5'-Nucleotidase, ecto modulators and uses thereof

Abstract

To solve the problem in which: in view of the role played by CD73 in cancer, as well as a diverse array of other diseases, disorders and conditions, and the current lack of CD73 inhibitors available to medical practitioners, new CD73 inhibitors, and compositions and methods associated therewith, are needed.SOLUTION: Compounds that modulate the conversion of AMP to adenosine by 5'-nucleotidase, ecto, and compositions containing the compounds and methods for synthesizing the compounds, are described herein. The use of such compounds and compositions for the treatment and / or prevention of a diverse array of diseases, disorders and conditions, including cancer- and immune-related disorders, that are mediated by 5'-nucleotidase, ecto is also provided.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] (Cross - Reference to Related Applications) This application claims the benefit of priority under 35 U.S.C. § 119(e) to U.S. Provisional Application No. 62 / 276,564, filed on January 8, 2016, and U.S. Provisional Application No. 62 / 324,077, filed on April 18, 2016, each of which is hereby incorporated by reference in its entirety for all purposes. to the United States, and United States Provisional Application No. 62 / 324,077 filed on April 18, 2016 (each of these is incorporated by reference in its entirety for the purpose of this specification). This is an application that claims the priority benefit of ).

[0002] Statement on the Rights of Inventions Made by Research and Development with Federal Government Support No application.

[0003] References such as "array lists", tables, or lists of computer programs submitted on compact discs References No application. (Field of the Invention)

[0004] For example, compounds and compositions for the inhibition of adenosine by 5'-nucleotidase, ecto, also known as CD73, and pharmaceutical compositions containing the same are provided herein. Also provided herein are methods for treating or preventing diseases, disorders, or conditions, and their symptoms, mediated by the inhibition of adenosine by, for example, 5'-nucleotidase, ecto. mediated by the inhibition of adenosine by 5'-nucleotidase, ecto are provided.

Background Art

[0005] (Background of the Invention) Purinoceptive signaling, a type of extracellular signaling mediated by purine nucleotides and nucleosides such as ATP and adenosine, involves the activation of purinoceptors in cells and / or neighboring cells and results in the regulation of cell function. Most cells are ​has the ability to release nucleotides, which generally occurs via regulated exocytosis (see Praetorius, H. A.; Leipziger, J. (1March 2010) Ann Rev Physiology 7 2(1): 377-393). The released nucleotides can then be hydrolyzed extracellularly by various cell membrane-bound enzymes called ectonucleotidases.

[0006] Ectonucleotides catalyze the conversion of ATP to adenosine, an endogenous regulator that affects multiple systems (including the immune, cardiovascular, central nervous, and respiratory systems). Adenosine also promotes fibrosis in various tissues. In the first step of adenosine production, ectonucleoside triphosphate diphosphohydrolase 1 (ENTPD1), also known as CD39 (cluster of differentiation 39), hydrolyzes ATP to ADP, and then ADP to AMP. In the next step, AMP is converted to adenosine by 5'-nucleotidase, ecto (NT5E or 5NT), also known as CD73 (cluster of differentiation 73).

[0007] The enzymatic activities of CD39 and CD73 play strategic roles in calibrating the duration, magnitude, and chemical nature of purinergic signaling sent to various cells (e.g., immune cells). Changes in these enzymatic activities can alter or determine the course of several pathophysiological events (including cancer, autoimmune diseases, infections, atherosclerosis, and ischemia-reperfusion injury), suggesting that these exogenous enzymes are novel therapeutic targets for managing various disorders.

[0008] ​​​​​​​​​​Inhibition of CD73 by monoclonal antibodies, siRNA, or small molecules delays tumor growth and metastasis (Stagg, J. (2010) PNAS U.S.A. 107:1547 - 52). For example, anti - CD73 antibody therapy has been shown to inhibit breast cancer growth and metastasis in animal models (Stagg, J. (26 Jan 2010) PNAS U.S.A, 107(4):1547 - 52). Furthermore, the use of antibodies that specifically bind to CD73 is being evaluated for the treatment of bleeding disorders (e.g., hemophilia) (U.S. Patent No. 9,090,697). In recent years, several attempts have been made to develop therapeutically useful small - molecule CD73 inhibitors. For example, Bhattarai et al. ((2015) J Med Chem 58:6248 - 63) have studied derivatives and analogs of α,β - methylene - ADP (AOPCP), one of the most metabolically stable, potent, and selective CD73 inhibitors known to date, and purine CD73 derivatives have been reported in the patent literature (International Publication No. WO 2015 / 164573). However, the development of small molecules is hampered, for example, by low metabolic stability. SUMMARY OF THE INVENTION PROBLEMS TO BE SOLVED BY THE INVENTION

[0009] Given the role of CD73 in cancer, as well as in a variety of other diseases, disorders, and conditions, and the lack of CD73 inhibitors available to medical practitioners, new CD73 inhibitors, and related compositions and methods are needed. MEANS FOR SOLVING THE PROBLEM

[0010] (SUMMARY OF THE INVENTION) ​​​​​​The present invention relates to compounds that modulate the conversion of AMP to adenosine by 5'-nucleotidase, ecto (NT5E or 5NT; also known as CD73), and to compositions (e.g., pharmaceutical compositions) containing such compounds. Such compounds, methods for their synthesis, and compositions are described in detail below. The present invention also relates to the use of such compounds and compositions for the treatment and / or prevention of a variety of diseases, disorders, and conditions that are mediated in whole or in part by CD73. CD73 inhibitors are relevant to the treatment of a variety of disorders including cancer, fibrosis, neurological and neurodegenerative disorders (e.g., depression and Parkinson's disease), cerebral and cardiac ischemia, immune-related disorders, and disorders with an inflammatory component. [See, for example, Sorrentino et al (2013) OncoImmunol, 2:e22448, doi: 10.4161 / onci.22448; and Regateiro et al.(2012) Clin. Exp. Immunol, 171:1-7 for reference]. In specific embodiments, the compounds described herein act to inhibit the immunosuppressive and / or anti-inflammatory activity of CD73 and are useful as a treatment or

[0011] preventive therapy when such inhibition is desirable. Unless otherwise specified, when the use of a compound of the present invention is described herein, it should be understood that such compound may be in the form of a composition (e.g., a pharmaceutical composition). As used herein, the terms "CD73 inhibitor", "CD73 blocker", "adenosine by 5'-nucleotidase, ecto inhibitor", "NT5E inhibitor", "5NT inhibitor", and other related terms accepted in the art refer to compounds that inhibit the conversion of AMP to adenosine by 5'-nucleotidase, ecto in in vitro assays, in vivo assays, or in a clinical setting. For example, in one embodiment, the compound inhibits the activity of 5'-nucleotidase, ecto by at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% compared to a control. In another embodiment, the compound inhibits the conversion of AMP to adenosine by 5'- nucleotidase, ecto by at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% compared to a control. In yet another embodiment, the compound reduces the level of adenosine produced by 5'- nucleotidase, ecto by at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95% compared to a control. It should be understood that the above percentages are exemplary and that other percentages may also be suitable depending on the specific application and the desired level of inhibition.

[0012] The terms "CD73 inhibitor", "CD73 blocker", "adenosine by 5'-nucleotidase, ecto inhibitor", "NT5E inhibitor", "5NT inhibitor", and other related terms accepted in the art refer to compounds that inhibit the conversion of AMP to adenosine by 5'-nucleotidase, ecto in in vitro assays, in vivo assays, or in a clinical setting. In a bomo model and / or other means for indicating therapeutic efficacy, it refers to a compound that can directly or indirectly regulate the CD73 receptor. This term also refers to a compound that exhibits at least some therapeutic benefit in a human subject.

[0013] The compounds of the present invention are thought to exert their activity by inhibiting CD73, but an exact understanding of the mechanism of action underlying the compounds is not required to practice the present invention. For example, the compounds may also exert their activity at least in part by modulating (e.g., inhibiting) other components of the purinergic signaling pathway, such as CD39. The purinergic signaling system involves transporters, enzymes, and receptors involved in the synthesis, release, action, and extracellular inactivation of (primarily) ATP and its extracellular degradation product adenosine (Sperlagh, B. et al. (Dec 2012) Neuropsychopharmacologia Hungarica 14(4):231-38). Inhibition of CD73 results in a decrease in adenosine, so CD73 inhibitors can be used for the treatment of diseases or disorders mediated by adenosine and its action on adenosine receptors, including A1, A2A, A2B, and A3 [see Yegutkin, GG (May 2008) Biochimica Biophysica Acta 1783(5):673-94). For the purposes of the present disclosure, the purinergic signaling process can be described as including the following components. The first component, purinergic receptors (P1, P2X, and P2Y), respond to the release of ATP or adenosine with various physiological functions (e.g., intestinal smooth muscle contraction, neurotransmitter release, and regulation of cell proliferation and differentiation) (Sperlagh, B. et al. (Dec 2012) Neuropsychopharmacologia Hungarica 14(4):231-38). The second component, enzymes involved in the synthesis and degradation of purines, such as CD39 and CD73, convert ATP to adenosine and vice versa. The third component, transporters

[0014] For the purposes of the present disclosure, the purinergic signaling process can be described as including the following components. The first component, purinergic receptors (P1, P2X, and P2Y), respond to the release of ATP or adenosine with various physiological functions (e.g., intestinal smooth muscle contraction, neurotransmitter release, and regulation of cell proliferation and differentiation). The second component, enzymes In general, all cells often express regulated estrogen receptors that mediate relaxation of smooth muscles; They have the ability to release nucleotides into the extracellular environment via exocytosis. The component nucleoside transporters (NTs) transport nucleoside substrates (e.g., adenosine adenosine is a membrane transport protein that transports adenosine across the cell membrane; the extracellular concentration of adenosine is Possibly a feedback loop connecting receptor signaling and transporter function As previously mentioned, ectonucleotidase (CD7 3 and CD39) hydrolyze nucleotides released into the extracellular environment and release them into further components. Another component of the purinergic signaling process includes pannexins; In particular, pannexin-1 channels (PANX1) mediate P2X / P2Y purinergic signaling. It is an essential component of the pathway and makes an important contribution to pathophysiological ATP release.

[0015] In one particular embodiment, the present invention provides a compound of formula (I): [ka] or a pharma- ceutically acceptable salt, hydrate, or solvate thereof, So, each R 1 is hydrogen, optionally substituted C1-C6 alkyl, optionally substituted Aryl, and -C(R 2 R 2 )-OC(O)-OR 3 Independently selected from the group consisting of Or two R's 1 groups may be optionally joined to form a 5- to 7-membered ring; each R 2 H and in some cases each R 3 is H, C1~ Independently selected from the group consisting of C6 alkyl and optionally substituted aryl; R 5 is selected from the group consisting of H and optionally substituted C1-C6 alkyl; X is O, CH 2 and S; A is

Chemical formula

Chemical formula

[0016] Compounds excluded from the above are those in which the combination of X, A, and Het is

Chemical formula

[0017] In some embodiments, the invention is a method of treating or preventing cancer in a subject (e.g., a human), the method comprising administering to the subject a therapeutically effective amount of at least one CD73 inhibitor described herein. The invention contemplates a method of treating or preventing cancer in a subject by administering to the subject an amount of a CD73 inhibitor effective to reverse or halt the progression of CD73-mediated immunosuppression. In some embodiments, CD73-mediated immunosuppression is mediated by antigen-presenting cells (APCs). Examples of cancers that can be treated using the compounds and compositions described herein include, but are not limited to, prostate, colorectal, pancreatic, cervical, gastric, endometrial, brain,

[0018] liver, bladder, ovary, testis, head, neck, skin (including melanoma and basal cell carcinoma), mesothelium, white blood cells (including lymphoma and leukemia), esophagus, breast, muscle, connective tissue, lung (including small cell lung cancer and ​​​​​cancers of the adrenal gland, thyroid, kidney, or bone (including non-small cell carcinoma); glioblastoma, mesothelioma, renal cell carcinoma , gastric cancer, sarcoma, choriocarcinoma, basal cell carcinoma of the skin, and testicular seminoma. 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, or Kaposi sarcoma. Cancers that are candidates for treatment with the compounds and compositions of the

[0019] invention are discussed further below. The present invention contemplates a method of treating a subject undergoing bone marrow transplantation or peripheral blood stem cell transplantation by administering a therapeutically effective amount of a CD73 inhibitor sufficient to increase a delayed type hypersensitivity

[0020] response to tumor antigens, delay the time to recurrence of post-transplant malignancy, prolong the relapse-free survival period after transplantation, and / or prolong the long-term survival period after transplantation. In certain embodiments, the invention contemplates a method of treating or preventing an infectious disorder (e.g., viral infection) in a subject (e.g., a human) by administering a therapeutically effective amount of at least one CD73 inhibitor (e.g., a novel inhibitor of the

[0021] invention). In some embodiments, the infectious disorder is a viral infection (e.g., chronic viral infection), bacterial infection, fungal infection, or parasitic infection. In certain embodiments, the viral infection is human immunodeficiency virus or cytomegalovirus. In yet another embodiment, the invention contemplates a method of treating and / or

[0022] Other diseases that can be treated or prevented, in whole or in part, by modulation of CD73 activity. The diseases, disorders, and conditions are potential indications for the CD73 inhibitor compounds of the present invention.

[0023] The present invention further relates to a method for treating a CD73-inhibiting agent as described herein in combination with one or more additional agents. The one or more additional agents may have any CD73 modulating activity. In some embodiments, the agonists may act via different mechanisms of action. Such agents may be used in combination with radiation (e.g., localized or systemic radiation therapy), and and / or other therapeutic modalities of a non-pharmacological nature. The agent and said one additional agent may be in the form of a single composition or multiple compositions, The therapeutic modalities may be administered simultaneously, sequentially, or via other formulations. Treatment regimens are contemplated in which a radiation phase is followed by a chemotherapy phase. Combination therapy may be additive or synergistic. Other advantages of combination therapy are described below.

[0024] In some embodiments, the present invention relates to a method for transplanting a patient with a bone marrow transplant, a peripheral blood stem cell transplant, or other type of The present invention further includes the use of a CD73 inhibitor as described herein in combination with a transplant therapy.

[0025] In a particular embodiment, the present invention relates to a method for treating a pulmonary circulation disorder comprising administering to a subject the method of the present invention in combination with an immune checkpoint inhibitor. The present invention contemplates the use of inhibitors of CD73 function as described in the above document to enhance the amplification of antigen-specific T cell responses. Blockade of immune checkpoints is a promising approach for treating human cancers. It has been shown that some of the candidates for blocking are selective in various types of tumor cells. Examples of immune checkpoint (ligand and receptor) upregulated thereto include P D1 (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); A2a R (adenosine A2a receptor A2aR); and killer inhibitory receptor. Immune check point inhibitors, and combination therapies therewith, are discussed in detail elsewhere in this specification .

[0026] In another embodiment, 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 CD73 inhibitor and at least one chemotherapeutic agent, wherein the chemotherapeutic agent includes, but is not particularly limited to, alkylating agents (e.g., nitrogen mustards such as chlorambucil, cyclophosphamide, ifosfamide, mechlorethamine, melphalan, and uracil mustard; aziridines such as thiotepa; methanesulfonic acid esters such as busulfan ; nucleoside analogs (e.g., gemcitabine) ; nitrosoureas such as carmustine, lomustine; topoisomerase 1 inhibitors (e.g., irinotecan ); platinum complexes such as cisplatin and carboplatin; bioreductive alkylating agents such as mitomycin, procarbazine, dacarbazine, and altretamine; DNA strand breakers (e.g., bleomycin); topoisomerase II inhibitors (e.g., amsacrine actinomycin D, daunorubicin, idarubicin, mitoxantrone, doxorubicin etoposide, and teniposide); DNA minor groove binders (e.g., pirarubicin); metabolism ​​Antagonists (e.g., methotrexate and trimethoprim; pyrimidine antagonists, e.g., fluorouracil, floxuridine, CB3717, azacitidine, cytarabine, and fludarabine; purine antagonists, e.g., mercaptopurine, 6 -thioguanine, fludarabine, pentostatin; asparaginase; and ribonucleotide reductase inhibitors, e.g., hydroxyurea); tubulin interacting agents (e.g., vincristine, estramustine, vinblastine, docetaxel, epothilone derivatives, and paclitaxel); hormonal agents (e.g., estrogen; conjugated estrogen; ethinyl estradiol; diethylstilbestrol; chlorotrianisene; idenestrol; progestins, e.g., hydroxyprogesterone caproate, medroxyprogesterone, and megestrol; and androgens, e.g., testosterone, testosterone propionate, fluoxymesterone, and methyltestosterone); corticosteroids (e.g., prednisone, dexamethasone, methylprednisolone, and prednisolone); luteinizing hormone releasing agents or gonadotropin releasing hormone antagonists (e.g., leuprolide acetate and goserelin acetate); and antihormonal antigens (e.g., antiandrogen agents such as tamoxifen, flutamide, etc., and antiadrenal agents such as mitotane and aminoglutethimide) are included. The present invention also contemplates the use of a CD73 inhibitor in combination with other agents known in the art (e.g., arsenic trioxide) and other chemotherapeutic agents that may be developed in the future.

[0027] In some embodiments of a method of treating cancer, at least one chemotherapeutic agent and Administration of a therapeutically effective amount of a CD73 inhibitor in combination results in a cancer survival rate greater than that observed by administration of either agent alone. In a further embodiment related to a method of treating cancer, administration of a therapeutically effective amount of a CD7 3 inhibitor in combination with at least one chemotherapeutic agent results in a reduction in tumor size or a delay in tumor growth greater than that observed by administration of either agent alone. In a further embodiment, the invention contemplates a method of treating or preventing cancer in a subject comprising administering to the subject a therapeutically effective amount of at least one CD73 inhibitor and at least one signal transduction inhibitor (STI). In certain embodiments, the at least one

[0028] STI is selected from the group consisting of a bcr / abl kinase inhibitor, an epidermal growth factor (EGF) receptor inhibitor, a he r-2 / neu receptor inhibitor, and a farnesyl transferase inhibitor (FTI). Other candidate STI agents are described elsewhere herein. In a further embodiment, the invention contemplates a method of enhancing the rejection of tumor cells in a subject comprising administering a CD73 inhibitor in combination with at least one chemotherapeutic agent and / or radiation therapy, wherein the resulting rejection of tumor cells is greater than that obtained by administration of any one alone of the CD73 inhibitor, chemotherapeutic agent, or radiation therapy. In a further embodiment, the invention contemplates a method of treating or preventing cancer in a subject comprising administering to the subject a therapeutically effective amount of at least one CD73 inhibitor and at least one immunomodulatory agent other than a CD73 inhibitor.

[0029] The invention also contemplates a method of enhancing the rejection of tumor cells in a subject comprising administering a CD73 inhibitor in combination with at least one chemotherapeutic agent and / or radiation therapy, wherein the resulting rejection of tumor cells is greater than that obtained by administration of any one alone of the CD73 inhibitor, chemotherapeutic agent, or radiation therapy. Here, the resulting rejection of tumor cells is greater than that obtained by administration of any one alone of the CD73 inhibitor, chemotherapeutic agent, or radiation therapy.

[0030] In a further embodiment, the invention contemplates a method of treating or preventing cancer in a subject comprising administering to the subject a therapeutically effective amount of at least one CD73 inhibitor and at least one immunomodulatory agent other than a CD73 inhibitor. ​​Provided is a method for treating cancer in a subject.

[0031] The present invention includes administering to a subject (e.g., a human) a therapeutically effective amount of at least one CD73 inhibitor and a therapeutically effective amount of an anti-infective agent, e.g., one or more antibacterial agents, to treat or prevent an infectious disorder (e.g., a viral infection) in the subject.

[0032] In a further embodiment, treatment of the infectious disease is effected by co-administering a therapeutically effective amount of the CD73 inhibitor of the present invention with a vaccine. In some embodiments the vaccine is an anti-viral vaccine, e.g., an anti-HIV vaccine. In another embodiment the aforementioned vaccine is effective against tuberculosis or malaria. In yet another embodiment the vaccine is a tumor vaccine (e.g., a vaccine effective against melanoma); the tumor vaccine can comprise a genetically modified tumor cell or cell line transfected to express granulocyte macrophage colony stimulating factor (GM-CSF), a genetically modified tumor cell or cell line. In certain embodiments the vaccine comprises one or more immunogenic peptides and / or dendritic cells.

[0033] In certain embodiments regarding the treatment of an infectious disease by administering a CD73 inhibitor and at least one additional therapeutic agent, the state of infection observed after administering both the CD73 inhibitor and the additional therapeutic agent is improved compared to the same state of infection observed after administering either alone. In some embodiments, the state of infection observed is a decrease in viral load, an increase in CD4+ T cell count, a decrease in opportunistic infection, an increase in survival time, eradication of chronic infection, or a combination thereof.​​​​​​

Brief Description of the Drawings

[0034]

Figure 1

Modes for Carrying Out the Invention

[0035] (Detailed Description of the Invention) Before further describing the present invention, it should be understood that the present invention is not limited to the specific embodiments described herein, and that the terms used herein are for the purpose of describing specific embodiments and are not intended to limit the present invention. When a range of values is provided, each intervening value, up to and including 1 / 10 of the unit of the lower limit, between the upper and lower limits of that range (unless otherwise specified), as well as any other stated value or intervening value within the stated range, is encompassed by the present invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and any specific excluded limits within the stated range are also encompassed by the present invention. When the stated range includes one or both of the limits, ranges excluding one or both of those included limits are also included in the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the technical field to which the present invention pertains. As used in this specification and the appended claims, it should be noted that the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. It should further be noted that the claims may be drafted to exclude any element.

[0036] When a range of values is provided, each intervening value, up to and including 1 / 10 of the unit of the lower limit, between the upper and lower limits of that range (unless otherwise specified), as well as any other stated value or intervening value within the stated range, is encompassed by the present invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and any specific excluded limits within the stated range are also encompassed by the present invention. When the stated range includes one or both of the limits, ranges excluding one or both of those included limits are also included in the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the technical field to which the present invention pertains. As used in this specification and the appended claims, it should be noted that the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. It should further be noted that the claims may be drafted to exclude any element. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the technical field to which the present invention pertains. As used in this specification and the appended claims, it should be noted that the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. It should further be noted that the claims may be drafted to exclude any element.

[0037] As used in this specification and the appended claims, it should be noted that the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. It should be noted that the claims may be drafted to exclude any element. It should further be noted that the claims may be drafted to exclude any element. Yes. Accordingly, this description is intended to serve as a premise for the use of exclusive terms such as "only", "solely", etc. in connection with the recitation of claim elements or the use of "negative" limitations. It is not intended to limit the scope of the invention in any way, but rather to provide a context for understanding the invention.

[0038] The publications discussed in this specification are provided solely for their disclosure prior to the filing date of the present application. Further, the publication date provided may be different from the actual date of publication, and it may be necessary to independently verify that.

[0039] General Introduction The number of subjects diagnosed with cancer and the number of deaths due to cancer continue to increase. Traditional treatment approaches, including chemotherapy and radiotherapy, generally become difficult for patients to tolerate and less effective as cancer (e.g., tumors) evolves to avoid such treatments. Recent experimental evidence indicates that CD73 inhibitors are important new treatments for cancer (e.g., breast cancer).

[0040] Promising data also support the role of inhibitors of CD73 function that inhibit the anti-inflammatory activity and / or immunosuppressive activity of CD73. Thus, CD73 inhibitors may be useful for treating, for example, immunosuppressive diseases (e.g., HIV and AIDS). Inhibition of CD73 may also be an important treatment strategy for patients with neurological or neuropsychiatric diseases or disorders such as depression.

[0041] The present invention particularly relates to small molecule compounds having CD73 inhibitory activity, and compositions thereof, and methods of using said compounds and compositions for the treatment and prevention of the diseases, disorders, and conditions described herein. ​​​​​​​​​​​​​Definitions

[0042] Unless otherwise specified, the following terms are intended to have the meanings indicated: The term is defined elsewhere herein.

[0043] The term "alkyl," by itself or as part of another substituent, means any of the following, unless otherwise stated. If not, it has the specified number of carbon atoms (i.e., C1-C8 means 1-8 carbons). Examples of alkyl groups include methyl, ethyl, n- Propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n- Examples include pentyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0044] The term "cycloalkyl" refers to a group having the indicated number of ring atoms (e.g., C3-C6 cycloalkyl). (cyl), and is either fully saturated or has one or less double bonds between the vertices of the ring. "Cycloalkyl" also refers to aryl groups such as bicyclo[2.2.1]heptane, Refers to bicyclic and polycyclic hydrocarbon rings such as bicyclo[2.2.2]octane.

[0045] The term "cycloheteroalkyl" refers to a heterocyclic ring having the indicated number of ring vertices (or members). 1 to 5 heteroatoms selected from N, O, and S (which may surround 1 to 5 carbon vertices) substituted), in which the nitrogen and sulfur atoms are optionally oxidized. and the nitrogen atom is optionally quaternized. Cycloheteroalkyl is a monocyclic, bicyclic, or polycyclic ring systems. Non-limiting examples of cycloheteroalkyl groups include pyrrolidinyl, ... , imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidino N, hydantoin, dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, etc. are exemplified. The cycloheteroalkyl group can be bonded to the rest of the molecule via a ring carbon or a heteroatom. When the term "optionally substituted" is used to describe either "cycloheteroalkyl" or "cycloheteroalkyl-alkyl", the cycloheteroalkyl or alkyl moiety is optionally substituted as defined below for the alkyl moiety. For example, an optionally substituted cycloheteroalkyl-alkyl group may be optionally substituted in either or both of the cycloheteroalkyl and alkyl moieties as in the following definition of alkyl substituents. Morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, etc. are exemplified. The cycloheteroalkyl group can be bonded to the rest of the molecule via a ring carbon or a heteroatom. When the term "optionally substituted" is used to describe either "cycloheteroalkyl" or "cycloheteroalkyl-alkyl", the cycloheteroalkyl or alkyl moiety is optionally substituted as defined below for the alkyl moiety. For example, an optionally substituted cycloheteroalkyl-alkyl group may be optionally substituted in either or both of the cycloheteroalkyl and alkyl moieties as in the following definition of alkyl substituents. Morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, etc. are exemplified. The cycloheteroalkyl group can be bonded to the rest of the molecule via a ring carbon or a heteroatom. When the term "optionally substituted" is used to describe either "cycloheteroalkyl" or "cycloheteroalkyl-alkyl", the cycloheteroalkyl or alkyl moiety is optionally substituted as defined below for the alkyl moiety. For example, an optionally substituted cycloheteroalkyl-alkyl group may be optionally substituted in either or both of the cycloheteroalkyl and alkyl moieties as in the following definition of alkyl substituents. Examples include tetrahydrofuran, tetrahydrothiophene, quinuclidine, etc. The cycloheteroalkyl group can be bonded to the rest of the molecule via a ring carbon or a heteroatom. When the term "optionally substituted" is used to describe either "cycloheteroalkyl" or "cycloheteroalkyl-alkyl", the cycloheteroalkyl or alkyl moiety is optionally substituted as defined below for the alkyl moiety. For example, an optionally substituted cycloheteroalkyl-alkyl group may be optionally substituted in either or both of the cycloheteroalkyl and alkyl moieties as in the following definition of alkyl substituents. The cycloheteroalkyl group can be bonded to the rest of the molecule via a ring carbon or a heteroatom. When the term "optionally substituted" is used to describe either "cycloheteroalkyl" or "cycloheteroalkyl-alkyl", the cycloheteroalkyl or alkyl moiety is optionally substituted as defined below for the alkyl moiety. For example, an optionally substituted cycloheteroalkyl-alkyl group may be optionally substituted in either or both of the cycloheteroalkyl and alkyl moieties as in the following definition of alkyl substituents. When the term "optionally substituted" is used to describe either "cycloheteroalkyl" or "cycloheteroalkyl-alkyl", the cycloheteroalkyl or alkyl moiety is optionally substituted as defined below for the alkyl moiety. For example, an optionally substituted cycloheteroalkyl-alkyl group may be optionally substituted in either or both of the cycloheteroalkyl and alkyl moieties as in the following definition of alkyl substituents. When the term "optionally substituted" is used to describe either "cycloheteroalkyl" or "cycloheteroalkyl-alkyl", the cycloheteroalkyl or alkyl moiety is optionally substituted as defined below for the alkyl moiety. For example, an optionally substituted cycloheteroalkyl-alkyl group may be optionally substituted in either or both of the cycloheteroalkyl and alkyl moieties as in the following definition of alkyl substituents. When the term "optionally substituted" is used to describe either "cycloheteroalkyl" or "cycloheteroalkyl-alkyl", the cycloheteroalkyl or alkyl moiety is optionally substituted as defined below for the alkyl moiety. For example, an optionally substituted cycloheteroalkyl-alkyl group may be optionally substituted in either or both of the cycloheteroalkyl and alkyl moieties as in the following definition of alkyl substituents. When the term "optionally substituted" is used to describe either "cycloheteroalkyl" or "cycloheteroalkyl-alkyl", the cycloheteroalkyl or alkyl moiety is optionally substituted as defined below for the alkyl moiety. For example, an optionally substituted cycloheteroalkyl-alkyl group may be optionally substituted in either or both of the cycloheteroalkyl and alkyl moieties as in the following definition of alkyl substituents. When the term "optionally substituted" is used to describe either "cycloheteroalkyl" or "cycloheteroalkyl-alkyl", the cycloheteroalkyl or alkyl moiety is optionally substituted as defined below for the alkyl moiety. For example, an optionally substituted cycloheteroalkyl-alkyl group may be optionally substituted in either or both of the cycloheteroalkyl and alkyl moieties as in the following definition of alkyl substituents. When the term "optionally substituted" is used to describe either "cycloheteroalkyl" or "cycloheteroalkyl-alkyl", the cycloheteroalkyl or alkyl moiety is optionally substituted as defined below for the alkyl moiety. For example, an optionally substituted cycloheteroalkyl-alkyl group may be optionally substituted in either or both of the cycloheteroalkyl and alkyl moieties as in the following definition of alkyl substituents.

[0046] As used herein, the wavy line "—" that intersects a single bond, double bond, or triple bond in any chemical structure shown herein represents the point of attachment of a single bond, double bond, or triple bond to the rest of the molecule. Further, a bond extending to the center of a ring (e.g., a phenyl ring) means attachment at any available ring vertex. One of ordinary skill in the art will understand that a plurality of substituents shown as attached to a ring will occupy ring vertices that provide a stable compound or are sterically compatible. In the case of a divalent moiety, the representation includes either orientation (forward or reverse). As used herein, the wavy line "—" that intersects a single bond, double bond, or triple bond in any chemical structure shown herein represents the point of attachment of a single bond, double bond, or triple bond to the rest of the molecule. Further, a bond extending to the center of a ring (e.g., a phenyl ring) means attachment at any available ring vertex. One of ordinary skill in the art will understand that a plurality of substituents shown as attached to a ring will occupy ring vertices that provide a stable compound or are sterically compatible. In the case of a divalent moiety, the representation includes either orientation (forward or reverse).

Chemical formula

[0047] The terms “alkoxy”, “alkylamino”, and “alkylthio” (or thioalkoxy ) are used in their ordinary meanings and refer to an alkyl group bonded to the rest of the molecule through an oxygen atom, an amino group, or a sulfur atom, respectively. Further, for a dialkylamino group , the alkyl moieties may be the same or different and may together form a 3- to 7-membered ring with the nitrogen atom to which each is attached . Thus, a group represented as dialkylamino or -NR R a R b and includes piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl, etc. means.

[0048] The terms “arylalkyl” and “heteroarylalkyl” are used in their conventional meanings and an aryl group or a heteroaryl group refers to a group bonded to the rest of the molecule through a C1-C4 alkylene linker. Exemplary embodiments of “arylalkyl” are phenylmethyl (or benzyl). Similarly, an exemplary embodiment of “heteroarylalkyl” is , for example, 3-pyridylpropyl. When “optionally substituted” is used to describe either of the terms “arylalkyl” or “heteroarylalkyl”, this refers to a group in which the aryl or heteroaryl moiety is optionally substituted as defined below and means that the alkyl moiety is optionally substituted as defined below. ​​

[0049] The term "halo" or "halogen", by itself or as part of another substituent, particularly in the absence of any other designation, means a fluorine, chlorine, bromine, or iodine atom. Further, terms such as "halo alkyl" are meant to include both monohaloalkyl and polyhaloalkyl. For example, the term "C1-C4 haloalkyl" is meant to include trifluoromethyl, 2,2,2-trif luoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0050] The term "aryl", in the absence of any other designation, means a polyvalent, unsaturated, typically aromatic hydrocarbon group which can be a fused or covalently bonded single ring or polycyclic ring (up to 3 rings). Non-limiting examples of aryl groups include phenyl, naphthyl, and biphenyl.

[0051] The term "heteroaryl" refers to an aryl group (or ring) containing 1-5 heteroatoms selected from N, O, and S, where the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atoms are optionally quaternized. A heteroaryl group can be bonded to the rest of the molecule through a heteroatom. Non-limiting examples of heteroaryl groups include pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, quinolinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl, purinyl, benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl, isobenzofuryl, i soindolyl, indolizinyl, benzotriazinyl, thienopyridinyl, thienopyrimi dinyl, pyrazolopyrimidinyl, imidazopyridine, benzothiaxolyl (benzothiaxol yl), and the like. yl), and the like. yl), benzofuranyl, benzothienyl, indolyl, quinolyl, isoquinolyl, isoth azolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl, triazolyl, te traazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl, thiazolyl, furyl, thienyl, and the like. The substituents of the heteroaryl ring can be selected from the group of acceptable substituents described below.

[0052] In some embodiments, the above terms (e.g., "alkyl", "aryl", and "heteroaryl") are optionally substituted. The selected substituents for each type of group are provided below.

[0053] Any substituent for an alkyl group (often including groups called alkylene, alkenyl, alkynyl, and cycloalkyl) is a number from 0 to (2m'+1), where m' in this case is the total number of carbon atoms in the group, of various groups selected from halogen, -OR', -NR'R", -SR', -SiR'R"R''', -OC(O)R', -C(O)R', -CO2R', -CON R'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)2N R'R", -NR'S(O)2R", -CN, and -NO2. R', R", and R''' are each independently hydrogen, unsubstituted C1-C8 alkyl , unsubstituted aryl, aryl substituted with 1-3 halogens, unsubstituted C1-C8 alkyl , C1-C8 alkoxy, or C1-C8 thioalkoxy group, or unsubstituted aryl-C1-C4 refers to an alkyl group. When R’ and R” are attached to the same nitrogen atom, they may combine with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring. For example, -NR’R ” means including 1-pyrrolidinyl and 4-morpholinyl.

[0054] Similarly, any substituents for the aryl group and heteroaryl group may vary and generally include - 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 selected from, and is a number in the range from 0 to the total number of open valences on the aromatic ring system, where R’, R’’, and R’’’ are independently selected from hydrogen, C1-C8 alkyl, C1-C8 haloalkyl, C3 -C6 cycloalkyl, C2-C8 alkenyl, and C2-C8 alkynyl. Other suitable substituents include each of the aryl substituents attached to the ring atoms by an alkylene tether of 1 to 4 carbon atoms bonded thereto. Two substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be substituted with a substituent of the formula -TC(

[0055] O)-(CH2) O)-(CH2) q -U-, 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 an aryl or heteroaryl ring are of the formula -A-(CH2) r -B - and may be substituted with a substituent of, where A and B are independently -CH2-, -O-, - NH-, -S-, -S(O)-, -S(O)2-, -S(O)2NR’, or a single bond , and r is an integer from 1 to 3. One of the single bonds of the newly formed ring may, in some cases be replaced by a double bond. Alternatively, two of the substituents on adjacent atoms of an aryl or heteroaryl ring are of the formula -(CH2) s -X-(CH2) t - and may be substituted with a substituent of, 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’ of -NR’- and -S(O )2NR’- is selected from hydrogen or unsubstituted C1-C6 alkyl. )2NR’- is selected from hydrogen or unsubstituted C1-C6 alkyl.

[0056] As used herein, the term “heteroatom” means including oxygen (O), nitrogen (N), sulfur (S), and silicon (Si).

[0057] The term “pharmaceutically acceptable salt” means salts of the active compounds prepared with relatively non-toxic acids or bases, depending on the particular substituents found in the compounds described herein. When the compounds of the present invention contain relatively acidic functional groups, the base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base in pure or an appropriate inert solvent. Examples of salts derived from pharmaceutically acceptable inorganic bases include ammonium, sodium, potassium, calcium, magnesium, and lithium salts. ammonium, sodium, potassium, calcium, magnesium, and lithium salts. Aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium , manganese (II), manganese (I), potassium, sodium, zinc, etc. are included. Pharmaceutically acceptable salts derived from organic bases include primary, secondary, and tertiary amines containing substituted amines, cyclic amines, natural amines, etc., such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethyl morpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. are included. When the compound of the present invention contains a relatively basic functional group, the acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired acid in a pure or 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, bicarbonic acid, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, or phosphorous acid, and relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, p -toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc. Also included are salts of amino acids such as alginate, and glucuronate . Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonic acid, phosphoric acid, phosphoric acid monohydride, phosphoric acid dihydride, sulfuric acid, sulfuric acid monohydride, hydroiodic acid, or phosphorous acid, and relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, p -toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc. Also included are salts of amino acids such as alginate, and glucuronate . Salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonic acid, phosphoric acid, phosphoric acid monohydride, phosphoric acid dihydride, sulfuric acid, sulfuric acid monohydride, hydroiodic acid, or phosphorous acid, and relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, p -toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc. are included. Also included are salts of amino acids such as alginate, and glucuronate . Salts of amino acids such as alginate, and glucuronate Salts of organic acids such as acids or galacturonic acid are also included (see, for example, Berge, S.M., etal, “Ph armaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66, 1-19). Some specific compounds of the present invention contain both basic functionality and acidic functionality that allow the compound to be converted into either a base addition salt or an acid addition salt.

[0058] The neutral form of the compound can be regenerated by contacting the salt with a base or an acid and isolating the parent compound by conventional methods. The parent form of the compound differs from the various salt forms in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to the parent form of the compound for the purposes of the present invention.

[0059] In addition to the salt forms, the present invention provides compounds in prodrug form. The prodrugs of the compounds described herein are compounds that readily undergo chemical change under physiological conditions to provide the compounds of the present invention. Further, the prodrugs can be converted into the compounds of the present invention by chemical or biochemical methods. For example, the prodrug can be slowly converted into the compound of the present invention when placed in a transdermal patch reservoir with an appropriate enzyme or chemical reagent.

[0060] Specific compounds of the present invention can exist in unsolvated forms as well as solvated forms including hydrated forms. Generally, the solvated forms are equivalent to the unsolvated forms and are intended to be encompassed within the scope of the present invention. Specific compounds of the present invention can exist in polymorphic or amorphous forms. Generally, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be included within the scope of the present invention.​​​​​​​​​ It is intended to be within the scope of the present invention.

[0061] The specific compounds of the present invention have asymmetric carbon atoms (optical centers) or double bonds. Racemates , diastereoisomers, geometric isomers, positional isomers, and individual isomers (e.g., separate mirror image isomers) are all intended to be encompassed within the scope of the present invention. When a stereochemical depiction is shown, it means a compound in which one of the isomers is present and substantially no other isomers are present. "Substantially free of other isomers" means at least an 80 / 20 ratio, more preferably 90 / 10, or 95 / 5, or more, of the two isomers. In some embodiments, one of the isomers is present in an amount of at least 99%.

[0062] The compounds of the present invention can also contain different ratios of atomic isotopes from the natural ones in one or more of the atoms constituting such compounds. Different ratios from the natural ones of the isotopes are defined as ranging from the amounts found in nature to amounts constituting 100% of the atom in question. For example, a compound can incorporate radioactive isotopes such as tritium ( 3H), iodine-125 ( 3 125I), or 125 carbon-14 ( 14C), or non-radioactive isotopes such as deuterium ( 14 2H) or carbon-13 ( 2 13C). Such isotope variations can provide additional utility over those described elsewhere in this application. For example, isotope variants of the compounds of the present invention are not particularly limited, but can find additional utility as diagnostic and / or imaging reagents 13 or as cytotoxic / radioactive toxicity therapeutic agents. Further The isotope variants of the compounds of the present invention can have changes in pharmacokinetic and pharmacodynamic properties that contribute to improved safety, tolerability, or efficacy during treatment. All isotope changes of the compounds of the present invention, whether radioactive or non-radioactive, are intended to be encompassed within the scope of the present invention.

[0063] The terms "patient" or "subject" are used interchangeably to refer to a human or non-human animal (e.g., a mammal).

[0064] The terms "administer", "administering", etc. refer to the contact of, for example, an inhibitor of CD73, a pharmaceutical composition containing the same, or a diagnostic agent with a subject, cell, tissue, organ, or biological fluid when they are applied to the same. In the context of cells, administration includes the contact of a reagent with the cells (e.g., in vitro or ex vivo), as well as the contact of the reagent with the fluid when the fluid is in contact with the cells.

[0065] The terms "treat", "treating", "treatment", etc. refer to a series of actions (e.g., administration of CD73 or a pharmaceutical composition containing the same) that are initiated after a disease, disorder, or condition has been diagnosed, observed, etc., to eliminate, reduce, suppress, relieve, or improve at least one of the underlying causes of the disease, disorder, or condition that afflicts the subject, or at least one of the symptoms associated with the disease, disorder, or condition that afflicts the subject, either temporarily or permanently. Thus, treatment includes inhibiting an active disease (e.g., preventing the progression or further progression of a disease, disorder, or condition or the clinical symptoms associated therewith).

[0066] As used herein, the term "in need of treatment" refers to a determination made by a physician or other caregiver that a subject is in need of treatment or would benefit from treatment. This determination is made based on a variety of factors within the realm of the physician's or caregiver's expertise. As used herein, the term "in need of treatment" refers to a determination made by a physician or other caregiver that a subject is in need of treatment or would benefit from treatment. This determination is made based on a variety of factors within the realm of the physician's or caregiver's expertise. As used herein, the term "in need of treatment" refers to a determination made by a physician or other caregiver that a subject is in need of treatment or would benefit from treatment. This determination is made based on a variety of factors within the realm of the physician's or caregiver's expertise. As used herein, the term "in need of treatment" refers to a determination made by a physician or other caregiver that a subject is in need of treatment or would benefit from treatment. This determination is made based on a variety of factors within the realm of the physician's or caregiver's expertise.

[0067] The terms "prevent," "preventing," "prevention," etc., in the context of a subject susceptible to a particular disease, disorder, or condition, refer to a series of actions (e.g., administration of an inhibitor of CD73 or a pharmaceutical composition comprising the same) initiated to temporarily or permanently prevent, suppress, arrest, or reduce the risk (determined by the absence of a clinical condition) of a subject developing the disease, disorder, condition, etc., or to delay its onset (e.g., prior to the onset of the disease, disorder, condition, or its symptoms). In some cases, the term also refers to slowing the progression of a disease, disorder, or condition, or suppressing its progression to a detrimental or undesirable state. The terms "prevent," "preventing," "prevention," etc., in the context of a subject susceptible to a particular disease, disorder, or condition, refer to a series of actions (e.g., administration of an inhibitor of CD73 or a pharmaceutical composition comprising the same) initiated to temporarily or permanently prevent, suppress, arrest, or reduce the risk (determined by the absence of a clinical condition) of a subject developing the disease, disorder, condition, etc., or to delay its onset (e.g., prior to the onset of the disease, disorder, condition, or its symptoms). In some cases, the term also refers to slowing the progression of a disease, disorder, or condition, or suppressing its progression to a detrimental or undesirable state. The terms "prevent," "preventing," "prevention," etc., in the context of a subject susceptible to a particular disease, disorder, or condition, refer to a series of actions (e.g., administration of an inhibitor of CD73 or a pharmaceutical composition comprising the same) initiated to temporarily or permanently prevent, suppress, arrest, or reduce the risk (determined by the absence of a clinical condition) of a subject developing the disease, disorder, condition, etc., or to delay its onset (e.g., prior to the onset of the disease, disorder, condition, or its symptoms). In some cases, the term also refers to slowing the progression of a disease, disorder, or condition, or suppressing its progression to a detrimental or undesirable state. The terms "prevent," "preventing," "prevention," etc., in the context of a subject susceptible to a particular disease, disorder, or condition, refer to a series of actions (e.g., administration of an inhibitor of CD73 or a pharmaceutical composition comprising the same) initiated to temporarily or permanently prevent, suppress, arrest, or reduce the risk (determined by the absence of a clinical condition) of a subject developing the disease, disorder, condition, etc., or to delay its onset (e.g., prior to the onset of the disease, disorder, condition, or its symptoms). In some cases, the term also refers to slowing the progression of a disease, disorder, or condition, or suppressing its progression to a detrimental or undesirable state. The terms "prevent," "preventing," "prevention," etc., in the context of a subject susceptible to a particular disease, disorder, or condition, refer to a series of actions (e.g., administration of an inhibitor of CD73 or a pharmaceutical composition comprising the same) initiated to temporarily or permanently prevent, suppress, arrest, or reduce the risk (determined by the absence of a clinical condition) of a subject developing the disease, disorder, condition, etc., or to delay its onset (e.g., prior to the onset of the disease, disorder, condition, or its symptoms). In some cases, the term also refers to slowing the progression of a disease, disorder, or condition, or suppressing its progression to a detrimental or undesirable state. The terms "prevent," "preventing," "prevention," etc., in the context of a subject susceptible to a particular disease, disorder, or condition, refer to a series of actions (e.g., administration of an inhibitor of CD73 or a pharmaceutical composition comprising the same) initiated to temporarily or permanently prevent, suppress, arrest, or reduce the risk (determined by the absence of a clinical condition) of a subject developing the disease, disorder, condition, etc., or to delay its onset (e.g., prior to the onset of the disease, disorder, condition, or its symptoms). In some cases, the term also refers to slowing the progression of a disease, disorder, or condition, or suppressing its progression to a detrimental or undesirable state. The terms "prevent," "preventing," "prevention," etc., in the context of a subject susceptible to a particular disease, disorder, or condition, refer to a series of actions (e.g., administration of an inhibitor of CD73 or a pharmaceutical composition comprising the same) initiated to temporarily or permanently prevent, suppress, arrest, or reduce the risk (determined by the absence of a clinical condition) of a subject developing the disease, disorder, condition, etc., or to delay its onset (e.g., prior to the onset of the disease, disorder, condition, or its symptoms). In some cases, the term also refers to slowing the progression of a disease, disorder, or condition, or suppressing its progression to a detrimental or undesirable state. The terms "prevent," "preventing," "prevention," etc., in the context of a subject susceptible to a particular disease, disorder, or condition, refer to a series of actions (e.g., administration of an inhibitor of CD73 or a pharmaceutical composition comprising the same) initiated to temporarily or permanently prevent, suppress, arrest, or reduce the risk (determined by the absence of a clinical condition) of a subject developing the disease, disorder, condition, etc., or to delay its onset (e.g., prior to the onset of the disease, disorder, condition, or its symptoms). In some cases, the term also refers to slowing the progression of a disease, disorder, or condition, or suppressing its progression to a detrimental or undesirable state.

[0068] As used herein, the term "in need of prevention" refers to a determination made by a physician or other caregiver that a subject is in need of preventive treatment or would benefit from treatment. This determination is made based on a variety of factors within the realm of the physician's or caregiver's expertise. As used herein, the term "in need of prevention" refers to a determination made by a physician or other caregiver that a subject is in need of preventive treatment or would benefit from treatment. This determination is made based on a variety of factors within the realm of the physician's or caregiver's expertise. As used herein, the term "in need of prevention" refers to a determination made by a physician or other caregiver that a subject is in need of preventive treatment or would benefit from treatment. This determination is made based on a variety of factors within the realm of the physician's or caregiver's expertise. As used herein, the term "in need of prevention" refers to a determination made by a physician or other caregiver that a subject is in need of preventive treatment or would benefit from treatment. This determination is made based on a variety of factors within the realm of the physician's or caregiver's expertise.

[0069] The phrase "therapeutically effective amount" refers to an amount that, when administered to a subject, can have any detectable positive effect on any symptom, aspect, or feature of a disease, disorder, or condition, alone or as part of a pharmaceutical composition, and either as a single dose or as part of a series of doses. The phrase "therapeutically effective amount" refers to an amount that, when administered to a subject, can have any detectable positive effect on any symptom, aspect, or feature of a disease, disorder, or condition, alone or as part of a pharmaceutical composition, and either as a single dose or as part of a series of doses. The phrase "therapeutically effective amount" refers to an amount that, when administered to a subject, can have any detectable positive effect on any symptom, aspect, or feature of a disease, disorder, or condition, alone or as part of a pharmaceutical composition, and either as a single dose or as part of a series of doses. 、refers to administering to a subject. A therapeutically effective amount can be confirmed by measuring the relevant physiological effects and can be adjusted in relation to, for example, the dosing regimen and the diagnostic analysis of the subject's condition. As an example, measurement of the serum level of a CD73 inhibitor (or its metabolite, etc.) at a specific time after administration can

[0070] indicate whether a therapeutically effective amount has been used. The term "in an amount sufficient to cause a change" means that there is a detectable difference between the level of an indicator measured before the implementation of a particular therapy (e.g., baseline level) and after

[0071] administration. The indicator includes any objective parameter (e.g., serum concentration) or subjective parameter (e.g., the subject's sense of well-being). The term "small molecule" refers to a compound having 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 particularly limited to, inorganic molecules,

[0072] organic molecules, organic molecules containing inorganic components, molecules containing radioactive atoms, and synthetic molecules. Therapeutically, small molecules may be more permeable to cells, less prone to degradation, and less likely to induce an immune response than larger molecules. The term "ligand" refers to, for example, a peptide, polypeptide, For example, it can bind to a receptor without significantly affecting signal transduction or adhesion. It includes agents. Furthermore, this term includes soluble versions of membrane-bound ligands, such as membrane-bound ligands modified by chemical or recombinant methods. The ligand or receptor may be entirely intracellular, i.e., it may be present in the cytosol, nucleus, or any of several other intracellular compartments. The complex of ligand and receptor is called a "ligand-receptor complex".

[0073] The terms "inhibitor" and "antagonist" or "activator" and "agonist" each refer to inhibitory or activating molecules, respectively, for example, for the activation of ligands, receptors, cofactors, genes, cells, tissues, or organs. An inhibitor is a molecule that reduces, blocks, prevents, delays activation, inactivates, desensitizes, or down-regulates, for example, a gene, protein, ligand, receptor, or cell. An activator is a molecule that increases, activates, promotes, enhances activation, sensitizes, or up-regulates, for example, a gene, protein, ligand, receptor, or cell. 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 and causes or promotes an increase in the activation of the target. An "antagonist" is a molecule that counteracts the action of an agonist. An antagonist prevents, reduces, inhibits, or neutralizes the activity of an agonist, and an antagonist can also prevent, inhibit, or reduce the constitutive activity of a target (e.g., a target receptor) even in the absence of an identified agonist.

[0074] The terms "modulate", "modulation", etc. refer to the ability of a molecule (e.g., an activator or inhibitor) to increase or decrease, directly or indirectly, the function or activity of CD73. A modulator - may act alone or may use cofactors, such as proteins, metal ions, or small molecules . Examples of modulators include small molecule compounds and other bioorganic molecules . A number of libraries of small molecule compounds (e.g., combinatorial libraries) are commercially available and can serve as a starting point for identifying modulators. One of ordinary skill in the art can develop one or more assays (e.g., biochemical or cell-based assays) that can screen such compound libraries to identify one or more compounds having the desired properties. Thereafter, skilled medicinal chemists can optimize such one or more compounds, for example, by synthesis and evaluation of their analogs and derivatives. Synthesis and / or molecular modeling studies can also be utilized to identify activators .

[0075] The "activity" of a molecule describes or refers to the binding of the molecule to a ligand or receptor; catalytic activity; the ability to stimulate gene expression, or cell signaling, differentiation, or maturation; antigenic activity; the modulation of the activity of other molecules . The term "proliferative activity" encompasses the activity that promotes, is required for, or is specifically associated with normal cell division, as well as cancer, tumor formation, dysplasia, cell transformation, metastasis, and angiogenesis.

[0076] As used herein, "equivalent", "equivalent activity", "activity equivalent to", "equivalent effect ", "effect equivalent to", etc. are relative terms that can be viewed quantitatively and / or qualitatively This is the case. The meanings of these terms often depend on the context in which they are used. As an example, two agents that activate a receptor can be considered to have comparable effects from a qualitative perspective, but when measured in an assay recognized in the art (e.g., a dose-response assay) or in an animal model recognized in the art, if one agent can only achieve 20% of the activity of the other agent, the two agents do not have equivalent effects from a quantitative perspective. When comparing one result with another (e.g., comparing one result with a reference standard), "equivalent" often (but not always) means that one 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 specific embodiment, if one result deviates from the reference standard by less than 15%, less than 10%, or less than 5%, it is equivalent to the reference standard. For example, without being particularly limited, activity or effect can mean efficacy, stability, solubility, or immunogenicity. When comparing one result with another (e.g., comparing one result with a reference standard), "equivalent" often (but not always) means that one 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 specific embodiment, if one result deviates from the reference standard by less than 15%, less than 10%, or less than 5%, it is equivalent to the reference standard. For example, without being particularly limited, activity or effect can mean efficacy, stability, solubility, or immunogenicity. In a specific embodiment, if one result deviates from the reference standard by less than 15%, less than 10%, or less than 5%, it is equivalent to the reference standard. For example, without being particularly limited, activity or effect can mean efficacy, stability, solubility, or immunogenicity. In a specific embodiment, if one result deviates from the reference standard by less than 15%, less than 10%, or less than 5%, it is equivalent to the reference standard. For example, without being particularly limited, activity or effect can mean efficacy, stability, solubility, or immunogenicity. In a specific embodiment, if one result deviates from the reference standard by less than 15%, less than 10%, or less than 5%, it is equivalent to the reference standard. For example, without being particularly limited, activity or effect can mean efficacy, stability, solubility, or immunogenicity. In a specific embodiment, if one result deviates from the reference standard by less than 15%, less than 10%, or less than 5%, it is equivalent to the reference standard. For example, without being particularly limited, activity or effect can mean efficacy, stability, solubility, or immunogenicity. In a specific embodiment, if one result deviates from the reference standard by less than 15%, less than 10%, or less than 5%, it is equivalent to the reference standard. For example, without being particularly limited, activity or effect can mean efficacy, stability, solubility, or immunogenicity.

[0077] "Substantially pure" indicates that a certain component constitutes more than about 50% of the total content of the composition, and typically more than about 60% of the total content of the composition. More typically, "substantially pure" refers to a composition in which at least 75%, at least 85%, at least 90%, or more of the total composition is the component of interest. In some cases, the component of interest will constitute more than about 90% or more than about 95% of the total content of the composition. "Substantially pure" indicates that a certain component constitutes more than about 50% of the total content of the composition, and typically more than about 60% of the total content of the composition. More typically, "substantially pure" refers to a composition in which at least 75%, at least 85%, at least 90%, or more of the total composition is the component of interest. In some cases, the component of interest will constitute more than about 90% or more than about 95% of the total content of the composition. "Substantially pure" indicates that a certain component constitutes more than about 50% of the total content of the composition, and typically more than about 60% of the total content of the composition. More typically, "substantially pure" refers to a composition in which at least 75%, at least 85%, at least 90%, or more of the total composition is the component of interest. In some cases, the component of interest will constitute more than about 90% or more than about 95% of the total content of the composition. "Substantially pure" indicates that a certain component constitutes more than about 50% of the total content of the composition, and typically more than about 60% of the total content of the composition. More typically, "substantially pure" refers to a composition in which at least 75%, at least 85%, at least 90%, or more of the total composition is the component of interest. In some cases, the component of interest will constitute more than about 90% or more than about 95% of the total content of the composition. In some cases, the component of interest will constitute more than about 90% or more than about 95% of the total content of the composition.

[0078] ​When referring to a ligand / receptor, antibody / antigen, or other binding pair, the terms "specifically binds" " or "selectively binds" denote a binding reaction that determines the presence of a protein in a heterogeneous population of other biological substances of the protein. Thus, under the specified conditions, a particular ligand binds to a particular receptor and does not bind in significant amounts to other proteins present in the sample. A binding composition derived from the antigen-binding site of an antibody of the intended method binds to the antigen, or a variant or mutant protein thereof, with an affinity at least 2-fold, at least 10-fold, at least 20-fold, or at least 10 0-fold greater than the affinity for any other antibody or binding composition obtained therefrom. In certain embodiments, the antibody has an affinity greater than about 1 0 liter / mole as measured, for example, by Scatchard analysis (Munsen, et al. 1980 Analyt. Biochem.107:220-239). 0 9 For example, the term "response" of a cell, tissue, organ, or organism encompasses changes in biochemical or physiological

[0079] behavior such as concentration, density, adhesion, or migration within a biological compartment, gene expression rate, or state of differentiation, where said changes correlate with activation, stimulation, or treatment and with internal mechanisms such as genetic programming. In certain contexts, terms such as "activation," "stimulation," etc. refer to cell activation regulated by internal mechanisms and external or environmental factors. On the other hand, terms such as "inhibition," "downregulation," etc. refer to the opposite effects. The terms "polypeptide," "peptide," and "protein," which are used interchangeably herein, refer to a polymeric form of amino acids of any length, which is genetically encoded. One, terms such as "activation," "stimulation," etc. refer to cell activation regulated by internal mechanisms and external or environmental factors. On the other hand, terms such as "inhibition," "downregulation," etc. refer to the opposite effects. refer to the opposite effects.

[0080] The terms "polypeptide," "peptide," and "protein," which are used interchangeably herein, refer to a polymeric form of amino acids of any length, which is genetically encoded, and this is the amino acids and non-genetically encoded amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides having a modified polypeptide backbone. These terms include, but are not particularly limited to, fusion proteins with heterologous amino acid sequences and other fusion proteins with heterologous and homologous leader sequences with or without an N-terminal methionine residue, fusion proteins such as immunologically labeled proteins, and the like.

[0081] As used herein, the terms "variants" and "homologs" are used interchangeably to refer to amino acid sequences or DNA sequences similar to a reference amino acid sequence or nucleic acid sequence, respectively. This term includes naturally occurring variants and non-naturally occurring variants. Naturally occurring variants include homologs (polypeptides and nucleic acids with different amino acid sequences or nucleotide sequences from one species to another), and allelic variants (polypeptides and nucleic acids with different amino acid sequences or nucleotide sequences from one individual to another within a species). Thus, variants and homologs include naturally occurring DNA sequences and the proteins encoded thereby, and their isoforms, as well as splice variants of proteins or genes. This term also includes nucleic acid sequences in which one or more bases have changed from a naturally occurring DNA sequence, but which are translated into an amino acid sequence corresponding to a naturally occurring protein due to the degeneracy of the genetic code. Non-naturally occurring variants and homologs include polypeptides and nucleic acids containing changes in the amino acid sequence or nucleotide sequence, respectively. Although one or more bases have changed in the naturally occurring DNA sequence, it is translated into an amino acid sequence corresponding to the naturally occurring protein due to the degeneracy of the genetic code. Non-naturally occurring variants and homologs include polypeptides and nucleic acids containing changes in the amino acid sequence Rarely, here, changes in the array are introduced artificially (e.g., mutant proteins). For example the changes are generated in the laboratory by human intervention ("human hand"). Thus, variants and homologs that do not exist in nature may refer to those that are different from the naturally occurring sequences by one or more conservative substitutions and / or tags and / or conjugates.

[0082] As used herein, the term "mutein" broadly refers to a recombinantly mutated protein. These proteins usually have single or multiple amino acid substitutions and are often derived from cloned genes that have been subjected to site-directed or random mutagenesis or complete mutagenesis, or from completely synthesized genes.

[0083] Terms such as "DNA", "nucleic acid", "nucleic acid molecule", "polynucleotide" are used interchangeably to refer to a polymeric form of deoxyribonucleotides or ribonucleotides, or analogs thereof, of any length. Non-limiting examples of polynucleotides include linear and circular nucleic acids, messenger RNA (mRNA), complementary DNA (cDNA), recombinant polynucleotides, vectors, probes, primers, etc.

[0084] 5'-Nucleotidase, ecto and its inhibitors Human CD73 (also called 5'-nucleotidase, ecto; NT5E; or 5NT ) is a protein of 574 amino acid residues (accession number AAH6593). Eukaryotic CD73 functions as a non-covalently linked homodimer with two structural domains, and the N-terminal and C-terminal domains are linked by a hinge region, which allows the enzyme to have a large domain Upon receiving the in-migration, it can switch between an open conformation and a closed conformation (Knapp, K. et al. (2012) Structure 20:2161-73).

[0085] As used herein, the terms "CD73 inhibitor", "CD73 blocker", "5'-nucleo tidase, ecto inhibitor of adenosine", "NT5E inhibitor", "5NT inhibitor ", and all other related terms accepted in the art refer to compounds that can directly or indirectly regulate the CD73 receptor in in vitro assays, in vivo models, and / or other means demonstrating therapeutic efficacy. This term also refers to compounds that exhibit at least some therapeutic potency in human subjects. CD73 inhibitors can be competitive, non-competitive, or irreversible CD73 inhibitors. A "competitive CD73 inhibitor" is a compound that reversibly inhibits CD73 enzyme activity at the catalytic site; a "non-competitive CD73 inhibitor" is a compound that reversibly inhibits CD73 enzyme activity at a non-catalytic site; an "irreversible CD7 3 inhibitor" is a compound that irreversibly eliminates CD73 enzyme activity by forming a covalent bond with the enzyme (or other stable means of inhibiting enzyme function). A "competitive CD73 inhibitor" is a compound that reversibly inhibits CD73 enzyme activity at the catalytic site; a "non-competitive CD73 inhibitor" is a compound that reversibly inhibits CD73 enzyme activity at a non-catalytic site; an "irreversible CD7 3 inhibitor" is a compound that irreversibly eliminates CD73 enzyme activity by forming a covalent bond with the enzyme (or other stable means of inhibiting enzyme function). A "competitive CD73 inhibitor" is a compound that reversibly inhibits CD73 enzyme activity at the catalytic site; a "non-competitive CD73 inhibitor" is a compound that reversibly inhibits CD73 enzyme activity at a non-catalytic site; an "irreversible CD7

[0086] CD73 inhibitors can regulate purinergic signaling, a type of extracellular signaling mediated by purine nucleotides and nucleosides such as ATP and adenosine. Purinergic signaling involves the activation of purine receptors in cells and / or nearby cells, leading to the regulation of cell functions. The enzymatic activity of CD73 affects the duration, intensity, and chemical nature of purinergic signals sent to various cells (e.g., immune cells). CD73 inhibitors can regulate purinergic signaling, a type of extracellular signaling mediated by purine nucleotides and nucleosides such as ATP and adenosine. Purinergic signaling involves the activation of purine receptors in cells and / or nearby cells, leading to the regulation of cell functions. The enzymatic activity of CD73 affects the duration, intensity, and chemical nature of purinergic signals sent to various cells (e.g., immune cells). The enzymatic activity of CD73 affects the duration, intensity, and chemical nature of purinergic signals sent to various cells (e.g., immune cells). Play a strategic role in calibrating quality. Changes in these enzyme activities are related to cancer, autoimmune diseases and inflammatory diseases, infectious diseases, atherosclerosis, and ischemia-reperfusion injury, and can determine the results of several pathophysiological events, suggesting that these extracellular enzymes are novel therapeutic targets for managing various disorders.

[0087] Studies using tissues overexpressing CD73 and CD73 knockout mice have provided evidence that CD73 inhibitors have potential usefulness against melanoma, lung cancer, prostate cancer, and breast cancer (for example, see Sadej R. (2006) Melanoma Res 16:213-22). Higher expression levels of CD73 are associated with tumor angiogenesis, invasiveness, resistance to chemotherapy, and metastasis, so tumor progression and metastasis can be controlled using CD73 inhibitors. Other potential usefulness is described elsewhere in this specification. As described above, the compounds of the present invention are thought to exert their activity by inhibiting CD73, but an

[0088] exact understanding of their mechanism of action is not necessary to practice the present invention. For example, the compounds can also exert their activity, at least in part, by modulating (for example, inhibiting) other components of the purinergic signaling pathway (for example, CD39). The purinergic signaling system consists of transporters, enzymes, and receptors involved in the synthesis, release, action, and extracellular inactivation of (mainly) ATP and its extracellular degradation product adenosine (Sperlagh, B. et al. (Dec 2012)Neuropsychopharmacologia Hungarica 14(4):23 ). 1-38). Figure 1 shows a simplified representation of extracellular purinergic signaling (see, for example, North RA (Oct 2002) Physiological Reviews 82(4):1013-67). As shown therein there are several potential opportunities for the regulation of the signaling process. However as will be apparent to those skilled in the art, some of these opportunities are more tractable than others .

[0089] Identification of CD73 inhibitors with desirable properties The present invention relates, in part, to the identification of CD73 inhibitors having at least therapeutically relevant properties or characteristics. Candidate inhibitors can be identified, for example, using assays or models recognized in the art, examples of which will be apparent to those skilled in the art. The assays used to determine the CD73 inhibitory activity of the compounds described herein are set forth in the experimental section . After identification, candidate inhibitors can be further evaluated by using techniques that provide data on the characteristics of the inhibitor (e.g., pharmacokinetic parameters). Comparison of a candidate inhibitor with a reference standard substance (which may be the "best in class" of current inhibitors) indicates the potential of such a candidate.

[0090] CD73 inhibitors that can function as reference or benchmark compounds include α,β-methylene-ADP (AOPCP) described by Bhattarai et al. ((2015) J Med Chem 58:6248-63) and its derivatives and analogs, and the purinergic CD73 derivatives reported in PCT Publication No. 2015 / 164573 . Subsequently identified by those skilled in the art .

[0091] CD73 inhibitors that can function as reference or benchmark compounds include α,β-methylene-ADP (AOPCP) described by Bhattarai et al. ((2015) J Med Chem 58:6248-63) and its derivatives and analogs, and the purinergic CD73 derivatives reported in PCT Publication No. 2015 / 164573 . Subsequently identified by those skilled in the art . The viability of the candidate CD73 inhibitors can also be evaluated using other reference compounds.

[0092] Compounds of the present invention Formula (I):

Chem.

Chem.

Chemical formula

Chemical formula

[0093] For the above formula, the term "optionally substituted" is used in relation to an alkyl group, a cycloalkyl group, a cycloheteroalkyl group, an aryl group, and a heteroaryl group. Among each of these groups, some selected optional substituents are as follows: an alkyl group, a halogen, -OR’, -NR’R”, -SR’, -SiR’R”R”’, -OC(O)R’, -C(O)R’, -CO2R’, -CONR’R”, -OC(O)N R’R”, -NR”C(O)R’, -NR’-C(O)NR”R”’, -NR”C(O) 2R', -CN, and -NO2. R', R'', and R''' are each independently hydrogen, R' and R" are bonded to the same nitrogen atom. When R" and R'" are bonded to the same nitrogen atom, they When combined with the alkyl group, it forms a 3-, 4-, 5-, 6-, or 7-membered ring. For example, -NR'R" is It is meant to include 1-pyrrolidinyl and 4-morpholinyl. Cycloalkyl and cycloheteroalkyl groups: The above-listed options for "alkyl group" are Selected substituents are also useful for cycloalkyl and cycloheteroalkyl groups. Additionally, each of the cycloalkyl and cycloheteroalkyl groups may be oxo (=O). It may be further substituted. Alkyl and heteroaryl groups: -halogen, -OR', -OC(O)R', -NR 'R', -R', -CN, -NO2, -CO2R', CONR'R', C(O)R', -O C(O)NR'R', -NR'C(O)R', -NR'C(O)2R', -NR'-C( O)NR”R”’, -S(O)2R’, S(O)2NR’R”, NR’S(O)2R”, and and perfluoro(C1-C4)alkyl, where R', R'', and R''' are hydrogen, C1 independently selected from C1-C4 alkyl, C1-C4 haloalkyl, and C3-C6 cycloalkyl will be done.

[0094] In one selected group of embodiments, compounds of formula (I) are provided wherein A is ,formula [ka] (This is 1 to 5 R 6 (optionally substituted with).

[0095] In another alternative group of embodiments, compounds of formula (I) are provided, where A is

Chemical formula

[0096] In some selected embodiments, any one of a1 to a16 is combined with any one of b1 to b9 to provide a selected embodiment of formula (I). For example, compounds of formula (I) having the following Het-A- combinations in this specification are provided: a1 / b1; a1 / b2; a1 / b3; a1 / b4; a1 / b5; a1 / b6; a1 / b7; a 1 / b8; a1 / b9; a2 / b1; a2 / b2; a2 / b3; a2 / b4; a2 / b5 ; a2 / b6; a2 / b7; a2 / b8; a2 / b9; a3 / b1; a3 / b2; a3 / b3; a3 / b4; a3 / b5; a3 / b6; a3 / b7; a3 / b8; a3 / b9; a 4 / b1; a4 / b2; a4 / b3; a4 / b4; a4 / b5; a4 / b6; a4 / b7 ; a4 / b8; a4 / b9; a5 / b1; a5 / b2; a5 / b3; a5 / b4; a5 / b5; a5 / b6; a5 / b7; a5 / b8; a5 / b9; a6 / b1; a6 / b2; a 6 / b3; a6 / b4; a6 / b5; a6 / b6; a6 / b7; a6 / b8; a6 / b9 ; a7 / b1; a7 / b2; a7 / b3; a7 / b4; a7 / b5; a7 / b6; a7 / b7; a7 / b8; a7 / b9; a8 / b1; a8 / b2; a8 / b3; a8 / b4; a 8 / b5; a8 / b6; a8 / b7; a8 / b8; a8 / b9; a9 / b1; a9 / b2 ; a9 / b3; a9 / b4; a9 / b5; a9 / b6; a9 / b7; a9 / b8; a9 / b9; a10 / b1; a10 / b2; a10 / b3; a10 / b4; a10 / b5; a10 / b6; a10 / b7; a10 / b8; a10 / b9; a11 / b1; a11 / b2 b9;a10 / b1;a10 / b2;a10 / b3;a10 / b4;a10 / b5;a1 0 / b6;a10 / b7;a10 / b8;a10 / b9;a11 / b1;a11 / b2; a11 / b3;a11 / b4;a11 / b5;a11 / b6;a11 / b7;a11 / b 8;a11 / b9;a12 / b1;a12 / b2;a12 / b3;a12 / b4;a12 / b5;a12 / b6;a12 / b7;a12 / b8;a12 / b9;a13 / b1;a 13 / b2;a13 / b3;a13 / b4;a13 / b5;a13 / b6;a13 / b7 ;a13 / b8;a13 / b9;a14 / b1;a14 / b2;a14 / b3;a14 / b4;a14 / b5;a14 / b6;a14 / b7;a14 / b8;a14 / b9;a1 5 / b1;a15 / b2;a15 / b3;a15 / b4;a15 / b5;a15 / b6; a15 / b7;a15 / b8;a15 / b9;a16 / b1;a16 / b2;a16 / b 3;a16 / b4;a16 / b5;a16 / b6;a16 / b7;a16 / b8; or a 16 / b9.

[0097] In yet another selected embodiment, a compound of formula (I) is provided, wherein He t is of the formula:

Chemical formula

[0098] In yet another selected embodiment, a compound of formula (I) is provided, which is represented by one of the following subordinate formulas:

Chemical formula

[0099] In another group of selected embodiments, a compound of formula (I) is provided, where Het is selected from the following:

Chemical formula

[0100] Another further selected embodiment of formula (I) is a compound having a dependent formula selected from: the following:

Chemical formula

[0101] Also in one group of embodiments, the following formula:

Chemical formula

Chemical formula

Chemical formula

[0102] In one selected group of embodiments, the compound of formula (IVa) is one in which Het is

Chemical formula

[0103] In another selected group of embodiments, the compound of formula (IVa) is one in which Het is

Chem.

[0104] In yet another selected group of embodiments, the compound has the formula:

Chem.

[0105] In one selected group of embodiments, the compound of formula (IVb) has R a being NH2, N HR 7 NR 7 R 7 SR 7 and OR 7 selected from the group consisting of. In one selected group of embodiments, the compound of formula (Ib) has R c being halogen, R 7 OR 7 S R 7 SO2R 7 -X 1 -NH2, -X 1 -NHR 7 -X 1 -NR 7 R 7 -X 1 -OH, -X 1 -OR 7 -X 1 -SR 7 and -X 1 -SO2R 7 selected from the group consisting of.

[0106] In yet another group of embodiments, the compound of formula (IVb) has R e being H .

[0107] Synthesis methods Generally, the compounds provided herein can be prepared by conventional methods such as those described in the following examples.

[0108] Modifications to enhance the properties of inhibitors Improving one or more physical properties of the treatment modalities disclosed herein, and / or the manner in which they are administered, is often beneficial and sometimes essential. Improving physical properties includes, for example, methods of increasing water solubility, bioavailability, serum half-life, and / or therapeutic half-life; and / or methods of modulating biological activity.

[0109] Modifications known in the art include pegylation, Fc fusion, and albumin fusion. Generally associated with large molecular substances (e.g., polypeptides), such modifications have recently been evaluated using certain small molecules. As an example, Chiang, M. et al. (J.Am. Chem. Soc., 2014, 136(9):3370-73) describe small molecule agonists of the adenosine 2a receptor conjugated to the immunoglobulin Fc domain. The small molecule-Fc conjugate retained strong interactions with Fc receptors and the adenosine 2a receptor and exhibited superior properties compared to the unbound small molecule. Covalent attachment of PEG molecules to small molecule therapeutics has also been described (Li, W. et al., Progress in Polymer Science, 2013 38:421-44).

[0110] Therapeutic and prophylactic uses The present invention contemplates the use of the CD73 inhibitors described herein in the treatment or prevention of a wide range of diseases, disorders, and / or conditions, and / or their symptoms. Specific uses are described below. ​​​​​​​​​​​​ It should be understood that, although described in detail, the present invention is not limited thereto. Further, general categories of specific diseases, disorders, and conditions are described below, but some diseases, disor ders, and conditions may be members of two or more categories, and others may not be members of any of the disclosed categories.

[0111] Tumor-related disorders . According to the present invention, CD73 inhibitors can be used to treat or prevent proliferative conditions or disorders, including cancer, such as uterine cancer, cervical cancer, breast cancer, prostate cancer, testicular cancer, gastrointestinal cancers (e.g., esophageal cancer, oropharyngeal cancer, gastric cancer, small intestine cancer or large intestine cancer, colon, rectal cancer), kidney cancer, renal cell carcinoma, bladder cancer, bone cancer, myeloma, skin cancer, head cancer 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), ganglioneuroma, cancers of the central nervous system (CNS) and peripheral nervous system (PNS), as well as cancers of the hematopoietic and immune systems (e.g., spleen or thymus). The present invention also includes, for example, immunogenic tumors, non-immunogenic tumors, dormant tumors, virus-induced cancers (e.g., epithelial cell carcinoma, endothelial cell carcinoma, squamous cell carcinoma, and papillomavirus), adenocarcinoma, lymphoma, carcinoma, melanoma, leukemia, myeloma, sarcoma, teratocarcinoma, chemically induced cancers, metastasis, and angiogenesis. The present invention provides methods for treating or preventing other cancer-related diseases, disorders, or conditions. The present invention is intended to reduce resistance to tumor cells or cancer cell antigens, for example, by modulating the activity of regulatory T cells and / or CD8+ T cells (e.g., Ramirez-Montagut, et al. (2003)Oncogene 22:3180-87; and (see Sawaya, et al. (2003) New Engl. J. Med. 349:1501-09). In certain embodiments the tumor or cancer is colon cancer, ovarian cancer, breast cancer, melanoma, lung cancer, glioblastoma, or leukemia . The use of the terms cancer-related diseases, disorders, and conditions is meant to broadly encompass conditions that are directly or indirectly related to cancer, including pre-cancerous

[0112] conditions such as angiogenesis and dysplasia. In certain embodiments, the cancer is metastatic or at risk of becoming metastatic, or may be present in a diffuse tissue including cancer of the blood or bone marrow (e.g., leukemia). In some further embodiments, the compounds of the invention can be used to overcome T cell tolerance.

[0113] In some embodiments, the invention provides a method of treating a proliferative condition, cancer, tumor, or pre-cancerous condition with a CD73 inhibitor and at least one additional therapeutic or diagnostic agent (examples of which are described elsewhere in this specification).

[0114] Immune-related disorders and disorders caused by inflammatory components . As used herein, the terms “immune disease,” “immune condition,” “immune disorder,” “inflammatory disease,” “inflammatory condition,” “inflammatory disorder,” etc. mean to broadly encompass any immune-mediated condition (e.g., autoimmune disease), or a disorder with an inflammatory component that can be treated by a CD73 inhibitor described herein such that some therapeutic benefit is obtained. Such conditions are often closely intertwined with other diseases, disorders, and conditions. By way of example, an “immune condition” refers to a proliferative condition such as cancer, tumor, and angiogenesis (infections (acute and chronic), tumors, and cancers that are resistant to eradication by the immune system).

[0115] The CD73 inhibitor of the present invention can be used to increase or enhance the immune response; to improve vaccination, including enhancing the effectiveness of vaccines; and to increase inflammation. Immunodeficiency diseases, immunosuppressive therapies, acute and / or chronic infections, and immunodeficiencies associated with aging can be treated with the compounds disclosed herein. CD73 inhibitors can also be used to stimulate the immune system of patients suffering from medically induced immunosuppression (including patients who have received bone marrow transplantation, chemotherapy, or radiotherapy). In certain embodiments of the present disclosure, the CD73 inhibitor is used to increase or enhance the immune response to an antigen by providing adjuvant activity. In certain embodiments, at least one antigen or vaccine is administered to a subject in combination with at least one

[0116] CD73 inhibitor of the present invention to extend the immune response to the antigen or vaccine. Without being particularly limited, at least one antigenic substance or vaccine composition comprising a virus, bacterium, and fungus, or a portion thereof, protein, peptide, tumor specific antigen, and nucleic acid vaccine can also be used in combination with at least one CD73 inhibitor of the present invention. 1 CD73 inhibitor. The present invention contemplates the use of the CD73 inhibitors described herein in the treatment and / or prevention of any viral, bacterial, fungal, parasitic, or other infectious disease, disorder, or condition, where treatment with a CD73 inhibitor may be beneficial by inhibiting the immunosuppressive and anti-inflammatory activities of CD73. Examples of such diseases and disorders include HIV and A and the like.

[0117] Microorganism-related disorders By inhibiting the immunosuppressive and anti-inflammatory activities of CD73, the present invention contemplates the use of the CD73 inhibitors described herein in the treatment and / or prevention of any viral, bacterial, fungal, parasitic, or other infectious disease, disorder, or condition, where treatment with a CD73 inhibitor may be beneficial. Examples of such diseases and disorders include HIV and A IDS, Staphylococcus and Streptococcus infections (e.g., Staphylococcus aureus and Streptococcus oralis), Leishmania, Toxoplasma, Trichomonas, Giardia, Candida albicans, Bacillus anthracis, and Pseudomonas aeruginosa are included. The compounds of the present invention can be used for the treatment of sepsis, reduction or inhibition of bacterial growth, and reduction or inhibition of inflammatory cytokines.

[0118] CNS-related and neurological disorders Inhibition of CD73 is also an important therapeutic strategy for patients with neurological, neuropsychiatric, neurodegenerative, or other diseases, disorders, and conditions associated with the central nervous system, including disorders related to impairment of cognitive and motor functions. Examples include Parkinson's disease, extrapyramidal syndrome (EPS), dystonia, akathisia, tardive dyskinesia, restless legs syndrome (RLS), epilepsy, periodic limb movements during sleep (PL MS), attention deficit disorder, depression, anxiety, dementia, Alzheimer's disease, Huntington's disease, multiple sclerosis, cerebral ischemia, hemorrhagic stroke, subarachnoid hemorrhage, and traumatic brain injury.

[0119] Other disorders Embodiments of the present invention contemplate administration of the CD7 3 inhibitors described herein to a subject for the treatment or prevention of any other disorder that can benefit from at least a certain level of CD73 inhibition. Such diseases, disorders, and conditions include, for example, those of the cardiovascular system (e.g., cardiac ischemia), gastrointestinal (e.g., Crohn's disease), metabolic (e.g., diabetes), liver (e.g., liver fibrosis, NASH, and NAFLD), lung (e.g., COPD and asthma), eye (e.g., diabetic retinopathy), and kidney (e.g., renal insufficiency) disorders.

[0120] ​In some embodiments, the CD73 inhibitor of the present invention inhibits statin-induced adenosine production or reduces or decreases the increase in blood glucose caused by statins in subjects taking statins (e.g., lovastatin and pravastatin), and can be used for this purpose. The CD73 inhibitor of the present invention can be in the form of a composition suitable for administration to a subject. Generally, such a composition is a "pharmaceutical composition" comprising a CD73 inhibitor and one or more pharmaceutically acceptable or physiologically acceptable diluents, carriers, or excipients. In certain embodiments the CD73 inhibitor is present in a therapeutically acceptable amount. The pharmaceutical composition can be used in the methods of the present invention. Thus, for example, the pharmaceutical composition can be administered to a subject ex vivo or in vivo to carry out the treatment

[0121] Pharmaceutical compositions methods, prophylactic methods, and uses described herein. The pharmaceutical composition of the present invention can be formulated to be compatible with the intended method or route of administration. Exemplary routes of administration are described herein. Further, the pharmaceutical composition can be used in combination with other therapeutically active agents or compounds described herein for treating or preventing the diseases, disorders, and conditions contemplated by the present invention. The pharmaceutical composition containing an active ingredient (e.g., an inhibitor of CD73 function) can be in a form suitable for oral use, such as tablets, capsules, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, hard or soft capsules, or syrups, solutions, microbeads, or The pharmaceutical composition of the present invention can be formulated to be compatible with the intended method or route of administration. Exemplary routes of administration are described herein. Further, the pharmaceutical composition can be used in combination with other therapeutically active agents or compounds described herein for treating or preventing the diseases, disorders, and conditions contemplated by the present invention. methods, prophylactic methods, and uses described herein. can be administered to a subject ex vivo or in vivo to carry out the treatment

[0122] The pharmaceutical composition of the present invention can be formulated to be compatible with the intended method or route of administration. Exemplary routes of administration are described herein. Further, the pharmaceutical composition can be used in combination with other therapeutically active agents or compounds described herein for treating or preventing the diseases, disorders, and conditions contemplated by the present invention. Exemplary routes of administration are described herein. Further, the pharmaceutical composition can be used in combination with other therapeutically active agents or compounds described herein for treating or preventing the diseases, disorders, and conditions contemplated by the present invention. The pharmaceutical composition of the present invention can be formulated to be compatible with the intended method or route of administration. Exemplary routes of administration are described herein. Further, the pharmaceutical composition can be used in combination with other therapeutically active agents or compounds described herein for treating or preventing the diseases, disorders, and conditions contemplated by the present invention. The pharmaceutical composition containing an active ingredient (e.g., an inhibitor of CD73 function) can be in a form suitable for oral use, such as tablets, capsules, troches, lozenges, aqueous or oily suspensions, dispersible powders

[0123] The pharmaceutical composition containing an active ingredient (e.g., an inhibitor of CD73 function) can be in a form suitable for oral use, such as tablets, capsules, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, hard or soft capsules, or syrups, solutions, microbeads, or powders, or granules, hard or soft capsules, or syrups, solutions, microbeads, or It may also be an elixir. Pharmaceutical compositions intended for oral use can be prepared according to any method known in the art, and such compositions may contain one or more agents such as, for example, sweetening agents, flavoring agents, coloring agents, and preservatives in order to provide elegant and palatable preparations. Tablets, capsules, etc. contain the active ingredient in a mixture with non-toxic pharmaceutically acceptable excipients suitable for the manufacture of tablets. These excipients may include, for example, diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrating agents such as corn starch or alginic acid; binding agents such as starch, gelatin, or acacia, and lubricants such as magnesium stearate, stearic acid, or talc. Tablets, capsules, etc. suitable for oral administration are not coated or may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action. For example, time-delay materials such as glyceryl monostearate or glyceryl distearate can be used. They may also be coated by techniques known in the art to form osmotic therapeutic tablets for controlled release.

[0124] Additional agents include polyesters, polyamino acids, hydrogels, polyvinylpyrrolidone, polyanhydrides, polyglycolic acid, ethylene-vinyl acetate, methylcellulose, carboxymethylcellulose, protamine sulfate, or lactide / glycolide copolymers, polylactide / glycolide copolymers, or ethyl to control the delivery of the administered composition. ​​​​Contain biodegradable or biocompatible particles or polymeric substances such as vinyl acetate copolymers For example, oral preparations can be prepared by the use of coacervation technology or interfacial polymerization, hydroxymethylcellulose or gelatin - microcapsules or poly(methyl methacrylate) microcapsules respectively, or encapsulated in microcapsules prepared in a colloidal drug delivery system. Colloidal dispersion systems include oil - in - water emulsions, micelles , mixed micelles, and liposomes, macromolecular complexes, nanocapsules, microspheres , microbeads, and lipid - based systems. The methods for preparing the above - mentioned formulations will be apparent to those skilled in the art .

[0125] Formulations for oral use may also be provided as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate, or kaolin, or microcrystalline cellulose, or as soft gelatin capsules in which the active ingredient is mixed with a water or oil medium such as peanut oil, liquid paraffin, or olive oil.

[0126] Aqueous suspensions contain the active substance mixed with excipients suitable for their manufacture. Such excipients include suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and gum arabic; dispersing or wetting agents such as natural phosphatides (such as lecithin), or condensation products of alkylene oxides and fatty acids (such as polyoxyethylene stearate), or condensation products of ethylene oxide and long - chain aliphatic alcohols . a substance (e.g., heptadecaethyleneoxycetanol), or a condensation product of ethylene oxide with a partial ester obtained from a fatty acid and a hexitol (e.g., polyoxyethylene sorbitol monooleate), or a condensation product of ethylene oxide with a partial ester obtained from a fatty acid and a hexitol anhydride (e.g., polyethylene sorbitan monooleate) may also be used. The aqueous suspension may also contain one or more preservatives.

[0127] The oily suspension may be formulated by suspending the active ingredient in a vegetable oil such as peanut oil, olive oil, sesame oil, or coconut oil, or a mineral oil such as liquid paraffin. The oily suspension may contain a thickening agent such as beeswax, hard paraffin, or cetyl alcohol. To provide an orally acceptable oral formulation, sweetening agents and flavoring agents as described above may be added.

[0128] Dispersible powders and granules suitable for preparing an aqueous suspension by adding water provide the active ingredient in a mixture with a dispersing agent or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above.

[0129] The pharmaceutical composition of the present invention may also be in the form of an oil-in-water emulsion. The oily 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 emulsifying agents are natural gums such as gum arabic or tragacanth; natural phospholipids such as soy lecithin, and esters or partial esters derived from fatty acids; hexitol anhydrides such as sorbitan monooleate; and partial esters and ethylene oxide condensation products thereof. Condensation products with renoxides, such as polyoxyethylene sorbitan monooleate are possible.

[0130] The pharmaceutical composition typically comprises a therapeutically effective amount of a CD73 inhibitor contemplated by the present invention and one or more pharmaceutically and physiologically acceptable formulations. Suitable pharmaceutically and physiologically acceptable diluents, carriers, or excipients are not particularly limited, but include antioxidants (e.g., ascorbic acid and sodium bisulfite), preservatives (e.g., benzyl alcohol , methylparaben, ethyl or n-propyl, p-hydroxybenzoic acid), emulsifiers, suspending agents, dispersing agents, solvents, fillers, bulking agents, surfactants, buffers, vehicles, diluents, and / or adjuvants. For example, suitable vehicles may be saline or citrate-buffered saline supplemented with other substances common to pharmaceutical compositions for parenteral administration . Neutral buffered saline or saline mixed with serum albumin are further exemplary excipients. Those skilled in the art will readily recognize the various buffers that can be used in the pharmaceutical compositions and dosage forms contemplated herein . Typical buffers include, but are not particularly limited to, pharmaceutically acceptable weak acids, weak bases, or mixtures thereof. As an example, buffer components may be water-soluble substances 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, Tris buffer, N-(2-hydroxyethyl)piperazine -N’-(2-ethanesulfonic acid) (HEPES), 2-(N-morpholino)ethanesulfonic acid (MES), sodium salt of 2-(N-morpholino)ethanesulfonic acid (MES), and the like. Acceptable buffers include, for example, Tris buffer, N-(2-hydroxyethyl)piperazine -N’-(2-ethanesulfonic acid) (HEPES), 2-(N-morpholino)ethanesulfonic acid (MES), sodium salt of 2-(N-morpholino)ethanesulfonic acid (MES), 3-(N-Morpholino)propanesulfonic acid (MOPS), and N-tris[hydroxy methyl]methyl-3-aminopropanesulfonic acid (TAPS) are included.

[0131] After formulating the pharmaceutical composition, it can be stored in a sterile vial as a solution, suspension, gel, emulsion, solid, or de hydrated or lyophilized powder. Such formulations can be stored in any form that is ready for use, in a lyophilized form that needs to be reconstituted before use, in a liquid form that needs to be diluted before use, or in any other acceptable form. In some embodiments, the pharmaceutical composition is provided in a single-use container (e.g., a single-use vial, amp ule, syringe, or auto-injector (similar to EpiPen®)), and in other embodiments, a multi-use container (e.g., a multi-use vial) is provided. The formulation can also include a carrier to protect the composition from rapid degradation or removal from the body, such as a controlled-release formulation including liposomes, hydrogels, prodrugs, and microencapsulation delivery

[0132] systems. For example, a time-delay material such as glyceryl monostearate or glyceryl stearate can be used alone or in combination with a wax. Any drug delivery device can be used to deliver the CD73 inhibitor, including implants (e.g., implantable pumps) and catheter systems, low-rate infusion pumps, and devices (all of which are well known to those skilled in the art). Depot injections, which are generally administered subcutaneously or intramuscularly, can also be used to deliver the present disclosed agent over a defined period of time.

[0133] ​​It can be used to release the CD73 inhibitor disclosed in the detailed description. Depot injections are usually solid or oily and generally contain at least one of the formulation components described herein . Those skilled in the art are proficient in possible formulations and the use of depot injections.

[0134] The pharmaceutical composition may be in the form of a sterile injectable aqueous or oily suspension. This suspension can be prepared according to known techniques using the appropriate dispersing or wetting agents and suspending agents described above. Sterile injectable preparations may be sterile injectable solutions or suspensions in a non toxic parenterally acceptable diluent or solvent such as a solution in 1,3 - butanediol. . Acceptable diluents, solvents, and dispersion media that can be used include water, Ringer's solution, isotonic sodium chloride solution, Cremophor EL® (BASF, Parsippany, NJ), or phosphate buffered physiological saline (PBS), ethanol, polyols (e.g., glycerol, propylene glycol col, and liquid polyethylene glycol), and suitable mixtures thereof. Further , sterile non - volatile oils have conventionally been used as solvents or suspension media. For this purpose , any low - irritation fixed oil containing synthetic monoglycerides or diglycerides can be used. Furthermore, fatty acids such as oleic acid are used in the preparation of injections. By including agents that delay absorption (e.g., aluminum monostearate or gelatin), sustained absorption of certain injectable formulations can be achieved.

[0135] The present invention contemplates the administration of the CD73 inhibitor in the form of a suppository for rectal administration. The suppository is a mixture of the drug with a suitable non - irritating excipient that is solid at normal temperature but liquid at rectal temperature. ​It can be prepared by, and thus dissolves in the rectum to release the drug. Thus Such substances are not particularly limited, but include cocoa butter and polyethylene glycol is included.

[0136] The CD73 inhibitors contemplated by the present invention can be in the form of any other suitable pharmaceutical composition (e.g., nasal or inhalation spray) that is currently known or will be developed in the future. It can be in the form of.

[0137] Routes of administration The present invention contemplates the administration of CD73 inhibitors and their compositions by any suitable method. Suitable routes of administration include oral, parenteral (e.g., intramuscular, intravenous, subcutaneous (e.g., injection or implantation), intraperitoneal, intracapsular, intra-articular, intraperitoneal, intracerebral (intracerebral parenchyma, intraventricular), intranasal, intravaginal, sublingual, intraocular, rectal, topical (e.g., transdermal), oral, and inhalation). Generally, depot injections administered subcutaneously or intramuscularly can also be utilized to release the CD 73 inhibitors disclosed herein over a defined period. is released.

[0138] Certain embodiments of the present invention contemplate oral administration.

[0139] Combination therapies The present invention contemplates the use of CD73 inhibitors in combination with one or more active therapeutic agents (e.g., chemotherapeutic agents) or other prophylactic or therapeutic methods (e.g., radiation). Such combination therapies often have different complementary mechanisms of action. Such combination therapies can be particularly advantageous by allowing for dose reduction of one or more agents, thereby reducing or eliminating adverse effects associated with one or more agents. Furthermore, such combination therapies can be particularly advantageous by allowing for dose reduction of one or more agents, thereby reducing or eliminating adverse effects associated with one or more agents. Furthermore, such The combination therapy may have a synergistic therapeutic or prophylactic effect on the underlying disease, disorder, or condition. It may have.

[0140] As used herein, "combination" means treatments that can be administered separately, such as treatments that are separately formulated for separate administrations (e.g., as may be provided in a kit), and treatments that are administered together in a single formulation (e.g., as may be provided in a kit), and treatments that are administered together in a single formulation (e.g., "co-prescription").

[0141] In certain embodiments, the CD73 inhibitor is administered or applied continuously, for example when one agent is administered before one or more other agents. In other embodiments, the CD73 inhibitor is administered simultaneously. For example, when two or more agents are administered simultaneously or almost simultaneously, the two or more agents may be present in two or more separate formulations or may be combined in a single formulation (i.e., a co-formulation). Whether the two or more agents are administered continuously or simultaneously, they are considered to be administered in combination for the purposes of the present invention. They are considered to be administered in combination for the purposes of the present invention.

[0142] The CD73 inhibitors of the present invention can be used in combination with at least one other (active) agent in any suitable manner depending on the circumstances. In one embodiment, treatment with at least one active agent and at least one CD73 inhibitor of the present invention is maintained over a period of time. In another embodiment, treatment with at least one active agent is reduced or discontinued (e.g., when the subject is stable), while treatment with the CD 73 inhibitor of the present invention is maintained at a certain dosing regimen. In a further embodiment, treatment with at least one active agent is reduced or discontinued (e.g., when the subject is stable), while treatment with the CD 73 inhibitor of the present invention is maintained at a certain dosing regimen. In a further embodiment, treatment with at least ​in which case), on the other hand, treatment with the CD73 inhibitor of the present invention is reduced (e.g., low dose , low frequency of administration, or shorter duration of administration). In a further embodiment, treatment with at least one active agent is reduced or discontinued (e.g., when the subject is stable) , treatment with the CD73 inhibitor of the present invention is enhanced (e.g., high dose, high frequency of administration, long -term treatment regimen). In a further embodiment, treatment with at least one active agent is maintained , and treatment with the CD73 inhibitor of the present invention is reduced or discontinued (e.g., low dose, low frequency of administration, or shorter-term treatment regimen). In a further embodiment, treatment with at least one active agent and treatment with the CD73 inhibitor of the present invention are reduced or discontinued (e.g., low dose, low frequency of administration, or shorter-term treatment regimen).

[0143] Cancer-related diseases . The present invention provides a method of treating and / or preventing a proliferative state, cancer, tumor, or precancerous disease, disorder, or condition using a CD73 inhibitor and at least one additional therapeutic or diagnostic agent . In certain embodiments, the present invention provides a method of tumor suppression of tumor growth comprising administering the CD73 inhibitor described herein in combination with a signal transduction inhibitor (STI) to achieve an 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 of a signal transduction pathway

[0144] . The signal transduction inhibitor (STI) of the present invention includes (i) a bcr / abl kinase inhibitor (e.g GLEEVEC); (ii) epidermal growth factor (EGF) including kinase inhibitors and antibodies . . . e.g., GLEEVEC); (ii) epidermal growth factor (EGF) including kinase inhibitors and antibodies Receptor inhibitor; (iii) her-2 / neu receptor inhibitor (e.g., HERCEPTIN ); (iv) Inhibitor of Akt family kinase or Akt pathway (e.g., rapamycin) ; (v) Cell cycle kinase inhibitor (e.g., flavopiridol); and (vi) Phosphati idylinositol kinase inhibitor. Agents involved in immunomodulation can also be used in combination with the CD73 inhibitors described herein for the suppression of tumor growth in cancer patients as described herein and

[0145] Examples of chemotherapeutic agents include, but are not particularly limited to, alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carbocon, meturedopa, and uredopa; ethyleneimines and methylmelamines such as altretamine, triethylenemelamine, trimethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; nitrogen mustards such as chlorambucil, chloronaphazine, choline phosphamide, estramustine, ifosfamide, mechlorethamine, mephloretin oxide hydrochloride, melphalan, novembicin, phenesterine prednimustine, tropophosphamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine ; antibiotics such as aclacinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, caminomycin, cardinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin ; ethyleneimines and methylmelamines such as altretamine, triethylenemelamine, trimethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; nitrogen mustards such as chlorambucil, chloronaphazine, choline phosphamide, estramustine, ifosfamide, mechlorethamine, mephloretin oxide hydrochloride, melphalan, novembicin, phenesterine ; triethylenethiophosphoramide, and trimethylolmelamine; nitrogen mustards such as chlorambucil, chloronaphazine, choline phosphamide, estramustine, ifosfamide, mechlorethamine, mephloretin oxide hydrochloride, melphalan, novembicin, phenesterine prednimustine, tropophosphamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine ; antibiotics such as aclacinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, caminomycin, cardinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin ; prednimustine, tropophosphamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine ; antibiotics such as aclacinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, caminomycin, cardinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin ; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine ; antibiotics such as aclacinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, caminomycin, cardinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin ; antibiotics such as aclacinomycin, actinomycin, autramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, caminomycin, cardinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin ; carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin Bicinchon, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin ceramycin, rhodomycin, streptozocin, streptozotocin, tubericidin, ubenimex, dinostatin, zorubicin; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs, such as denopterin, methotrexate, pteropterin, trimethoprim; purine analogs, such as fludarabine, 6-mercaptopurine, thiampurine, thioguanine; pyrimidine analogs, such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine doxifluridine, enocitabine, floxuridine, 5-FU; androgens, such as calusterone, drostanolone propionate, epithiostanol, mepithiostane, testolactone; antiadrenal agents, such as aminoglutethimide, mitotane, trilostane ; folic acid supplements, such as folic acid; aceglatone; aldophosphamide glycoside; amorebric acid; amsacrine; bestrabucil; bisantrene; edatrexate; defofamine; decortin; diadizoone; elformithine; etoposide acetate; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllinic acid; 2-ethylhydrazide; procarbazine ; razoxane; sizofiran; spirogermanium; tenuazonic acid; triadizoone; 2 , 2’,2’’-trichloro-triethylamine; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gasitocin; ara binoside (Ara-C); cyclophosphamide; thiotepa; taxoids, such as pacli taxel and docetaxel; chlorambucil; gemcitabine; 6-thioguanine; mercap topurin; methotrexate; platinum and platinum coordination complexes, such as cisplatin and carbop latins; vinblastine; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; te niposide; daunomycin; aminopterin; zeloda; ibandronate; CPT11 ; topoisomerase inhibitors; difluoromethylornithine (DMFO); retinoic acid; esperamicin; capecitabine; and pharmaceutically acceptable salts, acids, or derivatives of any of the above are included.

[0146] The chemotherapeutic agent also acts to modulate or inhibit the hormonal action on tumors, such as anti-ho rmone agents, such as anti-estrogens (e.g., tamoxifen, raloxifene, aromata se inhibitor 4(5)-imidazole, 4-hydroxytamoxifen, trioxifene, ke oxifen, onapristone, and toremifene); and anti-androgen agents, such as f lutamide, nilutamide, bicalutamide, leuprorelin, leuprorelin, and goserelin; and pharmaceutically acceptable salts, acids, or derivatives of any of the above are included. In certain embodiments the combination therapy includes the administration of a hormone or related hormone agent.

[0147] Additional therapeutic methods that can be used in combination with a CD73 inhibitor include radiation therapy, tumor antigens monoclonal antibodies against, conjugates of monoclonal antibodies and toxins, T cell adjuvants , bone marrow transplantation, or antigen-presenting cells (e.g., dendritic cell therapy).

[0148] Immune checkpoint inhibitors . The present invention contemplates the use of an inhibitor of CD73 function as described herein in combination with an immune checkpoint inhibitor. The large number of genetic and epigenetic changes characteristic of all cancers provides a diverse set of antigens that the immune system can use to distinguish tumor cells from their normal counterparts.

[0149] In the case of T cells, the ultimate strength (e.g., cytokine production or proliferation levels) and quality (e.g., type of immune response generated such as cytokine production pattern) of the response initiated by antigen recognition by the T cell receptor (TCR) are regulated by the balance between co-stimulatory signals and inhibitory signals (immune checkpoints). Under normal physiological conditions, immune checkpoints are extremely important for preventing autoimmunity (i.e., maintaining self-tolerance) and protecting tissues from damage when the immune system is responding to pathogen infection. The expression of immune checkpoint proteins can be dysregulated by tumors as an important immune escape mechanism. In various types of tumor cells that are candidates for blockade, some examples of immune checkpoints (ligands and receptors) that are selectively upregulated include PD1 (programmed cell death protein 1); PDL1 (PD1 ligand); BTLA (B lymphocyte and T lymphocyte attenuator) T cell attenuator); etc.

[0150] ​​​​​​​​​​NK cell attenuator); CTLA4 (cytotoxic T lymphocyte-associated antigen 4); TIM3 (T cell membrane protein 3); LAG3 (lymphocyte activation gene 3); A2aR (adenosine A2a receptor A2aR); and, based on its structural characteristics, two classes: 1) killer cell immunoglobulin-like receptors (KIR), and ii) C-type lectin receptors (members of the type II transmembrane receptor family) including killer inhibitory receptors that can be divided. Other relatively less well-defined immune checkpoint s have been described in the literature, and both 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)) are present. [See Pardoll, (April 2012) Nature Rev. Cancer12:252-64].

[0151] The present invention contemplates the use of the inhibitors of the CD73 function described herein in combination with inhibitors of the above immune checkpoint receptors and ligands, as well as immune checkpoint receptors and ligands not yet described. Certain modulators of immune checkpoints are currently available, while others are in the late stages of development. To explain, when approved for the treatment of melanoma in 201 1, ipilimumab (YERVOY; Bristol-Myers Squibb), a fully humanized CTLA4 monoclonal antibody, became the first immune checkpoint inhibitor to receive regulatory approval in the United States. CTLA4 and an antibody (CTLA4-Ig ; A fusion protein comprising abatcept (ORENCIA; Bristol-Myers Squibb)) has been used for the treatment of rheumatoid arthritis, and other fusion proteins have been shown to be effective in kidney transplant patients dried with Epstein-Barr virus. PD1 antibodies are under development (e.g., nivolumab (Bristol-Myers Squibb)), and pembrolizumab (lambrolizumab) (Merck)) and anti-PDL1 antibodies are also being evaluated (e.g., MPDL3280A (Roche)). Nivolumab has been shown to be promising in patients with melanoma, lung, and kidney cancer .

[0152] The present invention includes any of the above pharmaceutically acceptable salts, acids, or derivatives.

[0153] Metabolic and cardiovascular diseases . The present invention provides a method for treating and / or preventing specific cardiovascular and / or metabolic related diseases, disorders, and conditions, as well as disorders associated therewith, using a CD73 inhibitor and at least one additional therapeutic agent or diagnostic agent.

[0154] Useful in combination therapies for the treatment of hypercholesterolemia (and atherosclerosis) Examples of therapeutic agents include statins that inhibit the enzymatic synthesis of cholesterol (e.g., Crestor, Lescol, Lipitor, Mevacor, Pravachol, and Zocor); bile acid resins that sequester cholesterol and prevent its absorption (e.g., cholestyramine, Locostor, Prevalite, Questran, and Welchol); ezetimibe that prevents cholesterol absorption (Zetia); fibrates that lower triglycerides and moderately increase HDL For example, tricholesterol; moderately lowers LDL cholesterol and triglycerides Iacin (e.g., Niacol); and / or combinations of the above (e.g., Vitrin (Ezetrin) timibe and simvastatin). Alternative cholesterol treatments that can be used include various supplements and herbs (e.g. For example, garlic, policosanol, and guggul.

[0155] The present invention includes pharma- ceutically acceptable salts, acids, or derivatives of any of the above.

[0156] Disorders with immune-related and inflammatory components The present invention relates to a CD73 inhibitor and at least one One additional therapeutic or diagnostic agent is used to treat immune-related diseases, disorders, and conditions; and inflammatory diseases. The present invention provides methods for treating and / or preventing diseases, disorders, and conditions having a molecular weight of about 1,000 to 1,000.

[0157] Examples of therapeutic agents useful in combination therapy are specific to the underlying disease, disorder, or condition and include those is known to the traders.

[0158] Microbial diseases The present invention relates to a method for treating a disease comprising administering to a patient a therapeutic agent or a diagnostic agent comprising administering to said patient a therapeutic agent or a diagnostic agent. (e.g., one or more additional therapeutic or diagnostic agents (e.g., one or more other antiviral agents and and / or one or more drugs not related to viral therapy) to treat viral, bacterial, or fungal infections. Treating and preventing fungal and parasitic diseases, disorders, and conditions, and disorders related thereto and / or a method for preventing the same.

[0159] Such combination therapies involve targeting different stages of the viral life cycle and acting with different mechanisms of action. Antiviral agents with mechanisms of action include, but are not limited to: Inhibitors of influenza (e.g., amantadine and rimantadine); reverse transcriptase inhibitors (e.g., , acyclovir, zidovudine, and lamivudine); agents targeting integrase; agents that block the binding of transcription factors to viral DNA; agents that affect translation (e.g., , antisense molecules) (e.g., formylsen); agents that modulate translation / ribozyme function; protease inhibitors; viral assembly modulators (e.g., rifampicin); antiretroviral agents, such as nucleoside analog reverse transcriptase inhibitors (e.g., azidothymidine (AZT), ddl, ddC, 3TC, d4T); non-nucleoside reverse transcriptase inhibitors (e.g., efavirenz, nevirapine); nucleotide analog reverse transcriptase inhibitors; and agents that prevent the release of viral particles (e.g., zanamivir and oseltamivir). The treatment and / or prevention of certain viral infections (e.g., HIV) often involves a combination of antiviral agents (a "cocktail").

[0160] Other antiviral agents contemplated for combination with a CD73 inhibitor are not particularly limited, but include: abacavir, adefovir, amantadine, amprenavir, amprenavir, arbidol, atazanavir, atripla, boceprevir, cidofovir, combivir, darunavir, delavirdine, didanosine, docosanol, edoxudine, emtricitabine, enfuvirtide, entecavir, famciclovir, fosamprenavir, foscarnet, phosphonete, http: / / en.wikipedia.org / wiki / Fusion_inhibitor ganciclovir, ibacitabine, immunovir, idoxuridine, i ; Mikimodo, Indinavir, Inosine, various interferons (e.g., Peginterferon α-2a), Ritonavir, Roviride, Maraviroc, Moroxydine, Methylthiazone, Nelfinavir, Nexavir, Penciclovir, Peramivir, Preconalyl, Podophyllo Toxin, Raltegravir, Ribavirin, Ritonavir, Pyrimidine, Saquinavir, Stub din, Telaprevir, Tenofovir, Tipranavir, Trifluridine, Tridivir, Tro mantadine, Truvada, Valaciclovir, Valganciclovir, Vicriviroc, Vidara bin, Viramidine, and Zalcitabine.

[0161] The present invention contemplates the use of an inhibitor of the CD73 function described herein in combination with an anti-parasitic agent. Such agents include, but are not particularly limited to, thiabendazole, pyrantel pamoate, mebendazole, praziquantel, niclosamide, bithionol, o xamniquine, metrifonate, ivermectin, albendazole, eflornithine, melarsoprol, pentamidine, benznidazole, nifurtimox, and nit romidazole. Those skilled in the art are aware of other agents that may be useful in the treatment of parasitic diseases.

[0162] Embodiments of the present invention contemplate the use of the CD73 inhibitor described herein in combination with an agent useful for the treatment or prevention of bacterial disorders. Antibacterial agents can be classified in various ways, such as based on their mechanism of action, chemical structure, and spectrum of activity. Examples of antibacterial agents include those that target the bacterial cell wall (e.g., cephalosporins and penicillins ) or the cell membrane (e.g., polymyxin), or those that interfere with essential bacterial enzymes. ) or those that target the cell membrane (e.g., polymyxin), or those that interfere with essential bacterial enzymes. It includes substances (e.g., sulfonamides, rifamycins, and quinolines). Protein Most antibacterial agents that target protein synthesis (e.g., tetracyclines and macrolides ) are bacteriostatic, while agents such as aminoglycosides are bactericidal. Another means of classifying antibacterial agents is based on their target specificity; "narrow-spectrum" agents target specific types of bacteria (e.g., gram-positive bacteria such as streptococci), and "broad-spectrum" agents have activity against a wider range of bacteria. Those skilled in the art recognize the appropriate type of antibacterial agent for use in a particular bacterial infection. Another means of classifying antibacterial agents is based on their target specificity; "narrow-spectrum" agents target specific types of bacteria (e.g., gram-positive bacteria such as streptococci), and "broad-spectrum" agents have activity against a wider range of bacteria. Those skilled in the art recognize the appropriate type of antibacterial agent for use in a particular bacterial infection. Another means of classifying antibacterial agents is based on their target specificity; "narrow-spectrum" agents target specific types of bacteria (e.g., gram-positive bacteria such as streptococci), and "broad-spectrum" agents have activity against a wider range of bacteria. Those skilled in the art recognize the appropriate type of antibacterial agent for use in a particular bacterial infection. Another means of classifying antibacterial agents is based on their target specificity; "narrow-spectrum" agents target specific types of bacteria (e.g., gram-positive bacteria such as streptococci), and "broad-spectrum" agents have activity against a wider range of bacteria. Those skilled in the art recognize the appropriate type of antibacterial agent for use in a particular bacterial infection. Another means of classifying antibacterial agents is based on their target specificity; "narrow-spectrum" agents target specific types of bacteria (e.g., gram-positive bacteria such as streptococci), and "broad-spectrum" agents have activity against a wider range of bacteria. Those skilled in the art recognize the appropriate type of antibacterial agent for use in a particular bacterial infection.

[0163] Embodiments of the present invention contemplate the use of the CD73 inhibitors described herein in combination with agents useful for the treatment or prevention of fungal diseases. Antifungal agents include polyenes (e.g., amphotericin, nystatin, and pimaricin); azoles (e.g., fluconazole, itraconazole, and ketoconazole); allylamines (e.g., naftifine and terbinafine), and morpholines (e.g., amorolfine); and antimetabolites (e.g., Embodiments of the present invention contemplate the use of the CD73 inhibitors described herein in combination with agents useful for the treatment or prevention of fungal diseases. Antifungal agents include polyenes (e.g., amphotericin, nystatin, and pimaricin); azoles (e.g., fluconazole, itraconazole, and ketoconazole); allylamines (e.g., naftifine and terbinafine), and morpholines (e.g., amorolfine); and antimetabolites (e.g., Embodiments of the present invention contemplate the use of the CD73 inhibitors described herein in combination with agents useful for the treatment or prevention of fungal diseases. Antifungal agents include polyenes (e.g., amphotericin, nystatin, and pimaricin); azoles (e.g., fluconazole, itraconazole, and ketoconazole); allylamines (e.g., naftifine and terbinafine), and morpholines (e.g., amorolfine); and antimetabolites (e.g., Embodiments of the present invention contemplate the use of the CD73 inhibitors described herein in combination with agents useful for the treatment or prevention of fungal diseases. Antifungal agents include polyenes (e.g., amphotericin, nystatin, and pimaricin); azoles (e.g., fluconazole, itraconazole, and ketoconazole); allylamines (e.g., naftifine and terbinafine), and morpholines (e.g., amorolfine); and antimetabolites (e.g., Embodiments of the present invention contemplate the use of the CD73 inhibitors described herein in combination with agents useful for the treatment or prevention of fungal diseases. Antifungal agents include polyenes (e.g., amphotericin, nystatin, and pimaricin); azoles (e.g., fluconazole, itraconazole, and ketoconazole); allylamines (e.g., naftifine and terbinafine), and morpholines (e.g., amorolfine); and antimetabolites (e.g., 5-fluorocytosine).

[0164] The present invention includes pharmaceutically acceptable salts, acids, or derivatives of the above agents (and members of classes of agents).

[0165] Administration The CD73 inhibitors of the present invention are administered to a subject in an amount that depends, for example, on the goal of administration (e.g., the degree of desired resolution); the age, weight, sex, and health and physical condition of the subject to whom the formulation is being administered; the route of administration; and The CD73 inhibitors of the present invention are administered to a subject in an amount that depends, for example, on the goal of administration (e.g., the degree of desired resolution); the age, weight, sex, and health and physical condition of the subject to whom the formulation is being administered; the route of administration; and The CD73 inhibitors of the present invention are administered to a subject in an amount that depends, for example, on the goal of administration (e.g., the degree of desired resolution); the age, weight, sex, and health and physical condition of the subject to whom the formulation is being administered; the route of administration; and It can be done. The dosing regimen can also take into account the presence, nature, and extent of adverse effects associated with the administered agent. An effective dose and dosing regimen can be readily determined, for example, from safety and dose escalation trials, in vivo tests (e.g., animal models), and other methods known to those of skill in the art.

[0166] Generally, dosing parameters indicate that the dose is less than the amount that can be irreversibly toxic to the subject (maximum tolerated dose (MTD)) and not less than the amount necessary to produce a measurable effect on the subject. Such amounts are determined, for example, by pharmacokinetic and pharmacodynamic parameters related to ADME, taking into account the route of administration and other factors.

[0167] An effective dose (ED) is the dose or amount of an agent that produces a therapeutic response or desired effect in a certain percentage of the subjects who take it. The "median effective dose" or ED 50 of an agent is the dose or amount of the agent that produces a therapeutic response or desired effect in 50% of the population administered. ED 50 is generally used as a measure of the reasonable expectation of the effect of an agent, but is not necessarily the dose that a physician considers appropriate taking into account all relevant factors. Thus, in some situations, the effective amount may be greater than the calculated ED 50 and in other situations the effective amount may be less than the calculated ED 50 and in still other situations the effective amount may be the same as the calculated ED 50

[0168] Furthermore, an effective amount of the CD73 inhibitor of the present invention can be an amount that produces a desired result in a healthy subject when administered to the subject one or more times. For example, a subject experiencing a particular disorder ​​​​​​​​​​​​​ In this case, the effective dose is 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 a dose that improves by 90% or more, where 1 00% is defined as the diagnostic parameters, metrics, markers, etc. exhibited by normal subjects .

[0169] In certain embodiments, the CD73 inhibitors contemplated by the present invention are administered to a subject one or more times per day (e.g., orally) at a dosage level of about 0.01 mg / kg body weight to about 50 mg / kg body weight per day, or about 1 mg / kg body weight to about 25 mg / kg body weight in order to obtain the desired therapeutic effect.

[0170] For oral administration, the composition can be provided in the form of tablets, capsules, etc. containing from 1.0 to 1000 milligrams of the active ingredient, particularly in the form containing 1.0, 3.0, 5.0, 10.0, 15 .0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0 .0, 250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800 .0, 900.0, and 1000.0 milligrams of the active ingredient. .

[0171] In certain embodiments, the dosage of the desired CD73 inhibitor is included in a "unit dosage form". The phrase "unit dosage form" refers to physically distinct units, each unit containing a predetermined amount of CD7 3 inhibitor, alone or in combination with one or more additional agents, to produce the desired effect. contain in an amount sufficient to. The parameters of the unit dosage form will be understood to depend on the particular agent and the effect to be achieved It will be appreciated that. Kits

[0172] The present invention also contemplates a kit comprising a CD73 inhibitor and a pharmaceutical composition thereof. The kit is generally in the form of a physical structure containing various components as described below, and can be used, for example, in the practice of the above methods.

[0173] The kit can be in the form of a pharmaceutical composition suitable for administration to a subject and can contain one or more of the CD73 inhibitors disclosed herein (e.g., provided in a sterile container). The CD73 inhibitor can be provided in a ready-to-use form (e.g., tablets or capsules) or in a form that requires reconstitution or dilution, for example, before administration (e.g., powder). If the CD73 inhibitor is in a form that needs to be reconstituted or diluted by the user, the kit can also contain a diluent (e.g., sterile water), buffer, pharmaceutically acceptable excipient, etc., packaged with or separately from the CD73 inhibitor. When combination therapy is contemplated, the kit can contain several agents separately or already combined in the kit. Each component of the kit can be enclosed in a separate container, and all of the various containers can be within a single package cage. The kit of the present invention can be designed for the conditions (e.g., refrigeration or freezing) necessary to properly maintain the components contained therein. The kit can contain several agents separately or already combined in the kit. Each component of the kit can be enclosed in a separate container, and all of the various containers can be within a single package cage. The kit of the present invention can be designed for the conditions (e.g., refrigeration or freezing) necessary to properly maintain the components contained therein.

[0174] The kit includes a label or insert containing identification information of the components therein, and instructions for their use ( e.g., administration parameters of the active ingredient, clinical pharmacology, e.g., pharmacokinetics and pharmacodynamics, adverse effects It can include usage, taboos, etc. Labels or inserts can include manufacturing source information such as lot numbers and expiration dates. The label or insert can be incorporated into the physical structure that houses the components, separately housed within the physical structure, or fixed to the components of the kit (such as ampoules, tubes, or vials). The label or insert can be incorporated into the physical structure that houses the components, separately housed within the physical structure, or fixed to the components of the kit (such as ampoules, tubes, or vials). The label or insert can be incorporated into the physical structure that houses the components, separately housed within the physical structure, or fixed to the components of the kit (such as ampoules, tubes, or vials).

[0175] The label or insert can further include computer-readable media such as disks (e.g., hard disks, cards, memory disks), optical disks such as CD-ROMs or DVD-ROM / RAMs, DVDs, MP3s, next-generation tapes, or electrical storage media such as RAM and ROM, or hybrids thereof, such as magnetic / optical storage media, flash media, or memory-type cards. In some embodiments, although the actual instructions do not exist in the kit, means are provided for obtaining instructions from a remote source via, for example, the Internet. CD-ROM or DVD-ROM / RAM and other optical discs, DVD, MP3, next-generation tape, or electrical storage media such as RAM and ROM, or hybrids thereof, such as magnetic / optical storage media, flash media, or memory-type cards. CD-ROM or DVD-ROM / RAM and other optical discs, DVD, MP3, next-generation tape, or electrical storage media such as RAM and ROM, or hybrids thereof, such as magnetic / optical storage media, flash media, or memory-type cards. CD-ROM or DVD-ROM / RAM and other optical discs, DVD, MP3, next-generation tape, or electrical storage media such as RAM and ROM, or hybrids thereof, such as magnetic / optical storage media, flash media, or memory-type cards. CD-ROM or DVD-ROM / RAM and other optical discs, DVD, MP3, next-generation tape, or electrical storage media such as RAM and ROM, or hybrids thereof, such as magnetic / optical storage media, flash media, or memory-type cards. In some embodiments, although the actual instructions do not exist in the kit, means are provided for obtaining instructions from a remote source via, for example, the Internet. CD-ROM or DVD-ROM / RAM and other optical discs, DVD, MP3, next-generation tape, or electrical storage media such as RAM and ROM, or hybrids thereof, such as magnetic / optical storage media, flash media, or memory-type cards. In some embodiments, although the actual instructions do not exist in the kit, means are provided for obtaining instructions from a remote source via, for example, the Internet.

Examples

[0176] Examples The following examples are presented to provide a complete disclosure and description to those skilled in the art of how to make and use the present invention, and are not intended to limit the scope of what the inventor regards as the invention, nor are they intended to indicate that the following experiments have been performed or that all of the experiments that could be performed are. The exemplary descriptions presented in the present tense are not necessarily those that have been performed, and it should be understood that the descriptions can be made to generate data of the nature described. Efforts have been made to maintain accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.). The following examples are presented to provide a complete disclosure and description to those skilled in the art of how to make and use the present invention, and are not intended to limit the scope of what the inventor regards as the invention, nor are they intended to indicate that the following experiments have been performed or that all of the experiments that could be performed are. The exemplary descriptions presented in the present tense are not necessarily those that have been performed, and it should be understood that the descriptions can be made to generate data of the nature described. Efforts have been made to maintain accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.). The following examples are presented to provide a complete disclosure and description to those skilled in the art of how to make and use the present invention, and are not intended to limit the scope of what the inventor regards as the invention, nor are they intended to indicate that the following experiments have been performed or that all of the experiments that could be performed are. The exemplary descriptions presented in the present tense are not necessarily those that have been performed, and it should be understood that the descriptions can be made to generate data of the nature described. Efforts have been made to maintain accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.). The following examples are presented to provide a complete disclosure and description to those skilled in the art of how to make and use the present invention, and are not intended to limit the scope of what the inventor regards as the invention, nor are they intended to indicate that the following experiments have been performed or that all of the experiments that could be performed are. The exemplary descriptions presented in the present tense are not necessarily those that have been performed, and it should be understood that the descriptions can be made to generate data of the nature described. Efforts have been made to maintain accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.). The following examples are presented to provide a complete disclosure and description to those skilled in the art of how to make and use the present invention, and are not intended to limit the scope of what the inventor regards as the invention, nor are they intended to indicate that the following experiments have been performed or that all of the experiments that could be performed are. The exemplary descriptions presented in the present tense are not necessarily those that have been performed, and it should be understood that the descriptions can be made to generate data of the nature described. Efforts have been made to maintain accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.). The following examples are presented to provide a complete disclosure and description to those skilled in the art of how to make and use the present invention, and are not intended to limit the scope of what the inventor regards as the invention, nor are they intended to indicate that the following experiments have been performed or that all of the experiments that could be performed are. The exemplary descriptions presented in the present tense are not necessarily those that have been performed, and it should be understood that the descriptions can be made to generate data of the nature described. Efforts have been made to maintain accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.). The following examples are presented to provide a complete disclosure and description to those skilled in the art of how to make and use the present invention, and are not intended to limit the scope of what the inventor regards as the invention, nor are they intended to indicate that the following experiments have been performed or that all of the experiments that could be performed are. The exemplary descriptions presented in the present tense are not necessarily those that have been performed, and it should be understood that the descriptions can be made to generate data of the nature described. Efforts have been made to maintain accuracy with respect to the numbers used (e.g., amounts, temperatures, etc.). Some experimental errors and deviations should be taken into account.

[0177] Unless otherwise specified, parts are parts by weight, molecular weight is weight-average molecular weight, temperature is in degrees Celsius (°C), and pressure is atmospheric pressure or approximately atmospheric pressure. The following standard abbreviations are used : wt = wild type; bp = base pair; kb = kilobase. nt = nucleotide; aa = amino acid; s or sec = second; min = minute; h or hr = hour; ng = nanogram; μg = microgram; mg = milligram; g = gram; kg = kilogram; dl or dL = de ciliter; μl or μL = microliter; ml or mL = milliliter; l or L = liter; μM = micromolar concentration; mM = millimolar concentration; M = molar concentration; kDa = ki lodalton; i.m. = intramuscularly; i.p. = intraperitoneally; SC or SQ = subcutaneously ( ); QD = daily; BID = twice a day; QW = weekly; QM = monthly; HPLC = high-performance liquid chro matography; BW = body weight; U = unit; ns = not statistically significant; PBS = phos phate-buffered saline; IHC = immunohistochemistry; DMEM = Dulbecco's modified Eagle's medium ; EDTA = ethylenediaminetetraacetic acid.

[0178] LC: Agilent 1100 series; mass spectrometer: Agilent G6120BA, single quadrupole type; L C-MS method: Agilent Zorbax Eclipse Plus C18, 4.6 × 100 mm, 3.5 μM, 3 5 °C, flow rate of 1.5 mL / min, gradient of 0% - 100% B for 2.5 minutes, washing with 100% B for 0 .5 minutes; A = 0.1% formic acid / 5% acetonitrile / 94.9% water; B = formic acid 0. 1% / water 5% / acetonitrile 94.9%. Flash column: ISCO Rf + Reverse phase HPLC: ISCO-EZ; Column: Kinetex 5μm EVO C18 100A; 250×21.2 mm (Phenomenex) [Example 1] [({[(2R,3S,4R,5R)-5-[6-(cyclopentylamino)-2- [N-(3,4-dihydroxyoxolan-2-yl)-9H-purin-9-yl]methacrylate Synthesis of bis(hydroxy)phosphorylmethyl)phosphonic acid [ka]

[0179] Step a: 2,6-dichloropurine riboside (321 mg, 1 mmol), cyclopentylamine (103 μL, 1.05 mmol, 1.05 equiv.) A mixture of 6 and triethylamine (146 μL, 1.05 mmol, 1.05 equiv.) was added to 6 The mixture was stirred at 0° C. overnight. The solvent was removed from the reaction mixture, and the crude product was used in the next step without purification. It was used in about. 15 H 21 ESI MS of ClNO4 [M+H] + , calculated value 370.8, measured value 370.2.

[0180] Step b: The product from step a (370 mg, 1 mmol) was dissolved in trimethyl phosphate (5 mL ), cooled to 0° C. (ice bath), and then dissolved in methylene bis(trimethyl phosphate) (2 mL) Add dropwise a cold solution of phosphonic acid dichloride (1.25 g, 5 mmol, 5 equiv.) The reaction mixture was stirred at 0° C. for 3 h and then diluted with 0.5 M triethylammonium bicarbonate solution. (7 mL) and stirred at 0° C. for 15 min and then at room temperature for 2 h. The combined solution was purified by reverse-phase HPLC (C18 column, 0 - 30% gradient of acetonitrile and water containing 0.1% TF A), and the product was obtained as a white solid in 28% yield (181 mg ): 1 1H NMR (400 MHz, DMSO) δ 8.45 - 8.32 (m, 2H), 5.85 (d, J = 5.5 Hz, 1H), 4.5 5 - 4.36 (m, 2H), 4.23 - 4.07 (m, 4H), 2.26(t, J = 20.5 Hz, 2H), 2.04 - 1.85 (m , 2H), 1.77 - 1.46 (m, 6H). 13C 16 1H 25 ESIMS of C14H18ClN5O9P2 [M+H] + , calculated value 528.8, measured value 528.1. [Example 2] ((((2R,3S,4R,5R)-5-(6-((4-(tert-Butyl)ben zyl)amino)-2-chloro-9H-purin-9-yl)(3,4-dihydroxytetra hydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid Synthesis [Chemical formula]

[0181] In the same manner as in Example 1, using 4-tert-butylbenzylamine instead of cyclopentylamine, the title compound was synthesized: 1H NMR (400 MHz, DMSO-d6) δ 8.91 (t 1 , J = 6.3 Hz, 1H), 8.43 (s, 1H), 7.33 (d, J = 8.2 Hz, 2H), 7.26 (d, J = 8.2 Hz, , 2H), 2H), 5.86 (d, J = 5.8 Hz, 1H), 4.68 - 4.56(m, 2H), 4.52 (t, J = 5.4 Hz, 1H), 4. 23 - 4.03 (m, 4H), 2.26 (t, J = 20.5 Hz,2H), 1.25 (s, 9H). C 22 H 31 ESI of C MS [M+H] + , calculated value 606.1, measured value 606.2. [Example 3] ((((2R,3S,4R,5R)-5-(2-chloro-6-(isopropylamino )-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl) methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0182] In the same manner as in Example 1, instead of cyclopentylamine, isopropylbenzylamine was used to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.40 (s, 1H), 8 .23 (d, J = 8.1 Hz, 1H), 5.85 (d, J = 5.9Hz, 1H), 4.51 (t, J = 5.5 Hz, 1H), 4.3 6 (s, 1H), 4.24 - 4.03 (m, 4H), 2.25 (t, J= 20.5 Hz, 2H), 1.21 (dd, J = 6.6, 2. 0 Hz, 5H). C 14 H 22 ESIMS of C + , calculated value 502.1, measured value 502.1. [Example 4] (((2R,3S,4R,5R)-5-(2-chloro-6-(cyclopropylamino )-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl )(methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical formula

[0183] Using the same method as in Example 1, cyclopropylamine was used instead of cyclopentylamine to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.42 (s , 1H), 5.86 (d, J = 5.8 Hz, 1H), 4.52 (t, J = 5.4 Hz, 1H), 4.28 - 4.03 (m, 4H), 2.97 (s, 1H), 2.25 (t, J = 20.5 Hz, 2H), 0.75 (s, 2H), 0.64 (s, 3H). C 14 H 20 ClN5 O9P2 ESI MS [M+H] + , calculated value 500.1, measured value 500.1. [Example 5] (((2R,3S,4R,5R)-5-(2-chloro-6-(neopentylamino )-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl) (methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical formula

[0184] Using the same method as in Example 1, neopentylamine was used instead of cyclopentylamine to synthesize the title compound: 11H NMR (400 MHz, DMSO-d6) δ 8.42 (s, 1H), 8.32 (t, J = 6.4 Hz, 1H), 5.85 (d, J = 5.7 Hz, 1H),4.52 (t, J = 5.4 Hz, 1H), 4.31 - 4.04 (m, 4H), 3.82 (d, J = 7.0 Hz, 1H), 3.42 -3.17 (m, 2H), 2.26 (t, J = 20.5 Hz, 2 H), 0.91 (s, 9H). C 16 H 26 ESI MS of C10H14ClN5O9P2 [M+H] + , calculated value 530.1, measured value 530.2. [Example 6] Synthesis of (((((2R,3S,4R,5R)-5-(2-chloro-6-(isopropyl(methyl amino)-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran- 2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid [Chemical formula]

[0185] The title compound was synthesized in the same manner as in Example 1, using N-methylisopropylamine instead of cyclopentylamine: 1H NMR (400 MHz, DMSO-d6) δ 8.42 (s, 1H), 5 1 .88 (d, J = 5.9 Hz, 1H), 4.50 (t, J = 5.4Hz, 1H), 4.22 - 4.17 (m, 1H), 4.11 (d, J = 6.4 Hz, 3H), 3.03 (s, 3H), 2.26 (t, J= 20.5 Hz, 2H), 1.23 (s, 6H). C 1H NMR of C10H14ClN5O9P2 15 H 24 ESI MS of C10H14ClN5O9P2 [M+H] +, Calculated value: 516.1, Measured value: 516.1. [Example 7] ((((2R,3S,4R,5R)-5-(6-((3,5-Bis(trifluoromethyl)benzyl)amino)-9H-purin-9-yl)-3,4-dihydroxytetrahydro furan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis Synthesis

Chemical Structure

[0186] Using the same method as in Example 1, except that 6-chloropurine riboside and 3,5-bis(trifluoromethyl)benzylamine were used in step a, the title compound was synthesized: 1 H NMR (400 MHz, DMSO-d6) δ 8.67 (s,1H), 8.47 (s, 1H), 8.26 (s, 1H), 8.07 (s, 2H), 7 .99 (s, 1H), 5.94 (d, J = 5.7 Hz, 1H), 4.88(s, 2H), 4.61 (t, J = 5.4 Hz, 1H), 4 .23 (t, J = 4.2 Hz, 1H), 4.20 - 4.04 (m,3H), 2.25 (t, J = 20.5 Hz, 2H). C 20 H 20 ESIMS of F6N5O9P2 [M-H] - , Calculated value: 650.1, Measured value: 650.2. [Example 8] ((((2R,3S,4R,5R)-5-(6-((4-Bromobenzyl)amino) -9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)meth oxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical Structure

[0187] In the same manner as in Example 1, except that in step a, 6-chloropurine riboside and the corresponding amine were used to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.60 (s, 1H ), 8.44 (s, 1H), 8.24 (s, 1H), 7.49 (d, J = 8.3 Hz, 2H), 7.29 (d, J = 8.3 Hz, 2H ), 5.94 (d, J = 5.7 Hz, 1H), 4.67 (s, 2H), 4.61 (t, J = 5.3 Hz, 1H), 4.23 (t, J = 4.2 Hz, 1H), 4.19 - 4.05 (m, 3H), 2.25 (t, J = 20.5 Hz, 2H). C 18 H 21 BrN5O9P2 The ESI MS [M-H] - , calculated value 592.0, measured value 592.1. [Example 9] ((((2R,3S,4R,5R)-5-(6-((4-(tert-Butyl)ben zyl)amino)-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran -2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid Synthesis [Chemical formula]

[0188] In the same manner as in Example 1, except that in step a, 6-chloropurine riboside and the corresponding amine were used to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.62 (s, 1H ), 8.45 (s, 1H), 8.26 (s, 1H), 7.37 - 7.22(m, 4H), 5.94 (d, J = 5.7 Hz, 1H), 4. 67 (s, 2H), 4.60 (t, J = 5.4 Hz, 1H), 4.23(t, J = 4.1 Hz, 1H), 4.20 - 4.05 (m, 3H), 2.25 (t, J = 20.5 Hz, 2H), 1.24 (s,9H). C 22 H 30 ESIMS of C5H9N5O9P2 [M-H] - , calculated value 570.1, measured value 570.3. [Example 10] ((((2R,3S,4R,5R)-5-(6-(([1,1'-Biphenyl]-4 -ylmethyl)amino)-9H-purin-9-yl)-3,4-dihydroxytetrahydro furan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0189] Using the same method as in Example 1, but using 6-chloropurine riboside and the corresponding amine in step a, the title compound was synthesized: H NMR (400 MHz, DMSO-d6) δ 8.67 (s, 1H) , 8.46 (s, 1H), 8.27 (s, 1H), 7.66 - 7.57(m, 4H), 7.49 - 7.40 (m, 4H), 7.37 - 7 .30 (m, 1H), 5.95 (d, J = 5.7 Hz, 1H), 4.76(s, 2H), 4.61 (t, J = 5.3 Hz, 1H), 4 .24 (t, J = 4.1 Hz, 1H), 4.20 - 4.06 (m,3H), 2.25 (t, J = 20.5 Hz, 2H). C 24 ​H 26 ESIMS of N5O9P2 [M-H] - , calculated value 590.1, measured value 590.2. [Example 11] (((((2R,3S,4R,5R)-3,4-Dihydroxy-5-(6-((4-( (trifluoromethyl)benzyl)amino)-9H-purin-9-yl)tetrahydrofura n-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical Structure]

[0190] In the same manner as in Example 1, except that in step a, 6-chloropurine riboside and the corresponding amine were used to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.68 (s, 1H ), 8.46 (s, 1H), 8.25 (s, 1H), 7.67 (d, J = 8.1 Hz, 2H), 7.54 (d, J = 8.1 Hz, 2H ), 5.95 (d, J = 5.8 Hz, 1H), 4.79 (s, 2H), 4.61 (t, J = 5.3 Hz, 1H), 4.24 (t, J = 4.1 Hz, 1H), 4.20 - 4.06 (m, 3H), 2.25 (t, J = 20.5 Hz, 2H). C 19 H 21 F3N5O9P2 ESI MS of [M-H] - , calculated value 582.1, measured value 582.2. [Example 12] (((((2R,3S,4R,5R)-3,4-Dihydroxy-5-(6-((4-me (thylbenzyl)amino)-9H-purin-9-yl)tetrahydrofuran-2-yl)me Synthesis of (Toxy)(Hydroxy)Phosphoryl)(Methyl)Phosphonic Acid [Chemical formula]

[0191] In the same manner as in Example 1, except that in step a, 6-chloropurine riboside and the corresponding amine were used to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.61 (s, 1H ), 8.45 (s, 1H), 8.26 (s, 1H), 7.22 (d, J = 7.8 Hz, 2H), 7.10 (d, J = 7.8 Hz, 2H ), 5.94 (d, J = 5.7 Hz, 1H), 4.67 (s, 2H), 4.60 (t, J = 5.4 Hz, 1H), 4.23 (t, J = 4.2 Hz, 1H), 4.19 - 4.04 (m, 3H), 2.31 - 2.18 (m, 5H). C 19 H 24 ESI MS of C [M-H] - , calculated value 528.1, measured value 528.2. [Example 13] ((((2R,3S,4R,5R)-5-(6-((3,5-Dichlorobenzyl)a mino)-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl methoxy)(Hydroxy)Phosphoryl)(Methyl)Phosphonic Acid Synthesis [Chemical formula]

[0192] In the same manner as in Example 1, except that in step a, 6-chloropurine riboside and the corresponding amine were used to synthesize the title compound: 11H NMR (400 MHz, DMSO-d6) δ 8.61 (s, 1H ), 8.46 (s, 1H), 8.26 (s, 1H), 7.48 (t, J = 2.0 Hz, 1H), 7.39 (s, 2H), 5.95 (d, J = 5.7 Hz, 1H), 4.70 (s, 2H), 4.61 (t, J = 5.4 Hz, 1H), 4.24 (t, J = 4.2 Hz, 1H ), 4.20 - 4.05 (m, 3H), 2.26 (t, J = 20.5 Hz, 2H). C 18 H 20 ESI MS of Cl2N5O9P2 [M - H] - , calculated value 582.1, measured value 582.2. [Example 14] Synthesis of (((2R,3S,4R,5R)-5-(6-(benzylamino)-2-methyl- 9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid Step a: To a nitrogen-purged reaction mixture of the iodo derivative (1.03 g, 1.7 mmol) and trimethyltin (470 μL, 3.34 mmol) in NMP (10 mL), Pd(

Chemical Structure

[0193] PPh3)4 (196 mg, 0.17 mmol, 10 mol%) was added, and the reaction mixture was heated at 120 ℃ overnight. LCMS indicated the formation of the product. This was cooled to room temperature, diluted with water and extracted with ethyl acetate, dried (MgSO4), filtered, and concentrated. The residue was purified by flash column to obtain the product (1 g). ESI MS of C H N5O7 [M + H] 24 H 27 + ​, Calculated value 498. 2, Measured value 498.3.

[0194] Step b: To a solution of the acetate derivative (1 g, 2.01 mmol) from Step a in methanol (5 mL) was added K2CO3 (276 mg, 2 mmol), and the reaction mixture was stirred at room temperature for 1 hour. Next, this was diluted with dichloromethane and filtered through a pad of silica. The filtrate was concentrated and purified by flash column (ISCO, 40 g column, 0 - 20 % methanol in dichloromethane, 20 minutes) to give the compound as an off - white solid (45 0 mg, 60%). C H 18 H 21 ESI MS of C + HN5O4 [M + H]

[0195] Step c: The product from Step b (150 mg, 0.4 mmol) was dissolved in trimethyl phosphate (3 mL), cooled to 0 °C (ice bath), and then an ice - cold solution of methylenebis(phosphonic acid dichloride) (504 mg, 2 mmol, 5 equiv) in trimethyl phosphate (1 mL) was added dropwise. The reaction mixture was stirred at 0 °C for 3 hours, then carefully quenched with 0.5 M triethylammonium bicarbonate solution (8 mL) and stirred at 0 °C for 15 minutes and then at room temperature for 2 hours . The reaction mixture was purified by reverse - phase HPLC (C18 column, gradient of 0 - 30% acetonitrile and water containing 0.1% TFA) to give the product as a white solid: 1H NMR (40 1 0 MHz, DMSO - d6) δ 8.48 - 8.32(m, 2H), 7.38 - 7.18 (m, 5H), 5.92 (d, J = 6.0 Hz ​, 1H), 4.71 (s, 2H), 4.55 (t, J = 5.5 Hz, 1H), 4.19 - 3.98 (m, 4H), 2.44 (s, 3H) , 2.23 (t, J = 20.5 Hz, 2H). C 19 H 25 ESI MS of C5H9N5O9P2 [M-H] - , calculated value 528.1, measured value 528.2 [Example 15] ((((2R,3S,4R,5R)-5-(6-(Benzylamino)-2-vinyl- 9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical Structure

[0196] Step a: A mixture of N-benzyl-2-chloropurine riboside (783 mg, 2 mmol), vinylboronic acid pinacol ester (462 mg, 3 mmol, 1.5 equiv), K2CO3 (828 mg, 6 mmol, 3 equiv), and Pd(PPh3)4 in 1,2-dimethoxyethane:H2O (9:1, 10 mL) was stirred under N2 at 85 °C for 1 day. The reaction mixture was cooled to room temperature, diluted with EtOAc (100 mL), and washed with H2O (50 mL). The organic layer was separated, dried over MgSO4, filtered, and the solvent was evaporated to give a yellow solid. The crude product was washed with MTBE (50 mL) and used for the next step (550 mg, 72%). 6 -benzyl- 2-chloropurine riboside (783 mg, 2 mmol), vinylboronic acid pinacol ester (462 mg, 3 mmol, 1.5 equiv), K2CO3 (828 mg, 6 mmol, 3 equiv), and Pd(PPh3)4 in 1,2-dimethoxyethane:H2O (9:1, 10 mL) was stirred under N2 at 85 °C for 1 day. The reaction mixture was cooled to room temperature, diluted with EtOAc (100 mL), and washed with H2O (50 mL). The organic layer was separated, dried over MgSO4, filtered, and the solvent was evaporated to give a yellow solid. The crude product was washed with MTBE (50 mL) and used for the next step (550 mg, 72%). The organic layer was separated, dried over MgSO4, filtered, and the solvent was evaporated to give a yellow solid. The crude product was washed with MTBE (50 mL) and used for the next step (550 mg, 72%). The crude product was washed with MTBE (50 mL) and used for the next step (550 mg, 72%).

[0197] Step b: The title compound was synthesized in the same manner as in Example 1. 1 H NMR (400 MHz, DMSO-d6) ​δ 8.39 (s, 1H), 7.38 (d, J = 7.0 Hz, 2H), 7.29 (t, J = 7.6 Hz, 2H), 7.25 - 7.15 (m, 1H), 6.64 (dd, J = 17.2, 10.4 Hz, 1H), 6.39 (dd, J = 17.2, 2.4 Hz, 1H), 5.9 4 (d, J = 6.0 Hz, 1H), 5.55 (d, J = 10.5 Hz, 1H), 4.73 (s, 2H), 4.63 (t, J = 5.5 Hz, 1H), 4.28 - 4.00 (m, 4H), 2.25 (t, J = 20.4 Hz, 2H). C 20 H 26 ESI MS of C H + N5O9P2 [M+H], calculated 542.1, found 542.2.

[0198] Step c: The product from step b (40 mg, 0.06 mmol) was dissolved in MeOH (10 mL) and purged with N2, then 10% Pd / C (50% wet, 30 mg) was added. The reaction mixture was stirred vigorously for 2 h under H2 (balloon), filtered, and the product was purified by RP18 HPLC (H2O + 0.1% TFA / acetonitrile + 0.1% TFA) to give a white solid (14 mg, 35%): 1H NMR (400 MHz, DMSO-d6) δ 8.52 - 8.18 (m, 2H) 1 H NMR (400 MHz, DMSO-d6) δ 8.52 - 8.18 (m, 2H) , 7.33 - 7.27 (m, 2H), 7.27 - 7.18 (m, 2H), 7.15 (t, J = 7.2 Hz, 1H), 5.86 (d, J = 6.0 Hz, 1H), 4.64 (s, 2H), 4.55 (t, J = 5.5 Hz, 1H), 4.19 - 3.98 (m, 4H), 2.7 0 - 2.61 (m, 2H), 2.16 (t, J = 20.5 Hz, 2H), 1.16 (t, J = 7.6 Hz, 3H). C 20 H 27 N ESI MS of 5O9P2 [M+H] + , calculated value 544.1, measured value 544.2. [Example 16] ((((2R,3S,4R,5R)-5-(2-Allyl-6-(benzylamino)- 9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)metho xy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0199] The title compound was synthesized in the same manner as in Example 15. 1 H NMR (400 MHz, DMSO-d6) δ 8.4 6 (s, 1H), 8.37 (s, 1H), 7.41 - 7.34 (m, 2H), 7.30 (t, J = 7.5 Hz, 2H), 7.25 - 7 .18 (m, 1H), 6.17 - 6.03 (m, 1H), 5.92 (d, J = 6.0 Hz, 1H), 5.28 - 5.00 (m, 2H), 4.70 (s, 2H), 4.60 (t, J = 5.6 Hz, 1H), 4.27 - 4.02 (m, 4H), 3.49 (d, J = 6.8 H z, 2H), 2.24 (t, J = 20.5 Hz, 2H). ESI MS of [M+H] + , calcd C 21 H 28 ESI MS of C N5O9P2 + , calculated value 556.1, measured value 556.3. [Example 17] (((2R,3S,4R,5R)-5-(6-(benzylamino)-2-propyl -9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)meth oxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chem.

[0200] The title compound was synthesized in the same manner as in Example 15: 1 H NMR (400 MHz, DMSO-d6) δ 8.4 0 (s, 2H), 8.29 (s, 1H), 7.29 (d, J = 7.6Hz, 2H), 7.23 (t, J = 7.5 Hz, 2H), 7.1 5 (t, J = 7.2 Hz, 1H), 5.86 (d, J = 6.0 Hz,1H), 4.64 (s, 2H), 4.54 (t, J = 5.5 Hz, 1H), 4.19 - 3.94 (m, 4H), 2.71 - 2.55(m, 2H), 2.17 (t, J = 20.5 Hz, 2H), 1. 66 (q, J = 7.4 Hz, 2H), 0.93 - 0.70 (m,3H). C 21 H 30 ESIMS of C + H N5O9P2 [M+H] , calculated value 558.1, measured value 558.2. [Example 18] [({[(2R,3S,4R,5R)-5-[6-(benzylamino)-2-methoxy -9H-purin-9-yl]-3,4-dihydroxyoxolane-2-yl]methoxy} (hydroxy)phosphoryl)methyl]phosphonic acid synthesis

Chem.

[0201] Step a: A known riboside (250 mg, 0.64 mmol) was dissolved in 25% NaOMe in MeOH solution (2 mL) and stirred at 60 °C overnight. The reaction mixture was concentrated under reduced pressure, and then the residue was diluted with H2O (15 mL) and acetic acid until neutral pH. It was filtered to collect the product (white solid, 180 mg, 73%). C H N5O5 ESI MS [M+H] 18 H 22 , calculated + value 388.4, found 388.1.

[0202] Step b: Using the same method as in Example 1, the title compound was obtained as a white solid (37 mg, 14 %): 1 H NMR (400 MHz, DMSO) δ 8.48 (s, 1H), 8.20 (s, 1H), 7.37 - 7.1 7 (m, 5H), 5.82 (d, J = 5.9 Hz, 1H), 4.64(d, J = 5.0 Hz, 3H), 4.28 - 4.00 (m, 4 H), 3.80 (s, 3H), 2.23 (t, J = 20.5 Hz,2H). C 19 H 26 N5O 10 P2 ESIMS [M+H] + , calculated value 546.4, found 546.1. [Example 19] [({[(2R,3S,4R,5R)-5-[6-(benzylamino)-2-(methyl amino)-9H-purin-9-yl]-3,4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid synthesis [Chemical formula]

[0203] ​​Project a: A known riboside (250 mg, 0.64 mmol) was dissolved in 40% MeNH2 in an H2O solution (2 mL) and stirred at 60 °C overnight. Next, the reaction mixture was concentrated under reduced pressure , and the residue was diluted with H2O (15 mL). The product was collected by filtration (white solid, 21 0 mg, 85%). C H 18 H 23 N6O4 ESI MS [M+H] + , calculated value 387.4, measured value 387.3.

[0204] Project b: Using the same method as in Example 1, the title compound was obtained as a white solid (38 mg, 15 %): 1 H NMR (400 MHz, DMSO) δ 8.08 (s, 1H), 7.42 - 7.19 (m, 5H), 5.7 9 (d, J = 6.1 Hz, 1H), 4.75 - 4.45 (m, 3H), 4.24 - 4.02 (m, 4H), 2.81 (s, 3H), 2 .22 (t, J = 20.4 Hz, 2H). C 19 H 26 N6O9P2 ESI MS [M-H] - , calculated value 543.4, measured value 54 3.2. [Example 20] ((((2R,3S,4R,5R)-5-(6-(benzylamino)-2-(dimethyl lamino)-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2 -yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical Structure

[0205] Using the same method as in Example 19, but using dimethylamine in Project a, the title compound was synthesized: 1 ​HNMR (400 MHz, DMSO-d6) δ 8.15 (s, 1H), 8.09 (s, 1H), 7.36 (d, J = 7. 2 Hz, 2H), 7.29 (t, J = 7.5 Hz, 2H), 7.21(t, J = 7.2 Hz, 1H), 5.81 (d, J = 5.5 Hz, 1H), 4.68 - 4.57 (m, 3H), 4.26 - 4.20(m, 1H), 4.20 - 4.00 (m, 3H), 3.06 (s, 6H), 2.24 (t, J = 20.4 Hz, 2H). C 20 H 29 ESI MS of C + H 1 N6O9P2 [M+H] + , calculated value 559.1, measured value 559.2. [Example 21] Synthesis of ((2R,3S,4R,5R)-5-(6-(benzylamino)-2-(pyrrol idin-1-yl)-9H-purin-9-yl)-3,4-dihydroxytetrahydrofura n-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid [Chemical Structure Diagram]

[0206] The title compound was synthesized in the same manner as in Example 19, except that pyrrolidine was used in step a: Synthesis: 1 HNMR (400 MHz, DMSO-d6) δ 8.15 (s, 2H), 7.42 - 7.14 (m, 5H), 5.82 ( d, J = 5.5 Hz, 1H), 4.71 - 4.51 (m, 3H),4.26 (t, J = 4.3 Hz, 1H), 4.21 - 4.00 ( m, 3H), 3.46 (s, 4H), 2.23 (t, J = 20.4 Hz,2H), 1.89 (s, 4H). C 22 H31 of N6O9P2 ESI MS [M+H] + , calculated value 585.1, measured value 585.2. [Example 22] (((((2R,3S,4R,5R)-5-(6-(benzylamino)-2-(piperi din-1-yl)-9H-purin-9-yl)-3,4-dihydroxytetrahydrofura n-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical Structure Diagram]

[0207] Using the same method as in Example 19, but using piperidine in step a, the title compound was synthesized: 1 HNMR (400 MHz, DMSO-d6) δ 8.20 (s, 1H), 8.10 (s, 1H), 7.38 - 7.33 ( m, 2H), 7.33 - 7.25 (m, 2H), 7.25 - 7.16(m, 1H), 5.81 (d, J = 5.6 Hz, 1H), 4.66 - 4.52 (m, 3H), 4.20 (t, J = 4.3 Hz, 1H),4.17 - 4.00 (m, 3H), 3.74 - 3.62 (m, 4H), 2.24 (t, J = 20.5 Hz, 2H), 1.64 - 1.38(m, 6H). C 23 H 31 ESIMS of N6O9P2 [M-H - , calculated value 597.2, measured value 597.3. [Example 23] (((((2R,3S,4R,5R)-5-(6-(benzylamino)-2-morpho no-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl) ​Synthesis of (Methoxy)(Hydroxy)Phosphoryl)(Methyl)Phosphonic Acid

Chem.

[0208] The title compound was synthesized in the same manner as in Example 19, except that morpholine was used in Step a: Synthesized: 1 HNMR (400 MHz, DMSO-d6) δ 8.24 - 8.02 (m, 2H), 7.37 - 7.17 (m, 5H), 5.79 (d, J = 5.9 Hz, 1H), 4.72 - 4.51 (m,3H), 4.23 - 3.99 (m, 4H), 3.61 (s, 8H ), 2.23 (t, J = 20.5 Hz, 2H). C 22 H 29 N6O 10 ESI MS of P2 [M-H] - , calculated value 599.2, measured value 599.3 [Example 24] ((((2R,3S,4R,5R)-5-(6-(Benzylamino)-2-(isop ropylthio)-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran- 2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic Acid Synthesis

Chem.

[0209] Step a: A solution of Compound X (5 g, 10.2 mmol) in MeOH (60 ml) was treated with ammonia gas at -20 °C for 10 minutes. The mixture was then warmed to room temperature and stirred until the reaction was complete. Next, nitrogen was bubbled through the reaction solution to remove excess ammonia gas. The mixture was concentrated and purified by preparative HPLC to give the desired product (750 mg, 20%) ​​ Obtained.

[0210] Step b: The product from step a (0.36 g, 1 mmol) in anhydrous EtOH (3.3 mL), benzylamine (0.115 mL, 1.05 mmol, 1.05 eq), and Et3N (0.15 mL, 1.1 mmol, 1.1 eq) were stirred at 70 °C for 4 h. Next, the reaction mixture was cooled to room temperature, concentrated, and used without further purification.

[0211] Step c: The product from step b was dissolved in trimethyl phosphate (5 mL), cooled to 0 °C (ice bath), and then a cold solution of methylenebis(phosphonic dichloride) (1.2 g, 15 mmol, 5 eq) in trimethyl phosphate (3 mL) was added dropwise. The reaction mixture was stirred at 0 °C for 3 h, then carefully quenched with 0.5 M triethylammonium bicarbonate solution (6 mL), stirred at 0 °C for 15 min, and then at room temperature for 2 h. The reaction mixture was purified by reverse-phase HPLC (C18 column, 0 - 40% gradient of acetonitrile and water containing 0.1% TFA), and the product was obtained as a white solid in 6% yield (38 mg): 1H NMR (400 MHz, DMSO-d6 1 ) δ 8.53 (s, 1H), 8.27 (s, 1H), 7.37 - 7.17(m, 5H), 5.84 (d, J = 5.8 Hz, 1H), 4.65 (s, 2H), 4.56 (t, J = 5.5 Hz, 1H),4.24 - 4.17 (m, 1H), 4.17 - 4.01 (m, 3H) , 3.82 - 3.71 (m, 1H), 2.24 (t, J = 20.5Hz, 2H), 1.28 (d, J = 6.8 Hz, 6H).. C2 1H 1H 29 ESI MS of C21H1N5O9P2S [M+H] +, Calculated value 590.1, Measured value 590.2 [Example 25] ((((2R,3S,4R,5R)-5-(6-(Benzylamino)-2-(isop ropylsulfonyl)-9H-purin-9-yl)-3,4-dihydroxytetrahydrof ran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0212] Step a: The product from step a of Example 24 (4.5 g, 12.5 mmol) in methylene chloride (50 mL) was treated portionwise with m-CPBA (2.2 g, 38.2 mmol). The reaction solution was stirred at room temperature until the reaction was complete. The mixture was diluted with methylene chloride (200 mL), washed twice with aqueous NaHSO3 solution, dried over Na2SO4, and concentrated. The residue was purified by preparative HPLC to give the desired product as a white solid (780 mg, 16%).

[0213] Step b: The product from step a (0.393 g, 1 mmol) in anhydrous EtOH (3.3 mL), benzylamine (0.115 mL, 1.05 mmol, 1.05 eq), and Et3N (0.15 mL, 1.1 mmol, 1.1 eq) were stirred at 70 °C for 4 h. Next, the reaction mixture was cooled to room temperature, concentrated, and used without further purification.

[0214] Step c: The product from step b was dissolved in trimethyl phosphate (4 mL), cooled to 0 °C (ice bath), and then a cold solution of methylenebis(phosphonic acid dichloride) ( 1.2 g, 5 mmol, 5 eq) in trimethyl phosphate (2 mL) was added dropwise. The reaction mixture was stirred at 0 °C for 3 h. ​Stir and then carefully quench with 0.5 M triethylammonium bicarbonate solution (6 mL). It was stirred at 0 °C for 15 minutes and then at room temperature for 2 hours. The reaction mixture was purified by reverse phase HPLC (C18 column, 0 - 40% gradient of acetonitrile and water containing 0.1% TFA). The product was obtained as a white solid in 22% yield (50 mg): 1 1H NMR (400 MHz, DMSO-d6 ) δ 9.21 (t, J = 6.2 Hz, 1H), 8.66 (s, 1H), 7.42 - 7.15 (m, 5H), 5.97 (d, J = 6 .1 Hz, 1H), 4.74 - 4.66 (m, 2H), 4.60 (dd, J = 6.1, 5.0 Hz, 1H), 4.26 - 4.22 (m, 1H), 4.19 - 4.07 (m, 4H), 3.78 (p, J = 6.8 Hz, 1H), 2.26 (t, J = 20.5 Hz, 2H), 1.12 (dd, J = 6.8, 2.4 Hz, 6H). C 21 H 29 N5O 11 ESI MS of P2S [M+H] + , calculated 622.1, found 622.2. [Example 26] (((2R,3S,4S,5R)-5-(6-Amino-2-fluoro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical Structure

[0215] The title compound was synthesized using the corresponding alcohol in the same manner as in step b of Example 1. 1 ​​​​1H NMR (400 MHz, DMSO-d6) δ 8.17 (s, 1H), 8.02 - 7.72 (m, 2H), 6.15 (d, J = 4.3 Hz, 1H), 4.30 - 4.09 (m, 4H), 4.00 - 3.88 (m, 1H), 2.24 (t, J = 20.5 Hz, 2H). C 11 H 17 ESI MS of C9H5FN5O9P2 [M+H] + , calculated 444.0, found 444.1. [Example 27] ((((2R,3R,4R,5R)-5-(6-Amino-9H-purin-9-yl) -4-fluoro-3-hydroxyoxolan-2-yl)methoxy)(hydroxy )(phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0216] The title compound was synthesized in the same manner as in Step b of Example 1 using the corresponding alcohol: : 1 1H NMR (400 MHz, DMSO-d6) δ 8.45 (s, 1H), 8.26 (s, 1H), 7.92 (s, 2H), 6.2 7 (dd, J = 17.2, 2.8 Hz, 1H), 5.50 (ddd, J = 52.5, 4.5, 2.8 Hz, 1H), 4.64 - 4.52 (m, 1H), 4.29 - 4.08 (m, 3H), 2.25 (t, J = 20.4 Hz, 2H). C 11 H 16 ESI MS of C9H6FN5O8P2 [M+H] + , calculated 428.1, found 428.1. [Example 28] [({[(2R,3R,4S,5R)-5-(6-Amino-2-chloro-9H-purin -9-yl)-4-fluoro-3-hydroxyoxolan-2-yl]methoxy}hydro Synthesis of xy)phosphoryl)methyl]phosphonic acid

Chemical formula

[0217] The title compound was synthesized using a commercially available alcohol in the same manner as in Example 1: 1 H N MR (400 MHz, DMSO-d6) δ 8.28 (d, J = 2.2 Hz, 1H), 7.92 (s, 2H), 6.36 (dd, J = 1 4.3, 4.6 Hz, 1H), 5.26 (dt, J = 52.5, 4.3Hz, 1H), 4.51 (dt, J = 18.6, 4.7 Hz, 1 H), 4.19 (t, J = 6.0 Hz, 2H), 4.04 (t, J =5.0 Hz, 1H), 2.26 (t, J = 20.5 Hz, 2H ); C 11 H 15 MS of ClFN5O8P2: (ES) m / z [M-H] - Calculated value 460.1, Measured value 460.1. [Example 29] ((((2R,3R,4S,5R)-5-(6-(Benzylamino)-2-chloro- 9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical formula

[0218] Step a: 2,6-Dichloropurine (3.6 g, 18.8 mmol) was dissolved in 90 mL of acet onitrile and treated with Cs2CO3 (7.5 g, 23 mmol, 1.2 equiv). The mixture The combined solution was stirred at room temperature for 30 minutes. A known bromo derivative (8.75 g, 21 mmol, 1. 1 eq) was dissolved in 100 mL of acetonitrile and added dropwise to the mixed solution via an addition funnel. The mixed solution was stirred at room temperature overnight. The mixed solution was filtered through a pad of silica gel and concentrated. The residue was adsorbed onto silica and purified by column chromatography (hexane / ethyl acetate) to obtain the product as a white solid in 77% yield (7.72 g). 1 H NMR ( 400 MHz, Chloroform-d) δ 8.39 (d, J = 3.0Hz, 1H), 8.10 (ddt, J = 8.5, 3.1, 0.9 Hz, 4H), 7.74 - 7.36 (m, 6H), 6.64 (dd, J= 21.8, 2.8 Hz, 1H), 5.83 - 5.69 (m, 1H), 5.40 (ddd, J = 49.9, 2.8, 0.8 Hz, 1H),4.89 - 4.77 (m, 2H), 4.62 (q, J = 4. 0 Hz, 1H). C 24 H 17 ESI MS of Cl2FN4O5 [M+H] + , calculated value 531.1, measured value 531.1.

[0219] Step b: The product from step a (9.0 g, 17 mmol ) in anhydrous EtOH (60 mL), benzylamine (3 mL, 26 mmol, 1.5 eq), and Et3N (5 mL, 3 4 mmol, 2.0 eq) were stirred at 70 °C for 4 hours. Next, the reaction mixture was cooled to room temperature, the product was collected by filtration and used without further purification (white solid, 8.9 g, 8 7%). C 31 H 25 ESI MS of ClFN5O5 [M+H] + , calculated value 602.2, measured value 602.0.

[0220] The above product (10.2 g, 17 mmol) and K2CO3 (7 g, 51 mmol, 3 equivalents ) were dissolved in 170 mL of methanol and stirred at room temperature for 4 hours. Next, the reaction mixture was filtered through a pad of silica gel and concentrated. The reaction mixture was purified by column chromatography (methylene chloride / methanol) to obtain the product as a white solid in 80% yield (5. 3 g): 3 g): 1 1H NMR (400 MHz, DMSO-d6) δ 8.97 (t, J = 6.3 Hz, 1H), 8.31 (d, J = 2.0 Hz, 1H), 7.36 - 7.18 (m, 5H), 6.34 (dd, J =13.6, 4.7 Hz, 1H), 5.23 (dt, J = 52. 6, 4.3 Hz, 1H), 4.66 (q, J = 7.3, 5.7 Hz,2H), 4.43 (dt, J = 19.0, 4.8 Hz, 1H), 3.84 (q, J = 4.9 Hz, 1H), 3.65 (tq, J =12.0, 6.2, 5.2 Hz, 2H). ). C 17 H 18 ClFN 5O3 ESI MS [M+H] + , calculated value 394.1, measured value 394.1.

[0221] Step c: The product from step b (800 mg, 2 mmol) was dissolved in trimethyl phosphate (15 m L) and cooled to 0 °C (ice bath). Next, a cold solution of methylene bis (phosphonic acid dichloride) (2.5 g, 10 mmol, 5 equivalents) in trimethyl phosphate (5 mL) was added dropwise. The reaction mixture was stirred at 0 °C for 3 hours, then carefully quenched with 0.5 M triethylammonium bicarbonate solution (15 mL), stirred at 0 °C for 15 minutes, and then at room temperature for 2 hours. The reaction mixture was quenched carefully with 0.5 M triethylammonium bicarbonate solution (15 mL), stirred at 0 °C for 15 minutes, and then at room temperature for 2 hours. The reaction The reaction mixture was purified by reverse-phase HPLC (C18 column, 0 - 40% gradient of acetonitrile and water containing 0.1% TFA), and the product was obtained as a white solid in 22% yield (290 mg): 1 1H NMR (400 MHz, DMSO-d6) δ 8.99 (t, J = 6.3 Hz, 1H), 8.30 (d, J = 2.2 Hz, 1H), 7.40 - 7.18 (m, 5H), 6.38 (dd, J =14.3, 4.6 Hz, 1H), 5.45 - 5.04 (m, 1 H), 4.65 (t, J = 5.5 Hz, 2H), 4.54 - 4.42(m, 1H), 4.19 (t, J = 6.1 Hz, 2H), 4.0 4 (t, J = 5.1 Hz, 1H), 2.26 (t, J = 20.5Hz, 2H). C 18 H 21 ESI MS of C l - F N5O8P2 [M-H , calcd 550.8, found 550.2 [Example 30] [Chemical formula]

[0222] The title compound was synthesized in the same manner as in Example 29 using the product of step a of Example 29 and the corresponding amine : 1 1H NMR (400 MHz, DMSO-d6) as a mixture of rotamers δ ​8.32 (d, J = 2.1 Hz, 1H), 7.40 - 7.19 (m,5H), 6.42 (dd, J = 14.5, 4.6 Hz, 1H), 5.55 (s, 1H), 5.27 (dt, J = 52.4, 4.2 Hz,1H), 4.95 (s, 1H), 4.50 (dt, J = 18.4, 4.5 Hz, 1H), 4.19 (t, J = 6.1 Hz, 2H),4.05 (q, J = 5.0 Hz, 1H), 3.65 (s, 1H), 3.11 (s, 2H), 2.26 (t, J = 20.5 Hz, 2H). C 19 H 23 ESI MS of ClFN5O8P2 [M+H] + , calculated value 566.1, found 566.2. [Example 31a] ((((2R,3R,4S,5R)-5-(2-Chloro-6-(methylamino)-9 H-purin-9-yl)-4-fluoro-3-hydroxyoxolane-2-yl )methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical Structure Diagram]

[0223] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 the title compound was synthesized: 1 H NMR (400 MHz, DMSO-d6) δ 8.36 (q, J = 4.6 Hz, 1H), 8.27 (s, 1H), 6.45 (brs, 2H), 6.37 (dd, J= 14.3, 4.6 Hz, 1H), 5.25 (dt, J = 52 .4, 4.3 Hz, 1H), 4.50 (dt, J = 18.6, 4.6Hz, 1H), 4.19 (t, J = 5.9 Hz, 2H), 4.04 (q, J = 5.2 Hz, 1H), 3.33 (brs, 1H), 2.93(d, J = 4.5 Hz, 3H), 2.26 (t, J = 20. 4 Hz, 2H). C 12 H 17 ESI MS of ClFN5O8P2 [M-H] - , calculated value 474.7, measured value 474.1. [Example 31b] ((((2R,3R,4S,5R)-5-(2-chloro-6-(ethylamino)-9 H-purin-9-yl)-4-fluoro-3-hydroxyoxolane-2-yl )methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0224] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.43 (t, J = 5.7 Hz, 1H), 8.26 (s, 1H), 7.28 (brs, 2H), 6.37 (dd, J= 14.3, 4.6 Hz, 1H), 5.25 (dt, J = 52 .4, 4.3 Hz, 1H), 4.50 (dt, J = 18.5, 4.6Hz, 1H), 4.19 (t, J = 6.1 Hz, 2H), 4.03 (q, J = 5.1 Hz, 1H), 3.87 (brs, 1H), 3.45(m, 1H), 2.27 (t, J = 20.5 Hz, 2H), 1 .17 (t, J = 7.2 Hz, 3H). C13 H 19 ESI MS of ClFN5O8P2 [M+H] + , calculated value 490.7, measured value 490.1。 [Example 32] (((((2R,3R,4S,5R)-5-(2-Chloro-6-(isopropylamino )-9H-purin-9-yl)-4-fluoro-3-hydroxyoxolane-2 -yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical Structure

[0225] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 the title compound was synthesized: 1 H NMR (400 MHz, DMSO-d6) δ 8.27 (m, 2H), 6.37 (d, J = 13.9 Hz, 1H), 5.28 (brs, 2H), 5.25 (d, J = 52.1 Hz, 1H), 4.98 (brs, 1H), 4.51 (d, J = 18.3 Hz, 1H), 4.35 (sept, J = 7.9Hz, 1H), 4.19 (m, 2H), 4.04 (m, 1H), 2 .26 (t, J = 20 Hz, 2H), 1.21 (dd, J = 6.6,2.1 Hz, 6H). C 14 H 21 ESI MS of ClFN5O8P2 [M-H] - , calculated value 502.7, measured value 502.2。 [Example 33] (((((2R,3R,4S,5R)-5-(2-Chloro-6-(cyclopropylamino no)-9H-purin-9-yl)-4-fluoro-3-hydroxyoxolane- Synthesis of ((2R,3R,4S,5R)-5-(2-chloro-6-((cyclopropylmethyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxyoxolan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid [Chemical formula]

[0226] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 the title compound was synthesized: 1 H NMR (400 MHz, DMSO-d6) δ 8.59 (s, 1H), 8.28 (d, J = 2.1 Hz, 1H), 6.38 (dd, J = 14.2, 4.6 Hz,1H), 5.26 (ddd, J = 52.5, 4.3, 4.3 Hz , 1H), 4.51 (dt, J = 18.5, 4.5 Hz, 1H),4.19 (t, J = 6.1 Hz, 2H), 4.03 (q, J = 5 .0 Hz, 1H), 2.98 (s, 1H), 2.36 - 2.15 (m,2H), 0.82 - 0.48 (m, 4H). C 14 H 18 ClFN5O 8P2 ESI MS [M-H] - , calculated value 500.03, measured value 500.0. [Example 34] ((((2R,3R,4S,5R)-5-(2-chloro-6-((cyclopropylmethyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxyoxolan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis tetrahydro Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 [Chemical formula]

[0227] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 the title compound was synthesized: 11H NMR (400 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.28 (s, 1H ), 6.37 (dd, J = 14.2, 4.6 Hz, 1H), 5.25(ddd, J = 52.5, 4.3, 4.3 Hz, 1H), 4.54 - 4.47 (m, 1H), 4.19 (t, J = 6.3 Hz, 2H),4.05 - 4.01 (m, 1H), 3.81 - 3.74 (m, 1 H), 3.30 - 3.27 (m, 1H), 2.26 (dd, J =20.5, 20.5 Hz, 2H), 1.1 - 1.3 (m, 1H), 0 .48 - 0.37 (m, 2H), 0.28 - 0.26 (m, 2H). C 15 H 20 ESI MS of ClFN5O8P2 [M-H] - , calculated value 514.1, found 514.0. [Example 35] ((((2R,3R,4S,5R)-5-(2-Chloro-6-(cyclopentylamino -9H-purin-9-yl)-4-fluoro-3-hydroxyoxolane- 2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid Synthesis [Chemical Structure Diagram]

[0228] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 , the title compound was synthesized: 1 1H NMR (400 MHz, DMSO-d6) δ 8.41 (d, J = 7.8 Hz, 1H), 8.27 (s, 1H), 6.37 (dd, J = 14.4, 4.6 Hz,1H), 5.25 (dt, J = 52.4, 4.3 Hz, 1H), 4.55 - 4.37 (m, 2H), 4.19 (t, J = 6.1 Hz,2H), 4.03 (q, J = 5.1 Hz, 1H), 2.26 ( t, J = 20.5 Hz, 2H), 1.93 (s, 2H), 1.64 (d,J = 62.5 Hz, 6H). C 16 H 23 ClFN5O8P2 The ESI MS [M+H] of + , calculated value 530.1, measured value 530.2. [Example 36] ((((2R,3R,4S,5R)-5-(2-chloro-6-(((S)-tetrah ydrofuran-3-yl)amino)-9H-purin-9-yl)-4-fluoro-3-hyd roxy tetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl) Synthesis of phosphonic acid [Chemical formula]

[0229] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.66 (d, J = 6.2 Hz, 1H), 8.31 (s, 1H), 6.38 (dd, J = 14.3, 4.6 Hz,1H), 5.26 (ddd, J = 52.4, 4.2, 2.4 Hz , 1H), 4.61 - 4.67 (m, 1H), 4.57 - 4.45 (m,1H), 4.19 (t, J = 6.1 Hz, 2H), 4.04 (q, J = 5.0 Hz, 1H), 3.89 (dt, J = 15.3,7.8 Hz, 2H), 3.73 (q, J = 7.8 Hz, 1H), 3.61 (dd, J = 8.9, 4.4 Hz, 1H), 2.36 - 1.99 (m, 4H). C 15 H 20 ESI MS of ClFN5O9P2 M-H] - , calculated value 530.04, measured value 530.1. [Example 37] ((((2R,3R,4S,5R)-5-(2-chloro-6-(((R)-tetrahydro furan-3-yl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxy tetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl) Synthesis of phosphonic acid

Chemical formula

[0230] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.65 (d, J = 6.8 Hz, 1H), 8.31 (s, 1H), 6.39 (dd, J = 14.2, 4.6 Hz, 1H), 5.26 (ddd, J = 52.4, 4.3, 4.3 Hz , 1H), 4.69 - 4.56 (m, 1H), 4.51 (dt, J = 18.6, 4.6 Hz, 1H), 4.20 (t, J = 6.1 Hz , 2H), 4.04 (q, J = 5.0 Hz, 1H), 3.89 (dt, J = 18.6, 7.9 Hz, 2H), 3.74 (q, J = 7 .8 Hz, 1H), 3.67 - 3.54 (m, 1H), 2.35 - 1.90 (m, 4H). C 15 H 20 ESI MS of ClFN5O9P2 M-H]- , Calculated value 530.04, measured value 530.1. [Example 38] (((((2R,3R,4S,5R)-5-(2-chloro-6-((tetrahydro-2 H-pyran-4-yl)amino)-9H-purin-9-yl)-4-fluoro-3-hyd roxyoxytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl) Synthesis of phosphonic acid [Chem.]

[0231] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.47 - 8.34 (m, 1H), 8.30 (s, 1H), 6.37 (dd, J = 14.1, 4.8 Hz, 1H), 5.25 (ddd, J = 52.4, 4.3, 4.3 Hz, 1H) , 4.92 - 4.65 (m, 1H), 4.59 - 4.39 (m, 1H), 4.19 (t, J = 6.2 Hz, 2H), 4.03 (q, J = 5.1 Hz, 1H), 3.89 (d, J = 11.3 Hz, 2H), 3.41 (t, J = 11.4 Hz, 2H), 2.26 (dd, J = 20.5 Hz, 2H), 1.92 - 1.45 (m, 4H). C 16 H 22 ESI MS [M-H] of ClFN5O9P2 - , calculated value 5 44.06, measured value 544.1. [Example 39] (((((2R,3R,4S,5R)-5-(2-chloro-6-(pyrrolidin-1-yl ((2R,3R,4S,5R)-5-(2-Chloro-6-(piperidin-1-yl)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran- 2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chem.

[0232] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 the title compound was synthesized: 1 H NMR (400 MHz, DMSO-d6) δ 8.27 (d, J = 2.1 Hz, 1H), 6.39 (dd, J = 14.1, 4.7 Hz, 1H), 5.26 (dt,J = 52.5, 4.3 Hz, 1H), 4.50 (dt, J = 18.5, 4.6 Hz, 1H), 4.19 (t, J = 5.9 Hz,2H), 4.05 (q, J = 5.3, 4.1 Hz, 3H), 3.6 0 (t, J = 6.8 Hz, 2H), 2.27 (t, J = 20.5Hz, 2H), 2.01 (p, J = 6.7 Hz, 2H), 1.92 (q, J = 6.7 Hz, 2H). C 15 H 21 ESI MS of C11H16ClFN5O8P2 [M+H] + , calculated value 516.1, measured value 516. 1. [Example 40] ((((2R,3R,4S,5R)-5-(2-Chloro-6-(piperidin-1-yl)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran- 2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chem.

Chem.

[0233] In the same manner as in Example 29, the product of step a of Example 29 and the corresponding amine were used to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.30 (d, J = 2.2 Hz, 1H), 6.39 (dd, J = 14.3, 4.6 Hz, 1H), 5.26 (dt,J = 52.4, 4.3 Hz, 1H), 4.50 (dt, J = 18.4, 4.6 Hz, 1H), 4.19 (t, J = 6.0 Hz,2H), 4.04 (q, J = 5.0 Hz, 1H), 3.88 (m, 2H), 2.27 (t, J = 20.5 Hz, 2H), 1.64 (d, J= 31.0 Hz, 8H). C 16 H 23 ES of ClFN5O8P2 I MS [M+H] + , calculated value 530.1, measured value 530.2. [Example 41] ((((2R,3R,4S,5R)-5-(2-chloro-6-morpholino-9H-p urin-9-yl)-4-fluoro-3-hydroxyoxolane-2-yl)metho (hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0234] In the same manner as in Example 29, the product of step a of Example 29 and the corresponding amine were used to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.34 (d, J = 2.1 Hz, 1H), 6.41 (dd, J = 13.9, 4.6 Hz, 1H), 5.27 (dt,J = 52.5, 4.3 Hz, 1H), 4.51 (dt, J = 18.5, 4.6 Hz, 1H), 4.19 (t, J = 5.8 Hz, 2H), 4.04 (q, J = 5.1 Hz, 1H), 3.79 - 3 .67 (m, 5H), 2.26 (t, J = 20.5 Hz, 2H). C 15 H 21 ESI MS of C 15 H 21 ClFN5O9P2 [M+H] + , calculated value 532.1, found 532.1. [Example 42] ((((2R,3R,4S,5R)-5-(2-chloro-6-(isoindolin-2 -yl)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofura n-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0235] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 , the title compound was synthesized: 1 H NMR (400 MHz, DMSO-d6) δ 8.37 (d, J = 2.1 Hz, 1H), 7.48 (dt, J = 9.9, 4.7 Hz, 2H), 7.43 - 7.28 (m, 2H), 6.44 (dd, J = 13.8, 4.7 Hz , 1H), 5.41 (s, 2H), 5.29 (dt, J = 52.6, 4.4 Hz, 1H), 4.98 (s, 2H), 4.54 (dt, J = 18.7, 4.7 Hz, 1H), 4.21 (t, J = 5.9 Hz, 2H), 4.05 (q, J = 4.9 Hz, 1H), 2.27 (t , J = 20.5 Hz, 2H). C 19 H 21 ESI MS of C 19 H 21 ClFN5O8P2 [M+H]+ , Calculated value 564.1, measured value 564.1 . [Example 43] ((((2R,3R,4S,5R)-5-(2-chloro-6-((4-chlorobenz yl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydro furan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0236] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 9.01 (t, J = 6.2 Hz, 1H), 8.32 (d, J = 2.1 Hz, 1H), 7.64 - 7.08 (m,4H), 6.38 (dd, J = 14.3, 4.6 Hz, 1H), 5.26 (dt, J = 52.5, 4.3 Hz, 1H), 4.64 (q,J = 7.3, 5.4 Hz, 2H), 4.51 (dt, J = 1 8.7, 4.6 Hz, 1H), 4.28 - 4.11 (m, 2H), 4.04(q, J = 5.1 Hz, 1H), 2.27 (t, J = 20 .5 Hz, 2H). C 18 H 20 ESI MS [M-H] of C - H [Example 44] ((((2R,3R,4S,5R)-5-(2-chloro-6-((4-fluoroben zyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydro Synthesis of ((2R,3R,4S,5R)-5-(2-chloro-6-((3-methylbenzyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxyoxolan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid

Chem.

[0237] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 the title compound was synthesized: 1 H NMR (400 MHz, DMSO-d6) δ 9.00 (t, J = 6.3 Hz, 1H), 8.31 (d, J = 2.2 Hz, 1H), 7.52 - 7.24 (m,3H), 7.23 - 7.01 (m, 2H), 6.38 (dd, J = 14.3, 4.6 Hz, 1H), 5.26 (dt, J = 52.4,4.3 Hz, 1H), 4.72 - 4.55 (m, 2H), 4.20 (t, J = 6.0 Hz, 3H), 4.04 (q, J = 5.1 Hz,1H), 2.27 (t, J = 20.5 Hz, 2H). C 18 H2 ESI MS of C12H2ClF2N5O8P2 [M-H] - , calculated value 568.0, measured value 568.2. [Example 45] ((((2R,3R,4S,5R)-5-(2-chloro-6-((3-methylbenzyl yl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxyoxolan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29

Chem.

[0238] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 Thus, the title compound was synthesized as a white solid (87.1 mg; 31%): 1 H NMR (400 MH z, DMSO-d6) δ 8.96 (t, J = 6.3Hz, 1H), 8.30 (d, J = 2.2 Hz, 1H), 7.20 (t, J = 7.5 Hz, 1H), 7.17 - 7.10 (m, 2H), 7.04 (d,J = 7.4 Hz, 1H), 6.38 (dd, J = 14.3, 4.6 Hz, 1H), 5.25 (dt, J = 52.4, 4.3 Hz,1H), 4.68 - 4.56 (m, 2H), 4.51 (dt, J = 18.4, 4.6 Hz, 1H), 4.19 (t, J = 6.0 Hz,2H), 4.03 (q, J = 5.1 Hz, 1H), 2.35 - 2 .17 (m, 2H). C 19 H 22 ClFN5O8P2 ESI MS [M-H] - , calculated value 564.1, measured value 564.2. [Example 46] (((((2R,3R,4S,5R)-5-(2-chloro-6-((3-fluoroben zyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxy-tetrahydro furan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical Structure

[0239] In the same manner as in Example 29, using the product of step a of Example 29 and the corresponding amine Thus, the title compound was synthesized as a white solid (65.1 mg; 23%): 1 H NMR (400 MH z, DMSO-d6) δ 9.02 (t, J = 6.3Hz, 1H), 8.32 (d, J = 2.2 Hz, 1H), 7.42 - 7.32 ( m, 1H), 7.17 (t, J = 9.2 Hz, 2H), 7.07 (td,J = 8.4, 2.2 Hz, 1H), 6.39 (dd, J = 14.4, 4.6 Hz, 1H), 5.26 (dt, J = 52.5, 4.3Hz, 1H), 4.74 - 4.60 (m, 2H), 4.51 (d t, J = 18.5, 4.7 Hz, 1H), 4.26 - 4.13 (m,2H), 4.04 (q, J = 5.0 Hz, 1H), 2.27 (t , J = 20.5 Hz, 2H). C 18 H 19 ESI MS of ClF2N5O8P2 [M-H] - , calculated value 568.0, measured value 568.2 。 [Example 47] (((((2R,3R,4S,5R)-5-(2-chloro-6-((3-chlorobenzyl amino)-9H-purin-9-yl)-4-fluoro-3-hydroxy-tetrahydro furan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical Structure

[0240] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29, the title compound was synthesized as a white solid (70.6 mg; 24%): 1 H NMR (400 MH z, DMSO-d6) δ 9.03 (t, J = 6.2Hz, 1H), 8.33 (d, J = 2.2 Hz, 1H), 7.45 - 7.27 ( m, 4H), 6.39 (dd, J = 14.4, 4.6 Hz, 1H), 5.26 (dt, J = 52.4, 4.2 Hz, 1H), 4.74 - 4.58 (m, 2H), 4.51 (dt, J = 18.5, 4.6 Hz, 1H), 4.20 (t, J = 6.1 Hz, 2H), 4.04 ( q, J = 5.1 Hz, 1H), 2.27 (t, J = 20.5 Hz, 2H). C 18 H 19 ESI MS of Cl2FN5O8P2 [M-H] - , Calculated value: 584.0, Measured value: 584.0. [Example 48] ((((2R,3R,4S,5R)-5-(2-chloro-6-((2-chlorobenzyl amino)-9H-purin-9-yl)-4-fluoro-3-hydroxy-tetrahydro furan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical Structure Diagram]

[0241] Using the product of step a in Example 29 and the corresponding amine in the same method as in Example 29 the title compound was synthesized: 1 H NMR (400 MHz, DMSO-d6) δ 8.99 (t, J = 6.1 Hz, 1H), 8.35 (s, 1H), 7.47 (dd, J = 6.0, 3.3 Hz, 1H), 7.35 - 7.22 (m, 3H), 6.40 (dd, J = 14.2, 4.6 Hz, 1H), 5.27 (dt, J = 52.4, 4.3 Hz, 1H), 4.73 (d, J = 5.2 Hz, 2H), 4.52 (d, J = 18.5 Hz, 1H), 4.20 (t, J = 6.2Hz, 2H), 4.05 (q, J = 5.1 Hz, 1H), 2 .27 (t, J = 20.5 Hz, 2H). C 18 H 20 ESI MS of C 18 H 20 Cl2FN5O8P2 [M+H] + , calculated value 586.0, measured value 586.1. [Example 49] ((((2R,3R,4S,5R)-5-(2-chloro-6-((2-chlorobenzyl )(methyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxyte trahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphon ic acid synthesis [Chemical Structure Diagram]

[0242] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 , the title compound was synthesized: 1 H NMR (400 MHz, DMSO-d6) δ 8.32 (d, J = 37.7 Hz, 1 H ), 7.55 - 7.46 (m, 1 H), 7.31 (bs, 2 H),7.15 (bs, 1 H), 6.41 (d, J = 14.4 Hz, 1 H), 5.61 (bs, 1 H), 5.26 (d, J = 52.6 Hz,1 H), 5.00 (b, 1 H), 4.49 (bs, 1 H), 4.17 (bs, 2 H), 4.03 (bs, 1 H), 3.70 (bs, 1H), 3.18 (bs, 2 H), 2.25 (t, J = 20. 4 Hz, 2 H). C 19 H 24 ESI MS of C 19 H 24 Cl2N5O9P2 [M+H]+ , Calculated value: 600.0, Measured value: 600.1. [Example 50] ((((2R,3R,4S,5R)-5-(2-Chloro-6-((pyridin-4-yl methyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxytetra hydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid Synthesis [Chemical Structure]

[0243] Using the product of step a of Example 29 and the corresponding amine in the same method as in Example 29 , the title compound was synthesized: 1 H NMR (400 MHz, DMSO-d6) δ 9.13 (s, 1H), 8.66 (d, J = 5.7 Hz, 2H), 8.37 (s, 1H), 7.65 (d, J = 5.6 Hz, 2H), 6.40 (dd, J = 14.0, 4.6 H z, 1H), 5.40 - 5.08 (m, 1H), 4.80 (d, J = 6.1 Hz, 2H), 4.53 (d, J = 18.3 Hz, 1H) , 4.19 (s, 2H), 4.04 (d, J = 5.2 Hz, 1H), 2.25 (t, J = 20.4 Hz, 2H). C 17 H 20 ClFN 6O8P2 ESI MS [M+H] + , Calculated value: 553.1, Measured value: 553.2. [Example 51] ((((2R,3R,4S,5R)-5-(2-Chloro-6-(phenethylamino) -9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2- yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid Synthesis [Chem.]

[0244] Using the product of step a of Example 29 and the corresponding amine in the same manner as in Example 29 the title compound was synthesized: 1 H NMR (400 MHz, DMSO-d6) δ 8.50 (t, J = 5.7 Hz, 1H), 8.27 (s, 1H), 7.38 - 7.13 (m, 5H), 6.37(dd, J = 14.4, 4.7 Hz, 1H), 5.25 (dt, J = 52.4, 4.2 Hz, 1H), 4.51 (dt, J = 18.5,4.6 Hz, 1H), 4.19 (t, J = 6.1 Hz, 2H), 4.04 (t, J = 5.1 Hz, 1H), 3.66 (d, J = 7.2Hz, 2H), 2.92 (t, J = 7.5 Hz, 2H), 2 .26 (t, J = 20.5 Hz, 2H). C 19 H 23 ESI MS of C15H16ClFN5O8P2 [M+H]+ + , calculated value 566.1, found 566.1. [Example 52] ((((2R,3R,4S,5R)-5-(6-(Benzylamino)-2-methyl- 9H-purin-9-yl)-4-fluoro-3-hydroxyoxolan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis Using the same method as in Example 29 and using 6-chloro-2-methylpurine instead of 2,6-dichloropurine, the title compound was synthesized: [Chem.]

[0245] Using the same method as in Example 29, using 6-chloro-2-methylpurine instead of 2,6-dichloropurine, the title compound was synthesized: ​1 1H NMR (400 MHz, DMSO-d6) δ 8.51 (s, 1H), 8.23 (s, 1H), 7.44 - 7.19 (m, 5H), 6.44 (dd, J = 15.0, 4.6 Hz, 1H), 5.41 - 5.13 (m, 1H), 4.72 (s, 2H), 4.53 (dd, J = 18.4, 4.7 Hz, 1H), 4.19 (t, J = 6.1 Hz , 2H), 4.04 (t, J = 5.1 Hz, 1H), 2.46 (s, 3H), 2.26 (t, J = 20.5 Hz, 2H). C 19 H2 ESI MS of 4FN5O8P2 [M+H] + , calculated value 532.1, measured value 532.2. [Example 53] ((((2R,3R,4S,5R)-5-(6-(Cyclopentylamino)-2-methyl- 9H-purin-9-yl)-4-fluoro-3-hydroxyoxolane- 2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical Structure

[0246] In the same manner as in Example 29, using 6-chloro-2-methylpurine instead of 2,6-dichloropurine and cyclopentylamine instead of benzylamine, the title compound was synthesized: 1 1H NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1H), 8.30 (s, 1H), 6.45 (dd, J = 14.4, 4.6 Hz, 1H), 5.25 (dt, J = 52.5, 4.3 Hz, 1H), 4.53 (dt, J = 18.3, 4.5 Hz, 1H), 4.20 (t, J = 6.1 Hz, 2H), 4.04(q, J = 5.0 Hz, 1H), 2.26 (t, J = 20.5 Hz, 2H), 1.98 (s, 2H), 1.82 - 1.46 (m, 6H).C 17 H 26 ESI MS of FN5O8P2 [M+H] + , calculated value 510.1, found 510.2. [Example 54] ((((2R,3R,4S,5R)-5-(6-(Benzylamino)-2-(trif luoromethyl)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydro furan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0247] In the same manner as in Example 29, using 6-chloro-2-trif luoromethylpurine instead of 2,6-dichloropurine, the title compound was synthesized: 1 H NMR (400 MHz, DMSO-d6) δ 9.11 (d, J = 6.3 Hz, 1 H), 8.49 (d, J =2.1 Hz, 1 H), 7.39 - 7.35 (m, 2 H), 7 .34 - 7.27 (m, 2 H), 7.25 - 7.20 (m, 1 H),6.48 (dd, J = 14.0, 4.7 Hz, 1 H), 5.3 0 (dt, J = 52.4, 4.3 Hz, 1 H), 5.20 (bs, 1H), 4.70 (t, J = 5.7 Hz, 1 H), 4.56 ( dt, J = 18.6, 4.7 Hz, 1 H), 4.21 (t, J =6.2 Hz, 2 H), 4.06 (q, J = 5.1 Hz, 1 H) , 2.26 (t, J = 20.5 Hz, 2 H). C 19 H 21 ESI MS of F4N5O8P2 [M+H] + , calculated value 586.1, measured value 586.2. [Example 55] (((((2R,3R,4S,5R)-5-(6-(Cyclopentylamino)-2-( trifluoromethyl)-9H-purin-9-yl)-4-fluoro-3-hydroxy-tet rahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid Synthesis [Chemical formula]

[0248] In the same manner as in Example 29, 6-chloro-2-trifluoromethylpurine was used instead of 2,6-dichloropurine, and cyclopentylamine was used instead of benzylamine to synthesize the title compound: 1 H NMR (400 MHz, DMSO-d6) δ 8.59 - 8.40 (m, 2 H), 6.47 (dd, J = 13.9, 4.7 Hz, 1 H), 5.30 (dt, J =52.4, 4.3 Hz, 1 H), 5.11 (bs, 1 H), 4 .52 (dd, J = 28.1, 14.1 Hz, 2 H), 4.21 (t,J = 6.0 Hz, 2 H), 4.06 (q, J = 5.2 Hz , 1 H), 2.26 (t, J = 20.4 Hz, 2 H), 2.08 -1.90 (m, 2 H), 1.80 - 1.50 (m, 6 H). C 17 H 23 ESI MS of F4N5O8P2 [M+H] + , calculated value 564.2, measured value 564.1.​ [Example 56] ((((2R,3R,4S,5R)-5-(6-(Benzylamino)-2-phenyl -9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2- yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0249] Step a: The product (750 mg, 1.25 mmol) from Step b(1) of Example 29, phen ylboronic acid (229 mg, 1.88 mmol), and potassium carbonate (518 mg, 3. 75 mmol) were suspended in 3:1 THF:H2O (10.3 mL). This mixture was degassed by N2 sparge for 10 minutes. Subsequently, Pd(PPh3)4 (144 mg, 0.13 mmol) was added, and the resulting mixture was further degassed for 5 minutes, then sealed and heated at 80 °C overnight . After cooling to room temperature, the reaction solution was diluted with EtOAc and washed with water and brine. The organic layer was dried over M gSO4, filtered, and concentrated under reduced pressure. This crude material consisted of a mixture of mono- and di-debenzoylated products used directly in Step b .

[0250] Step b: The product from Step a was dissolved in methanol (12.5 mL), and potassium carbonate ( 518 mg, 3.75 mmol) was added. The resulting suspension was stirred overnight at room temperature and then partitioned between E tOAc and water. The organic layer was washed with brine and then dried (MgSO4), concentrated under reduced pressure. The desired product was obtained as a white solid after column chromatography (SiO2, 0 - 10% gradient of MeOH and CH2Cl2) (41 mg, 8%, 2 steps) . C​23 H 22 ESI MS of FN5O3 [M+H] + , calculated value 436.2, measured value 436.3

[0251] Step c: Using the same method as in Example 1, the title compound was obtained as a white solid: 1 H N MR (400 MHz, DMSO-d6) δ 8.58 (s,1H), 8.44 - 8.32 (m, 2H), 8.29 (d, J = 2.4 Hz, 1H), 7.40 - 7.50 (m, 5H), 7.31 (dd, J =8.3, 6.9 Hz, 2H), 7.24 - 7.15 (m, 1H), 6.59 (dd, J = 15.4, 4.6 Hz, 1H), 5.30 (dt,J = 52.4, 4.1 Hz, 1H), 4.82 (s, 2H), 4.69 - 4.48 (m, 1H), 4.22 (d, J = 6.6 Hz,2H), 4.08 (q, J = 5.1 Hz, 1H), 2.27 (t , J = 20.5 Hz, 2H). C 24 H 26 ESI MS of FN5O8P2 [M-H] - , calculated value 592.1, measured value 592.2 [Example 57] ((((2R,3R,4S,5R)-5-(2-Benzyl-6-(benzylamino) -9H-purin-9-yl)-4-fluoro-3-hydroxyoxolane-2- yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical Structure

[0252] Step a: The product (391 mg, 0.659 mmol) of Step b(1) in Example 29, ben Potassium n-butyltrifluoroborate (391 mg, 1.98 mmol), and cesium carbonate (1.07 g, 3.30 mmol) were suspended in THF:H2O (6.5 mL) at a ratio of 20:1. This mixture was degassed by N2 sparging for 10 minutes. Subsequently, Pd(PPh3)2Cl2 (96 mg, 0.132 mmol) was added, and the resulting mixture was further degassed for 5 minutes and then sealed and heated at 80 °C for 48 hours. After cooling to room temperature, the reaction mixture was diluted with EtOAc and washed with water and brine. The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The desired product was obtained as a beige solid (174 mg, 40%) after column chromatography (SiO2, EtOAc / hexane).

[0253] Step b: The product from step a (174 mg, 0.265 mmol) was dissolved in methanol (2. 65 mL), and potassium carbonate (110 mg, 3.75 mmol) was added. The resulting suspension was stirred at room temperature for 1.5 hours and then partitioned between EtOAc and water. The organic layer was washed with brine, then dried (MgSO4), and concentrated under reduced pressure. The desired product was obtained as a white solid (102 mg, 86%) after column chromatography (SiO2, 0 - 10% gradient of MeOH and CH2Cl2). 24 H 24 ESI MS [M+H] of C + , calculated value 450 .2, measured value 450.3.

[0254] Step c: Using the same method as in Example 1, the title compound was obtained as a white solid: 1 H N 1H NMR (400 MHz, DMSO-d6) δ 8.54 (s,1H), 8.22 (s, 1H), 7.61 - 6.94 (m, 10H), 6.44 (dd, J = 15.1, 4.6 Hz, 1H), 5.23 (dt, J = 52.4, 4.1 Hz, 1H), 4.82 - 4.40 (m, 3H) , 4.18 (t, J = 6.5 Hz, 2H), 4.03 (dd, J = 10.9, 5.9 Hz, 3H), 2.26 (t, J = 20.5 H z, 2H). C 25 H 28 ESI MS of FN5O8P2 [M-H] - , calculated value 606.1, measured value 606.3. [Example 58] ((((2R,3R,4S,5R)-5-(6-(Cyclopentylamino)-2-( piperidin-1-ylmethyl)-9H-purin-9-yl)-4-fluoro-3-hydroxy xytetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl)ho Synthesis of sulfonic acid

Chemical Structure

[0255] Step a: The product of Step a in Example 35 (10.0 g, 17.24 mmol), phenyl vinylboronic acid (3.83 g, 25.86 mmol), and sodium carbonate (5.44 m g, 51.72 mmol) were suspended in 3:1 THF:H2O (100 mL). This mixture was degassed by N2 sparging for 10 minutes. Subsequently, Pd(PPh3)4 (1.99 g, 1.72 mmol) was added, and the resulting mixture was degassed for an additional 5 minutes and then heated to reflux overnight . After cooling to room temperature, the reaction mixture was diluted with EtOAc and washed with water and brine. The organic layer was dried over M gSO4, filtered, and concentrated under reduced pressure. The desired product was obtained by column chromatography After filtration (SiO2, 5% - 50% EtOAc / hexane), a colorless solid (8.06 g, 72%) was obtained.

[0256] Step b: To a suspension of the product from step a (8.0 6 g, 12.04 mmol) in THF:H2O (127.5 mL) in a 2:1 ratio, sodium periodate (15.5 g, 72.4 mmol) , and 2,6-lutidine (2.80 mL, 24.1 mmol), potassium osmate dihydrate (100 mg, 0.30 mmol) was added. The resulting thick suspension was stirred overnight at room temperature and then partitioned between EtOAc and water. The organic layer was washed sequentially with water and brine, dried over MgSO4, and concentrated under reduced pressure. The title compound was obtained as an off-white oil (6.74 g, 97 %) after column chromatography (SiO2, EtOAc / hexane). C H 30 H 28 FN5O6 ESI MS [M + H] + , calculated value 574.2, measured value 574.4.

[0257] Step c: 1) To a solution of the product from step b (500 mg, 0.8 7 mmol) in dichloroethane (4.5 mL), piperidine (104 μL, 1.05 mmol) was added, followed by sodium triacetoxyborohydride (223 mg, 1.05 mmol) all at once. The reaction mixture was stirred overnight at room temperature and then partitioned between EtOAc and water. The organic layer was washed with brine , dried over MgSO4, and concentrated under reduced pressure to obtain the title compound, which was used without further purification. C H 35 H 39 FN6O5 ESI MS [M + H] + , calculated value 643.3, measured value 643.3.

[0258] Step c: 2) Dissolve the crude product in methanol (8.7 mL), and add potassium carbonate (36 2 mg, 2.62 mmol). Stir the resulting suspension overnight at room temperature, and then partition it between EtO Ac and water. Wash the organic layer with brine, then dry it (MgSO4), and concentrate it under reduced pressure. Obtain the desired product as a white solid after column chromatography (SiO2, 0 - 100% gradient of MeOH and CH2Cl2) (151 mg, 40%, 2 steps). C H 21 H 31 ESI MS of FN6O3 [M+H] + , calculated value 435.2, measured value 435.3.

[0259] Step d: Using the same method as in Example 1, obtain the title compound as a white solid: 1 H N 1H NMR (400 MHz, DMSO-d6) δ 9.40 (s,1H), 8.58 - 8.03 (m, 2H), 6.45 (dd, J = 14.1, 4.8 Hz, 1H), 5.25 (dt, J = 52.5, 4.3 Hz,1H), 4.59 (d, J = 16.3 Hz, 2H), 4.40 (s , 1H), 4.20 (t, J = 6.1 Hz, 2H), 4.04 (q, J= 5.1 Hz, 1H), 3.61 (s, 1H), 3.08 (s , 2H), 2.24 (t, J = 20.4 Hz, 2H), 2.06 -1.35 (m, 10H). C 22 H 35 ESI MS of FN6O8P2 M-H] - , calculated value 591.2, measured value 591.3. [Example 59] ((((2R,3R,4S,5R)-5-(6-(Cyclopentylamino)-2-( (Methoxymethyl)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydro (furan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis (Synthesis) [Chemical formula]

[0260] Step a: To a solution of the product of Step b of Example 58 (1.0 g, 1 .74 mmol) in dichloroethane (20 mL), sodium triacetoxyborohydride (443 mg, 2. 09 mmol) was added all at once. The reaction mixture was stirred at room temperature overnight and then partitioned between EtOAc and water. The organic layer was washed with brine, dried over MgSO4, and concentrated under reduced pressure to obtain the title compound, which was used without further purification. C H 30 H 30 FN5O6 ESI MS [M+H] + , Calculated value 576.2, measured value 576.3.

[0261] Step b: 1) To a solution of the product of Step a in dichloromethane (10 mL) at 0 °C, TsC l (436 mg, 2.29 mmol) and triethylamine (400 μL, 2.87 mmol) were added. The reaction mixture was warmed to room temperature and stirred overnight. The reaction mixture was diluted with EtOAc and washed with saturated NaHCO3, 10% citric acid, water, and brine. The organic layer was dried over MgSO4 and concentrated under reduced pressure to obtain the crude title compound (1.20 g, 94%, 2 steps), which was used directly in the next step.

[0262] Step b: 2) Methanol (10 ml) was added to a flask containing the crude tosylate (700 mg, 0.959 mmol) and potassium carbonate (6 62 mg, 4.8 mmol). The resulting The resulting suspension was stirred overnight, then diluted with EtOAc and washed with water and brine. The organic layer was dried over MgSO4 and concentrated under reduced pressure. The title compound (85 mg, 23%) was obtained after column chromatography (SiO2, 0 - 15% gradient of MeOH and CH2Cl2). C 17 H 24 ESI MS of C + H

[0263] Step c: Using the same method as in Example 1, the title compound was obtained as a white solid: 1 H N MR (400 MHz, DMSO-d6) δ 8.24 (s,1H), 6.46 (dd, J = 14.9, 4.6 Hz, 2H), 5.24 (dt , J = 52.5, 4.2 Hz, 1H), 4.54 (dt, J =18.3, 4.4 Hz, 2H), 4.40 (s, 2H), 4.20 (t, J = 6.1 Hz, 3H), 4.04 (t, J = 5.0 Hz, 1H),3.37 (s, 5H), 2.26 (t, J = 20.5 Hz, 2H), 1.96 (s, 3H), 1.81 - 1.41 (m, 10H). C 18 H 28 ESI MS of C - H FN5O9P2 [M-H], calculated value 5 38.1, measured value 538.2. [Example 60] ((((2R,3R,4S,5R)-5-(6-(Cyclopentylamino)-2-( (hydroxy(phenyl)methyl)-9H-purin-9-yl)-4-fluoro-3-hydroxy tetrahydrofuran-2-yl)methoxy)(hydroxy)phosphoryl)methyl) [Chemical Structure Diagram]

[0264] Step a: To a solution of the product of step b of Example 58 (330 mg, 0.58 mmol) in THF (6 mL) at -78 °C was added phenylmagnesium bromide (3.0 M / Et2O, 0.8 6 mL). The reaction mixture was stirred at this temperature for 1 h and then quenched with saturated NaHCO3. The crude reaction mixture was partitioned between EtOAc and water. The organic layer was washed with water and brine, dried over MgSO4, and 29 H 30 concentrated under reduced pressure. The crude material consisted of isomeric mono- + debenzoylated products, which were used directly in step b. C .2, found 548.3.

[0265] Step b: The product from step a was dissolved in methanol (5.8 mL) and potassium carbonate (2 40 mg, 1.74 mmol) was added. The resulting suspension was stirred at room temperature overnight and then partitioned between EtOAc and water. The organic layer was washed with brine, dried (MgSO4), and concentrated under reduced pressure. The desired product was obtained as a white solid (118 mg, 46%, 2 steps) 22 H 26 after column chromatography (SiO2, 0 - 10% gradient of MeOH and CH2Cl2). C + .2, found 444.3.

[0266] Step c: Using the same method as in Example 1, the title compound was obtained as a white solid (1:1 mixture of diastereomers): 11H NMR (400 MHz, DMSO-d6) δ 8.78 - 7.85 (m, 4H), 7.49 (s, 4H), 7.41 - 7.08 (m, 8H), 6.47(dd, J = 14.8, 4.6 Hz, 2H), 5.98 - 5.39 (m, 2H), 5.24 (dt, J = 52.4, 4.2 Hz, 1H),5.07 (s, 1H), 4.54 (d, J = 14.1 Hz, 0H ), 4.39 - 3.86 (m, 6H), 2.26 (t, J = 20.5Hz, 3H), 1.99 (d, J = 34.0 Hz, 5H), 1. 65 (d, J = 52.4 Hz, 13H). C 23 H 30 ESI MS of C - FN5O9P2 [M-H] , calculated value 600.2, measured value 60 [Example 61] Synthesis of (((2R,3R,4S,5R)-5-(6-(benzylamino)-2-(phen ylethynyl)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydro furan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid [Chemical formula]

[0267] Step a: The product (750 mg, 1.24 mmol) from Step b(1) of Example 29 was suspended in DM F (8.3 mL), Et3N (260 μL) was added, followed by phenylacetylene (205 μL). The mixture was degassed by N2 sparging for 10 minutes. Subsequently, C uI (24 mg) and Pd(PPh3)2Cl2 (44 mg) were added, and the resulting mixture was heated at 8 0 °C overnight. After cooling to room temperature, the reaction mixture was diluted with EtOAc, and 10% citric acid (aqueous It was washed with a solution, water, and brine. The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The desired product was obtained as a yellowish-brown oil (762 mg, 92%) after column chromatography (SiO2, EtOAc / hexane).

[0268] Step b: The product from step a (762 mg, 1.14 mmol) was dissolved in methanol (11. 4 mL), and potassium carbonate (473 mg, 3.42 mmol) was added. The resulting suspension was stirred overnight at room temperature and then partitioned between EtOAc and water. The organic layer was washed with brine and then dried (Na2SO4) and concentrated under reduced pressure. The desired product was obtained as a colorless oil after column chromatography (SiO2, 0 - 10% gradient of MeOH and CH2Cl2). C 25 H 22 ESI MS [M+H] of C + H FN5O3, calculated value 460.2, measured value 460.2.

[0269] Step c: Using the same method as in Example 1, the title compound was obtained as a white solid: 1 H N 1H NMR (400 MHz, DMSO-d6) δ 8.65 (s,1H), 8.37 (d, J = 2.3 Hz, 1H), 7.67 - 7.57 (m, 1H), 7.47 (td, J = 5.2, 2.1 Hz, 2H), 7.39 - 7.29 (m, 4H), 7.28 - 7.16 (m, 1H), 6.50 (dd, J = 15.2, 4.4 Hz, 1H), 5.28 (dt,J = 52.4, 4.1 Hz, 1H), 4.75 (s, 2H), 4.52 (d, J = 18.1 Hz, 1H), 4.20 (d, J = 6.4Hz, 2H), 4.06 (q, J = 5.0 Hz, 1H), 2 .28 (t, J = 20.5 Hz, 2H). C 26 H 26 ESI MS of FN5O8P2 [M-H] - , calculated value 616.1, measured value 61 6.3。 [Example 62] (((((2R,3R,4S,5R)-5-(6-(Benzylamino)-2-phenethyl -9H-purin-9-yl)-4-fluoro-3-hydroxyoxolane-2 -yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical Structure

[0270] Step a: To a solution of the product of Step b of Example 61 (2 03 mg, 0.44 mmol) in ethanol (4.4 mL) under a nitrogen atmosphere was added palladium-on-activated carbon (10 wt% wet, 20 mg). The nitrogen atmosphere was replaced with hydrogen, and the mixture was stirred at room temperature. After stirring overnight, the reaction mixture was diluted with EtOAc and filtered through celite. The filtrate was concentrated under reduced pressure to give the title compound (1 61 mg, 79%), which was used without further purification. C 25 H 26 ESI MS of FN5O3 [M+H] + , calculated value 464.2, measured value 464.4。

[0271] Step b: Using the same method as in Example 1, the title compound was obtained as a white solid: 1 1 H N MR (400 MHz, DMSO-d6) δ 8.60 -8.14 (m, 2H), 7.58 - 6.91 (m, 11H), 6.44 (d, J = 15.0 Hz, 1H), 5.22 (d, J = 52.4 Hz, 1H), 4.71 (s, 2H), 4.54 (dt, J = 18.4, 4.4 Hz, 1H), 4.19 (t, J = 6.2 Hz, 2H), 4.11 - 3.96 (m, 1H), 3.23 - 2.83 (m, 5H), 2.2 6 (t, J = 20.5 Hz, 2H). C 26 H 30 ESI MS of C5H8FN5O8P2 [M-H] - , calculated value 620.2, measured value 620 .2。 [Example 63] ((((2R,3R,4S,5R)-5-(6-(Benzylamino)-2-ethynyl -9H-purin-9-yl)-4-fluoro-3-hydroxyoxolane-2- yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0272] Step a: The product (2.0 g, 3.32 mmol) from Step b(1) of Example 29 was suspended in DMF (7.4 mL), diisopropylamine (2.3 mL) was added, followed by trimethyl silylacetylene (703 μL, 4.98) was added. The mixture was degassed by N2 sparging for 10 minutes. Subsequently, CuI (125 mg, 0.66 mmol) and Pd(PPh 3)2Cl2 (233 mg, 0.0.33 mmol) were added, and the resulting mixture was further degassed for 5 minutes , then sealed and heated at 80 °C for 36 hours. After cooling to room temperature, the reaction mixture was diluted with EtOAc and washed with saturated NH4Cl (aqueous), water, and brine. The organic layer was dried over MgSO4 and filtered, and concentrated under reduced pressure. The desired product was obtained by column chromatography (Si After (O2, 5% - 70% EtOAc / hexane), a beige solid (950 mg, 43%) was obtained.

[0273] Step b: The product from step a (950 mg, 1.43 mmol) was dissolved in methanol (14 m L), and potassium carbonate (592 mg, 4.29 mmol) was added. The resulting suspension was stirred at room temperature overnight and then partitioned between EtOAc and water. The organic layer was washed with brine , then dried (Na2SO4) and concentrated under reduced pressure. Column chromatography (SiO 2, 0 - 10% gradient of MeOH and CH2Cl2) gave the desired product, and the title compound was obtained as a white solid (230 mg, 42%). C 19 H 18 ESI MS [M + H] of FN5O3 + , calculated 384.1, found 384.2.

[0274] Step c: Using the same method as in Example 1, the title compound was obtained as a white solid: 1 H N 1H NMR (400 MHz, DMSO - d6) δ 8.65 (s,1H), 8.36 (d, J = 2.2 Hz, 1H), 7.39 - 7.26 (m, 5H), 7.28 - 7.17 (m, 1H), 6.44 (dd, J =14.8, 4.5 Hz, 1H), 5.25 (dt, J = 52.5, 4.1 Hz, 1H), 4.69 (s, 2H), 4.51 (d, J =18.1 Hz, 1H), 4.19 (d, J = 7.1 Hz, 2H), 2.27 (t, J = 20.5 Hz, 2H). C 20 H 22 ESI MS [M - H] of FN5O8P2 - , calculated 540.1, found 5 40.2. [Example 64] [({[(2R,3S,4S,5R)-5-[6-(benzyloxy)-2-chloro- 9H-purin-9-yl]-4-fluoro-3-hydroxyoxolan-2-yl]meth Synthesis of hydroxy(hydroxy)phosphorylmethyl)phosphonic acid [ka]

[0275] Step a: Under a nitrogen atmosphere, sodium hydride (90 mg, 2.26 mmol, 1.2 equiv.) was added A mixture of 10 mL of 1,2-dichlorophenyl ether (60% in oil) and benzyl alcohol (10 mL) was stirred at room temperature for 15 minutes. The product of step b(1) of Example 29 (1.00 g, 1.88 mmol) was added and the mixture was heated to room temperature. The reaction mixture was subjected to column chromatography (dichloromethane 0-10 % MeOH) to give the desired product as a white solid (721 mg, 97%). C 17 H 17 ESI MS of ClFN4O4 [M+H] + , calculated value 395.1, measured value 395.1.

[0276] Step b: The product from step a (197 mg, 0.5 mmol) was reacted with trimethyl phosphate (2 The mixture was dissolved in trimethyl phosphate (1.5 mL) and cooled to 0° C. A solution of phosphonic dichloride (624 mg, 2.5 mmol, 5 equiv.) was added dropwise to the solution. The reaction mixture was stirred at 0° C. for 3 h and then 0.5 M triethylammonium bicarbonate was added. The mixture was carefully quenched with 3.6 mL of 1H 2 SO 4 at -20°C for 15 min. The mixture was stirred at 0° C. for 15 min and then at room temperature for 15 min. The mixture was diluted with ethyl acetate It was washed three times with (10 mL). The aqueous layer was directly purified by reverse-phase HPLC (C18 column, water containing a gradient of 0 ~50% of acetonitrile and 0.1% TFA) to obtain the desired product as a white solid (40.2 mg, 15%): 1 1H NMR (400 MHz, DMSO-d6) δ 8.55 (d, J = 2.1 Hz, 1H), 7.57 - 7.51 (m, 2H), 7.46 - 7.35 (m, 3H), 6.49 (dd, J = 13.6, 4.7 Hz, 1H), 5.61 (s, 2H), 5.30 (dt, J = 52.4, 4.4 Hz, 1H), 4.53 (dt, J = 18.6, 4.7 Hz, 1H), 4.21 (t, J = 6.0 Hz, 2H), 4.06 (q, J = 5.0 Hz, 1H), 2.27 (t, J = 20.6 Hz, 2H). C 18 H 19 ESI MS of C - H [Example 65] Synthesis of ((((2R,3R,4S,5R)-5-(6-(benzylamino)-2-chloro- 9H-purin-9-yl)-4-fluoro-3-hydroxyoxolane-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid Step a: 2,4-Dichloro-7H-pyrrolo[2,3-d]pyrimidine (350 mg, 1

Chemical formula

[0277] 1.86 mmol) was dissolved in 15 mL of acetonitrile and treated with Cs2CO3 (788 mg, 2. .42 mmol, 1.3 equiv). The mixture was stirred at room temperature for 60 minutes. 2-Deoxy ribose 2-Fluoro-α-D-arabinofuranosyl bromide 3,5-dibenzoate (78 mg, 1.86 mmol, 1 equiv) was dissolved in 10 mL of acetonitrile and added dropwise to the mixture through a dropping funnel. The mixture was stirred overnight at room temperature. The mixture was filtered through a silica gel pad and concentrated. The residue was adsorbed onto silica and purified by column chromatography (hexane / ethyl acetate) to give the product as a white solid in 49% yield (480 mg).

[0278] Step b: A mixture of the product from step a (480 mg, 0.9 mmol), benzylamine (97 mg, 0.9 mmol), and Et3N (91 mg, 0.9 mmol) in anhydrous EtOH (4 mL) was stirred at 65 °C for 6 h. The excess solvent was removed under vacuum. The residue was dried under high vacuum for 30 min. Methanol (4 mL) and K2CO3 (249 mg, 1.8 mmol) were added and stirred at room temperature for 1 h. LCMS indicated completion of the reaction. This was filtered and the filtrate was concentrated. The residue was purified by flash column to give the product in quantitative yield.

[0279] Step c: The product from step b (360 mg, 0.91 mmol) was dissolved in trimethyl phosphate (4 mL), cooled to 0 °C (ice bath), and then a cold solution of methylenebis(phosphonic acid dichloride) (801 g, 3.2 mL, 3.5 equiv) in trimethyl phosphate (2 mL) was added dropwise. The reaction mixture was stirred at 0 °C for 1 h and then carefully quenched with ice-cold 0.5 M triethylammonium bicarbonate solution (11 mL), stirred at 0 °C for 15 min and then at room temperature for 1 h. The reaction mixture was purified by reverse-phase HPLC (C18 column, 0 - 40% gradient of acetonitrile) Purified with water containing 0.1% TFA to obtain the product as a white solid: 1 H N 1H NMR (400 MHz, DMSO-d6) δ 8.60 (t, J = 6.0 Hz, 1H), 7.59 - 7.13 (m, 6H), 6.72 (s, 1H), 6.49 (dd, J = 15.7, 4.5 Hz, 1H), 5.45 - 5.04 (m, 1H), 4.80 - 4.57 (m, 2H), 4.42 (dt, J = 18.6, 4.4 Hz, 1H), 4.19 - 4.15 (m, 2H), 3.98 (q, J = 5.0 Hz, 1H), 2 .26 (t, J = 20.5 Hz, 2H). C 18 H 22 ESI MS of ClFN4O8P2 [M-H] - , calculated value 549.1, measured value 549.2 [Example 66] ((((2R,3R,4S,5R)-5-(2-Chloro-6-(cyclopentylamino )-9H-purin-9-yl)-4-fluoro-3-hydroxyoxolane- 2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical Structure

[0280] In the same manner as in Example 65, using cyclopentylamine instead of benzylamine to synthesize the title compound: 1 1H NMR (400 MHz, DMSO-d6) δ 7.90 (d, J = 7.2 Hz, 1H), 7.24 (s, 1H), 6.72 (d, J = 3.6 Hz, 1H), 6.47 (dd, J = 15.9, 4.4 Hz, 1H), 5.15 ( dt, J = 52.6, 4.1 Hz, 1H), 4.52 - 4.35 (m, 2H), 4.15 (q, J = 6.3, 5.3 Hz, 2H), 3 .97 (q, J = 5.1 Hz, 1H), 2.23 (d, J = 20.5 Hz, 1H), 1.98 (d, J = 10.6 Hz, 2H), 1 .72 (s, 2H), 1.67 - 1.45 (m, 5H). C 17 H 24 ESI MS of ClFN4O8P2 [M-H] - , calculated value 527.1, measured value 527.2 [Example 67] ((((1-(6-(Benzylamino)-9H-purin-9-yl)propan-2-yl oxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis

Chemical formula

[0281] The title compound was synthesized using the corresponding alcohol in the same manner as in Step b of Example 1 : 1 HNMR (400 MHz, DMSO-d6) δ 8.79 (s, 1H), 8.31 (d, J = 15.8 Hz, 2H), 7.46 - 7.13 (m, 5H), 4.92 - 4.62 (m, 2H), 4.49 - 4.25 (m, 2H), 2.17 (td, J = 20.4, 4 .8 Hz, 2H), 1.14 (d, J = 6.3 Hz, 3H). C 16 H 22 ESI MS of N5O6P2 [M+H] + , calculated value 442 .1, measured value 442.1. [Example 68] ((((2-(6-(Benzylamino)-9H-purin-9-yl)propoxy)(hydro Synthesis of (Roxy)Phosphoryl)Methyl)Phosphonic Acid [Chemical formula]

[0282] The title compound was synthesized using the corresponding alcohol in the same manner as in step b of Example 1 as follows: 1 HNMR (400 MHz, DMSO-d6) δ 8.73 (s, 1H), 8.35 (s, 1H), 8.27 (s, 1H), 7.4 1 - 7.18 (m, 5H), 4.96 - 4.82 (m, 1H), 4.72(s, 2H), 4.39 - 4.19 (m, 2H), 2.18 ( t, J = 20.5, 1.6 Hz, 2H), 1.55 (d, J = 7.0Hz, 3H). C 16 H 22 ESIMS of C5H5N5O6P2 [M+H + , calculated value 442.0, measured value 442.1. [Example 69] ((((((2R,3R,4S,5R)-5-(2-Chloro-6-(Cyclopentyl Amino)-9H-Purin-9-yl)-4-Fluoro-3-Hydroxytetrahydrofura n-2-yl)Methoxy)(Hydroxy)Phosphoryl)Methyl)Phosphoryl)Bis(Oxy Bis(Methylene)Diisopropyl Bis(Carbonate) Synthesis [Chemical formula]

[0283] Methylenebisphosphonic acid (20 mg, 0.03 mmol, trifluoroacetate of Example 66) was dissolved in 0.5 mL of DMSO. Hunig's base (0.18 mL, 1 mmol , 30 equivalents) was added, followed by chloromethyl isopropyl carbonate (0.13 mL, 1 mmol, 30 equivalents).​ (1 mmol, 30 equiv) was added. The reaction mixture was stirred at room temperature for 5 days. The reaction mixture was reversed phase HPLC (C18 column, gradient of 0 - 40% acetonitrile and water containing 0.1% TFA) was used for purification to obtain the product as a white solid in a yield of 14% (3.6 mg). 1 H N MR (400 MHz, DMSO-d6) δ 8.42 (d, J = 7.7 Hz, 1H), 8.25 (s, 1H), 6.37 (dd, J = 1 5.2, 4.4 Hz, 1H), 5.67 - 5.43 (m, 4H), 5.24(ddt, J = 52.1, 7.7, 4.1 Hz, 1H), 4. 79 (pd, J = 6.2, 3.8 Hz, 2H), 4.57 - 4.38(m, 1H), 4.37 - 4.19 (m, 2H), 4.06 (q, J = 5.1 Hz, 1H), 2.68 (t, J = 21.2 Hz,2H), 1.92 (s, 2H), 1.80 - 1.47 (m, 6H), 1.28 - 1.15 (m, 12H). ). C 26 H 39 ClFN5O 14 ESI MS of P2 [M-H] - , calculated value 760.2, measured value 760.3. [Example 70] [({[(2S,3S,4R,5R)-5-{2-chloro-6-[cyclopentyl(methyl)amino]-9H-purin-9-yl}-3,4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid synthesis yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid was synthesized in the same manner as in Example 1.

Chemical Structure

[0284] The title compound was synthesized in the same manner as in Example 1. 11H NMR (400 MHz, DMSO-d6) δ 8.4 2 (s, 1H), 5.88 (d, J = 5.9 Hz, 1H), 4.53 -4.46 (m, 1H), 4.19 (dd, J = 5.0, 3.1 Hz, 1H), 4.15 - 4.06 (m, 3H), 3.17 (brs,3H), 2.26 (t, J = 20.5 Hz, 2H), 1.94 - 1.53 (m, 9H). C 17 H 27 ESI MS of C 17 H 27 ClN5O9P2 [M+H] + , calculated value 542.1, measured value 542.2. [Example 71] [({1-[(2S,3S,4R,5R)-5-[6-(Benzylamino)-2-chloro-9H-purin-9-yl]-3,4-dihydroxyoxolan-2-yl]ethoxy }(hydroxy)phosphoryl)methyl]phosphonic acid synthesis

Chemical Structure

[0285] Step a: Alcohol (4.8 g, 11.1 mmol) was dissolved in anhydrous CH2Cl2 (100 mL) and Dess-Martin periodinane (5.6 g, 13.3 mmol, 1.2 equiv) was added. The reaction mixture was stirred at room temperature for 3 h, then quenched with 10% Na2S2O3 (20 mL) and saturated NaHCO3 (50 mL). The organic layer was separated, dried over MgSO4, filtered and the solvent was evaporated. The crude aldehyde was purified by column chromatography (SiO2, CH2Cl2 → CH2Cl2:MeOH, 9:1) to give a yellow solid (4.8 g, quantitative). C H 20 H 21 ESI MS of C 20 H 21 ClN5O4 [M+H] +, Calculated value 430.1, measured value 430.2.

[0286] Step b: The product from step a (860 mg, 2.0 mmol) was dissolved in anhydrous THF (20 mL ), and cooled to -78 °C. A 3 M MeMgBr solution in Et2O (2 mL, 6 mi mol, 3 equivalents) was added dropwise, and the reaction mixture was stirred at -78 °C for 10 minutes, then slowly warmed to room temperature and stirred at room temperature for 2 hours. It was quenched with saturated NH4Cl (10 mL), the organic layer was separated, dried over MgSO4, filtered, and the solvent was distilled off. The crude product was used without further purification . C 21 H 25 ClN5O4's ESI MS [M+H] + , calculated value 446.2, measured value 446.3 .

[0287] Step c: The phosphonylation step was carried out in the same manner as in Example 1. 1 H NMR (400 MHz, DMSO- d6) δ 9.00 - 8.88 (m, 1H), 8.51(s, 1H), 7.37 - 7.17 (m, 5H), 5.81 (d, J = 7.0 Hz, 1H), 4.70 - 4.51 (m, 4H), 4.32 - 4.25(m, 1H), 3.83 (dd, J = 5.3, 2.6 Hz, 1H ), 2.22 (t, J = 20.5 Hz, 2H), 1.26 (d, J =6.4 Hz, 3H). C 19 H 25 ClN5O9P2's ESI M S [M+H] + , calculated value 564.1, measured value 564.1. [Example 72] [({1-[(2S,3S,4R,5R)-5-[6-(benzylamino)-2-chloro [[9H-Purin-9-yl]-3,4-dihydroxyoxolan-2-yl]propoxy Synthesis of [[(hydroxy)(phosphoryl)methyl]phosphonic acid [Chemical formula]

[0288] The title compound was synthesized in the same manner as in Example 71. 1 H NMR (400 MHz, DMSO-d6) δ 8.9 7 - 8.91 (m, 1H), 8.53 (s, 1H), 7.38 - 7.20(m, 5H), 5.79 (d, J = 7.4 Hz, 1H), 4 .65 (d, J = 6.3 Hz, 2H), 4.62 - 4.56 (m,1H), 4.54 - 4.46 (m, 1H), 4.34 (d, J = 5.5 Hz, 1H), 3.89 (dd, J = 6.1, 2.2 Hz,1H), 2.22 (t, J = 20.5 Hz, 2H), 1.69 (s, 1H), 1.58 (q, J = 7.1 Hz, 1H), 0.90 (t, J= 7.4 Hz, 3H). C 20 H 27 ESI of C lN5O9P2 MS [M+H] + , calculated value 578.1, measured value 578.2. [Example 73] [({[(2R,3R,4R,5R)-5-[2-Chloro-6-(cyclopentylamino )-9H-purin-9-yl]-3,4-dihydroxy-4-methyloxolan-2- yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid synthesis [Chemical formula]

[0289] Step a: (β-D-ribofuranose, 2-C-methyl-1,2,3,5-tetrabenzoate (4.0 g, 6.89 mmol, 1 eq)) and 2,6-dichloropurine (1.43 g, 7.58 mmol, 1.1 eq) in acetonitrile (23 mL) at 0 °C were added dropwise with 1,8-diazabicyclo[5.4.0]undec-7-ene (2.58 mL, 17.23 mmol, 2.5 eq), and then trimethylsilyl trifluoromethanesulfonate (5.11 mL, 28.25 mmol, 4.1 eq) over 5 minutes. The reaction mixture was stirred at 0 °C for 15 minutes and heated at 65 °C for 5 hours. After cooling to room temperature, the reaction solution was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate (x2) and brine (x1). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. After column chromatography (SiO2, 25% - 66% EtOAc / hexane), the desired product was obtained as a white solid (1.30 g, 97%). (β-D-ribofuranose, 2-C-methyl-1,2,3,5-tetrabenzoate (4.0 g, 6.89 mmol, 1 equivalent)) , and 2,6-dichloropurine (1.43 g, 7.58 mmol, 1.1 equivalent) were added dropwise with 1, 8-diazabicyclo[5.4.0]undec-7-ene (2.58 mL, 17.23 mmol, 2.5 equivalents), and then trimethylsilyl trifluoromethanesulfonate (5.11 mL, 28.25 mmol, 4.1 equivalents) over 5 minutes. The reaction mixture was stirred at 0 °C for 15 minutes and heated at 65 °C for 5 hours. After cooling to room temperature, the reaction solution was washed with dichloromethane diluted with saturated aqueous sodium bicarbonate solution (x2) and brine (x1). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. Column chromatography (SiO2 , 25% - 66% EtOAc / hexane) gave the desired product as a white solid (1.30 g, 97%).

[0290] Step b: 1) The product from step a (1.3 g, 2.01 mmol), cyclopentylamine (297 μL, 3.01 mmol, 1.5 equivalents), and triethylamine (560 μL, 4.02 mmol, 2.0 equivalents) were suspended in anhydrous EtOH (6.7 mL). The mixture was stirred at 70 °C for 4 hours. After cooling to room temperature, the mixture was concentrated under reduced pressure and the resulting material was used without further purification. 2) The above product was dissolved in methanol (20 mL), and potassium carbonate (1.06 g, 7. 63 mmol, 3.8 equivalents) was added. After stirring at ambient temperature for 2 hours, the residue was adsorbed on celite and column chromatography (SiO2, 0% - 10% DCM / MeOH) was performed.

[0291] 38 equivalents) was added. After stirring at ambient temperature for 2 hours, the residue was adsorbed on celite and column chromatography (SiO2, 0% - 10% DCM / MeOH) was performed. After stirring at ambient temperature for 2 hours, the residue was adsorbed on celite and column chromatography (SiO2, 0% - 10% DCM / MeOH) was performed. adsorbed on celite and column chromatography (SiO2, 0% - 10% DCM / MeOH) was performed. It was purified using and obtained as a colorless oily substance (612 mg, 79%, 2 steps).

[0292] Step c: The title compound was synthesized in the same manner as in Example 1. 1 H NMR (400 MHz, DMSO-d6) δ 8.38 (dd, J = 18.2, 8.1 Hz, 1 H), 8.26(d, J = 10.0 Hz, 1 H), 5.86 (s, 1 H), 4.42 (q, J = 7.2 Hz, 1 H), 4.27 (h, J =10.6, 10.0 Hz, 2 H), 4.06 (s, 3 H), 2.28 (t, J = 20.4 Hz, 2 H), 1.93 (d, J = 16.3Hz, 2 H), 1.78 - 1.43 (m, 6 H). C 17 H 25 ESIMS of ClN5O9P2 [M-H] - , calculated value 540.1, measured value 540.2. [Example 74] ((2R,3S,4R,5R)-5-(2-chloro-6-(cyclopentylamino)- 9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl hydrogen ((hydroxy(methoxy)phosphoryl)methyl)phosphonate synthesis

Chemical Structure

[0293] Step a: The nucleoside (2.0 g, 5.4 mmol) was dissolved in trimethyl phosphate (30 mL) and cooled to 0 °C (ice bath), and then a cold solution of methylenebis (phosphonic dichloride) (4.0 g, 16.2 mmol, 3 eq) in trimethyl phosphate (15 mL) was added dropwise and added. The reaction mixture was stirred at 0 °C for 2 hours, then cooled to about -40 °C, and anhydrous MeOH (3 0 mL) was added and it was slowly warmed to room temperature. The reaction mixture was neutralized with saturated NaHCO3 (80 mL ), diluted with water (150 mL) and EtOAc (150 mL). The organic layer was separated , dried over MgSO4, filtered, and the solvent was evaporated. The product was first purified by column chromatography (SiO2, EtOAc → EtOAc:MeOH, 8:2), and then by RP18 HP LC (H2O + 0.1% TFA / acetonitrile + 0.1% TFA) to obtain the desired product as a white solid in 11% yield (405 mg). C 19 H 31 ClN5O9P2 ESI MS [M + H] + , calculated value 570.1, measured value 570.2.

[0294] Step b: To a solution of the product from step a (75 mg, 0.13 mmol) in acetone (1 mL) , sodium iodide (50 mg, 0.33 mmol) was added. This solution was heated at 65 °C for 6 h. The solvent was evaporated, the residue was dissolved in water, and purified by reverse-phase HPLC (C18 column , 0 - 40% gradient of acetonitrile and water containing 0.1% TFA) to obtain the product as a white solid in 62% yield (51 mg). 1 H NMR (400 MHz, DMSO-d6) δ 8 .41 (d, J = 2.0 Hz, 1H), 8.36 (d, J = 7.8 Hz, 1H), 5.85 (d, J = 5.6 Hz, 1H), 5.0 4 (brs, 1H), 4.53 (t, J = 5.5 Hz, 1H), 4.47 - 4.34 (m, 1H), 4.24 - 3.95 (m, 4H), 3.58 (d, J = 11.3 Hz, 2H), 2.37 (dd, J = 20.5, 20.5 Hz, 2H), 2.07 - 1.36 (m, 8H ). C17 H 26 ESI MS of ClN5O9P2 [M+H] + , calculated value 542.8, measured value 542.2. [Example 75] [(2R,3S,4R,5R)-5-{2-chloro-6-[cyclopentyl(methyl) Amino]-9H-purin-9-yl}-3,4-dihydroxyoxolan-2-yl]me Synthesis of diphenyl[(diphenoxyphosphoryl)methyl]phosphonate. [ka]

[0295] The alcohol (380 mg, 1 mmol) was dissolved in trimethyl phosphate (5 mL) and incubated at 0 °C. (ice bath) and then distilled off methylenebis(diphosphonate) in trimethyl phosphate (3 mL). A cold solution of 1,2-dichloromethane (375 mg, 1.5 mmol, 1.5 equiv.) was added dropwise to the reaction mixture. The mixture was stirred at 0°C for 3 h. Solid phenol (470 mg, 5 mmol, 5 equiv.) was added. Once dissolved, TEA (835 μL, 6 mmol, 6 equiv.) was added dropwise. The mixture was stirred at 0° C. for 15 min and then at room temperature overnight. The product was diluted with H2O (15 mL) and the MT Extraction with BE (2 x 10 mL) was performed. The combined organic layers were dried over MgSO4, filtered, and the solvent The crude product was purified by column chromatography (SiO2, Hex → 100% EtO Ac) to give a white solid (80 mg, 10%). 1 H NMR (400 MHz, DMS O-d6) δ 8.38 (d, J = 4.3 Hz,1H), 7.41 - 7.33 (m, 4H), 7.32 - 7.25 (m, 2H), 7.2 5 - 7.11 (m, 9H), 5.89 (dd, J = 5.3, 3.2Hz, 1H), 5.63 (dd, J = 6.0, 4.4 Hz, 1H) , 5.47 - 5.41 (m, 1H), 4.62 - 4.54 (m, 1H),4.49 - 4.32 (m, 2H), 4.28 - 4.08 (m, 1H), 3.67 - 3.47 (m, 2H), 3.35 (s, 3H),1.90 - 1.52 (m, 8H). C 35 H 39 ClN5O9P2 The ESI MS [M+H] of + , calculated value 770.2, measured value 770.3. [Example 76] Bis(3-chlorophenyl)[({[(2R,3S,4R,5R)-5-{2-chloro -6-[cyclopentyl(methyl)amino]-9H-purin-9-yl}-3,4-dih droxyoxolan-2-yl]methoxy}(3-chlorophenoxy)phosphoryl)meth yl]phosphonate synthesis [Chemical formula]

[0296] The title compound was synthesized in the same manner as in Example 75. 1 H NMR (400 MHz, DMSO-d6) δ 8. 32 (s, 1H), 7.43 - 7.34 (m, 2H), 7.34 -7.05 (m, 10H), 5.88 (t, J = 4.7 Hz, 1H), 5.62 (s, 1H), 5.43 (s, 1H), 4.59 - 4.36(m, 3H), 4.26 - 4.10 (m, 2H), 3.86 - 3. 70 (m, 2H), 3.04 (s, 3H), 1.90 - 1.46 (m,8H). C 35 H 36ESI MS of Cl4N5O9P2 [M+H] + , calculated value 872.1, measured value 872.2. [Example 77] Bis(3,4-dichlorophenyl)[({[(2R,3S,4R,5R)-5-{2- chloro-6-[cyclopentyl(methyl)amino]-9H-purin-9-yl}3,4- dihydroxyoxolan-2-yl]methoxy}(3,4-dichlorophenoxy)phospho ryl)methyl]phosphonate synthesis

Chemical formula

[0297] The title compound was synthesized in the same manner as in Example 75. 1 H NMR (400 MHz, DMSO-d6) δ 8. 27 (s, 1H), 7.65 - 7.57 (m, 2H), 7.52 -7.40 (m, 3H), 7.32 - 7.04 (m, 4H), 5.87 (t, J = 5.0 Hz, 1H), 5.63 (t, J = 6.1 Hz,1H), 5.43 (dd, J = 5.6, 3.7 Hz, 1H), 4 .59 - 4.35 (m, 3H), 4.30 - 4.08 (m, 2H),3.85 (t, J = 22.0 Hz, 2H), 3.32 (s, 3H) , 1.89 - 1.45 (m, 8H). C 35 H 33 ESI MS of Cl7N5O9P2 [M+H] + , calculated value 974.0, measured value 9 74.2. [Example 78] Methyl 2-({[(2R,3S,4R,5R)-5-{2-chloro-6-[cyclopent yl(methyl)amino]-9H-purin-9-yl}-3,4-dihydroxyoxolane -2-yl]methoxy}[2-(methoxycarbonyl)phenoxy]phosphoryl)methyl Synthesis of [[2-(methoxycarbonyl)phenoxy]phosphoryl}oxy)benzoate

Chemical formula

[0298] The title compound was synthesized in the same manner as in Example 75. 1 H NMR (400 MHz, DMSO-d6) δ 8.3 1 - 8.27 (m, 1H), 7.82 - 7.74 (m, 3H), 7.57 - 7.49 (m, 2H), 7.46 - 7.39 (m, 1H), 7.36 - 7.17 (m, 6H), 5.83 (t, J = 5.9 Hz, 1H), 5.60 - 5.52 (m, 1H), 5.37 (s, 1H ), 4.55 - 4.29 (m, 3H), 4.15 - 4.04 (m, 2H), 3.81 - 3.74 (m, 2H), 3.72 - 3.65 (m , 3H), 3.32 (s, 9H), 1.88 - 1.47 (m, 8H). C 41 H 45 ClN5O 15 ESI MS [M+H] of P2 + , calculated value 944.2, found 944.3. [Example 79] [({[(2R,3R,4S,5R)-5-(2-chloro-6-{[(1S)-1-( 4-fluorophenyl)ethyl)amino]-9H-purin-9-yl)-4-fluoro- 3-hydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl)methyl] Synthesis of phosphonic acid

Chemical formula

[0299] The title compound was synthesized in the same manner as in Example 29. 1 H NMR (400 MHz, DMSO-d6) δ 8.9 2 (d, J = 8.3 Hz, 1H), 8.28 (s, 1H), 7.45(bs, 2H), 7.11 (td, J = 9.1, 1.4 Hz, 2 H), 6.34 (dd, J = 14.3, 4.6 Hz, 1H), 5.39(bs, 1H), 5.31 - 5.12 (m, 1H), 5.14 (b s, 1H), 4.48 (dt, J = 18.5, 4.5 Hz, 1H),4.17 (s, 3H), 4.01 (d, J = 5.2 Hz, 2H), 2.24 (t, J = 20.4 Hz, 3H), 1.51 (d, J =7.0 Hz, 3H). C 19 H 23 ESI MS of ClF2N5O8P2 [M+H] + , calculated value 584.1, measured value 584.2. [Example 80] [({[(2R,3R,4S,5R)-5-(2-chloro-6-{[(1R)-1-phenyl ethyl]amino}-9H-purin-9-yl)-4-fluoro-3-hydroxyoxo lan-2-yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid synthesis

Chemical Structure

[0300] The title compound was synthesized in the same manner as in Example 29. 1 H NMR (400 MHz, DMSO-d6) δ 8.9 0 (d, J = 8.3 Hz, 1H), 8.28 (s, 1H), 7.42(d, J = 7.6 Hz, 2H), 7.29 (t, J = 7.5 Hz, 2H), 7.19 (broad singlet, 1H), 6.34 (doublet of doublets, J = 14.8, 4.4 Hz, 1H), 5.39 (broad singlet, 1H), 5.23 (doublet, J = 52.6 Hz, 1H), 4.49 (doublet, J = 19.2 Hz, 2H), 4.17 (broad singlet, 2H), 4.01 (doublet, J = 5.1 Hz , 1H), 2.24 (triplet, J = 20.6 Hz, 2H), 1.52 (doublet, J = 7.0 Hz, 3H). C 19 H 24 E of ClFN5O8P2 SI MS [M+H] + , calculated value 566.1, measured value 566.1. [Example 81] [({[(2R,3R,4S,5R)-5-(2-chloro-6-{[(1S)-1-fluoro ethyl]amino}-9H-purin-9-yl)-4-fluoro-3-hydroxyox olan-2-yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid synthesis [Chemical formula]

[0301] The title compound was synthesized in the same manner as in Example 29. 1 H NMR (400 MHz, DMSO-d6) δ 8.9 1 (doublet, J = 8.3 Hz, 1H), 8.28 (singlet, 1H), 7.41 (doublet, J = 7.5 Hz, 2H), 7.29 (triplet, J = 7.6 Hz, 2H), 7.20 (doublet, J = 7.6 Hz, 1H), 6.34 (doublet, J = 14.1 Hz, 1H), 5.39 (broad singlet, 1H), 5.2 1 (doublet, J = 52.5 Hz, 1H), 4.47 (doublet, J = 18.3 Hz, 2H), 4.17 (singlet, 2H), 4.01 (singlet, 1H), 2 .24 (t, J = 20.6 Hz, 2H), 1.52 (d, J = 7.1Hz, 3H). C 19 H 24 ESI MS of ClFN5O8P2 [M +H] + , calculated value 566.1, measured value 566.1. [Example 82] [({[(2R,3R,4S,5R)-5-[6-(Cyclopentylamino)-2- hydroxy(oxan-4-yl)methyl]-9H-purin-9-yl]-4-fluoro -3-hydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl)methyl -phosphonic acid synthesis

Chemical Structure

[0302] Step a: The product of Step b in Example 58 (1.00 g, 1.75 mmol) was dissolved in THF (9 mL) and cooled to -78 °C. 4-Tetrahydropyranylmagnesium bromide (9 mL, 8.75 mmol, 0.2 M in THF) was added dropwise. The reaction mixture was warmed to room temperature and stirred at room temperature for 3 hours. The reaction mixture was cooled to 0 °C and methanol (50 mL) was added , and the mixture was stirred at room temperature for 14 hours. The reaction mixture was dried and packed onto silica gel, and purified by silica gel chromatography (0 - 10% MeOH in DCM) to obtain the desired product as a white solid (273 mg, 35%).

[0303] Step b: The title compound was synthesized as a white solid in the same manner as in Example 20 (44 m g; 29%). 1 HNMR (400 MHz, DMSO-d6) δ 6.60 - 6.40 (m, 1H), 5.26 (d, J = 53. 3 Hz, 1H), 4.63 - 4.39 (m, 2H), 4.30 - 4.13(m, 2H), 4.13 - 3.97 (m, 1H), 3.94 - 3.75 (m, 2H), 3.38 - 3.13 (m, 2H), 2.26(t, J = 20.4 Hz, 2H), 2.17 - 1.85 (m, 3 H), 1.85 - 1.22 (m, 12H). C 23 H 35 ESI MS of C - H FN5O9P2 [M-H], calculated value 606.2, measured value 60 [Example 83] (((((2R,3R,4S,5R)-5-(2-chloro-6-(cyclopentyl(m ethyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydro furan-2-yl)methoxy)(hydroxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0304] This compound was obtained in the same manner as in Example 29. 1 H NMR (400 MHz, DMSO-d6) δ 8.7 (br s, 2H), 8.30 (d, J = 2.1 Hz, 1H), 6.40 (dd,J = 14.3, 4.6 Hz, 1H), 6.09 (brs, 1H ), 5.25 (dt, J = 52.5, 4.3 Hz, 1H),4.53-4.43 (m, 1H), 4.23-4.14 (m, 2H), 4.09 - 3.98 (m, 1H), 2.28 (dd, J = 20.5 Hz, J =20.5 Hz, 2H), 2.5 (s, 3H), 1.96 - 1.44 (m, 9H). C 17 H 25ESI MS of ClFN5O8P2 [M+H] + , calculated: 544.8, found: 544.2. [Example 84] ((2R,3R,4S,5R)-5-(2-chloro-6-(cyclopentyl(methyl) Amino)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofura (hydroxy(methoxy)phosphoryl)methyl)phosphonate Synthesis of To [ka]

[0305] Step a: 2-Chloropurine fluororiboside (579 mg, 1.5 mmol) was dissolved in phosphoric acid The mixture was dissolved in trimethyl phosphate (7.5 mL), cooled to 0°C (ice bath), and trimethyl phosphate (4.5 Methylenebis(phosphonic dichloride) (1.87 g, 7.5 mmol, 5 equiv. mL) in The reaction mixture was stirred at 0° C. for 3 h and then methanol (7 1 mL) and incubated at 0° C. for 30 min, then at room temperature for 1 h, and then at 40° C. The mixture was stirred for 3 hours. The reaction mixture was concentrated under reduced pressure and dissolved in ethyl acetate (20 mL). The organic layer was washed with saturated aqueous NaHCO3, dried over sodium sulfate, and evaporated to dryness. The product was purified by column chromatography (gradient of 0-10% methanol in dichloromethane). Purification afforded the desired product as a pale yellow solid (701 mg, 80%). 1 H NMR (400 MHz, DMSO-d6) δ 8.32 (dd, J =12.0, 2.3 Hz, 1H), 6.42 (dd, J = 15.5, 4.4 Hz, 1H ), 6.15 (t, J = 4.8 Hz, 1H), 5.43 - 5.07(m, 1H), 4.60 - 4.39 (m, 1H), 4.27 (q, J = 7.3, 5.7 Hz, 2H), 4.12 - 4.03 (dq, J =9.6, 5.3 Hz, 1H), 3.69 - 3.59 (m, 9H) , 2.96 - 2.74 (m, 2H), 2.50 (s, 3H), 2.04 -1.42 (m,9H). C 20 H 31 ESI of ClFN5O8P2 MS [M+H] + , calculated value 586.9, measured value 586.2.

[0306] Step b: To a solution of the product from step a (58 mg, 0.1 mmol) in acetone (1 mL) was added sodium iodide (75 mg, 0.5 mmol). The solution was heated at 60 °C for 24 h. The solvent was evaporated, and the residue was dissolved in water and purified by reverse-phase HPLC (C18 column, 0 - 30% gradient of acetonitrile and water containing 0.1% TFA) to give the product as a white solid in 65% yield (42 mg). 1 H NMR (400 MHz, DMSO-d6) δ 8.31 (d , J = 2.2 Hz, 1H), 6.40 (dd, J = 14.6, 4.6Hz, 1H), 5.25 (dt, J = 52.4, 4.2 Hz, 1H), 4.48 (dt, J = 18.3, 4.4 Hz, 1H), 4.18(t, J = 6.1 Hz, 2H), 4.04 (m, 2H), 3. 58 (d, J = 11.2 Hz, 3H), 2.5 (s, 3H). 2.39(dd, J = 20.4 Hz, J = 20.4 Hz, 2H), 2 .00 - 1.42 (m, 9H). C 18 H 27ESI MS of ClFN5O8P2 [M+H] + , calculated value 558.8, measured value 558.2 。 [Example 85] ((2R,3R,4S,5R)-5-(2-Chloro-6-(cyclopentyl(methyl) amino)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofura n-2-yl)methyl hydrogen ((dimethoxyphosphoryl)methyl)phosphonate synthesis, and and methyl hydrogen ((((2R,3R,4S,5R)-5-(2-chloro-6-(cyclopen tyl(methyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxytetra hydrofuran-2-yl)methoxy)(methoxy)phosphoryl)methyl)phosphona to synthesis

Chemical Structure

[0307] Step a: Sodium iodide (40 mg, 0.26 mmol) was added to a solution of the product from Step a of Example 83 (150 mg, 0. 26 mmol) in acetone (3 mL). This solution was stirred at room temperature for 24 hours. The solvent was evaporated, and the residue was dissolved in water and purified by reverse-phase HPLC (C18 column, 0 - 30% gradient of acetonitrile and water containing 0.1% TFA) to obtain ((2R,3R,4S,5R)-5-(2-chloro-6-(cyclopentyl( methyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydro furan-2-yl)methyl hydrogen ((dimethoxyphosphoryl)methyl)phosphonate as a white solid in a yield of 20% (35 mg). 1 H NMR (400 MHz, DMSO-d6) δ 8.3 1 (d, J = 2.2 Hz, 1H), 6.41 (dd, J = 14.9,4.5 Hz, 1H), 5.25 (dt, J = 52.3, 4.1 Hz, 1H), 4.53-4.43 (m, 1H), 4.24 - 4.12 (m,2H), 4.08-4.02 (m, 1H), 3.66 (d, J = 2.0 Hz, 3H), 3.63 (d, J = 2.0 Hz, 3H),2.60 (dd, J = 20.8 Hz, J = 20.8 Hz, 2H), 2.50 (s, 3H), 2.01 - 1.55 (m, 9H). C 19 H 29 ESI MS [M+H] of ClF1N5O8P2 + , calculated value 572 .9, measured value 572.3.

[0308] Methyl hydrogen ((((2R,3R,4S,5R)-5-(2-chloro-6-(cyclopentyl(methyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxy tetrahydrofuran-2-yl)methoxy)(methoxy)phosphoryl)methyl)phosphonate was obtained as a white solid as a 1:1 mixture of diastereoisomers in a 30% yield (52 mg). 1 st nd HNMR (400 MHz, DMSO-d6) δ 8.34 (d, J = 2.3 Hz, 0.5H, 1 8.30 (d, J = 2.3 Hz, 0.5H, 2 nd dia), 6.54 - 6.32 (m, 1H), 5.38 - 5.11 (m, 1H), 4 .59 - 4.39 (m, 1H), 4.26 (m, 2H), 4.07 (m,1H), 3.64 (d, J = 11.3 Hz, 3H), 3.59 (d, J = 11.2, 1.5H, 1 stdia), 3.59 (d, J = 11.2, 1.5H, 1 st dia), 2.69 - 2.53 (m, 2 H), 2.5 (s, 3H), 1.97 - 1.52 (m, 9H). C 19 H 29 ESI MS of ClFN5O8P2 [M+H] + , calculated value 5 72.9, measured value 572.2. [Example 86] ((((2R,3R,4S,5R)-5-(2-chloro-6-(cyclopentyl(meth yl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydro furan-2-yl)methoxy)(methoxy)phosphoryl)methyl)phosphonic acid synthesis [Chemical formula]

[0309] This compound was obtained as a 1:1 mixture of diastereoisomers in the same manner as in Example 84. obtained. 1 1H NMR (400 MHz, DMSO-d6) δ 8.34 (d, J = 2.3 Hz, 0.5H, 1 st dia), 8.29 (d, J = 2.3 Hz, 0.5H, 2 nd dia), 6.52 - 6.32 (m, 1H), 6.07 (brs, 1H), 5.34 - 5.1 4 (m, 1H), 4.56 - 4.43 (m, 1H), 4.30 - 4.21 (m, 2H), 4.11 - 4.03 (m, 1H), 3.63 ( d, J = 11.2 Hz, 1.5H, 1 st dia), 3.63 (d, J = 11.2 Hz, 1.5H, 2 nd dia), 2.50 (s, 3 H), 2.48 - 2.34 (m, 2H), 1.92 - 1.53 (m, 9H). C 18 H 27 ESIMS of ClFN5O8P2 [M+H] + , Calculated value 558.3, measured value 558.2. [Example 87] [({[(2R,3S,4R,5R)-5-[6-Chloro-4-(cyclopentylamino )-1H-pyrazolo[3,4-d]pyrimidin-1-yl]dihydroxyoxolan- 2-yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid synthesis

Chemical formula

[0310] Step a: 4,6-Dichloro-1H-pyrazolo[3,4-d]pyrimidine (25 g, 13 2 mmol) and ammonium sulfate (0.20 g, 1.5 mmol) were dissolved in 150 mL of he xamethyldisilazane. Next, the mixture was heated to reflux and stirred for 3 hours. Next, the mi xture was concentrated to dryness. Next, the solid residue was taken up in 300 mL of acetonitrile, and protected ribose (50.6 g, 159 mmol) was added. The mixture was cooled to 0 °C, and TMS OTf (27 mL, 145 mmol) was added dropwise. Next, the mixture was warmed to room temperature and stirred overnight. Next, the mixture was concentrated and taken up in ethyl acetate. The organic layer was washed with saturated NaHCO3 and brine . The organic layer was dried over MgSO4, filtered, and concentrated. The crude residue was purified by column chromatography (hexane / ethyl acetate) to give the desired compound (48 g , 108 mmol) in an overall yield of 82%. 1 H NMR (400 MHz, DMSO-d6) δ 8.75 (s, 1H), 6.47 (d, J = 3.2 Hz, 1H), 5.82 (dd, J = 5.3, 3.2 Hz, 1H), 5.63 (t, J = 5.8 Hz, 1H), 4.47 - 4.40 (m, 1H), 4.37 - 4.30(m, 1H), 4.12 - 4.02 (m, 1H), 2.09 (s, 3H), 2.06 (s, 3H), 1.97 (s, 3H). C 16 H 16 ESI MS [M+H] of C + H Cl2N4NaO7 + , calculated value 469.0, measured value 469.0.

[0311] Step b: The product from step a (22 g, 49.3 mmol) was dissolved in MeOH (100 mL) and cooled to 0 °C. Cyclopentylamine (5.1 g, 51.8 mmol, 1. 05 eq) and triethylamine (7.2 mL, 51.8 mmol, 1.05 eq) were added and the reaction mixture was stirred at 0 °C for 15 min and then at room temperature for 4 h. 7M NH3 in MeOH (60 mL) was added and the reaction solution was stirred at room temperature for 1 day. The reaction mixture × solvent was distilled off and the crude product was used in the next step without purification. C 15 H 21 ESI MS [M+H] of C + lN5O4 , calculated value 370.1, measured value 370.2.

[0312] 1 H NMR (400 MHz, DMS O-d6) δ 8.68 (d, J = 7.2 Hz,1H), 8.24 (s, 1H), 6.00 (d, J = 4.2 Hz, 1H), 4.49 (t, J = 4.7 Hz, 1H), 4.41 (q, J = 6.7 Hz,1H), 4.26 (t, J = 4.7 Hz, 1H), 4.15 - 4.00 (m, 2H), 3.94 - 3.84 (m, 1H), 2.16 (t,J = 20.5 Hz, 2H), 2.04 - 1.91 (m, 2H ), 1.79 - 1.45 (m, 6H). C 16 H 25 ESI MS of C 16 H 25 ClN5O9P2 [M+H] + , calculated value 528.1, measured value 5 28.2。 [Example 88] [({[(2R,3S,4R,5R)-5-[4-(benzylamino)-6-chloro- 1H-pyrazolo[3,4-d]pyrimidin-1-yl]-3-,4-dihydroxyoxo ran-2-yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid synthesis [Chemical formula]

[0313] The title compound was synthesized in the same manner as in Example 87. 1 1H NMR (400 MHz, DMSO-d6) δ 9.3 8 - 9.18 (m, 1H), 8.35 - 8.16 (m, 1H), 7.39- 7.19 (m, 5H), 6.07 - 5.94 (m, 1H), 4.69 (d, J = 5.4 Hz, 2H), 4.58 - 4.44 (m,1H), 4.30 - 4.20 (m, 1H), 4.15 - 4.01 (m, 2H), 3.96 - 3.80 (m, 1H), 2.17 (t, J =20.9 Hz, 2H). C 18 H 22 ESI M S of C 18 H 22 ClN5O9P2 [M-H] -, Calculated value 548.1, measured value 548.1. [Example 89] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1S)-1-phenyl ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3, 4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl)methyl Synthesis of phosphonic acid

Chemical Structure

[0314] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 9.2 6 - 8.95 (m, 1H), 8.35 - 8.17 (m, 1H), 7.48 - 7.28 (m, 4H), 7.28 - 7.09 (m, 1H), 6.09 - 5.87 (m, 1H), 5.42 (q, J = 6.9 Hz, 1H), 4.60 - 4.33 (m, 1H), 4.33 - 4.16 (m, 1H), 4.13 - 3.96 (m, 2H), 3.97 - 3.80 (m, 1H), 2.35 - 1.95 (m, 2H), 1.62 - 1.36 (m, 3H). C 19 H 24 ESI MS [M-H] of C19H18ClN5O9P2 - , Calculated value 562.1, measured value 562.2. [Example 90] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1R)-1-phenyl ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3, 4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl)methyl Synthesis of phosphonic acid [Chemical formula]

[0315] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 9.1 6 (d, J = 8.4 Hz, 1H), 8.32 (s, 1H), 7.48 - 7.30 (m, 4H), 7.28 - 7.15 (m, 1H), 6 .09 - 5.79 (m, 1H), 5.47 - 5.36 (m, 1H), 4.58 - 4.42 (m, 1H), 4.32 - 4.19 (m, 1H ), 4.17 - 3.95 (m, 2H), 3.95 - 3.79 (m, 1H), 2.18 (t, J = 20.8 Hz, 2H), 1.71 - 1 .37 (m, 4H). C 19 H 24 ESI MS of HClN5O9P2 [M-H] - , calculated value 562.1, measured value 562.2. [Example 91] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(4-chlorophen nyl)methyl]amino}-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3, 4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl)methyl Synthesis of phosphonic acid [Chemical formula]

[0316] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 9.4 1 - 9.19 (m, 1H), 8.32 - 8.17 (m, 1H), 7.43 - 7.30 (m, 4H), 6.02 (d, J = 2.9 Hz, 1H), 4.68 (d, J = 4.4 Hz, 2H), 4.56 - 4.45 (m, 1H), 4.33 - 4.18 (m, 1H), 4.13 - 3.80 (m, 2H), 3.62 - 3.44 (m, 1H), 2.17 (t, J = 20.4 Hz, 1H). C 18 H 21 Cl2N5O9P2 of ESI MS [M-H] - , calculated value 582.0, measured value 582.0. [Example 92] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1S)-1-( 2-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)pho sphoryl)methyl]phosphonic acid synthesis

Chemical Structure

[0317] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 9.2 8 - 9.15 (m, 1H), 8.33 (dd, J = 1.5, 0.7 Hz, 1H), 7.43 (t, J = 7.8 Hz, 1H), 7.29 (dd, J = 7.8, 5.6 Hz, 1H), 7.23 - 7.08 (m, 2H), 6.00 (d, J = 4.2 Hz, 1H), 5.65 - 5.51 (m, 1H), 4.48 (t, J = 4.9 Hz, 1H), 4.26 (t, J = 4.5 Hz, 1H), 4.05 (dq, J = 10.1, 5.9, 5.2 Hz, 2H), 3.88 (dt, J = 11.3, 6.0 Hz, 1H), 2.29 - 2.08 (t, J = 2 0.4 Hz, 2H), 1.53 (d, J = 6.8 Hz, 3H). C 19 H 24 ESI MS of ClFN5O9P2 [M+H] + , calculated value 582.1, measured value 582.1 [Example 93] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1R)-1-( 2-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)pho sphoryl)methyl]phosphonic acid synthesis

Chemical Structure

[0318] The title compound was synthesized in the same manner as in Example 87. 1 1H NMR (400 MHz, DMSO-d6) δ 9.2 3 (d, J = 7.6 Hz, 1H), 8.34 (s, 1H), 7.44 (t, J = 7.8 Hz, 1H), 7.30 (q, J = 7.0 Hz, 1H), 7.18 (dt, J = 9.4, 6.4 Hz, 2H), 6.00 (d, J = 4.3 Hz, 1H), 5.60 (q, J = 7.1 Hz, 1H), 4.51 (t, J = 4.6 Hz, 1H), 4.26 (t, J = 4.6 Hz, 1H), 4.05 (tt, J = 1 0.1, 5.8 Hz, 2H), 3.88 (dd, J = 11.0, 6.2 Hz, 1H), 2.17 (t, J = 20.4 Hz, 2H), 1. 53 (d, J = 6.7 Hz, 3H). C 19 H 24 ESI MS of ClFN5O9P2 [M+H] + , calculated value 582.1, measured value 5 82.1。 [Example 94] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1S)-1-( 3-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)pho sphoryl)methyl]phosphonic acid synthesis

Chemical Structure

[0319] The title compound was synthesized in the same manner as in Example 87. 1 1H NMR (400 MHz, DMSO-d6) δ 9.1 7 (d, J = 7.9 Hz, 1H), 8.31 (s, 1H), 7.50 - 7.30 (m, 1H), 7.22 (d, J = 8.2 Hz, 2 H), 7.06 (td, J = 8.7, 2.5 Hz, 1H), 6.00 (d, J = 4.2 Hz, 1H), 5.41 (t, J = 7.3 H z, 1H), 4.48 (t, J = 4.7 Hz, 1H), 4.26 (t, J = 4.8 Hz, 1H), 4.05 (dq, J = 11.7, 6.5 Hz, 2H), 3.88 (dt, J = 11.2, 6.2 Hz, 1H), 2.17 (t, J = 20.5 Hz, 2H), 1.53 (d , J = 7.0 Hz, 3H). C 19 H 24 ESI MS of ClFN5O9P2 [M+H] + , calculated value 582.1, measured value 582.1 。 [Example 95] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1R)-1-( 3-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)pho sphoryl)methyl]phosphonic acid synthesis [Chemical Structure Diagram]

[0320] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 9.1 8 (d, J = 7.9 Hz, 1H), 8.31 (t, J = 0.9 Hz,1H), 7.43 - 7.32 (m, 1H), 7.23 (d, J = 8.8 Hz, 2H), 7.07 (t, J = 8.6 Hz, 1H),6.00 (d, J = 4.3 Hz, 1H), 5.42 (t, J = 7.3 Hz, 1H), 4.51 (t, J = 4.5 Hz, 1H),4.26 (t, J = 4.7 Hz, 1H), 4.11 - 3.98 (m , 2H), 3.88 (t, J = 8.6 Hz, 1H), 2.17 (t, J= 20.5 Hz, 2H), 1.52 (d, J = 7.0 Hz, 3H). C 19 H 24 ESI MS of C14H15ClFN5O9P2 [M+H] + , calculated value 582.1, measured value 582.1. [Example 96] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1S)-1-( 4-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin- {[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1R)-1-( Synthesis of {[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1R)-1-(

Chemical formula

[0321] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 9.1 6 (d, J = 7.9 Hz, 1H), 8.30 (d, J = 1.2 Hz,1H), 7.42 (dd, J = 8.4, 5.4 Hz, 2H), 7.15 (td, J = 8.9, 1.2 Hz, 2H), 6.00 (d, J= 4.2 Hz, 1H), 5.40 (t, J = 7.3 Hz, 1H), 4.48 (t, J = 4.8 Hz, 1H), 4.25 (t, J =4.5 Hz, 1H), 4.18 - 3.95 (m, 2H), 3. 95 - 3.82 (m, 1H), 2.16 (t, J = 20.4 Hz,2H), 1.52 (d, J = 7.2 Hz, 3H). C 19 H 24 C ESIMS of lFN5O9P2 [M+H] + , calculated value 582.1, measured value 582.1. [Example 97] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1R)-1-( 4-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)pho sphoryl)methyl]phosphonic acid synthesis

Chemical formula

[0322] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 9.1 6 (d, J = 7.9 Hz, 1H), 8.30 (t, J = 0.9 Hz,1H), 7.42 (dt, J = 6.1, 3.2 Hz, 2H), 7.23 - 7.08 (m, 2H), 6.00 (d, J = 4.3 Hz,1H), 5.40 (t, J = 7.2 Hz, 1H), 4.50 ( t, J = 4.5 Hz, 1H), 4.26 (t, J = 4.7 Hz,1H), 4.15 - 3.98 (m, 2H), 3.87 (q, J = 8.1, 5.5 Hz, 1H), 2.16 (t, J = 20.4 Hz,2H), 1.52 (d, J = 6.9 Hz, 3H). C 19 H 24 Cl ESIMS of FN5O9P2 [M+H] + , calculated value 582.1, measured value 582.1. [Example 98] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(2-chlorophen yl)methyl]amino}-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3, 4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl)methyl phosphonic acid synthesis

Chemical formula

[0323] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 9.2 9 (s, 1H), 8.29 (d, J = 1.9 Hz, 1H), 7.48(dd, J = 5.9, 3.1 Hz, 1H), 7.43 (d, J = 5.9 Hz, 1H), 7.32 (dt, J = 6.6, 2.5 Hz,2H), 6.07 - 6.00 (m, 1H), 4.84 - 4.69 (m, 2H), 4.51 (d, J = 5.0 Hz, 1H), 4.27 (s,1H), 4.06 (s, 2H), 3.89 (s, 1H), 2.1 6 (t, J = 20.6 Hz, 2H). C 18 H 22 ESI MS of Cl2N5O9P2 [M+H] + , calculated value 584.0, measured value 5 84.1。 [Example 99] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(2-chlorophen yl)methyl](methyl)amino}-1H-pyrazolo[3,4-d]pyrimidin-1-yl )-3,4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)phosphori yl)methyl]phosphonic acid synthesis

Chemical Structure

[0324] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 8.4 1 (s, 1H), 7.49 (d, J = 7.4 Hz, 1H), 7.38 -7.24 (m, 2H), 7.16 (d, J = 7.6 Hz, 1 H), 6.08 (bs, 1H), 5.04 (bs, 2H), 4.50 (d,J = 30.8 Hz, 1H), 4.24 (d, J = 39.5 H z, 1H), 4.06 (s, 2H), 3.89 (s, 1H), 3.37(d, J = 54.8 Hz, 3H), 2.15 (t, J = 20.8 Hz, 2H). C19 H 24 ESI MS of Cl2N5O9P2 [M+H] + , calculated value 598.0, measured value 598.1. [Example 100] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[2-(2-chloro phenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin-1-yl)- 3,4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl)meth Synthesis of til phosphonic acid

Chemical Structure

[0325] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 8.9 5 (t, J = 5.6 Hz, 1H), 8.28 - 8.10 (m, 1H),7.48 - 7.39 (m, 1H), 7.33 (d, J = 6. 4 Hz, 1H), 7.29 - 7.18 (m, 2H), 6.00 (d, J= 4.0 Hz, 1H), 4.50 (t, J = 4.5 Hz, 1 H), 4.25 (t, J = 4.5 Hz, 1H), 4.06 (d, J =14.9 Hz, 2H), 3.87 (t, J = 5.8 Hz, 1H ), 3.69 (q, J = 6.8 Hz, 2H), 3.10 - 3.00(m, 2H), 2.15 (t, J = 20.4 Hz, 2H). C1 9H 24 ESI MS of Cl2N5O9P2 [M+H] + , calculated value 598.0, measured value 598.2. [Example 101] [({[(2R,3S,4R,5R)-5-{4-[benzyl(methyl)amino]-6 -Chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl}3,4-dihydroxy oxolan-2-yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid synthesis Synthesis

Chemical formula

[0326] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 8.3 5 (s, 1H), 7.30 (dd, J = 20.4, 7.5 Hz, 5H), 6.07 (bs, 1H), 4.99 (bs, 1H), 4.53 ( bs, 1H), 4.28 (bs, 1H), 4.05 (s, 3H), 3.88 (s, 1H), 3.37 - 3.24 (m, 3H), 2.14 (t , J = 20.9 Hz, 3H). C 19 H 25 ESI MS [M+H] of C + H ClN5O9P2, calculated value 564.1, measured value 564.1 [Example 102] [({[(2R,3S,4R,5R)-5-{6-Chloro-4-[cyclopentyl(methyl amino]-1H-pyrazolo[3,4-d]pyrimidin-1-yl}3,4-dihydro xyoxolan-2-yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid synthesis

Chemical formula

[0327] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 8.3 1 (s, 1H), 6.06 (s, 1H), 4.51 (d, J = 4.7Hz, 1H), 4.28 (d, J = 5.1 Hz, 1H), 4.1 4 - 3.96 (m, 2H), 3.88 (s, 1H), 3.21 (s,3H), 2.14 (t, J = 19.9 Hz, 2H), 1.67 (b s, 8H). C 17 H 27 ESI MS of C 17 H 27 ClN5O9P2 [M+H] + , calculated value 542.1, measured value 542.2. [Example 103] [({[(2R,3S,4R,5R)-5-[6-chloro-4-(methylamino)-1 H-pyrazolo[3,4-d]pyrimidin-1-yl]-3,4-dihydroxyoxolan -2-yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid synthesis [Chemical formula]

[0328] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 8.8 2 (s, 1H), 8.16 (s, 1H), 6.01 (bs, 1H),4.50 (d, J = 5.9 Hz, 1H), 4.26 (bs, 1H), 4.05 (bs, 2H), 3.88 (bs, 1H), 2.95 (d, J =4.6 Hz, 3H), 2.15 (t, J = 20.4 Hz, 2 H). C 12 H 19 ESI MS of C 12 H 19 ClN5O9P2 [M+H] + , calculated value 474.0, measured value 474.2. [Example 104] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1R)-2,2 ,2-Trifluoro-1-phenylethyl]amino}-1H-pyrazolo[3,4-d]pyr rimidin-1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy}(hydro xy)phosphoryl)methyl]phosphonic acid synthesis

Chemical Structure

[0329] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 9.8 2 (d, J = 9.3 Hz, 1H), 8.51 (s, 1H), 7.65(d, J = 7.2 Hz, 2H), 7.56 - 7.31 (m, 3 H), 6.33 (p, J = 8.8 Hz, 1H), 6.04 (d, J =4.2 Hz, 1H), 4.50 (t, J = 4.4 Hz, 1H) , 4.26 (t, J = 4.6 Hz, 1H), 4.16 - 4.00 (m,2H), 3.90 (dd, J = 10.6, 5.8 Hz, 1H) , 2.18 (t, J = 20.5 Hz, 2H). C 19 H 21 ESI MS of ClF3N5O9P2 [M-H] - , calculated value 616.1, actual measurement 616.2. [Example 105] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(3S)-oxo lan-3-yl]amino}-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3 ,4-dihydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl)methy l]phosphonic acid synthesis

Chemical Structure

[0330] The title compound was synthesized in the same manner as in Example 87. 1 H NMR (400 MHz, DMSO-d6) δ 8.9 5 (d, J = 6.4 Hz, 1H), 8.26 (s, 1H), 6.01(d, J = 4.2 Hz, 1H), 4.69 - 4.59 (m, 1 H), 4.50 (t, J = 4.2 Hz, 1H), 4.26 (t, J =4.5 Hz, 1H), 4.15 - 3.99 (m, 2H), 3.9 5 - 3.81 (m, 3H), 3.74 (d, J = 7.9 Hz, 1H),3.67 - 3.58 (m, 1H), 2.35 - 2.06 (m, 3H), 1.98 - 1.80 (m, 1H). C 15 H 22 ClN5O 10 ESI MS of P2 [M-H] - , calculated value 528.1, found value 528.2. [Example 106] [({[(2R,3S,4R,5R)-5-[6-chloro-4-(cyclopentylamino no)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]-3,4-dihydroxyoxo xolan-2-yl]methoxy}({[(propan-2-yloxy)carbonyl]oxo xy}methoxy)phosphoryl)methyl]({[(propan-2-yloxy)carbonyl] oxy}methoxy)phosphinic acid synthesis [Chemical formula]

[0331] The title compound was synthesized in the same manner as in Example 69. 1 H NMR (400 MHz, DMSO-d6) δ 8.7 1 - 8.60 (m, 1H), 8.26 - 8.15 (m, 1H), 6.02 - 5.96 (m, 1H), 5.60 - 5.38 (m, 5H), 4.87 - 4.68 (m, 2H), 4.51 - 4.37 (m, 2H), 4.33 - 3.79 (m, 5H), 2.74 - 2.53 (m, 2H), 2.07 - 1.89 (m, 2H), 1.79 - 1.42 (m, 7H), 1.27 - 1.12 (m, 12H). C 26 H 40 ClN5O 15 ESI MS of P2 [M-H] - , calculated value 758.2, measured value 758.3. [Example 107] [({[(2R,3S,4R,5R)-5-(6-chloro-4-{[(1S)-1-phenyl ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3, 4-dihydroxyoxolan-2-yl]methoxy}({[(propan-2-yloxy )carbonyl]oxy}-methoxy)phosphoryl)methyl]({[(propan-2-yl oxy)carbonyl]oxy}methoxy)phosphinic acid synthesis [Chemical formula]

[0332] The title compound was synthesized in the same manner as in Example 69. 1 H NMR (400 MHz, DMSO-d6) δ 9.1 7 (d, J = 7.9 Hz, 1H), 8.32 - 8.27 (m, 1H), 7.42 - 7.28 (m, 4H), 7.26 - 7.20 (m, 1H), 6.01 (d, J = 3.7 Hz, 1H), 5.58 - 5.32 (m, 6H), 4.84 - 4.69 (m, 2H), 4.49 - 4.37 (m, 1H), 4.37 - 3.81 (m, 5H), 2.72 -2.52 (m, 1H), 1.53 (d, J = 6.9 Hz, 3H ), 1.28 - 1.17 (m, 12H). C 29 H 40 ClNO 15 ESI MS of P2 [MH] - , Calculated value 794.2, Measured value 7 94.2. [Example 108] [({[(2R,3R,4S,5R)-5-[4-(benzylamino)-6-chloro- 1H-Pyrazolo[3,4-d]pyrimidin-1-yl]-4-fluoro-3-hydroxy Oxolan-2-yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid Growth [ka]

[0333] Step a: 4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidine (1.0 g, 5 47.3 mmol) was dissolved in anhydrous CH3CN (10 mL) and cyclopentylamine (478.3 mmol) was added. mg, 5.6 mmol, 1.05 equiv.) was added, followed by TEA (779 μL, 5.6 mmol). The mixture was stirred at room temperature overnight and then anhydrous Cs2CO3 (3.4 g, 10.6 mmol, 2 eq.) and bromide (2.2 g, 5.3 mmol). The reaction mixture was stirred at room temperature overnight, then the solvent was evaporated. The crude residue was dissolved in MeOH (20 mL). The mixture was dissolved and anhydrous K2CO3 (2.2 g, 15.9 mmol, 3 equiv.) was added. The mixture was stirred at room temperature overnight, evaporated onto silica gel, and purified by column chromatography (SiO2, He x→100% EtOAc) to initially give product B (800 mg, 41% ) Next, product A (600 mg, 30%) was obtained. For B: C 15 H 20 ESI MS of ClFN5O3 [M+H] + , calculated value 372.1, measured value 372.2.

[0334] Step b: Using product B from step a, a phosphonylation step was carried out in the same manner as in Example 1 as follows: 1 1H NMR (400 MHz, DMSO-d6) δ 8.76 (d, J = 7.2 Hz, 1H), 8.29 (s, 1H), 6.52 (d, J = 6.5 Hz, 1H), 5.50 - 5.29 (m,1H), 4.75 (dt, J = 18.7, 7.5 Hz, 1H), 4.43 (h, J = 6.9 Hz, 1H), 4.31 - 4.22 (m,1H), 4.18 - 4.05 (m, 1H), 4.04 - 3.92 (m, 1H), 2.20 (t, J = 20.5 Hz, 2H), 2.05 -1.93 (m, 2H), 1.80 - 1.46 (m, 6H). C 16 H 24 ESI MS of ClFN5O8P2 [M+H] + , calculated value 530.1, measured value 530.2. [Example 109] [({[(2R,3R,4S,5R)-5-(6-chloro-4-{[(1S)-1-phenylethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-4- fluoro-3-hydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl )methyl]phosphonic acid synthesis ) [Chemical Structure Diagram]

[0335] The title compound was synthesized in the same manner as in Example 108. 1 H NMR (400 MHz, DMSO-d6) δ 9 .19 (d, J = 8.1 Hz, 1H), 8.33 (s, 1H), 7.40(d, J = 7.9 Hz, 2H), 7.33 (t, J = 7. 5 Hz, 2H), 7.23 (t, J = 7.4 Hz, 1H), ),6.34 (dd, J = 14.3, 4.6 Hz, 1H), 5.39 (b s, 1H), 5.31 - 5.12 (m, 1H), 5.14 (bs, 1H),4.48 (dt, J = 18.5, 4.5 Hz, 1H), 4.1 7 (s, 3H), 4.01 (d, J = 5.2 Hz, 2H), 2.24(t, J = 20.4 Hz, 3H), 1.51 (d, J = 7.0 Hz, 3H). C 19 H 24 ESI MS of ClFN5O8P2 [M+H] + , calculated value 566.1, measured value 566.1. [Example 110] [({[(2R,3R,4S,5R)-5-(6-chloro-4-{[(1R)-1-phenyl ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-4- fluoro-3-hydroxyoxolan-2-yl]methoxy}(hydroxy)phosphoryl )methyl]phosphonic acid synthesis

Chemical Structure

[0336] The title compound was synthesized in the same manner as in Example 108. 1 H NMR (400 MHz, DMSO-d6) δ 9 .18 (d, J = 8.0 Hz, 1H), 8.33 (d, J = 1.2Hz, 1H), 7.40 (d, J = 7.9 Hz, 2H), 7.3 3 (t, J = 7.3 Hz, 2H), 7.24 (t, J = 7.6 Hz,1H), 6.50 (d, J = 6.5 Hz, 1H), 5.51 - 5.23 (m, 2H), 4.82 - 4.66 (m, 1H), 4.22(bs, 1H), 4.13 - 4.02 (m, 1H), 3.94 (b s, 1H), 2.17 (t, J = 20.5 Hz, 2H), 1.53 (d,J = 7.1 Hz, 3H). C 19 H 24 of ClFN5O8P2 ESI MS [M+H] + , calculated value 566.1, measured value 566.2. [Example 111] [({[(2R,3R,4S,5R)-5-(6-chloro-4-{[(1S)-1-( 2-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)-4-fluoro-3-hydroxyoxolan-2-yl]methoxy}(hydro xy)phosphoryl)methyl]phosphonic acid synthesis [Chemical formula]

[0337] The title compound was synthesized in the same manner as in Example 108. 1 H NMR (400 MHz, DMSO-d6) δ 9 .27 (d, J = 7.6 Hz, 1H), 8.36 (s, 1H), 7.48 - 7.40 (m, 1H), 7.37 - 7.25 (m, 1H), 7.22 - 7.13 (m, 2H), 6.51 (d, J = 6.6 Hz,1H), 5.59 (p, J = 7.1 Hz, 1H), 5.49 - 5.26 (m, 1H), 4.74 (dt, J = 18.4, 7.6 Hz,1H), 4.30 - 4.17 (m, 1H), 4.15 - 4.02 (m, 1H), 3.99 - 3.90 (m, 1H), 2.17 (t, J =20.5 Hz, 2H), 1.54 (d, J = 7.0 Hz, 3 H). C 19 H 23 ESI MS of ClF2N5O8P2 [M+H] + , calculated value 584.1, measured value 584.2. [Example 112] [({[(2R,3R,4S,5R)-5-(6-chloro-4-{[(1R)-1-( 2-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)-4-fluoro-3-hydroxyoxolan-2-yl]methoxy}(hydro (xy)phosphoryl)methyl]phosphonic acid synthesis

Chemical Structure

[0338] The title compound was synthesized in the same manner as in Example 108. 1 H NMR (400 MHz, DMSO-d6) δ 9 .37 - 9.16 (m, 1H), 8.36 (d, J = 3.4 Hz,1H), 7.44 (d, J = 8.4 Hz, 1H), 7.29 (t, J = 7.0 Hz, 1H), 7.18 (dt, J = 10.6, 5.6Hz, 2H), 6.51 (t, J = 4.8 Hz, 1H), 5.6 0 (t, J = 6.8 Hz, 1H), 5.53 - 5.22 (m, 1H),4.84 - 4.64 (m, 1H), 4.31 - 4.16 (m, 1H), 4.16 - 4.00 (m, 1H), 3.94 (p, J = 3.7Hz, 1H), 2.18 (t, J = 20.4 Hz, 2H), 1.54 (d, J = 7.2 Hz, 3H). C 19 H 23 ESI MS of ClF2N5O8P2 [M+H] + , calculated value 584.1, measured value 584.2. [Example 113] [({[(2R,3R,4S,5R)-5-(6-chloro-4-{[(1S)-1-( 3-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)-4-fluoro-3-hydroxyoxolan-2-yl]methoxy}(hydro xy)phosphoryl)methyl]phosphonic acid synthesis

Chemical Structure

[0339] The title compound was synthesized in the same manner as in Example 108. 1 H NMR (400 MHz, DMSO-d6) δ 9 .21 (d, J = 7.8 Hz, 1H), 8.33 (s, 1H), 7.46 - 7.30 (m, 1H), 7.27 - 7.16 (m, 2H), 7.12 - 7.00 (m, 1H), 6.51 (d, J = 6.5 Hz,1H), 5.49 - 5.26 (m, 2H), 4.74 (dt, J = 18.4, 7.6 Hz, 1H), 4.29 - 4.18 (m, 1H),4.14 - 4.02 (m, 1H), 4.00 - 3.89 (m, 1H), 2.17 (t, J = 20.5 Hz, 2H), 1.54 (d, J= 7.0 Hz, 3H). C 19 H 23 ESI MS of ClF2N5O8P2 SI MS [M+H] + , calculated value 584.1, measured value 584.2. [Example 114] [({[(2R,3R,4S,5R)-5-(6-chloro-4-{[(1R)-1-( 2-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)-4-fluoro-3-hydroxyoxolan-2-yl]methoxy}(hydro xy)phosphoryl)methyl]phosphonic acid synthesis

Chemical Structure

[0340] The title compound was synthesized in the same manner as in Example 108. 1 H NMR (400 MHz, DMSO-d6) δ 9 .21 (d, J = 8.0 Hz, 1H), 8.34 (s, 1H), 7.42 - 7.32 (m, 1H), 7.27 - 7.20 (m, 2H), 7.12 - 7.03 (m, 1H), 6.51 (d, J = 6.5 Hz,1H), 5.49 - 5.29 (m, 2H), 4.74 (dt, J = 18.8, 7.8 Hz, 1H), 4.29 - 4.18 (m, 1H),4.15 - 4.03 (m, 1H), 3.99 - 3.90 (m, 1H), 2.17 (t, J = 20.5 Hz, 2H), 1.54 (d, J= 7.0 Hz, 3H). C 19 H 23 E of ClF2N5O8P2 SI MS [M+H] + , calculated value 584.1, measured value 584.2. [Example 115] [({[(2R,3R,4S,5R)-5-(6-chloro-4-{[(1S)-1-( 4-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidin- 1-yl)-4-fluoro-3-hydroxyoxolan-2-yl]methoxy}(hydro xy)phosphoryl)methyl]phosphonic acid synthesis

Chemical formula

[0341] The title compound was synthesized in the same manner as in Example 108. 1 H NMR (400 MHz, DMSO-d6) δ 9 .22 (d, J = 7.9 Hz, 1H), 8.34 (s, 1H), 7.45(dd, J = 8.7, 5.6 Hz, 2H), 7.17 (t, J = 8.9 Hz, 2H), 6.52 (d, J = 6.5 Hz, 1H),5.53 - 5.27 (m, 2H), 4.75 (dt, J = 18 .7, 7.6 Hz, 1H), 4.31 - 4.20 (m, 1H), 4.17- 4.04 (m, 1H), 3.97 (dd, J = 7.5, 3. 9 Hz, 1H), 2.19 (t, J = 20.5 Hz, 2H), 1.54(d, J = 7.0 Hz, 3H). C 19 H 23 ClF2N5O8P ESIMS [M+H] of 2 + , calculated value 584.1, measured value 584.2. [Example 116] [({[(2R,3R,4S,5R)-5-(6-chloro-4-{[(1R)-1-( 4-fluorophenyl)ethyl]amino}-1H-pyrazolo[3,4-d]pyrimidine- 1-yl)-4-fluoro-3-hydroxyoxolan-2-yl]methoxy}(hydro xy)phosphoryl)methyl]phosphonic acid synthesis

Chemical formula

[0342] The title compound was synthesized in the same manner as in Example 108. 1 H NMR (400 MHz, DMSO-d6) δ 9 .21 (d, J = 7.9 Hz, 1H), 8.34 (s, 1H), 7.54 - 7.37 (m, 2H), 7.17 (t, J = 8.8 Hz, 2H), 6.52 (d, J = 6.6 Hz, 1H), 5.60 - 5.23(m, 2H), 4.76 (dt, J = 18.7, 7.6 Hz, 1H), 4.24 (dt, J = 7.4, 4.8 Hz, 1H), 4.09(dt, J = 10.9, 7.4 Hz, 1H), 3.96 (dt, J = 10.7, 5.3 Hz, 1H), 2.19 (t, J = 20.5Hz, 2H), 1.54 (d, J = 7.1 Hz, 3H). C1 9H 23 ESI MS [M+H] of ClF2N5O8P2 + , calculated value 584.1, measured value 584.2. [Example 117] [({[(2R,3S,4R,5R)-5-[5-chloro-7-(cyclopentylamino no)-3H-imidazo[4,5-b]pyridin-3-yl]-3,4-dihydroxyox solan-2-yl]methoxy}(hydroxy)phosphoryl)methyl]phosphonic acid synthesis [Chemical formula]

[0343] Step a: To a solution of 5,7-dichloroimidazo[4,5-b]pyridine (376 mg, 2 mmol) in MeCN (14 mL) at room temperature, N,O-bis(trimethylsilyl)acet amide (0.523 mL, 2.14 mmol) was added dropwise, and the reaction mixture was heated at 85 °C for 1 hour. Heated for a certain time. The mixture was cooled to room temperature, and a solution of beta-D-ribofuranosyl 1,2,3,5-tetraacetate (726 mg, 2.28 mmol) and trimethylsilyl trifluoromethanesulfonate (0.471 mL, 2.60 mmol) in MeCN (7 mL) was added dropwise successively. The reaction mixture was heated at 85 °C for 4 hours. The mixture was cooled, saturated aqueous sodium bicarbonate solution (50 mL) was added, and then extracted with EtOAc (100 mL) three times, dried over sodium sulfate, and concentrated. -ose 1,2,3,5-tetraacetate (726 mg, 2.28 mmol) and trimethylsilyl trifluoromethanesulfonate (0.471 mL, 2.60 mmol) was added dropwise successively. The reaction mixture was heated at 85 °C for 4 hours. The mixture was cooled, saturated aqueous sodium bicarbonate solution (50 mL) was added, and then extracted with EtOAc (100 mL) three times, dried over sodium sulfate, and concentrated. The reaction mixture was heated at 85 °C for 4 hours. The mixture was cooled, saturated aqueous sodium bicarbonate solution (50 mL) was added, and then extracted with EtOAc (100 mL) three times, dried over sodium sulfate, and concentrated. The reaction mixture was heated at 85 °C for 4 hours. The mixture was cooled, saturated aqueous sodium bicarbonate solution (50 mL) was added, and then extracted with EtOAc (100 mL) three times, dried over sodium sulfate, and concentrated. The reaction mixture was heated at 85 °C for 4 hours. The mixture was cooled, saturated aqueous sodium bicarbonate solution (50 mL) was added, and then extracted with EtOAc (100 mL) three times, dried over sodium sulfate, and concentrated.

[0344] Step b: To the residue was added dioxane (2 mL) and cyclopentylamine (0.987 mL, 10 mmol). The mixture was heated at 100 °C for 16 hours. The reaction mixture was loaded onto silica gel, and purified by silica gel chromatography (0 - 10% MeOH in DCM) to obtain the desired product as a brown solid (298 mg, 40%). Step b: To the residue was added dioxane (2 mL) and cyclopentylamine (0.987 mL, 10 mmol). The mixture was heated...

Claims

1. A compound of the formula: 【Chemical 1】 or a pharmaceutically acceptable salt thereof, wherein Each R 1 is hydrogen; R 5 is selected from the group consisting of H and C 1 -C 6 alkyl; X is O; A is 【Chemical Formula 1-1】 selected from the group consisting of, and Het is 【Chemical 1-2】 selected from the group consisting of, wherein the wavy line indicates the point of attachment to the remainder of the compound, and wherein R a is selected from the group consisting of NHR 7 , NR 7 R 7 , and OR 7 ; R c is H, halogen, haloalkyl, NHR 7 , NR 7 R 7 , R 7 , OR 7 , SR 7 , SO 2 R 7 , or -X 1 -OR 7 ; R e is H; X 1 is C 1 to C 4 alkylene; and Each R 7 is C 1 to C 10 alkyl; C 2 to C 10 alkenyl; unsubstituted C 2 to C 10 alkynyl or C2-C10 alkynyl substituted with phenyl; C 3 to C 7 cycloalkyl; unsubstituted C 3 to C 7 cycloalkyl C 1 to C 4 alkyl or C3-C7 cycloalkyl C1-C4 alkyl substituted with OH; 4-7 membered cycloheteroalkyl; unsubstituted 4-7 membered cycloheteroalkyl C 1 to C 4 alkyl or 4-7 membered cycloheteroalkyl C1-C4 alkyl substituted with OH; aryl; unsubstituted aryl C 1 to C 4 alkyl or aryl C1-C4 alkyl substituted with 1 to 3 substituents independently selected from OH, C1-8 alkyl, C1-4 haloalkyl, phenyl and halo; and heteroaryl C 1 to C 4 alkyl, or two R 7 groups bonded to a nitrogen atom are joined together to form a 4-7 membered heterocyclic ring optionally fused to an aryl ring].

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein A is [Chemical Formula 4]

3. A compound of the formula: 【Chemical Formula 6】 or a pharmaceutically acceptable salt thereof, according to claim 1.

4. R c is halogen, R 7 , OR 7 , SR 7 , SO 2 R 7 or -X 1 -OR 7 The compound according to claim 3, or a pharmaceutically acceptable salt thereof, wherein R

5. A compound of the formula: 【Chemical Formula 18】 or a pharmaceutically acceptable salt thereof, according to claim 1.

6. A compound of the formula: 【Chemical Formula 19】 or a pharmaceutically acceptable salt thereof, according to claim 1.

7. 【Fig. 9-1】 【Chemical Formula 9-2】 【Chemical Formula 9-3】 【Chemical Formula 9-4】 【Chemical Formula 9-5】 【Chemical Formula 9-6】 【Chemical Formula 9-7】 【Chemical Formula 9-8】 【Chemical Formula 9-9】 ​ 【Chemical Formula 9-11】 A compound selected from

8. A pharmaceutical composition comprising the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

9. A pharmaceutical composition for treating a disease, disorder, or condition, at least a part of which is mediated by CD73, comprising an effective amount of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof, wherein the disease, disorder, or condition is cancer.

10. The pharmaceutical composition according to claim 9, wherein the compound or a pharmaceutically acceptable salt thereof is administered in an amount effective to reverse or halt the progression of CD73-mediated immunosuppression.

11. The pharmaceutical composition according to claim 9, wherein the cancer is cancer of the prostate, colon, rectum, pancreas, cervix, stomach, endometrium, brain, liver, bladder, ovary, testis, head, neck, skin, leukocyte, esophagus, breast, muscle, connective tissue, lung, adrenal gland, thyroid gland, kidney, or bone.

12. The pharmaceutical composition according to claim 9, wherein the cancer is glioblastoma, mesothelioma, renal cell carcinoma, gastric cancer, sarcoma, choriocarcinoma, basal cell carcinoma of the skin, or testicular seminoma.

13. The pharmaceutical composition according to claim 9, wherein the cancer is selected from the group consisting of melanoma, colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, leukemia, brain tumor, lymphoma, ovarian cancer, and Kaposi's sarcoma.

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