyne in a manner similar to Step 6 of Example 1. 1H NMR (400MHz, DMSO-d6) δ8.72(d,J=1.1Hz,1H),8.60(t,J=1.5Hz,1H),8.48(ddd,J=8.0,1.9,1. 2Hz,1H),8.01(dt,J=7.7,1.3Hz,1H),7.87(t,J=7.7Hz,1H),7.83(d,J=0.8Hz,1H),7.75(t,J=7 .9Hz,1H),7.41(dd,J=7.8,1.0Hz,1H),7.25(d,J=7.3Hz,1H),6.99(s,2H),5.84(s,2H),4.52(d ,J=1.8Hz,2H),4.30–4.21(m,1H),3.79–3.70(m,2H),3.70–3.61(m,2H),1.97–1.92(m,2H); ESI MS[M+H] + For C 24 H 22 N8O2, calculated value 455.2, measured value 455.3.

[0708] Example 90: m-(6-{1-[(6-{[(R)-tetrahydrofuran-3-yloxymethyl}-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-2-amino-4-pyrimidinyl)benzylnitrile

[0709]

[0710] The title compound was prepared from the corresponding azide and alkyne, similar to Example 89. 1 H NMR (400MHz, DMSO-d6) δ8.73(t,J=1.2Hz,1H),8.60(dt,J=1.8,1.0Hz,1H),8.48(ddd,J=8.0 ,1.9,1.2Hz,1H),8.01(dt,J=7.7,1.4Hz,1H),7.87(t,J=7.8Hz,1H),7.84(d,J=1.1Hz,1H), 7.76(t,J=7.9Hz,1H),7.41(dd,J=7.8,0.9Hz,1H),7.25(d,J=7.7Hz,1H),7.04(s,2H),5.84 (s,2H),4.53(d,J=1.8Hz,2H),4.31–4.19(m,1H),3.83–3.58(m,4H),1.97–1.92(m,2H); ESI MS[M+H] + For C 24 H 22 N8O2, calculated value 455.2, measured value 455.3.

[0711] Example 91: m-{2-amino-6-[1-({6-[(2-methoxyethoxy)methyl]-2-pyridyl}methyl)-1H-1,2,3-triazol-4-yl]-4-pyrimidinyl}benzylnitrile

[0712]

[0713] The title compound was prepared from the corresponding azide and alkyne, similar to Example 89. 1 H NMR (400MHz, DMSO-d6) δ8.70 (s, 1H), 8.59 (td, J=1.8, 0.6Hz, 1H), 8.47 (ddd, J=8 .0,1.8,1.2Hz,1H),8.00(ddd,J=7.7,1.7,1.1Hz,1H),7.87(t,J=7.8Hz,1H),7.8 2(s,1H),7.77–7.71(m,1H),7.43–7.39(m,1H),7.25–7.21(m,1H),6.93(s,2H), 5.83(s,2H),4.54(s,2H),3.66–3.57(m,2H),3.52–3.43(m,2H),3.24(s,3H);ESI MS[M+H] + For C 23 H 22 N8O2, calculated value 443.2, measured value 443.3.

[0714] Example 92: 3-{2-amino-6-[1-({6-[(2-methoxyethoxy)methyl]-2-pyridyl}methyl)-1H-1,2,3-triazol-4-yl]-4-pyrimidinyl}-2-methoxybenzonitrile

[0715]

[0716] The title compound was prepared from the corresponding azide and alkyne, similar to Example 89. 1 ¹H NMR (400MHz, chloroform-d) δ 8.30 (s, 1H), 8.01 (dd, J = 7.9, 1.8Hz, 1H), 7.91 (s, 1H), 7.78–7.62 (m, 2H), 7.48 (d, J = 8.0Hz, 1H), 7.35–7.26 (m, 1H), 7.12 (d, J = 7.7Hz, 1H), 5.71 (s, 2H), 5.12 (s, 2H), 4.70 (s, 2H), 3.94 (m, 3H), 3.77–3.71 (m, 2H), 3.65–3.59 (m, 2H), 3.41 (s, 3H); ESI MS [M+H] + For C24 H 24 N8O3, calculated value 473.2, measured value 473.3.

[0717] Example 93: 3-(2-amino-6-{1-[(6-cyclopropyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)-2-methoxybenzonitrile

[0718]

[0719] Step 1: Pd(OAc)₂ (516 mg, 2.3 mmol) was added to a mixture of 2-bromopyridine derivative (14 g, 46.4 mmol), cyclopropylboronic acid (8 g, 93 mmol), K₃PO₄ (34.5 g, 162.4 mmol), and PCy₃ (1.3 g, 4.64 mmol) in 210 mL of 20:1 toluene / H₂O. The reaction mixture was stirred in N₂ at 100 °C for 12 hours. Saturated NH₄Cl (50 mL) was added to quench the reaction, and the aqueous layer was extracted with EtOAc (2 x 70 mL). The combined organic layers were dried over Na₂SO₄, concentrated, and carried to the next step without further purification.

[0720] Step 2: Dissolve the crude TBS-ether from the previous step in 100 mL of THF and add 46.4 mL of a 1 M TBAF solution in THF dropwise. After 15 min, add 50 mL of saturated NH4Cl to quench the reaction and extract the aqueous layer with EtOAc (2 x 70 mL). Dry the combined organic layers with Na2SO4, concentrate, and purify by silica gel chromatography to obtain the desired alcohol (6.3 g, 91% in step 2).

[0721] Step 3: Synthesize the azide in a manner similar to step 5 in Example 1: colorless oil (6.2 g, 85%).

[0722] Step 4: Synthesize the title compound in a manner similar to step 6 in Example 1. 1H NMR (400MHz, DMSO-d6) δ8.62(s,1H),8.04(d,J=7.9Hz,1H),7.93(d,J=7.5Hz,1H),7.66(dd,J=7.7,7.7Hz,1H),7.62(s,1H),7.42(dd,J=7.5,7 .5Hz,1H),7.22(d,J=7.8Hz,1H),7.02(d,J=7.6Hz,1H),5.73(s,2H),3. 83(s,3H),2.09–2.02(m,1H),0.93–0.88(m,2H),0.82–0.78(m,2H).ESI MS[M+H] + For C 23 H 20 N8O, calculated value 425.2, measured value 425.3.

[0723] Example 94: m-(2-amino-6-{1-[(6-cyclopropyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)benzylnitrile

[0724]

[0725] The title compound was prepared from the corresponding azide and alkyne, similar to Example 93. 1 ¹H NMR (400MHz, chloroform-d) δ 8.49–8.39 (m, 1H), 8.35–8.23 (m, 2H), 7.93–7.81 (m, 1H), 7.80–7.68 (m, 1H), 7.63–7.45 (m, 2H), 7.14–7.03 (m, 1H), 7.02–6.89 (m, 1H), 5.63 (s, 2H), 5.25 (s, 2H), 2.08–1.95 (m, 1H), 1.05–0.92 (m, 4H); LC-MS retention time 3.15 min. LC-MS, Method A, ESI MS [M+H] + ] - For C 22 H 19 N8, calculated value 395.2, measured value 395.3.

[0726] Example 95: 3-(2-amino-6-{1-[(6-cyclopropyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)-2-fluorobenzylnitrile

[0727]

[0728] The title compound was prepared from the corresponding azide and alkyne, similar to Example 93.1 ¹H NMR (400MHz, chloroform-d) δ 8.38–8.18 (m, 2H), 7.91–7.82 (m, 1H), 7.76–7.64 (m, 1H), 7.57–7.46 (m, 1H), 7.42–7.31 (m, 1H), 7.14–7.04 (m, 1H), 7.00–6.91 (m, 1H), 5.62 (s, 2H), 5.29 (s, 2H), 2.09–1.95 (m, 1H), 1.07–0.89 (m, 4H); LC-MS retention time 3.15 min. LC-MS, Method A, ESI MS [M+H + For C 22 H 18 FN8, calculated value 413.2, measured value 413.3.

[0729] Example 96: 4-{1-[(6-cyclopropyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-6-(2,3-difluorophenyl)-2-pyrimidinylamine

[0730]

[0731] The title compound was prepared from the corresponding azide and alkyne, similar to Example 93. 1 ¹H NMR (400 MHz, chloroform-d) δ 8.29 (s, ¹H), 7.87 (s, ¹H), 7.78–7.69 (m, ¹H), 7.52 (dd, J = 7.8, 7.8 Hz, ¹H), 7.32–

[0732] 7.13(m,2H),7.09(d,J=7.8Hz,1H),6.95(d,J=7.6Hz,1H),5.62(s,2H),5.19(brs,2H),2.08–1.98(m,1H),1.05–0.94(m,4H).MS[M+H] + For C 21 H 17 F2N7, calculated value 406.2, measured value 406.3.

[0733] Example 97: 4-{1-[(6-cyclopropyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-6-(m-fluorophenyl)-2-pyrimidinylamine}

[0734]

[0735] The title compound was prepared from the corresponding azide and alkyne, similar to Example 93. 1H NMR (400MHz, CD3OD-d4)8.60(s,1H),7.93(d,J=8.0Hz,1H),7.89(d,J=12Hz,1H),7.77(s,1H),7.64(t,J=8Hz,1H),7.52(q,J=8 Hz,1H),7.25(t,J=8Hz,1H),7.17(d,J=4Hz,1H),7.09(d,J=8.0Hz,1H),5.72(s,1H),2.09-2.02(m,1H),0.98-0.91(m,4H).ESI MS[M+H] + For C 21 H 18 FN7, calculated value 388.4, measured value 388.3.

[0736] Example 98: 3-(2-amino-6-{1-[(6-isopropyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)-2-methoxybenzonitrile

[0737]

[0738] Step 1: 2-Bromo-6-({[dimethyl(2-methyl-2-propyl)silyl]oxy}methyl)pyridine (2.8 g, 9.2 mmol, 1.0 equivalent) and pinacol isopropylborate (2.3 g, 13.9 mmol, 1.5 equivalent) were dissolved in a solution of dioxane (37 mL, 0.25 M) and 2.0 M Na₂CO₃ aqueous solution (14 mL, 3.0 equivalent) and sprayed with N₂ for 10 min. After this, Pd(PPh₃)₄ (717 mg, 0.46 mmol, 0.05 equivalent) was added and the reaction mixture was heated to 95 °C for 18 h. After this, the reaction mixture was diluted with CH₂Cl₂ (100 mL), transferred to a separatory funnel, and washed with H₂O (100 mL). The organic phase was collected and the aqueous phase was extracted with CH₂Cl₂ (2 x 100 mL). The combined organic extracts were dried over MgSO₄ and concentrated under vacuum. The resulting oil was purified by column chromatography (0:1 EtOAc:hexanes → 1:9 EtOAc:hexane) to give the title compound (2.2 g, 90% yield) as a colorless oil.

[0739] Step 2: A solution of isopropylpyridine (2.2 g, 8.4 mmol, 1.0 equivalent) from Step 1 in methanol (20 mL, 0.5 M) and acetic acid (0.1 mL) was sprayed with N2 for 5 min, followed by the addition of PtO2 (117 mg, 0.52 mmol, 0.05 equivalent). The suspension was sprayed with an H2 balloon for 10 min, and the reaction was then stirred in an H2 atmosphere (balloon) for 20 h. After completion, the reaction mixture was filtered through a tinate filter, the filter cake was washed with methanol (2 x 10 mL), and the filtrate was concentrated under vacuum. The resulting oil was used for subsequent steps without further purification.

[0740] Step 3: The intermediate from the previous step was placed in a 1.0 M TBAF solution of THF (20 mL, 2.0 equivalents) and stirred at room temperature for 45 minutes. The reaction mixture was then directly loaded into SiO2 and purified by column chromatography (0:1 MeOH:CH2Cl2 → 1:9 MeOH:CH2Cl2) to obtain a colorless, oily (6-isopropyl-2-pyridyl)methanol (1.1 g, 87% yield).

[0741] Step 4: Diphenyl azidophosphate (1.8 mL, 8.4 mmol, 1.2 equivalence) was added to a solution of (6-isopropyl-2-pyridyl)methanol (1.1 g, 7.0 mmol, 1.0 equivalence) in toluene (14 mL, 0.5 M), followed by 1,8-diazabicyclo[5.4.0]undec-7-ene (1.3 mL, 8.4 mmol, 1.2 equivalence). The resulting mixture was heated to 60 °C for 1.5 h. The reaction mixture was then directly loaded into SiO2 and purified by column chromatography (0:1 EtOAc:hexane → 1:19 EtOAc:hexane) to obtain a colorless oily 2-(azidomethyl)-6-isopropylpyridine (890 mg, 72% yield).

[0742] Step 5: Synthesize the title compound in a manner similar to step 6 in Example 1. 1 H NMR(400MHz, DMSO-d6)δ8.71(s,1H),8.05(d,J=7.8Hz,1H)7.94(d,J=7.7Hz,1H),7.80–7.75(m,1H),7.63(s,1H),7.43(dd, J=7.7,7.7Hz,1H),7.29(d,J=7.9Hz,1H),7.09(d,J=7.6Hz,1H),5.81(s,2H),3.03–2.96(m,1H),1.19(d,J=6.9Hz,6H).ESI MS[M+H] + For C 23 H 22 N8O, calculated value 427.2, measured value 427.3.

[0743] Example 99: m-(2-amino-6-{1-[(6-isopropyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)benzylnitrile

[0744]

[0745] The title compound was prepared from the corresponding azide and alkyne, similar to Example 98. 1 H NMR(400MHz,CD3OD)δ8.82(s,1H),8.54(s,1H),8.45(d,J=8.1Hz,1H),7.94–7.86(m,overlap,3H),7.74(dd,J=8.0,8 .0Hz,1H),7.39(d,J=8.0Hz,1H),7.28,J=8.0Hz,1H),5.86(s,2H),3.10(sept,7.0Hz,1H),1.29(d,J=7.0Hz,6H).ESI MS[M+H] + For C 22 H 20 N8, calculated value 397.2, measured value 397.3

[0746] Example 100: 6-(m-fluorophenyl)-4-{1-[(6-isopropyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-2-pyrimidinylamine

[0747]

[0748] The title compound was prepared from the corresponding azide and alkyne, similar to Example 98. 1 H NMR (400MHz, CD3OD-d4)8.65(s,1H),7.94(d,J=8.0Hz,1H),7.89(d,J=8.0Hz,1H),7.73-7.78(m,3H),7.53(d,J=8Hz,1H),7.27(d,J= 8Hz,1H),7.16(d,J=4Hz,1H),7.20(dt,J=8,4Hz,1H),7.1(d,J=8.0Hz,1H),5.79(s,2H),3.04-3.07(m,1H),1.28(d,J=4Hz,6H)..ESI MS[M+H] + For C 21 H 20 FN7, calculated value 390.4, measured value 390.3.

[0749] Example 101: 3-(2-amino-6-{1-[(6-isopropyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)-2-fluorobenzyl nitrile

[0750]

[0751] The title compound was prepared from the corresponding azide and alkyne, similar to Example 98. 1 H NMR (400MHz, acetone-d6) δ8.59 (d, J=0.8Hz, 1H), 8.45–8.36 (m, 1H), 7.98 (dddd, J=7. 7,6.0,1.8,0.8Hz,1H),7.85(dd,J=2.7,0.8Hz,1H),7.74(td,J=7.8,0.7Hz,1H) ,7.58(t,J=7.8Hz,1H),7.25(d,J=7.8Hz,1H),7.15(dd,J=7.6,0.9Hz,1H),6.30 (s,2H),5.80(s,2H),3.03(hept,J=6.8Hz,1H),1.24(dd,J=6.9,0.8Hz,6H).ESI MS[M+H] + For C 22 H 19 FN8, calculated value 415.2, measured value 415.3.

[0752] Example 102: 6-(2,3-difluorophenyl)-4-{1-[(6-isopropyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-2-pyrimidinylamine

[0753]

[0754] The title compound was prepared from the corresponding azide and alkyne, similar to Example 98. 1 ¹H NMR (400MHz, acetone-d6) δ 8.72 (s, 1H), 7.89–7.87 (s, 2H), 7.79 (t, J = 7.9 Hz, 1H), 7.51 (q, J = 8.6 Hz, 1H), 7.39–7.34 (m, 1H), 7.29 (d, J = 7.8 Hz, 1H), 7.21 (d, J = 7.7 Hz, 1H), 5.85 (s, 2H), 3.07 (p, J = 7.2 Hz, 1H), 1.25 (dd, J = 6.9, 1.2 Hz, 6H). ESI MS [M+H] + For C 21 H 19 F2N7, calculated value 408.2, measured value 408.3.

[0755] Example 103: 6-(2-amino-6-{1-[(6-isopropyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)-2-methylbenzonitrile

[0756]

[0757] The title compound was prepared from the corresponding azide and alkyne, similar to Example 98. 1 H NMR (400MHz, acetone-d6) δ8.58(s,1H),7.86–7.70(m,3H),7.53(td,J=7.8,0.7Hz,1H),7.44(d,J=0.9Hz,1H),7.26(d,J= 7.8Hz,1H),7.15(d,J=7.7Hz,1H),6.22(s,2H),5.80(s,2H),3.04(p,J=6.9Hz,1H),1.25(dd,J=6.9,0.9Hz,6H).ESI MS[M+H] + For C 23 H 22 N8, calculated value 411.2, measured value 411.3.

[0758] Example 104: 3-(2-amino-6-{1-[(6-isopropyl-2-pyridyl)methyl]-1H-1,2,3-triazol-4-yl}-4-pyrimidinyl)-2-ethoxybenzyl nitrile

[0759]

[0760] The title compound was prepared from the corresponding azide and alkyne, similar to Example 98. 1 ¹H NMR (400MHz, acetone-d6) δ 8.72 (d, J = 1.7 Hz, 1H), 8.25–8.18 (m, 1H), 8.05 (d, J = 1.8 Hz, 1H), 7.89 (dd, J = 7.7, 2.0 Hz, 1H), 7.82–7.73 (m, 1H), 7.47 (td, J = 7.7, 1.8 Hz, 1H), 7.28 ( d,J=7.8Hz,1H),7.22(d,J=7.9Hz,1H),5.84(d,J=1.7Hz,3H),4.16(q,J=8.0Hz,2H) ,3.05(p,J=6.6Hz,1H),1.38(td,J=7.0,1.8Hz,4H),1.25(dd,J=6.9,1.9Hz,6H).ESI MS[M+H] + For C 24 H 24N8O, calculated value 441.2, measured value 441.3.

[0761] Example 105: m-[2-amino-6-(1-{[6-(tert-butyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0762]

[0763] Step 1: At -78°C, t-BuMgCl₂ (50.0 mL, 50.0 mmol, 1 M in THF) was added to a suspension of CuCN (2.24 g, 25.0 mmol) in THF (50 mL). The mixture was stirred at -78°C for 30 minutes. A bromopyridine derivative (1.51 g, 5.00 mmol) was added dropwise, and the mixture was stirred at -78°C for 2 hours. The mixture was warmed to room temperature for 14 hours, and NH₃ (50 mL, 25% aqueous solution) was added, followed by 50 mL of ethyl acetate. The mixture was stirred at room temperature for 30 minutes and filtered to remove any solids. The organic phase was dried over brine and MgSO₄ and passed through a silica-based stopper, then eluted with ethyl acetate. The organic phase was concentrated to provide the desired yellow oily product, which was used directly in the next step.

[0764] Steps 2-3: Synthesize the azide in a manner similar to Example 79. Colorless oil (253 mg, 27%, 3 steps).

[0765] Step 4: Synthesize the product in a manner similar to that in Example 1, Step 6: yellow solid (86 mg, 70%). 1 H NMR (400MHz, DMSO-d6) δ8.90(s,1H),8.63(s,1H),8.50(d,J=8.7Hz,1H),8.05(d,J=7.7Hz,1H),7.95(s ,1H),7.84–7.73(m,2H),7.53–7.33(m,1H),7.14(dd,J=7.7,0.9Hz,1H),5.88(s,2H),1.25(s,9H).ESI MS[M+H] + For C 23 H 23 N8, calculated value 411.2, measured value 411.3.

[0766] Example 106: 6-(3-chloro-2-methoxyphenyl)-4-(1-{[6-(tert-butyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0767]

[0768] The title compound was prepared from the corresponding azide and alkyne, similar to Example 105. 1 H NMR (400MHz, CDCl3) δ8.35(d,J=2.3Hz,1H),7.92(d,J=2.3Hz,1H),7.66(dd,J=7.8,1.9Hz,1H),7.62–7.55(m,1H),7.49–7.42 (m,1H),7.31–7.24(m,1H),7.19–7.11(m,1H),7.01(d,J=7.6Hz,1H),5.68(s,2H),5.23(s,2H),3.75(s,2H),1.34(s,9H).ESI MS[M+H] + For C 23 H 24 ClN7O, calculated value 450.2, measured value 450.3.

[0769] Example 107: 6-(3-fluoro-2-methoxyphenyl)-4-(1-{[6-(tert-butyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0770]

[0771] The title compound was prepared from the corresponding azide and alkyne, similar to Example 105. 1 H NMR(400MHz,DMSO-d6)δ8.65(s,1H),7.79–7.71(m,1H),7.65–7.57(m,2H),7.45–7.36(m,2H),7 .28–7.19(m,1H),7.07(d,J=7.4Hz,1H),6.80(s,2H),5.80(s,2H),3.85(s,3H),1.26(s,9H).ESI MS[M+H] + For C 23 H 25 FN7O, calculated value 434.2, measured value 434.4.

[0772] Example 108: 3-[2-amino-6-(1-{[6-(tert-butyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-2-methoxybenzonitrile

[0773]

[0774] The title compound was prepared from the corresponding azide and alkyne, similar to Example 105. 1H NMR(400MHz, DMSO-d6)δ8.76(s,1H),8.07(dd,J=7.9,1.7Hz,1H),7.99–7.95(m,1H),7.77(t,J=7.8Hz,1H),7.66(s, 1H),7.46(t,J=7.8Hz,1H),7.40(d,J=7.9Hz,1H),7.11(d,J=8.5Hz,1H),5.83(s,2H),3.86(s,3H),1.26(s,9H).ESI MS[M+H] + For C 24 H 25 N8O, calculated value 441.2, measured value 441.3.

[0775] Example 109: 3-[2-amino-6-(1-{[6-(tert-butyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-2-fluorobenzylnitrile

[0776]

[0777] The title compound was prepared from the corresponding azide and alkyne, similar to Example 105. 1 H NMR(400MHz, DMSO-d6)δ8.75(s,1H),8.31(td,J=7.8,1.8Hz,1H),8.13–8.07(m,1H),7.77(t,J=7.8Hz,1H),7.66( d,J=2.4Hz,1H),7.58(t,J=7.8Hz,1H),7.40(d,J=7.1Hz,1H),7.11(d,J=7.7Hz,1H),5.84(s,2H),1.25(s,9H).ESI MS[M+H] + For C 23 H 22 FN8, calculated value 429.2, measured value 429.3.

[0778] Example 110: 6-(2,3-difluorophenyl)-4-(1-{[6-(tert-butyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0779]

[0780] The title compound was prepared from the corresponding azide and alkyne, similar to Example 105. 1H NMR (400MHz, CDCl3) δ8.38(s,1H),7.87(s,1H),7.77–7.70(m,1H),7.60(dd,J=8.0,8.0Hz,1H),7.29(d,J =8.0Hz,1H),7.28–7.13(m,2H),7.02(d,J=7.8Hz,1H),5.69(s,2H),5.17(brs,2H),1.35(s,9H).MS[M+H] + For C 22 H 21 F2N7, calculated value 422.2, measured value 422.3.

[0781] Example 111: 6-[2-amino-6-(1-{[6-(tert-butyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-2-methylbenzonitrile

[0782]

[0783] The title compound was prepared from the corresponding azide and alkyne in a manner similar to Example 105, thereby providing 73 mg of brown solid. 1 H NMR(400MHz, DMSO-d6)8.70(s,1H),7.90(dt,J=7.7,1.3Hz,1H),7.75(td,J=7.8,1.2Hz,2H),7.52(t,J=7.7Hz,1H),7.46–7.33( m,1H),7.28(d,J=1.2Hz,1H),7.07(d,J=7.6Hz,1H),6.90(s,2H),5.81(s,2H),2.55(d,J=1.1Hz,3H),1.26(d,J=1.3Hz,9H).ESI MS[M+H] + For C 24 H 24 N8, calculated value 425.2, measured value 425.4.

[0784] Example 112: 6-(m-fluorophenyl)-4-(1-{[6-tert-butyl)2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-2-pyrimidinylamine

[0785]

[0786] The title compound was prepared from the corresponding azide and alkyne, similar to Example 105. 1H NMR (400MHz, CD3OD)8.64(s,1H),7.89(d,J=8.0Hz,1H),7.84(d,J=8.0Hz,1H),7.74(s,1H),7.67(t,J=8Hz,1H),7 .47(q,J=8Hz,1H),7.34(d,J=8Hz,1H),7.20(dt,J=8,4Hz,1H),7.1(d,J=8.0Hz,1H),5.75(s,2H),1.28(s,9H).ESI MS[M+H] + For C 22 H 22 FN7, calculated value 404.4, measured value 404.4.

[0787] Example 113: 2-[6-({4-[2-amino-6-(o-fluorophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]-2-methylpropiononitrile

[0788]

[0789] The title compound was prepared from the corresponding azide and alkyne, similar to Example 1. 1 H NMR (400MHz, chloroform-d) δ8.34(s,1H),8.01(ddd,J=7.8,7.8,1.9Hz,1H),7.92(d,J=2.2Hz,1H),7.74(dd,J=7.8,7.8Hz,1H),7.55(d,J=7 .8Hz,1H),7.48–7.42(m,1H),7.31–7.22(m,1H),7.22–7.14(m,2H),5.74(s,2H),5.30(s,2H),5.14(brs,2H),1.74(s,6H).MS[M+H] + For C 22 H 17 FN8, calculated value 415.2, measured value 415.2.

[0790] Example 114: 5-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-3-{[6-(tert-butyl)-2-pyridyl]methyl}-3H-1,2,3-triazole-4-carboxylic acid

[0791]

[0792] Step 1. Degas the mixture of chloride (500 mg, 2.17 mmol) and propargyl alcohol (0.5 mL) in DMF (3 mL) and Et3N (5 mL) for 5 minutes. Add PdCl2 (dppf) (79 mg, 5 mol%) and CuI (41 mg, 10 mol%) and heat the mixture to 75 °C for 1 hour. After routine treatment, purify by silica gel ((hexane / CH2Cl2)(1:1) / EtOAc 100:0 to 0:100) to provide the desired alkyne (210 mg, 39%).

[0793] Step 2: A mixture of the alkyne derivative (70 mg) and the azide derivative (60 mg, 1.1 eq.) was heated in toluene to 120 °C for 30 hours. Excess solvent was removed under vacuum and purified by rapid column chromatography to provide m-{2-amino-6-[5-(hydroxymethyl)-1-{[6-(tert-butyl)-2-pyridinyl]methyl}-1H-1,2,3-triazol-4-yl]-4-pyrimidinyl}benzylnitrile. 1 H NMR (400MHz, chloroform-d) δ8.42(s,1H),8.35–8.25(m,1H),8.02(s,1H),7.76(d,J=7.8Hz,1H),7.64–7.53(m,2H),7.32(d, J=7.8Hz,1H),7.17(d,J=7.8Hz,1H),6.63(brs,1H),5.73(s,2H),5.23(s,2H),5.07(brs,2H),1.24(s,9H).MS[M+H] + For C 24 H 24 N8O, calculated value 441.2, measured value 441.4.

[0794] Step 3: 35 mg (0.08 mmol) of m-{2-amino-6-[5-(hydroxymethyl)-1-{[6-(tert-butyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl]-4-pyrimidinyl}benzyl nitrile was placed in CH2Cl2 (3 mL) and MnO2 (1.05 g) was added. The resulting mixture was stirred at room temperature for 24 hours. The mixture was filtered through an inotropic salt and subsequently purified by silica gel chromatography (CH2Cl2 / EtOAc 90:10 to 20:80) to yield the corresponding aldehyde (15 mg, 43%) and acid (11 mg, 31%). 1H NMR (400MHz, DMSO-d6) δ8.71(s,1H),8.63(d,J=7.9Hz,1H),8.29(s,1H),8.02(d,J=7.9Hz,1H),7.84(dd,J=7.9,7.9Hz,1H), 7.71(dd,J=7.9,7.9Hz,2H),7.30(d,J=7.9Hz,1H),7.20(brs,2H),7.13(d,J=7.9Hz,1H),6.31(s,2H),1.18(s,9H).MS[M+H] + For C 24 H 22 N8O2, calculated value 455.2, measured value 455.3.

[0795] Example 115: m-[2-amino-6-(1-{[6-(1-hydroxycyclobutyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0796]

[0797] Step 1: Place 2.0 g (6.7 mmol) of a commercially available 2-bromopyridine derivative into a round-bottom flask. Add 13.0 mL of dry THF to the flask and cool to -78 °C in N2. Add 2.7 mL of nBuLi (2.5 M in THF) dropwise to the reaction mixture at -78 °C and stir for 30 min. Then add 0.58 mL (7.9 mmol) of cyclobutanone in one go and warm the reaction mixture to room temperature for 2 h (LCMS showed formation of the desired addition product). Cool the reaction mixture back to 0 °C and add 6.7 mL of TBAF (1 M in THF). After stirring the reaction mixture at 0 °C for 15 min, quench the reaction with 50.0 mL of saturated NH4Cl aqueous solution. Extract the aqueous layer with EtOAc (2 x 50 mL), dry with Na2SO4, and concentrate. Purify the starting material by silica gel chromatography to obtain the desired pyridine-diol (570 mg, 48% in step 2).

[0798] Step 2: At room temperature, diphenylphosphoazide (0.8 mL, 3.8 mmol) and DBU (0.6 mL, 3.8 mmol) were added to a solution of the diol (570.0 mg, 3.2 mmol) from Step 1 in CH2Cl2 (4.0 mL). The reaction mixture was stirred in N2 at room temperature for 10 h. After removing CH2Cl2, the residue was redissolved in EtOAc and subsequently washed with H2O (2 x 25 mL). The organic layer was dried over Na2SO4 and concentrated. The starting material was purified by silica gel chromatography to obtain the desired azide (450 mg, 69%).

[0799] Step 3: Prepare the title compound from the corresponding azide and alkyne in a manner similar to Example 1 (Step 6). 1 H NMR (400MHz, chloroform-d) δ8.43(td,J=1.8,0.6Hz,1H),8.33(s,1H),8.29(ddd,J=8.0,1.8,1.2Hz,1H),7.88(s,1H),7.81–7.72(m,2H),7.62–7.5 4(m,2H),7.16(dd,J=7.6,0.9Hz,1H),5.74(s,2H),5.37(s,2H),5.03(s,1H),2.68–2.38(m,4H),2.18–2.07(m,1H),1.93–1.75(m,1H).ESI MS[M+H] + For C 23 H 20 N8O, calculated value 425.2, measured value 425.3.

[0800] Example 116: m-[2-amino-6-(1-{[6-(1-hydroxycyclopentyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]benzylnitrile

[0801]

[0802] The title compound was prepared from the corresponding azide and alkyne, similar to Example 115. 1 H NMR (400MHz, chloroform-d) δ8.46(td,J=1.7,0.6Hz,1H),8.36–8.28(m,2H),7.91(s,1H),7.80–7.69(m,2H),7.61(td,J=7.8,0.6Hz, 1H),7.39(dd,J=8.0,0.9Hz,1H),7.13(dd,J=7.6,0.9Hz,1H),5.76(s,2H),5.18(s,2H),4.70(s,1H),2.10–1.78(m,8H).ESI MS[M+H] + For C 24 H 22 N8O, calculated value 439.2, measured value 439.3.

[0803] Example 117: 1-[6-({4-[2-amino-6-(2,3-difluorophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]cyclopentanol

[0804]

[0805] The title compound was prepared from the corresponding azide and alkyne, similar to Example 115. 1 H NMR (400MHz, chloroform-d) δ8.30(s,1H),7.89(d,J=2.1Hz,1H),7.78–7.71(m,1H),7.71(dd,J=7.9,7.9Hz,1H),7.37(d,J=7.9Hz ,1H),7.30–7.22(m,1H),7.22–7.14(m,1H),7.11(d,J=7.6Hz,1H),5.74(s,2H),5.16(s,2H),2.12–1.79(m,8H).MS[M+H] + For C 23 H 21 F2N7O, calculated value 450.2, measured value 450.3.

[0806] Example 118: 3-[2-amino-6-(1-{[6-(1-hydroxycyclopentyl)-2-pyridyl]methyl}-1H-1,2,3-triazol-4-yl)-4-pyrimidinyl]-2-fluorobenzyl nitrile

[0807]

[0808] The title compound was prepared from the corresponding azide and alkyne, similar to Example 115. 1 ¹H NMR (400MHz, chloroform-d) δ 8.34 (s, 1H), 8.33–8.22 (m, 1H), 7.92–7.85 (m, 1H), 7.81–7.61 (m, 2H), 7.47–7.34 (m, 2H), 7.12 (d, J = 7.6Hz, 1H), 5.75 (s, 2H), 5.24 (s, 2H), 4.63 (brs, 1H), 2.13–1.61 (m, 8H). MS [M+H] + For C 24 H 21 FN8O, calculated value: 457.2, measured value: 457.4.

[0809] Example 119: 1-[6-({4-[2-amino-6-(o-fluorophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]cyclopentanol

[0810]

[0811] The title compound was prepared from the corresponding azide and alkyne, similar to Example 115. 1H NMR (400MHz, chloroform-d) δ8.29(s,1H),8.01(dd,J=7.7,7.7Hz,1H),7.94(s,1H),7.71(dd,J=7.4,7.4Hz,1H),7.44(s,1H),7.37(d, J=7.7Hz,1H),7.22–7.14(m,1H),7.10(d,J=7.4Hz,1H),5.75(s,2H),5.10(s,2H),4.68(brs,1H),2.12–1.77(m,8H).MS[M+H] + For C 23 H 22 FN7O, calculated value 432.2, measured value 432.3.

[0812] Example 120: 3-{6-[1-({6-[(S)-3-hydroxy-1-pyrrolidinyl]-2-pyridyl}methyl)-1H-1,2,3-triazol-4-yl]-2-amino-4-pyrimidinyl}-2-methoxybenzonitrile

[0813]

[0814] Step 1: In an 8 mL glass vial equipped with a magnetic stirrer, add azide (96.2 mg, 0.451 mmol), alkyne (113 mg, 0.451 mmol), CuSO4·5H2O (6 mg, 22.6 μmol, 5 mol%), sodium ascorbate (17.9 mg, 90.3 μmol, 20 mol%), and 2:1 tBuOH / H2O (1.81 mL, 0.25 M). Heat the resulting mixture to 55 °C for 2 h. After completion, cool the reaction mixture to room temperature and dilute with CH2Cl2 (3 mL). Separate the phases and extract the aqueous phase again with CH2Cl2 (3 mL). Concentrate the combined organic extracts under vacuum. Purify the crude residue by silica gel rapid column chromatography (gradient CH2Cl2 / MeOH) to provide a beige solid product (176 mg, 84% yield).

[0815] Step 2: A 1-dram vial equipped with a magnetic stirrer was filled with bromopyridine substrate (20.0 mg, 43.1 μmol, 1.0 equivalent), (S)-3-hydroxypyrrolidine (4.50 mg, 51.7 μmol, 1.2 equivalent), NaOtBu (8.30 mg, 86.2 μmol, 2.0 equivalent), PdG3 precatalyst (300 μg, 0.431 μmol, 1 mol%), BrettPhos (200 μg, 0.431 μmol, 1 mol%), and dioxane (100 μL, 0.45 M). The resulting mixture was degassed by venting and backfilling with N2 (3x), and then stirred at 100 °C for 3 h. After completion, the reaction mixture was cooled to room temperature, diluted with EtOAc, and filtered through a tin-based salt. The filtrate was concentrated under vacuum and the crude residue was purified by silica gel rapid column chromatography (gradient CH2Cl2 / MeOH) to provide a product as a grayish-white solid (5 mg, yield 25%). 1 ¹H NMR (400MHz, chloroform-d) δ 8.38 (s, 1H), 8.03–7.97 (m, 1H), 7.90 (s, 1H), 7.70–7.63 (m, 1H), 7.46–7.35 (m, 1H), 7.32–7.26 (m, 1H), 6.50 (d, J = 7.2Hz, 1H), 6.32 (d, J = 8.5Hz, 1H), 5.52 (s, 2H), 5.16 (s, 2H), 4.67–4.55 (m, 1H), 3.94 (s, 3H), 3.65–3.50 (m, 4H), 2.22–2.06 (m, 2H); LC-MS retention time 2.32 min. LC-MS, Method A, ESI MS [M+H + For C 24 H 24 N9O2, calculated value 470.2, measured value 470.3.

[0816] Example 121: 3-{6-[1-({6-[(R)-3-hydroxy-1-pyrrolidinyl]-2-pyridyl}methyl)-1H-1,2,3-triazol-4-yl]-2-amino-4-pyrimidinyl}-2-methoxybenzonitrile

[0817]

[0818] The title compound was prepared from the corresponding azide and alkyne, similar to Example 120. 1 ¹H NMR (400 MHz, chloroform-d) δ 8.38 (s, ¹H), 8.01 (dd, J = 7.6, 1.7 Hz, ¹H), 7.89 (s, ¹H), 7.72–7.64 (m, ¹H), 7.45–

[0819] 7.38 (m, 1H), 7.31–7.26 (m, 1H), 6.50 (d, J = 7.1 Hz, 1H), 6.32 (d, J = 8.4 Hz, 1H), 5.52 (s, 2H), 5.16 (s, 2H), 4.65–4.59 (m, 1H), 3.93 (s, 3H), 3.64–3.51 (m, 4H), 2.20–2.07 (m, 2H); LC-MS retention time 2.32 min. LC-MS, Method A, ESI MS [M+H] + For C 24 H 24 N9O2, calculated value 470.2, measured value 470.4.

[0820] Example 122: 1-{[6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]carbonyl}-4-piperidinecarboxylic acid

[0821]

[0822] Step 1. Perform cycloaddition in a manner similar to step 6 of Example 1 to obtain methyl 6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinecarboxylic acid. 1 H NMR(400MHz, DMSO-d6)δ8.75(s,1H),8.59(dt,J=1.8,1.0Hz,1H),8.48(ddd,J=8.0,1.8,1.1Hz,1H),8.09–8.03(m,2H),8.00( dt,J=7.7,1.3Hz,1H),7.82(s,1H),7.78–7.72(m,1H),7.55(dd,J=6.5,2.3Hz,1H),6.95(s,2H),5.94(s,2H),3.88(s,3H).ESI MS[M+H] + For C 21 H 16 N8O2, calculated value 413.1, measured value 413.2

[0823] Step 2: At room temperature, methyl 6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinecarboxylate (10 mg, 0.024 mmol) in t-BuOH (0.2 mL) and H2O (0.1 mL) was added to LiOH·H2O (1.5 mg, 0.036 mmol, 1.5 equivalents). The mixture was stirred overnight and washed with MTBE. The reaction was quenched by the addition of 1 M HCl (approximately 50 μL), extracted with EtOAc, and concentrated to give 9.7 mg of the compound as a yellowish-brown solid. 1 H NMR (400MHz, DMSO-d6) δ8.61(td,J=1.8,0.6Hz,1H),8.49(ddd,J=8.0,1.9,1.1Hz,1H),8.13–7.99( m,3H),7.89(s,1H),7.84–7.71(m,1H),7.55(dd,J=6.8,2.1Hz,1H),5.95(s,2H),5.36(bs,3H).ESI MS[M+H] + For C 20 H 14 N8O2, calculated value 399.1, measured value 399.2

[0824] Step 3: The mixture of the above-mentioned acid (30 mg, 0.075 mmol), amine (50 mg, 0.15 mmol), pyridine (0.5 mL), and T3P (0.2 mL) was stirred at 50 °C for one hour. The mixture was then placed directly on silica gel for purification (CH2Cl2:MeOH 100:0 to 95:5) to provide the amide (35 mg, 90%). The ester was hydrolyzed to give the title acid (23 mg, 66%). 1 ¹H NMR (400 MHz, acetone-d6) δ 8.63 (s, 1H), 8.58 (s, 1H), 8.50 (d, J = 8.0 Hz, 1H), 7.98 (dd, J = 8.0, 8.0 Hz, 1H), 7.94–7.86 (m, 1H), 7.89 (s, 1H), 7.75 (dd, J = 8.0, 8.0 Hz, 1H), 7.60 (d, J = 8.0 Hz, 1H). 1H),7.54(d,J=8.0Hz,1H),6.30(brs,1H),5.91(s,2H),4.48–4.39(m,1H),3.84–3.75(m, 1H),3.15–2.95(m,2H),2.69–2.57(m,1H),2.06–1.84(m,3H),1.73–1.59(m,2H).MS[M+H] + For C 26 H23 N9O3, calculated value 510.2, measured value 510.3.

[0825] Example 123: 6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)nicotinic acid

[0826]

[0827] Step 1. Synthesize methyl 6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)nicotinic acid in a manner similar to Example 122. 1 ¹H NMR (400MHz, chloroform-d) δ 9.22 (dd, J = 2.1, 0.9Hz, 1H), 8.46 (s, 1H), 8.40 (s, 1H), 8.32 (dd, J = 8.1, 2.1Hz, 2H), 7.91 (s, 1H), 7.77 (dt, J = 7.7, 1.4Hz, 1H), 7.61 (t, J = 7.8Hz, 1H), 7.37–7.30 (m, 1H), 5.81 (s, 2H), 5.23 (s, 2H), 3.96 (s, 3H). ESI MS [M+H] + For C 21 H 16 N8O2, calculated value 413.1, measured value 413.2.

[0828] Step 2: Saponification of methyl 6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)nicotinic acid yields the title compound: 1 H NMR (400MHz, DMSO-d6) δ9.10–8.94(m,1H),8.81(s,1H),8.61(s,1H),8.49(d,J=8.1Hz,1H),8.33(dd,J=8.1,2.2Hz, 1H),8.02(dd,J=7.8,1.4Hz,1H),7.86(s,1H),7.76(t,J=7.9Hz,1H),7.49(d,J=8.2Hz,1H),5.98(s,2H).ESIMS[M+H] + For C 20 H 14 N8O2, calculated value 399.1, measured value 399.2.

[0829] Example 124: 2-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)isonicotinic acid

[0830]

[0831] Step 1. similar Example 122 synthesized methyl 2-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)isonicotinic acid, thereby providing 88 mg of brown solid. 1 H NMR (400MHz, DMSO-d6) δ8.83–8.71(m,2H),8.59(t,J=1.6Hz,1H),8.47(dt,J=8.2,1.3Hz,1H),8.00(dt,J=7.8,1.3 Hz,1H),7.88(t,J=1.2Hz,1H),7.84–7.80(m,2H),7.74(t,J=7.9Hz,1H),6.93(s,2H),5.98(s,2H),3.90(s,3H).ESI MS[M+H] + For C 21 H 16 N8O2, calculated value 413.1, measured value 413.2.

[0832] Step 2: Saponification of methyl 2-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)isonicotinic acid provides the title compound. 1 H NMR (400MHz, DMSO-d6) δ8.88(s,1H),8.75(d,J=5.0Hz,1H),8.62(t,J=1.7Hz,1H),8.50(d,J=8.0Hz,1 H),8.04(d,J=7.7Hz,1H),7.90(s,1H),7.85(s,1H),7.84–7.71(m,2H),6.00(s,2H),5.44(bs,3H).ESI MS[M+H] + For C 20 H 14 N8O2, calculated value 399.1, measured value 399.2.

[0833] Example 125: 3-[6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]propionic acid

[0834]

[0835] Step 1: The synthesis of the azide derivative was similar to step 5 of Example 1. Following silica gel chromatography (hexane / EtOAc 90:10 to 70:30), 500 mg (2.6 mmol) of alcohol provided the desired azide (265 mg, 46%).

[0836] Step 2: Perform cycloaddition of the azide and alkyne derivatives in a manner similar to step 6 of Example 1 to provide methyl 3-[6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]propionate. 1 ¹H NMR (400MHz, chloroform-d) δ 8.46 (ddd, J = 1.7, 1.7, 0.6Hz, 1H), 8.35 (s, 1H), 8.32 (ddd, J = 8.0, 1.9, 1.2Hz, 1H), 7.90 (s, 1H), 7.76 (ddd, J = 7.7, 1.7, 1.2Hz, 1H), 7.71–7.52 (m, 2H), 7.17 (d, J = 8.0Hz, 1H), 7.08 (m, J = 8.0Hz, 1H), 5.67 (s, 2H), 5.16 (s, 2H), 3.66 (s, 3H), 3.14 (t, J = 7.3Hz, 2H), 2.83 (t, J = 7.3Hz, 2H). MS [M+H] + For C 23 H 20 N8O2, calculated value 441.2, measured value 441.3.

[0837] Step 3: Add an aqueous solution of LiOH (0.2 mL, 1 M) to a solution of methyl ester (45 mg, 0.1 mmol) in THF (1 mL). Stir the resulting mixture vigorously at room temperature for 6 hours. Then quench the mixture by adding excess acetic acid and evaporate it onto silica. The residue was purified by silica gel chromatography (CH2Cl2 / MeOH 100:0 to 90:10) to provide 3-[6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]propionic acid (40 mg, 95%). 1H NMR (400MHz, DMSO-d6) δ8.76(s,1H),8.60(s,1H),8.48(d,J=8.0Hz,1H),8.02(d,J=8.0Hz,1H),7.86(s,1H),7.77(dd,J=7.7 ,7.7Hz,1H),7.30(d,J=7.7Hz,1H),7.14(d,J=7.7Hz,1H),5.82(s,1H),2.96(t,J=7.2Hz,2H),2.64(t,J=7.2Hz,2H).MS[M+H] + For C 22 H 18 N8O2, calculated value 427.2, measured value 427.2.

[0838] Example 126: 3-[6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]-2,2-dimethylpropionic acid

[0839]

[0840] Step 1. TBSCl (723 mg, 4.8 mmol) was added to a mixture of alcohol (781 mg, 4 mmol) and imidazole (465 mg, 8 mmol) in CH2Cl2 (15 mL). After complete conversion of the starting material, silica was added and the resulting mixture was evaporated to dryness. Purification by silica gel chromatography (95:5 to 80:20 hexane / EtOAc) yielded a pale yellow oily silyl ether (1.14 g, 92%).

[0841] Step 2: The mixture of ester (1.5 g, 5 mmol) in THF (8 mL) was treated with LiHMDS (1 M in THF, 12 mL) at -78 °C. The solution was stirred at this temperature for 20 min and MeI (13 mmol) was added. The mixture was stirred from -78 °C to room temperature overnight. After routine treatment, the residue was purified by silica gel chromatography (95:5 to 85:15 hexane / EtOAc) to yield the dialkylated ester (350 mg, 21%). The silyl ether (350 mg, 1.05 mmol) was dissolved in THF (2 mL) and acetic acid (20 μL) was added, followed by TBAF (1 M in THF, 2 mL). The mixture was stirred at room temperature for 2 h and after routine treatment, the residue was purified by silica gel chromatography (95:5 to 70:30 CH2Cl2 / hexane (1:1) / EtOAc) to provide the primary alcohol (95 mg, 40%).

[0842] Step 3: The synthesis of the azide derivative is similar to step 5 of Example 1 (100 mg, 94%).

[0843] Step 4: Cycloaddition was performed in a manner similar to step 6 of Example 1, and the subsequent ester was hydrolyzed in a manner similar to Example 125 to provide the title compound. 1 H NMR (400MHz, DMSO-d6) δ8.82(s,1H),8.62(s,1H),8.58(d,J=7.9Hz,1H),7.90–7.99(m,2H),7.70–7.82(m,2H) ,7.36(d,J=7.9Hz,2H),7.22(d,J=7.9Hz,1H),6.27(brs,2H),5.79(s,2H),2.63(s,2H),1.30(s,6H).MS[M+H] + For C 24 H 22 N8O2, calculated value 455.2, measured value 455.3.

[0844] Example 127: 3-[6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]butyric acid

[0845]

[0846] Step 1: The mixture of phosphonate (1.48 g, 6.6 mmol) in THF (10 mL) was treated with NaH (60% in mineral oil, 264 mg, 6.6 mmol) and after 10 minutes, a solution of ketone (1.59 g, 6 mmol) in THF (2 mL) was added. The resulting mixture was stirred overnight; a tinate salt was added and the mixture was evaporated to dryness, and then purified by silica gel chromatography (95:5 to 80:20 hexane / EtOAc) to provide a Z:E mixture (1.22 g, 61%) of α,β-unsaturated esters.

[0847] Pd / C (10%, 60 mg) was added to a solution of degassed α,β-unsaturated ester (660 mg, 1.96 mmol) in 10 mL of THF. The suspension was then placed in H₂O. 2(g) Stir for 4 hours. Filter through a tin-salt filter and evaporate the solvent to dryness to obtain thickened alkanes (660 mg, quantitative).

[0848] Step 2: The synthesis of the azide derivative was similar to that in Example 79 (468 mg, 96% of both steps).

[0849] Step 3: Perform cycloaddition and hydrolysis reactions in a manner similar to Example 125 to provide the title compound. 1 HNMR(400MHz,DMSO-d6)δ12.04(brs,1H),8.68(s,1H),8.56(s,1H),8.45(d,J=8.0Hz,1H ),7.97(d,J=8.0Hz,1H),7.79(s,1H),7.73(dd,J=7.8,7.8Hz,,2H),7.26(d,J=7.8Hz,1H) ,7.05(d,J=7.8Hz,1H),6.88(s,2H),5.79(s,2H),3.33–3.19(m,1H),2.67(dd,J=15.8,7. 2Hz,1H),2.44(dd,J=15.8,7.3Hz,1H),2.52–2.38(m,4H),1.16(d,J=7.0Hz,3H).MS[M+H] + For C 23 H 20 N8O2, calculated value: 441.2, measured value: 441.3.

[0850] Example 128: 3-[6-({4-[2-amino-6-(2,3-difluorophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]butyric acid

[0851]

[0852] Cycloaddition and hydrolysis reactions were carried out in a manner similar to Example 125 to provide the title compound. 1 ¹H NMR (400 MHz, acetone-d6) δ 8.61 (s, 1H), 7.93–7.85 (m, 1H), 7.84 (s, 1H), 7.75 (dd, J = 7.8, 7.8 Hz, 1H), 7.53–7.41 (m, 1H), 7.38–7.31 (m, 1H), 7.31 (d, J = 7.8 Hz, 1H), 7.19 (d, J = 7.8) Hz,1H),6.27(brs,2H),5.80(s,2H),3.40(qt,J=7.0,6.6Hz,1H),2.85(dd,J=15.8,7 .7Hz,1H),2.59(dd,J=15.8,6.8Hz,1H),2.08(s,2H),1.29(dd,J=7.0Hz,3H).MS[M+H] + For C 22 H 21 F2N7O2, calculated value 452.2, measured value 452.3.

[0853] Example 129: 2-[6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridyl]cyclopropanecarboxylic acid

[0854]

[0855] Step 1: The solution of Me3SOI (1.93 g, 8.75 mmol) in DMSO (8 mL) was treated with NaH (60% in mineral oil, 320 mg, 8 mmol). The resulting mixture was stirred for 30 minutes before adding an α,β-unsaturated ester (1.5 g, 4.9 mmol) in DMSO (4 mL) as solvent. The resulting mixture was stirred at 50 °C for 2 hours before treatment (EtOAc / H2O). The residue obtained after evaporation of the organic matter was purified by silica gel chromatography (95:5 to 85:15 hexane / EtOAc) to yield a cyclopropyl derivative (500 mg, a mixture of cis / trans isomers 65:35, 32%). The silyl ether (500 mg, 3.1 mmol) was dissolved in THF (3 mL) and acetic acid (40 μL) was added, followed by TBAF (1 M in THF, 3 mL). The mixture was stirred at room temperature for 2 hours and the residue was purified by silica gel chromatography (CH2Cl2 / hexane (1:1) / EtOAc 95:5 to 50:50) after routine treatment to provide primary alcohol (320 mg, a mixture of cis / trans isomers of 65:35, quantified).

[0856] Step 2: The synthesis of the azide derivative is similar to step 5 of Example 1 (275 mg, 76%, 65:35 isomer mixture).

[0857] Step 3: Perform cycloaddition and hydrolysis reactions in a manner similar to Example 125 to provide the title compound. 1 HNMR(400MHz,DMSO-d6)δ12.33(brs,1H),8.66(s,1H),8.57(s,1H),8.45(dd,J=8.0Hz,1H),7.98(d,J=8.0Hz,1H),7.80(s,1H),7.77–7.67(m,2 H),7.38(d,J=8.0Hz,1H),7.10(dt,J=8.0Hz,1H),6.88(brs,2H),5.75( s,2H),2.59–2.51(m,1H),1.94–1.84(m,1H),1.43–1.28(m,1H).MS[M+H] + For C 23 H 18N8O2, calculated value: 439.2, measured value: 439.3.

[0858] Example 130: 3-[6-({4-[2-amino-6-(m-cyanophenyl)-4-pyrimidinyl]-1H-1,2,3-triazol-1-yl}methyl)-2-pyridinyl]-3-methylbutyric acid

[0859]

[0860] Step 1: Cool a solution of n-butyllithium (144 mL, 360 mmol, 2.5 M in hexane) in 120 mL of diethyl ether to -78 °C and add dropwise 2-bromo-6-methylpyridine (41.0 mL, 360 mmol). Warm the reaction mixture to 0 °C and stir at this temperature for 15 minutes. In separate flasks, combine dibutyl sulfide (54.5 mL, 312 mmol) and copper iodide (I) (34.3 g, 180 mmol) and stir the mixture for 5 minutes until homogeneous. Add ether (240 mL), cool the solution to 0 °C, and add dropwise the pyridine solution from the previous step. Stir the mixture at 0 °C for another 20 minutes, at which point add a solution of acrylate (16.7 g, 120 mmol) in 120 mL of diethyl ether. Warm the reaction mixture to room temperature for more than 14 hours. The mixture was quenched with a saturated ammonium chloride solution and extracted with ethyl acetate (2 x 200 mL), washed with brine, and dried over sodium sulfate. The crude product was purified by silica gel chromatography (0 to 20% EtOAc in hexane) to provide the desired Michael addition product (16.44 g; 59%) as a brown oil.

[0861] Step 2: A mixture of the product from Step 1 (16.44 g, 70.8 mmol), sodium chloride (1.24 g, 21.2 mmol), water (1.42 mL), and DMSO (71 mL) was stirred at 160 °C for 3 hours. The reaction mixture was cooled, MTBE (500 mL) was added, the organic phase was washed with water (4 x 400 mL) and dried over sodium sulfate. The starting material was dissolved in 3.0 M hydrochloric acid-methanol (236 mL) and stirred at 50 °C for 60 hours. The reaction mixture was then rinsed with sodium bicarbonate. (s) The product was slowly quenched, filtered, and concentrated. The crude product was purified by silica gel chromatography (7.5% EtOAc in hexane) to provide the desired product as a colorless oil (8.73 g; 59%).

[0862] Step 3: At 0°C, m-CPBA (19.9 g, 84.2 mmol, 75% in water) was slowly added to a solution of the product from Step 2 (8.73 g, 42.1 mmol) in dichloromethane (168 mL) over a period of more than 5 minutes. solidThe reaction mixture was stirred at 0°C for 1 hour and then at room temperature for 14 hours. The organic layer was washed with 0.1M NaOH solution, dried over sodium sulfate, and concentrated. The starting material was redissolved in dichloromethane (84 mL), cooled to 0°C, and TFAA (59 mL) was added dropwise. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was slowly quenched with saturated Na₂CO₃ solution and extracted with ethyl acetate (3 x 200 mL). The starting material was purified by silica gel chromatography (0 to 75% EtOAc in hexane solution) to provide the desired product (5.55 g; 59%) as a red oil.

[0863] Step 4: DBU (4.46 mL, 29.8 mmol) was ad...

Claims

1. A compound of Formula (I) or a pharmaceutically acceptable salt thereof, wherein, G 1 is N or CR 3a ; G 2 is N or CR 3b ; G 3 for CR 3c ; R 3a , R 3b , and R 3c are each independently H, deuterium, or C 1-3 alkyl; R 1a and R 1b are each independently selected from the group consisting of i) H or deuterium, ii) optionally substituted by 1-3 R 5 C substituted by 1-3 R 1-8 alkyl, iii) Optionally placed by 1-3 R 5 -X replaced by substituent 1 -OC 1-8 alkyl, iv) -C(O)-R 6 , v) optionally substituted by 1-3 R 7 Y, and vi) optionally substituted by 1-3 R 7 substituted by 1-3 R 1 -Y; or vii)R 1a and R 1b Together with the nitrogen attached to them, they are optionally formed by 1-3 R 8 The 5-6 membered heterocyclic alkyl ring substituted by the substituent, wherein the heterocyclic alkyl ring has 0-2 heteroatom ring vertices selected from the group consisting of O, N and S; each Y is C 3-8 cycloalkyl or 4- to 6-membered heterocycloalkyl having 1-3 heteroatom ring vertices selected from the group consisting of O, N, and S; R 2 is H or deuterium; R 4 H, deuterium or C 1-3 alkyl; Ar 1 substituted by 1-3 R 9 substituted by 1-3 R Ar 2 substituted by 1-3 R 10 substituted by 1-3 R each X 1 is C 1-6 alkylene; each R is independently selected from the group consisting of hydroxy, C 5 independently selected from the group consisting of hydroxy, C 3-8 cycloalkyl, phenyl, -O-phenyl, -C(O)OR a , and oxo; each R 6 is C 1-8 alkyl or Y, wherein each is optionally substituted with 1-3 substituents selected from the group consisting of hydroxy, -O-phenyl, phenyl, and -O-C 1-8 alkyl; Each R 7 Select independently from the following group: C 1-8 Alkyl, hydroxyl, -OC 1-8 Alkyl, oxo, and C(O)OR a ; Each R 8 Select independently from the following group: C 1-8 Alkyl, hydroxyl, and oxo; Each R 9 Select independently from the following group: C 1-8 Alkyl, C 1-8 Deuterated alkyl, -OC 1-8 Alkyl, -OC 1-8 Deuterated alkyl groups, -X 1 -OC 1-8 Alkyl, -OX 1 -OC 1-8 Alkyl, -X 1 -OX 1 -OC 1-8 Alkyl, -C(O)OR a Halogen, cyano, -NR b R c Y, -X 1 -C 3-8 cycloalkyl and -X 2 -Z, where X 2 Selected from the following group: C 1-6 Alkylene, -C 1-6 Alkylenes -O-, -C(O)-, and -S(O)2-, Z being 4- to 6-membered heterocyclic alkyl groups having 1-3 heteroatom ring vertices selected from the group consisting of O, N, and S, and wherein each of the R 9 The substituents are optionally replaced by 1-3 Rs 11 Replaced; Each R 10 Select independently from the following group: C 1-8 Alkyl, C 1-8 Deuterated alkyl, halogen, cyano, -OC 1-8 Alkyl, -OC 1-8 Deuterated alkyl groups, -X 1 -OC 1-8 Alkyl, -OX 1 -OC 1-8 Alkyl, -S(O)2-C 1-6 Alkyl, -C(O)NR d R e and 4-6 membered heteroaryl groups having 1-3 heteroatom ring vertices selected from the group consisting of O, N and S, wherein each of the R 10 The substituents are optionally replaced by 1-3 Rs 12 Replaced, or in Ar 2 Two R's at adjacent ring vertices 10 Optionally combined to form a 5-membered heterocycle optionally replaced by 1-2 halogens; Each R 11 Independently selected from the following group: hydroxyl, halogen, cyano, -NR d R e -C(O)OR a , phenyl, C 3-8 cycloalkyl, and optionally C(O)OR a The C that was replaced 1-4 alkyl; each R is independently selected from the group consisting of halogen, cyano, hydroxyl, -C(O)OR 12 is independently selected from the group consisting of halogen, cyano, hydroxyl, -C(O)OR a ; and each R is independently H, deuterium, or C1-6alkyl; a H, deuterium, or C1-6alkyl; 1-6 C1-6alkyl; each R b and R c is independently selected from the group consisting of H, deuterium, C 1-8 alkyl, -S(O)2-C 1-6 alkyl, -C(O)OR a , and -X 1 -C(O)OR a ; each R d and R e is independently selected from the group consisting of H, deuterium, C 1-8 alkyl, -S(O)2-C 1-6 alkyl.

2. The compound of claim 1, wherein at least one R 10 is cyano.

3. The compound of claim 1, wherein each R 9 Select independently from the following group: C 1-8 Alkyl, C 1-8 Deuterated alkyl, -OC 1-8 Alkyl, -OC 1-8 Deuterated alkyl groups, -X 1 -OC 1-8 Alkyl, -OX 1 -OC 1-8 Alkyl and -X 1 -OX 1 -OC 1-8 Alkyl, wherein each of the R 9 The substituents are optionally replaced by 1-3 Rs 11 What it replaced.

4. The compound of claim 1, wherein each R 9 Independently selected from the following group: -C(O)OR a -NR b R c Y, -X 1 -C 3-8 cycloalkyl 、 and -X 2 -Z, where X 2 Selected from the following group: C 1-6 Alkylene, -C 1-6 Alkylenes -O-, -C(O)-, and –S(O)2-, Z being 4- to 6-membered heterocyclic alkyl groups having 1-3 heteroatom ring vertices selected from the group consisting of O, N, and S, and wherein each of the R 9 The substituents are optionally replaced by 1-3 Rs 11 What it replaced.

5. The compound of claim 1, wherein G 2 is N.

6. The compound of claim 1, wherein G 1 is N.

7. The compound of claim 1, wherein G 1 is CR 3a。 8. The compound of claim 1, wherein R 4 is H.

9. The compound of claim 1, wherein R 1b is H.

10. The compound of claim 1, wherein each R 10 Select independently from the following group: C 1-8 Alkyl, halogen, cyano, -OC 1-8 Alkyl, -X 1 -OC 1-8 Alkyl and -OX 1 -OC 1-8 Alkyl, wherein each of the R 10 The substituents are optionally replaced by 1-3 Rs 12 What it replaced.

11. The compound of claim 10, wherein each R 10 is independently selected from the group consisting of C 1-8 alkyl, halo, cyano, and -O-C 1-8 alkyl.

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