CN110885352A

CN110885352A - CD73 inhibitor and pharmaceutical application thereof

Info

Publication number: CN110885352A
Application number: CN201811057145.XA
Authority: CN
Inventors: 吕佳声; 顾家敏; 吴冬冬; 陈刚; 孙成勇; 吉祥; 王林; 周峰; 张秀春; 孔宪起
Original assignee: Risen Suzhou Pharma Tech Co Ltd
Current assignee: Risen Suzhou Pharma Tech Co Ltd
Priority date: 2018-09-11
Filing date: 2018-09-11
Publication date: 2020-03-17
Anticipated expiration: 2038-09-11
Also published as: JP7192130B2; WO2020051686A1; KR20210068036A; CN110885352B; JP2021527139A; CA3111450A1; CN112771061A; CN112771061B; BR112021004435A2

Abstract

The present invention provides compounds of formula (I) and pharmaceutically acceptable salts and esters thereof, as well as pharmaceutical compositions thereof, the use of the compounds and pharmaceutical compositions thereof for the treatment and/or prevention of CD 73-related diseases, disorders, and conditions, including cancer-related and immune-related disorders.

Description

CD73 inhibitor and pharmaceutical application thereof

Technical Field

The present invention relates to compounds and compositions that inhibit CD73 (extracellular-5' -nucleotidase) and their use for the treatment and/or prevention of CD73 related or associated diseases, disorders, and conditions, including cancer-related and immune-related disorders.

Background

Extracellular nucleotidases are a group of extracellular enzymes located on the cell surface. Members of the extracellular nucleotidase family include extracellular-nucleotide pyrophosphatase/phosphodiesterase (E-NPP), extracellular-nucleoside triphosphate diphosphohydrolases (E-NTPDases), extracellular-5' -nucleotidase (E5NT, also known as CD73), and Alkaline Phosphatase (AP). These enzymes hydrolyze various extracellular nucleotides to nucleosides including adenosine. Extracellular nucleotides are important signaling molecules that trigger cellular responses by acting on their respective receptors (e.g., adenosine activates the P1 receptor, and nucleotides (ADP, ATP) activates the P2 receptor). Adenosine 5' -monophosphate (AMP) is the primary substrate for CD73, and its hydrolysis product is adenosine. Adenosine is ubiquitous in the body and is an important regulator of purinergic cell signaling that is critical to many physiological and pathophysiological processes.

A number of data indicate that CD73 is enzymatically active in cancer development and metastasis CD73 is upregulated in many cancer cell types and tumors, indicating that expression of CD73 is associated with tumor neovascularization, invasion and metastasis the hydrolytic cascade from extracellular ATP to adenosine is an important immunosuppressive regulatory pathway in the tumor microenvironment CD73 overexpresses impairing adaptive anti-tumor immune responses and promotes tumor growth and metastasis extracellular adenosine is also involved in the regulation of adaptive responses to hypoxia it has been shown that e5NT activity is reduced with monoclonal antibodies, sirnas and small molecule inhibitors including AMPCP (adenosine [ (α) -methylene ] diphosphate to attenuate tumor growth and metastasis (see, e.g., Zhou et al, oncol. rep.17(2007):1 1341346; Stagg and Smyth, 2010 cogene,29 (5346): 5358) tumor growth and metastasis is attenuated in CD cheuchi et al 73 deficient mice and that these effects have been demonstrated to be primarily due to the development of cancer as well as a potential inhibitor in these mice (see, e.g. eurein et al, 2016).

Tumor cells overcome the anti-tumor response, in part by immunosuppressive mechanisms. Several such immune-regulatory mechanisms are known; among them, adenosine is a key factor. Adenosine can be produced by cancer cells and immune cells in the tumor microenvironment and used to suppress anti-tumor responses. Adenosine Triphosphate (ATP) is catalyzed by two cell surface proteins, CD73 and CD39, to produce adenosine, and this process is enhanced under conditions of metabolic stress, such as tumor hypoxia. Adenosine exerts its immunomodulatory functions through four adenosine receptors (ARs, referred to as a1, A2A, A2B, and A3, respectively) expressed in various immune cells. Overexpression of adenosine producing enzymes (e.g., CD73 and AR) has been associated with tumor development in a variety of cancer types. Since signaling of ARs increases tumor progression, modulation of this signaling represents a promising therapeutic approach for cancer (m.h. kazemi, et al., j.cell. physiol.,233(2018): 2032-.

As described above, there are four different forms of extracellular nucleotide diphosphohydrolases (CD39), extracellular nucleotide pyrophosphatase/phosphodiesterase, alkaline phosphatase and extracellular-5' -nucleotidase (e5NT, also known as CD73) the family of extracellular nucleotide enzymes, CD73 is a glycosyl phosphatidylinositol-anchored di-zinc metal phosphatase CD73 catalyzes dephosphorylation of extracellular Adenosine Monophosphate (AMP) to generate adenosine, which cooperates with CD39 to form an extracellular enzyme cascade, converting ATP to adenosine, the process of converting AMP to adenosine, catalyzed by CD73, is believed to be the major factor of increased extracellular adenosine levels in tumor cells (Stagg, J.et ah, Natl.Acad.Sci.USA.: 107: 2010: 7) CD 73. the expression of extracellular adenosine levels in tumor cells (Stagg. J.10. 10. nat. J.10. 12. A. C. Acd. Sci. USA.:107, 2010: 37: 11) and the like, and the development of tumor cells with the potential of tumor cells mediated by TNF-mediated hypoxia-mediated tumor cells, tumor proliferation, tumor cells expressing TNF-mediated by TNF-mediated proliferation of the extracellular tumor-mediated proliferation of the TNF-receptor type, TNF-7, the extracellular adenosine receptor, the extracellular tyrosine kinase (TNF-7, the gene of the gene, the gene of the TNF-7, the gene of the tumor, the gene of.

The adenosine pathway has also been considered to be a major immunosuppressive component of many human tumors (see, e.g., Whiteside, t.l., Expert rev.anticancer ther.,17(2017): 527-535). Adenosine and inosine are key immune checkpoints in cancer. The cooperation of adenosine and the PGE2 pathway in the tumor microenvironment contributes to the suppression of anti-tumor immune effector cells. Thus, targeting the adenosine pathway with pharmacological inhibitors or antibodies is a promising therapeutic strategy in cancer.

In preclinical in vivo studies, activity that blocks extracellular nucleotidase or adenosine receptor signaling has been successful in inhibiting tumor growth and metastasis. The use of adenosine pathway blockade alone or in combination with other immunotherapies (including checkpoint inhibitors) is now in the initial phase I clinical trial in patients with advanced malignancies.

Small molecule inhibitors of CD73 have been reported. For example, Adams et al (international PCT application publication WO2017/098421) describe substituted benzothiadiazine derivatives, which are CD73 inhibitors, pharmaceutical compositions thereof, and their use in treating cancer, precancerous syndrome, and diseases associated with CD73 inhibition.

Debien et al (International PCT application publication WO 2017/120508; U.S. patent application publication US2017/0267710) describe compounds that modulate the conversion of AMP to adenosine by extracellular-5 '-nucleotidase, compositions containing the compounds, methods of synthesizing the compounds, and the use of the compounds and compositions for the treatment and/or prevention of various diseases mediated by extracellular-5' -nucleotidase.

Cacataian et al (International PCT application publication WO2015/164573) describe purine derivatives and pharmaceutical compositions thereof, which are inhibitors of CD73 and are useful for the treatment of cancer.

Chen et al (International PCT application publication WO 2018/049145) disclose the preparation of nucleotides as extracellular nucleotidase inhibitors and the use of these compounds in the treatment or prevention of cancer.

The contents of all articles and references cited herein are incorporated by reference in their entirety.

Disclosure of Invention

The present invention relates to compounds and compositions that: which comprises a compound that inhibits the activity of extracellular-5' -nucleotidase (also known as e5NT, CD73, NT5E and 5 NT). Inhibition of CD73 enzymatic activity results in the inhibition or modulation of extracellular adenosine levels, thereby modulating the physiological environment of cells and tissues.

The invention also relates to the use of such compounds and compositions in the treatment and/or prevention of diseases, disorders, and conditions mediated in whole or in part by CD 73. CD73 inhibitors have been used in the treatment of a number of diseases, including cancer, fibrosis, neurological and neurodegenerative diseases (e.g., depression and parkinson's disease), ischemic cardiovascular and cerebrovascular diseases, immune related diseases and inflammatory related diseases. In particular embodiments, the CD73 inhibitor compounds and compositions described herein may act to inhibit the immunosuppressive and/or anti-inflammatory activity of CD73, and may be used as therapeutic agents or prophylactic therapies when such inhibition is desired.

In a first broad aspect, the invention provides compounds of formula I, and pharmaceutically acceptable salts or esters thereof:

wherein W is oxygen, sulfur, nitrogen or methylene; x is selected from phosphoryl (-P (═ O) (OR)-), sulfonyl (-S (═ O)₂-) and a carbonyl (-C (═ O) -) moiety, wherein R is hydrogen, an ester-forming group, or a protecting group; y is selected from phosphonate (-PO)₃R₂) Sulfonate (-SO)₃R) and a carboxylate (-CO)₂R), wherein R is hydrogen, an ester-forming group or a protecting group; r¹Selected from hydroxyl and hydrogen; r²Is chlorine or hydrogen; and R³And R⁴Independently selected from hydrogen, alkyl, alkenyl and alkynyl, wherein R³And R⁴Has 11 to 30 carbon atoms.

In one embodiment, R³And R⁴Independently selected from hydrogen, alkyl, alkenyl and alkynyl, wherein R³And R⁴has-C (═ O) R⁵OR-C (═ O) OR⁵Wherein R is⁵Is alkyl, alkenyl or alkynyl having 11 to 30 carbon atoms;

in another embodiment, R³And R⁴Independently selected from hydrogen and ring systems containing bicyclic, tricyclic, spirocyclic, fused or bridged ring carbocyclic (aromatic or non-aromatic) or heterocyclic ring systems, and if R is³And R⁴When not simultaneously hydrogen, the ring system is substituted or unsubstituted.

In yet another embodiment, R³Is hydrogen or lower alkyl (e.g. C)_1-6) And R is⁴is-C (═ O) R⁵OR-C (═ O) OR⁵Wherein R is⁵Is C_11-30Alkyl radical, C_11-30Alkenyl or C_11-30Alkynyl.

In some embodiments, R³Is hydrogen or lower alkyl, and R⁴is-C (═ O) R⁵OR-C (═ O) OR⁵Wherein R is⁵Is a ring system which is a carbocyclic (aromatic or non-aromatic) or heterocyclic ring system containing bicyclic, tricyclic, spirocyclic, fused, or bridged rings, and which is substituted or unsubstituted.

In other embodiments, R³Is hydrogen or lower alkyl, and R⁴Is an oligoethylene glycol residue or a derivative thereof. For exampleBut is not limited to, — (CH)₂CH₂O)_n─R⁶Where n is equal to 3 to 10, R⁶Selected from hydrogen or lower alkyl (e.g., methyl, ethyl, propyl, butyl, etc.).

In still other embodiments, R³Is hydrogen or lower alkyl, and R⁴Is a substituted or unsubstituted arylalkyl, alkylaryl, or a group consisting of an aromatic hydrocarbon group (aryl or heteroaryl) and an aliphatic hydrocarbon group.

In one embodiment, the present invention provides compounds of formula II and/or formula III, and pharmaceutically acceptable salts or esters thereof:

wherein, W, X, Y, R¹、R³And R⁴As defined above.

In another embodiment, the present invention provides a compound of formula IV, and pharmaceutically acceptable salts or esters thereof:

wherein, X, Y, R¹、R²、R³And R⁴As defined above.

In another embodiment, the present invention provides a compound of formula V and/or formula VI, and pharmaceutically acceptable salts or esters thereof:

wherein, X, Y, R¹、R³And R⁴As defined above.

In another embodiment, the present invention provides compounds of formula VII and/or formula VIII, and pharmaceutically acceptable salts or esters thereof:

wherein R is hydrogen, an ester-forming group, or a protecting group; and R is¹、R²、R³And R⁴As defined above.

In another embodiment, the invention provides a compound of formula IX, and pharmaceutically acceptable salts or esters thereof:

wherein R is hydrogen, an ester-forming group, or a protecting group; and R¹、R²、R³And R⁴As defined above.

In one embodiment, R¹Is a hydroxyl group (i.e., the carbohydrate moiety in the compound is a D-ribose moiety). In yet another embodiment, R¹Is hydrogen (i.e., the carbohydrate moiety in the compound is a 2-deoxy-D-ribose moiety).

In yet another embodiment, R²Is hydrogen. In yet another embodiment, R²Is hydrogen and R¹Is hydroxy (i.e., the compound is an adenosine derivative). In another embodiment, R²Is hydrogen and R¹Is hydrogen (i.e., the compound is a deoxyadenosine derivative). In yet another embodiment, R²Is hydrogen and R³And R⁴None are hydrogen (i.e., the compound is an adenosine derivative or deoxyadenosine derivative having a substituent on the amino group of the adenine moiety). In other embodiments, R²Is chloro and the compound is a 2-chloro-D-adenosine derivative or a 2-chloro-D-deoxyadenosine derivative.

In some embodiments, R³Is hydrogen or lower alkyl (e.g. C)_1-6) And R is⁴Is an alkyl, alkenyl or alkynyl group having 11 to 30 carbon atoms (i.e., C)_11-30Alkyl radical, C_11-30Alkenyl or C_11-30Alkynyl).

In another embodiment, R³Is hydrogen or lower alkyl and R⁴Is a substituent of a bicyclic, tricyclic or polycyclic ring system, wherein the ring isThe system is a fused, spiro, bridged or parallel (parallel) ring system, and the ring system is a carbocyclic ring, an aliphatic ring, an aromatic ring, a heterocyclic ring, or a combination thereof.

In yet another embodiment, R³Is hydrogen or lower alkyl, and R⁴is-C (═ O) R⁵OR-C (═ O) OR⁵Wherein R is⁵Is an alkyl, alkenyl or alkynyl group having 11 to 30 carbon atoms.

In some embodiments, R³Is hydrogen or lower alkyl, and R⁴is-C (═ O) R⁵OR-C (═ O) OR⁵Wherein R is⁵Is a substituent comprising a bicyclic, tricyclic, or polycyclic ring system, wherein the ring system is a fused ring system, a spiro ring system, a bridged ring system, or a parallel (parallel) ring system, and wherein the ring system is a carbocyclic ring, an aliphatic ring, an aromatic ring, a heterocyclic ring, or a combination thereof.

In one embodiment, R⁴Is a group containing adamantyl moieties. In yet another embodiment, R⁴Is a substituted or unsubstituted 1-adamantyl or 2-adamantyl group. In another embodiment, R⁴Is substituted or unsubstituted 1-methyladamantane. In some embodiments, R⁴Is 1-ethyl adamantane, 1-propyl adamantane or 1-butyl adamantane, where the adamantyl moiety can be substituted or unsubstituted.

In another embodiment, R⁴Is a group containing a naphthyl moiety. In yet another embodiment, R⁴Is a substituted or unsubstituted α -naphthyl or β -naphthyl in other embodiments, R is a substituted or unsubstituted naphthyl⁴Is α -naphthylmethyl or β -naphthylmethyl, with or without additional substituents⁴Selected from the group consisting of naphthylethyl, naphthylpropyl and naphthylbutyl, wherein the naphthyl moiety can be unsubstituted or substituted.

In yet another embodiment, R³、R⁴Together with the nitrogen to which they are attached form a ternary fused ring system such as, but not limited to, a substituted or unsubstituted carbazolyl moiety.

In one embodiment, formula (II) isIn the formula I, W is oxygen or methylene; x is selected from-P (═ O) (OR) -and-C (═ O) -; y is selected from-PO₃R₂、-SO₃R and-CO₂R; r is H or C1-C10 alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, hexyl, etc.).

Preferably, in formula I, W is oxygen; x is selected from-P (═ O) (OH) -and-C (═ O) -; y is selected from-PO₃R₂、-SO₃H and-CO₂H; r is H, methyl or ethyl.

In one embodiment, in formula I, R¹Is a hydroxyl group.

In one embodiment, in formula I, R²Is chlorine.

In one embodiment, in formula I, R³And R⁴Independently selected from hydrogen; amino protecting groups (e.g., Boc); C11-C30 alkyl; C11-C30 alkenyl containing at least one ethylenic linkage; -C (═ O) R⁵；-C(＝O)OR⁵；-C(＝O)NR⁵(ii) a A C6-C30 aryl group; a C5-C30 heteroaryl group containing a N atom; C7-C20 aralkyl (benzyl, naphthylmethyl); bicyclic, tricyclic, spiro, or bridged C6-C30 cycloalkyl; C6-C10 heterocycloalkyl containing at least one N atom which is bicyclic, tricyclic, spirocyclic or bridged; -CH (CH)₂CH₂O)_n─R⁶Where n is equal to 3 to 10 (e.g., n-4, 5, 6, 7, 8, etc.).

The C11-C30 alkyl group is preferably a C11-C20 alkyl group, more preferably a C12-C18 alkyl group. Specific examples thereof include dodecyl, tetradecyl, hexadecyl, and octadecyl.

The C11-C30 alkenyl containing at least one olefinic bond is preferably C11-C20 alkenyl containing at least one olefinic bond, and more preferably C12-C18 alkenyl containing at least one olefinic bond.

The C6-C30 aryl group is preferably a C6-C20 aryl group, more preferably a C10-C18 aryl group. Specific examples thereof include phenyl, naphthyl, anthryl, fluorenyl, fluorenylketo, pyrenyl and the like.

The N-containing C5-C30 heteroaryl group is preferably a C5-C20 nitrogen heteroaryl group, and more preferably a C5-C14 nitrogen heteroaryl group. Specific examples include indolyl, carbazolyl and the like.

C7-C20 aralkyl is preferably C7-C14 aralkyl. Specific examples are benzyl, phenethyl, phenylpropyl, naphthylmethyl, naphthylethyl and the like.

Bicyclic, tricyclic, spirocyclic or bridged C6-C30 cycloalkyl is preferably C6-C20 cycloalkyl, more preferably C7-C15 cycloalkyl. Among them, cycloalkyl groups of bridged rings are more preferable. Specific examples are adamantyl

And the like.

C6-C10 heterocycloalkyl containing at least one N atom which is bicyclic, tricyclic, spirocyclic or bridged is preferably bridged. Examples are

And the like.

Or, R³And R⁴Combined together with the commonly attached N atom to form a bicyclic, tricyclic, spirocyclic, or bridged ring (optionally further containing at least one N atom) C4-C10 heterocycloalkyl. Preferred are nitrogen-containing spiro cycloalkyl groups having C4-C10, as exemplified by

Or R³And R⁴Taken together with the commonly attached N atom, form a C4-C12 heteroaryl group. Specific examples thereof include pyrrolyl, pyridyl, pyrimidinyl, pyrazinyl, indolyl, quinolyl, isoquinolyl, purinyl, carbazolyl and the like.

Wherein R is⁵Is C1-C15 alkyl (preferably C1-C12 alkyl such as methyl, ethyl, propyl, butyl, pentyl, hexyl, octyl, decyl, dodecyl, etc.); C11-C30 alkenyl (preferably C12-C20 alkenyl such as dodecenyl, heptadecadienyl, etc.); bicyclic, tricyclic, spirocyclic or bridged C11-C30 cycloalkyl or heterocycloalkyl containing at least one N atom (said C11-C30 cycloalkyl is preferably C6-C20 cycloalkyl, more preferably C7-C15 cycloalkyl, with bridged cycloalkyl being more preferred; said C11-C30 heterocycloalkyl containing at least one N atom is preferably bridged; examples are

Etc.);

R⁶selected from hydrogen or C1-C4 alkyl (e.g., methyl, ethyl, propyl, butyl).

Preferably, R³And R⁴At least one of which is H.

R is as defined above³And R⁴May be further substituted with at least one of the following substituents: C1-C10 alkyl or cycloalkyl (methyl, ethyl, propyl, butyl, pentyl, cyclopentyl, hexyl, cyclohexyl, adamantyl), C1-C10 alkoxy (e.g., methoxy, ethoxy, etc.), halo C1-C10 alkyl, hydroxy, amino, halogen, and the like.

Preferably, the substituent may be methyl, ethyl, propyl, butyl, hexyl, cyclohexyl, methoxy, fluoro, chloro, hydroxy, amino, trifluoromethyl, hexafluoroethyl, and the like.

In a further embodiment, in formula I:

w is oxygen, X is-P (═ O) (OH) -, and Y is-PO₃H₂；

R¹Is a hydroxyl group;

R²is chlorine;

R³and R⁴Independently selected from hydrogen; C11-C16 alkyl; -C (═ O) R⁵；-C(＝O)OR⁵(ii) a C6-C15 aryl (e.g., naphthyl, anthracenyl); a C5-C12 heteroaryl group containing a N atom; aralkyl of C7 to C12; C6-C12 cycloalkyl bridged;

or, R³And R⁴Taken together with the commonly attached N atom, form a C4-C12 heteroaryl group.

R⁵Is C5-C20 alkyl; C12-C20 alkenyl; a bridged C11-C30 cycloalkyl or heterocycloalkyl group containing at least one N atom;

the R is³And/or R⁴May be further substituted by a substituent, which may be methyl.

In still further embodiments, in formula I:

w is oxygen, X is-P (═ O) (OH) -Y is-PO₃H₂；

R¹Is a hydroxyl group;

R²is chlorine;

R³and R⁴Selected from hydrogen, dodecyl, -C (═ O) R⁵，-C(＝O)OR⁵Naphthyl, anthracenyl, indolyl, benzyl, naphthylmethyl,

or, R³And R⁴Combined with a commonly bonded N atom to form a carbazolyl group,

Preferably, R³And R⁴One of which is hydrogen.

In some embodiments, compounds of table 1 and pharmaceutically acceptable salts or esters thereof are provided.

Table 1: exemplary Compounds

In a second broad aspect, the present invention provides a pharmaceutical composition comprising: a compound as defined herein, or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable carrier. In some embodiments, a pharmaceutical composition is provided that includes at least one of the compounds represented by formula I through formula IX, or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable carrier. In some embodiments, a pharmaceutical composition is provided that includes at least one of the compounds represented by formula I through formula IX, or a pharmaceutically acceptable salt or ester thereof, wherein, in the compound, R³And R⁴One of which is not hydrogen or C1-C10 alkyl, alkene or alkynyl. In some embodiments, there is provided a pharmaceutical composition comprising a compound shown in table 1, or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable carrier.

In a third broad aspect, the present invention provides compounds, compositions and methods for inhibiting CD73 activity in a subject in need thereof, comprising administering to the subject an effective amount of a compound and/or pharmaceutical composition described herein.

In particular embodiments, the compounds of the present invention act to inhibit CD73 immunosuppressive and/or anti-inflammatory activity, and are useful as therapeutic or prophylactic treatments when such inhibition is desired. Unless otherwise indicated, when describing the use of the compounds of the present invention herein, it is to be understood that these compounds may be in the form of compositions (i.e., pharmaceutical compositions). As used herein, the terms "CD 73 inhibitor", "CD 73 blocker", "extracellular-5' -nucleotidase inhibitor of adenosine", "NT 5E inhibitor", "5 NT inhibitor" and all other art-relevant acceptable terms are used interchangeably to refer to compounds capable of directly or indirectly inhibiting the CD73 receptor in vitro tests, in vivo models, and/or other tests that represent CD73 inhibition and potential therapeutic or prophylactic efficacy. The term also refers to compounds that exhibit at least some therapeutic or prophylactic benefit in a human subject.

Although the compounds of the present invention are believed to act by inhibiting CD73, the practice of the present invention does not require an accurate understanding of the mechanism of action of the compounds. For example, the compounds may also have effects at least in part by modulating (e.g., inhibiting) other components of the purinergic signaling pathway (e.g., CD 39). The purinergic signaling system consists of transporters, enzymes, and receptors for (primarily) synthesis, release, action, and extracellular inactivation of ATP and its extracellular breakdown product adenosine. Because inhibition of CD73 reduces adenosine production, CD73 inhibitors are useful for treating diseases or disorders mediated by adenosine, and which act on adenosine receptors, including a1, A2A, A2B, and A3.

In the present invention, the purinergic signaling process described comprises the following components. Purinergic receptors (P1, P2X, and P2Y) are the first components, membrane receptors that mediate various physiological functions (e.g., relaxation of intestinal smooth muscle) in response to ATP or adenosine release; in general, all cells have the ability to release nucleotides into the extracellular environment by regulating exocytosis. The second component is Nucleoside Transporters (NTs), which are membrane transporters that transport nucleoside substrates (e.g., adenosine) across the cell membrane; the extracellular concentration of adenosine can be regulated by NTs, possibly in the form of a feedback loop linking receptor signaling with transporter function. As previously mentioned, the nucleotides released into the extracellular environment by hydrolysis of extracellular nucleotidases (CD73 and CD39) also contain additional components.

In some embodiments, the present invention provides methods for treating or preventing cancer in a subject (e.g., a human) comprising administering to the subject a therapeutically effective amount of at least one CD73 inhibitor compound or composition described herein. In some embodiments of such methods, at least one CD73 inhibitor compound or composition is administered to the subject in an amount effective to reverse, slow, or prevent the progression of CD 73-mediated immunosuppression. In some embodiments, CD 73-mediated immunosuppression is mediated by Antigen Presenting Cells (APCs).

The type of cancer or tumor that can be treated or prevented using the compounds and compositions of the present invention is not particularly limited. Examples of cancers and tumors that may be treated or prevented using the compounds and compositions described herein include, but are not limited to, prostate, colorectal, pancreatic, cervical, gastric, endometrial, brain, liver, bladder, ovarian, testicular, head, neck, skin (including melanoma and basal carcinoma), epithelial membranes, leukocytes (including lymphoma and leukemia), esophagus, breast, muscle, connective tissue, lung (including small cell lung cancer and non-small cell lung cancer), adrenal, thyroid, kidney or bone cancers, gliomas, mesothelioma, renal cell carcinoma, gastric cancer, sarcoma, choriocarcinoma, skin basal cell carcinoma, and testicular seminoma. In some embodiments of the invention, the cancer is melanoma, colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, leukemia, brain tumor, lymphoma, sarcoma, ovarian cancer, or kaposi's sarcoma.

In some embodiments, the present invention provides methods of treating a subject receiving a bone marrow transplant or peripheral blood stem cell transplant comprising administering a therapeutically effective amount of a CD73 inhibitor compound or composition sufficient to increase delayed-type hypersensitivity to a tumor antigen, delay the time to recurrence of a malignant tumor after transplantation, increase the survival rate after transplantation without recurrence, and/or increase the long-term survival rate after transplantation.

In certain embodiments, the present invention provides methods for treating or preventing an infectious disorder (e.g., a viral infection) in a subject (e.g., a human), comprising administering to the subject a therapeutically effective amount of at least one CD73 inhibitor compound or composition of the present invention. In some embodiments, the infectious disorder is a viral infection (e.g., a chronic viral infection), a bacterial infection, a fungal infection, or a parasitic infection. In certain embodiments, the viral infection is human immunodeficiency virus or cytomegalovirus.

In other embodiments, the invention provides methods of treating and/or preventing immune-related diseases, disorders, and conditions, diseases having an inflammatory component, and disorders related thereto, using at least one CD73 inhibitor compound or composition provided herein.

Other diseases, disorders, and conditions that may be treated or prevented, in whole or in part, by inhibiting CD73 activity are also candidate indications for the CD73 inhibitor compounds and compositions provided herein.

In some embodiments, the present invention further provides the use of the CD73 inhibitor compounds and compositions described herein in combination with one or more additional agents. The one or more additional agents may have some CD 73-modulating activity and/or they may act through different mechanisms of action. In some embodiments, such agents comprise radiation (e.g., local or systemic radiotherapy) and/or other forms of treatment of a non-pharmacological nature. When a combination therapy is used, the CD73 inhibitor and one additional agent may be in the form of a single composition or multiple compositions, and the mode of treatment may be administered simultaneously, sequentially or by some other regimen. For example, in some embodiments, embodiments are provided in which a chemotherapy phase is performed after the irradiation phase. The combination therapy may have additive or synergistic effects.

In some embodiments, the present invention provides the use of a CD73 inhibitor compound or composition described herein in combination with bone marrow transplantation, peripheral blood stem cell transplantation, or other types of transplantation therapy.

In a specific embodiment, the present invention provides the use of an inhibitor of CD73 function in combination with an immune checkpoint inhibitor as described herein. Blocking immune checkpoints, which results in the expansion of antigen-specific T cell responses, is shown to be a promising approach in human cancer therapy. Non-limiting examples of immune checkpoints (ligands and receptors), some of which are selectively upregulated in various types of tumor cells, are candidates for blockade, include PD1 (programmed cell death protein 1), PDL1(PD1 ligand), BTLA (B and T lymphocyte attenuating agents), CTLA4 (cytotoxic T lymphocyte-associated antigen 4), TIM3(T cell membrane protein 3), LAG3 (lymphocyte activator gene 3), A2aR (adenosine A2a receptor A2aR), and killer inhibitory receptors.

In other embodiments, the present invention provides methods of treating cancer in a subject comprising administering to the subject a therapeutically effective amount of at least one CD73 inhibitor compound or composition thereof and at least one chemotherapeutic agent, including but not limited to: alkylating agents, (e.g., nitrogen mustards such as chlorambucil, cyclophosphamide, isoflutolamide, dichloromethyldiethylamine, melphalan, and uracil mustard, aziridines such as thiotepa, methanesulfonates such as busulfan, nucleoside analogs such as gemcitabine, nitrosoureas such as carmustine, lomustine, and streptozotocin, topoisomerase 1 inhibitors such as irinotecan, platinum complexes such as cisplatin and carboplatin, bioreductive alkylating agents such as mitomycin, procarbazine, dacarbazine, and hexamethylmelamine); DNA strand breaking agents (e.g., bleomycin); topoisomerase II inhibitors (such as amsacrine, dactinomycin, daunorubicin, idarubicin, mitoxantrone, daunorubicin, etoposide, and teniposide); DNA minor groove binding agents (e.g., Plicamydin); antimetabolites (e.g., folic acid antagonists such as methotrexate and trimetrexate, pyrimidine antagonists such as fluorouracil, fluorodeoxyuridine, CB3717, azacitidine, cytarabine, and floxuridine, purine antagonists such as mercaptopurine, 6-thioguanine, fludarabine, pentostatin, asparaginase, and ribonucleotide reductase inhibitors such as hydroxyurea); tubulin interacting agents (e.g., vincristine, estramustine, vinblastine, docetaxel, epothilone derivatives, and paclitaxel); hormones (e.g., estrogens; conjugated estrogens; ethinyl estradiol; diethylstilbestrol; chlormadinone; gestrel; progestins, such as hydroxyprogesterone caproate, medroxyprogesterone, and megestrol; and androgens, such as testosterone, testosterone propionate, fluoxymesterone, and methyltestosterone); adrenocortical steroids (e.g., prednisone; dexamethasone; methylprednisolone and prednisolone); luteinizing hormone releasing agents or gonadotropin releasing hormone antagonists (e.g., leuprolide acetate and goserelin acetate); and anti-hormonal antigens (e.g., tamoxifen, anti-androgens such as flutamide, and epinephrine such as mitotane and aminoglutethimide). The present invention also provides for the use of CD73 inhibitors in combination with other agents known in the art (e.g., arsenic trioxide) and other chemotherapeutic agents that may be developed in the future.

In some embodiments of the methods for treating cancer, the survival observed for cancer when a therapeutically effective amount of a CD73 inhibitor is administered in combination with at least one chemotherapeutic agent is greater than the survival observed for cancer when either agent is administered alone. In other embodiments directed to methods of treating cancer, administration of a therapeutically effective amount of a CD73 inhibitor in combination with at least one chemotherapeutic agent results in a decrease in tumor size or a decrease in tumor growth that is superior to the decrease in tumor size or decrease in tumor growth observed with administration of either agent alone.

In another embodiment, there is provided a method for treating or preventing cancer in a subject comprising administering to the subject a therapeutically effective amount of at least one CD73 inhibitor compound or composition, and at least one Signal Transduction Inhibitor (STI). In a specific embodiment, the at least one STI is selected from the group consisting of bcr/abl kinase inhibitors, Epidermal Growth Factor (EGF) receptor inhibitors, her-2/neu receptor inhibitors, and Farnesyl Transferase Inhibitors (FTIs).

In other embodiments, the present invention provides methods of enhancing rejection of tumor cells in a subject comprising administering a CD73 inhibitor compound or composition in combination with at least one chemotherapeutic agent and/or radiation therapy, wherein the resulting rejection of tumor cells is superior to that obtained by administering a CD73 inhibitor, chemotherapeutic agent or radiation therapy alone.

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 immunomodulatory agent that is different from a CD73 inhibitor. It is to be understood that as used herein, "CD 73 inhibitor" refers to a compound provided herein, for example, a compound of any one of formulas I-IX, a compound of table 1, or a pharmaceutically acceptable salt or ester thereof, as well as to pharmaceutical compositions thereof.

In some embodiments, the present invention provides methods for treating or preventing an infectious disorder (e.g., a viral infection) in a subject (e.g., a human), comprising administering to the subject 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 antimicrobial agents).

In additional embodiments, treatment of an infectious disease is achieved by the combined administration of a vaccine and the administration of a therapeutically effective amount of a CD73 inhibitor provided herein. In some embodiments, the vaccine is an antiviral vaccine, including, for example, an anti-HIV vaccine. In other embodiments, the vaccine is effective against tuberculosis or malaria. In yet another embodiment, the vaccine is a tumor vaccine (e.g., a vaccine effective against melanoma); the tumor vaccine may comprise genetically modified tumor cells or genetically modified cell lines, including genetically modified tumor cells or genetically modified cell lines that have been transfected to express granulocyte-macrophage stimulating factor (GM-CSF). In particular embodiments, the vaccine comprises one or more immunogenic peptides and/or dendritic cells.

In certain embodiments involving treatment of an infection by administration of a CD73 inhibitor and at least one additional therapeutic agent, the symptoms of infection observed after administration of both the CD73 inhibitor and the additional therapeutic agent are improved compared to the same symptoms of infection observed with either agent alone. In some embodiments, the observed symptom of the infection may be a reduction in viral load, an increase in CD4+ T cell count, a reduction in opportunistic infections, an increase in survival time, eradication of chronic infections, or a combination thereof.

Drawings

For a better understanding of the invention and to show more clearly how it may be carried into effect, reference will now be made, by way of example, to the accompanying drawings, which illustrate features of embodiments in accordance with the invention and in which:

figure 1 is a CD73 inhibition curve for compound 9; and

figure 2 is a plot of the CD73 inhibition rate of compound 22.

Detailed Description

The cases diagnosed with cancer and dying from cancer continue to increase. Traditional treatment methods include chemotherapy and radiation therapy; these traditional treatments, on the one hand, often make the subject intolerant and, on the other hand, cancers (e.g., tumors) evolve to circumvent such treatments, thereby rendering these treatments progressively ineffective. Recent experimental evidence suggests that CD73 inhibitors may represent an important novel therapeutic modality for cancer (e.g., breast cancer).

The data also indicate that inhibitors of CD73 function have the effect of inhibiting the anti-inflammatory activity of CD73 and/or the immunosuppressive activity of CD73, and therefore CD73 inhibitors may be useful in the treatment of, for example, immunosuppressive diseases (e.g., HIV and AIDs). Inhibition of CD73 may also be an important therapeutic strategy for subjects with neurological or neuropsychiatric disorders, or depression.

The invention provides, inter alia, small molecule compounds and compositions thereof having CD73 inhibitory activity, and methods of using the compounds and compositions for the treatment and prevention of the diseases, disorders, and conditions described herein. The compounds provided by the present invention are useful as inhibitors of CD73 and, therefore, are useful in the treatment of diseases, disorders, and conditions in which CD73 activity plays a role. In addition, the compounds provided by the present invention are useful as inhibitors of adenosine receptors (e.g., the A2A receptor). Accordingly, the compounds provided herein are useful for the treatment of diseases, disorders, and conditions associated with the activity of one or more adenosine receptors.

In one embodiment, the invention provides a method of treating a subject (e.g., a human) having cancer or a disorder mediated by CD73, comprising administering to the subject a therapeutically effective amount of a CD73 inhibitor provided herein, e.g., a compound provided herein or a pharmaceutically acceptable composition thereof.

It will be understood that the pharmaceutical compositions comprise a compound disclosed herein (or a pharmaceutically acceptable salt or ester thereof) and a pharmaceutically acceptable carrier, additive or vehicle. In certain embodiments, the amount of the compound in the composition is such that it is effective as a CD73 inhibitor in a biological sample (e.g., in vitro assay, in vivo model, etc.) or in a subject. In certain embodiments, the compositions are formulated for administration to a subject in need of such compositions. In some embodiments, the composition is an injectable formulation. In other embodiments, the composition is formulated for oral administration to a subject.

The invention also provides a method of treating a subject (e.g., a human) having cancer or an adenosine receptor (e.g., A2AR) -mediated disorder, comprising the step of administering to the subject a therapeutically effective amount of a CD73 inhibitor provided herein, e.g., a compound provided herein or a pharmaceutically acceptable composition thereof. In certain embodiments, the amount of the compound in the composition is such that it is effective for use as an inhibitor of an adenosine receptor (e.g., A2AR) in a biological sample (e.g., an in vitro assay, an in vivo model, etc.) or in a subject. In certain embodiments, the compositions are formulated for administration to a subject in need of such compositions. In some embodiments, the composition is an injectable formulation. In other embodiments, the composition is formulated for oral administration to a subject.

In yet another embodiment, the present invention provides a method for 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 signaling inhibitor (STI). In a specific embodiment, the at least one STI is selected from the group consisting of bcr/abl kinase inhibitors, Epidermal Growth Factor (EGF) receptor inhibitors, her-2/neu receptor inhibitors, and Farnesyl Transferase Inhibitors (FTIs). The invention also provides a method of enhancing 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 superior to that obtained by administering a CD73 inhibitor, chemotherapeutic agent, or radiation therapy alone. In yet 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 immunomodulatory agent that is different from a CD73 inhibitor.

In other embodiments, the present invention provides methods for treating or preventing an infectious disorder (e.g., a viral infection) in a subject (e.g., a human), comprising administering to the subject 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 antimicrobial agents).

Definition of

In order to provide a clear and consistent understanding of the terms used in the description of the invention, some definitions are provided below. Furthermore, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

The use of the words "a" or "an" when used in conjunction with the term "comprising" in the claims and/or the specification may mean "one", but it is also known with the meaning of "one or more", "at least one" and "one or more than one". Similarly, the word "another" may mean at least a second or a great number.

As used in this specification and claims, the words "comprise" (and any form of comprise, such as "comprises" and "comprising"), "have" (and any form of have, "having," "includes," and "containing") are inclusive and open-ended and do not exclude additional unrecited elements or process steps.

The term "about" or "approximately" is used to indicate that the value includes errors introduced by the instruments and methods used in determining the value.

The term "derivative" as used herein is understood to mean another compound which is structurally similar and differs in some fine structure.

This specification refers to a number of chemical terms and abbreviations used by those skilled in the art. However, for clarity and consistency, definitions of selected terms are provided.

"alkyl" generally refers to saturated hydrocarbons having 1 to 30 carbon atoms, including straight chain, branched chain, and cyclic alkyl groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, isopropyl, tert-butyl, sec-butyl, isobutyl, cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, dodecyl, hexadecyl, eicosyl, and the like. Under certain conditions, "alkyl" may also be capped with the number of carbon atoms for expression of a particular size of alkyl group. For example, the "substituent R^xIs an expression of "alkyl group having 11 to 30 carbon atoms", means that an alkyl group of 10 or less carbon atoms or more than 30 carbon atoms is excluded. The term alkyl includes both unsubstituted alkyl and substituted alkyl. The term "C₁-C_nAlkyl "and" C_1-nAlkyl groups "(where n is an integer from 2 to 30) are used interchangeably to represent alkyl groups having from 1 to the indicated" n "carbon atoms. The alkyl residue may be substituted or unsubstituted. In some embodiments, for example, an alkyl group can be substituted with a hydroxyl, amino, carboxyl, carboxylate, amide, carbamate, or aminoalkyl group, among others.

The term "alkenyl" refers to unsaturated hydrocarbons having 2 to 30 carbon atoms, including straight, branched, and cyclic nonaromatic alkenyl groups, and containing 1 to 6 carbon-carbon double bonds. Examples of alkenyl groups include, but are not limited to, vinyl, allyl, 1-propen-2-yl, 1-buten-3-yl, 1-buten-4-yl, 2-buten-4-yl, 1-penten-5-yl, 1, 3-pentadien-5-yl, cyclopentenyl, cyclohexenyl, ethylcyclopentenyl, ethylcyclohexenyl, and the like. The term alkenyl includes unsubstituted alkenyls and substituted alkenyls. The term "C₂-C_nAlkenyl "and" C_2-nAlkenyl "(where n is an integer from 3 to 30) is used interchangeably to mean alkenyl having from 2 to the indicated" n "carbon atoms. As with the foregoing definition of "alkyl," alkenyl "may also be capped with the number of carbon atoms for the purpose of expressing a particular size of alkenyl.

"alkynyl" refers to unsaturated hydrocarbons having 2 to 30 carbon atoms, including straight, branched, and cyclic non-aromatic alkynyl groups, and containing 1 to 6 carbon-carbon triple bonds. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propyn-3-yl, 1-butyn-4-yl, 2-butyn-4-yl, 1-pentyn-5-yl, 1, 3-pentadiyn-5-yl, and the like. The term alkynyl includes both unsubstituted alkynyl and substituted alkynyl groups. The term "C₂-C_nAlkynyl "and" C_2-nAlkynyl ", wherein n is an integer from 3 to 30, is used interchangeably to denote alkynyl having from 2 to the indicated" n "carbon atoms. As with the foregoing definition of "alkyl," alkynyl "may also be capped with the number of carbon atoms for expression of a particular size of alkynyl group.

As used herein, "low" in "lower aliphatic", "lower alkyl", "lower alkenyl" and "lower alkyl" means that the moiety has at least one (two alkenyl and alkynyl groups) and 6 or less carbon atoms, unless otherwise specified.

The terms "cycloalkyl", "alicyclic", "carbocyclic" and equivalents as used herein refer to groups comprising a saturated or partially unsaturated carbocyclic ring in a monocyclic, spiro (sharing one atom between two rings) or fused (sharing at least one bond between two rings) carbocyclic ring system, wherein the carbocyclic ring system has from 3 to 30 carbon atoms. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopenten-1-yl, cyclopenten-2-yl, cyclopenten-3-yl, cyclohexyl, cyclohexen-1-yl, cyclohexen-2-yl, cyclohexen-3-cycloheptyl, bicyclo [4,3, 0]]Nonyl, norbornyl, and the like. The term cycloalkyl includes both unsubstituted cycloalkyl and substituted cycloalkyl. The term "C₃-C_nCycloalkyl radicals "and" C_3-nCycloalkyl ", wherein n is an integer from 4 to 30, is used interchangeably to denote cycloalkyl having from 3 to the indicated" n "carbon atoms in the ring structure. As used herein, unless otherwise specified, an "oligocycloalkyl" group refers to a group having at least 3 and 8 or fewer carbon atoms in its ring structure.

The term cycloalkyl or cycloalkyl residue as used herein may be saturated or a group containing one or more double bonds within the ring. In particular, they may be saturated or contain a double bond in the ring system. In unsaturated cycloalkyl residues, the double bond may be present at any suitable position. Monocycloalkyl residues include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl or cyclotetradecyl, which may also be substituted by C_1-4An alkyl group. Examples of substituted cycloalkyl residues are 4-methylcyclohexyl and 2, 3-dimethylcyclopentyl. Examples of parent structures for bicyclic systems are norbornane, bicyclo [2.2.1]Heptane, bicyclo [2.2.2]Octane and bicyclo [3.2.1]Octane.

The term "heterocycloalkyl" and equivalent expressions as used herein refer to a group containing a saturated or partially unsaturated carbocyclic ring, having from 3 to 30 carbon atoms, including from 1 to 6 heteroatoms, in a monocyclic, spiro (sharing one atom) or fused (sharing at least one bond) carbocyclic ring system(e.g., N, O, S, P) or containing heteroatoms (e.g., NH, NRx (Rx is alkyl, acyl, aryl, heteroaryl, or cycloalkyl), PO₂、SO、SO₂Etc.). The heterocycloalkyl group may be attached to C or to a heteroatom (e.g., through a nitrogen atom). Examples of heterocycloalkyl include, but are not limited to, pyrrolidinyl, tetrahydrofuranyl, tetrahydrodithienyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, thiaxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxiraneyl, thietanyl, oxazetanyl, diazepinyl, thiazetanyl, 1,2,3, 6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1, 3-dioxolanyl, pyrazolinyl, dithianyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo [3 ], 1,0]Hexyl, 3-azabicyclo [4,1, 0] s]Heptyl, 3H-indolyl, quinolizinyl, and sugars, and the like. The term heterocycloalkyl includes both unsubstituted heterocycloalkyl and substituted heterocycloalkyl. The term "C₃-C_nHeterocycloalkyl "and" C_3-n-heterocycloalkyl ", wherein n is an integer from 4 to 30, is used interchangeably to denote a heterocycloalkyl group having from 3 to the indicated" n "atoms in the ring structure, including at least one heterogroup or atom as defined above. As used herein, unless otherwise specified, "lower heterocycloalkyl" means having at least 3 and equal to or less than 8 carbon atoms in its cyclic structure.

The terms "aryl" and "aryl ring" as used herein refer to an aromatic group having "4 n + 2" (pi) electrons in a conjugated mono-or polycyclic ring system (fused or non-fused) and having 6 to 30 ring atoms, wherein n is an integer from 1 to 7. Polycyclic ring systems include at least one aromatic ring. Aryl groups may be directly linked or through C₁-C₆An alkyl (also known as arylalkyl or aralkyl) linkage. Examples of aryl groups include, but are not limited to, phenyl, benzyl, phenethyl, 1-phenylethyl, tolyl, naphthyl, biphenyl, terphenyl, indenyl, benzocyclooctenyl, benzeneAnd cycloheptenyl, azulenyl, acenaphthenyl, fluorenyl, phenanthryl, anthracenyl and the like. The term aryl includes both unsubstituted aryl and substituted aryl. The term "C₆-C_nAryl "or" C_6-nAryl "(where n is an integer from 6 to 30) is used interchangeably to refer to aryl groups having from 6 to the" n "carbon atoms shown in the ring structure, including at least one heterocyclic group or atom as defined above.

The terms "heteroaryl" and "heteroaryl ring" as used herein refer to an aromatic group having "4 n + 2" (pi) electrons in a conjugated monocyclic or polycyclic ring system (fused or non-fused), wherein n is an integer from 1 to 7 and includes one to six heteroatoms (e.g., N, O, S, P) or includes heteroatoms (e.g., NH, NRx (Rx is alkyl, acyl, aryl, heteroaryl, or cycloalkyl), PO₂、SO、SO₂Etc.). Polycyclic ring systems include at least one heteroaromatic ring. Heteroaryl may be directly linked or through C₁-C₆Alkyl (also known as heteroarylalkyl or heteroaralkyl) linkages. The heteroaryl group can be attached to a carbon or to a heteroatom (e.g., through a nitrogen atom). Examples of heteroaryl groups include, but are not limited to, pyridyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, tetrazolyl, furanyl, thienyl; isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolidinyl, quinolinyl, isoquinolinyl, indolyl, isoindolyl, chromenyl, isochromenyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, pyrazinyl, triazinyl, isoindolyl, pteridinyl, furanyl, benzofuranyl, benzothiazolyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl, quinolinyl, quinolinonyl, isoquinolinyl, quinoxalinyl, naphthyridinyl, furopyridinyl, carbazolyl, phenanthridinyl, acridinyl, peryleneyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxazinyl, dibenzofuranyl, and the like. The term heteroaryl includes unsubstituted heteroaryl and substituted heteroaryl. The term "C₅-C_nHeteroaryl "and" C_5-nHeteroaryl ", wherein n is an integer from 6 to 30, are used interchangeably to denote a cyclic structure having from 5 to the indicatedAn "n" atomic heteroaryl group includes at least one heterocyclic group or atom as defined above.

Examples of heterocycles include, but are not limited to, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothienyl, benzoxazolyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, 4 α H-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H, 6H-1,5, 2-dithiazinyl, dihydrofuro, furyl, furanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, dihydroindolyl, 3H-indolyl, isoquinolyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolyl, oxadiazolyl, 1,2, 3-oxadiazolyl, 1,2, 4-oxadiazolyl, 1,2, 5-oxadiazolyl, 1,2, 3-oxadiazolyl, 1,2, 4-oxadiazolyl, 2, 5-dithiazolyl, 1, 2-thienyl, 3-pyridyl, 1, 4-pyrazolinyl, 1,2, 3-thiazolinyl, 2-thiazolinyl, 1, 2-thiazolinyl, 1, 2-thiazolinyl, 4-thiazolinyl, 1H-pyrazolinyl, 2-thiazolinyl, 2-pyrazolinyl, 4-thiazolinyl, pyrazolinyl, pyrazolidinyl, pyrazolinyl, 1H-pyrazolinyl, pyrazolidinyl, 1,2, 1,2, pyrazolinyl, 3-pyrazolidinyl, pyrazolinyl, 1,2, 3-pyrazolinyl, 2-pyrazolidinyl, 2, pyrazolinyl, 2-pyrazolinyl, 2, pyrazolinyl, 1, pyrazolinyl, pyrazol.

The term "amine" or "amino" as used herein refers to an unsubstituted or substituted group of the general formula-NR_aR_bA fragment of (1), wherein R_aAnd R_bEach independently is hydrogen, alkyl, aryl or heterocyclyl, or R_aAnd R_bTogether with the nitrogen atom to which they are attached form a heterocyclic ring. The term amino refers to a compound or fragment in which at least one carbon or heteroatom is covalently bonded to a nitrogen atom. Thus, the terms "alkylamino" and "dialkylamino" as used herein refer to a compound having one and at least two C, respectively₁-C₆An amine group in which an alkyl group is bonded to a nitrogen atom. The terms "arylamino" and "diarylamino" include groups in which at least one or two aryl groups are bound to a nitrogen atom. The term "amide" or "aminocarbonyl" refers to a structure of a compound or fragment in which the carbon of the carbonyl or thiocarbonyl group is attached to a nitrogen atom. The term acylamino refers to a structure in which an amino group is attached directly to an acyl group.

The term "bicyclic" or "bicyclic" refers to a two ring system sharing two ring carbon atoms, which may be located anywhere along either ring, and generally refers to bicyclic hydrocarbon groups, bicyclic aromatic carbon atom ring structures, and saturated or partially unsaturated bicyclic carbon atom ring structure groups in which one or more carbon atom ring members have been replaced with a heteroatom such as an O, S, or N atom that is permitted for structural stability. The bicyclic ring system may be a fused ring system, such as bicyclo [4.4.0] decane or naphthalene, or a bridged ring system, such as bicyclo [2.2.2] octane.

The term "tricyclic" or "tricyclic" refers to a ring system having three rings that share three ring carbon atoms, which are located at any position along each ring; the term generally refers to tricyclic hydrocarbon groups, tricyclic aromatic carbon atom ring structural groups and saturated or partially unsaturated tricyclic carbon atom ring structural groups in which one or more carbon atom ring members have been replaced with a heteroatom such as an O, S or N atom that is permitted for structural stability. The tricyclic system may have three rings in a fused ring arrangement, such as anthracene or tetradecahydroanthracene, or a bridged ring, such as in adamantane or tricyclo [3.3.1.1] decane.

The term "polycyclic" or "polycyclic" refers to ring systems having more than 3 rings that share more than three rings, with more than three ring carbon atoms located anywhere along any ring. The term generally refers to polycyclic hydrocarbon groups, polycyclic aromatic carbon atom ring structure groups, and saturated or partially unsaturated polycyclic carbon atom ring structure groups in which one or more carbon atom ring members have been substituted with heteroatoms such as O, S or N atoms as permitted by structural stability.

The term "fused ring" or "fused ring" refers to a polycyclic ring system containing fused rings. Typically, the fused ring system contains 2 or 3 rings and/or up to 18 ring atoms. As noted above, the cycloalkyl, aryl, and heterocyclyl groups can form a fused ring system. Thus, the fused ring system may be aromatic, partially aromatic or non-aromatic and may contain heteroatoms. According to this definition, a spiro ring system is not a fused polycyclic, but the fused polycyclic ring system of the invention may itself have a spiro ring attached thereto through a single ring atom of the system. Examples of fused ring systems include, but are not limited to, naphthyl (e.g., 2-naphthyl), indenyl, phenanthryl, anthryl, pyrenyl, benzimidazole, benzothiazole, and the like.

The term "spiro" or "spiro" refers to an organic compound that exhibits a distorted structure of two or more rings (ring systems) in which 2 or 3 rings are joined together by a common atom. Spirocyclic compounds may be fully carbocyclic (all carbon), such as spiro [5.5] undecane or heterocyclic (having one or more non-carbon atoms), including but not limited to carbocyclic spiro compounds, heterocyclic spiro compounds and polyspirocyclic compounds.

The term "bridged ring" or "bridged" refers to a carbocyclic or heterocyclic moiety in which two or more atoms are shared in two or more ring structures, wherein the shared atom is C, N, S or other heteroatoms arranged in a chemically sound substitution pattern. Alternatively, a "bridged ring" compound also refers to a carbocyclic or heterocyclic structure: one atom at any position of the main ring is bonded to a second atom on the main ring, which does not comprise part of the main ring structure, by a chemical bond or an atom other than a bond. The first and second atoms may or may not be adjacent to each other in the main ring. Illustrated below are specific non-limiting examples of bridged ring structures contemplated herein. Other carbocyclic or heterocyclic bridged ring structures are also contemplated, including bridged rings in which the bridging atoms are C or heteroatoms arranged in a chemically sound substitution pattern, as is known in the art.

The term "nitro" as used herein means-NO₂. The terms "halo" and "halogen" refer to a bromo, chloro, fluoro, or iodo substituent. The term "thiol", "thio" or "mercapto" refers to-SH. The term "hydroxy" or "hydroxyl" refers to-OH. The term "alkylthio" refers to a structure in which an alkyl group is attached to a mercapto group. Suitable alkylthio groups include groups having from 1 to about 12 carbon atoms, preferably from 1 to about 6 carbon atoms. The term "alkylcarboxy" as used herein refers to a structure in which an alkyl group is attached to a carboxyl group.

The term "alkoxy" as used herein refers to a structure in which an alkyl group is attached to an oxygen atom. Representative alkoxy groups include groups having from 1 to about 30 carbon atoms. "lower alkoxy" refers to a structure wherein an alkyl group is attached to an oxygen atom, wherein alkyl includes groups having from 1 to about 8 carbon atoms such as methoxy, ethoxy, propoxy, tert-butoxy, and the like. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, isopropoxy, propoxy, butoxy, pentyloxy, fluoromethoxy, difluoromethoxy, trifluoromethoxy, chloromethoxy, dichloromethoxy, trichloromethoxy, and the like. The term "alkoxy" includes unsubstituted or substituted alkoxy, as well as perhaloalkoxy and the like.

The term "carbonyl" or "carboxyl" as used herein includes structures in which the carbon of the compound or fragment is attached to an oxygen atom through a double bond. Examples of carbonyl containing moieties include aldehydes, ketones, carboxylic acids, amides, esters, anhydrides, and the like.

The term "acyl" as used herein is a carbonyl group having a carbon atom attached to hydrogen (i.e., formyl), an aliphatic radical (C)₁-C₃₀Alkyl radical, C₁-C₃₀Alkenyl radical, C₁-C₃₀Carbonyl on alkynyl radicals, e.g. acetyl), cycloalkyl (C)₃-C₈Cycloalkyl and a group composed of cycloalkyl and alkanyl), heterocyclic group (C)₃-C₈Heterocycloalkyl and C₅-C₆Heteroaryl), aryl (C)₆Aryl, such as benzoyl). The acyl group may be unsubstituted or substituted acylA group (e.g., salicyloyl).

It is to be understood that the term "substituted" or "substituted" as used herein includes the implicit proviso that such substitution results in a stable compound (e.g., the compound is not capable of undergoing spontaneous rearrangement, cyclization, elimination, etc.) as a function of the valency of the substituent atom and the substituent. The term "substituted" as used herein includes all permissible substituents of organic compounds. In a broad sense, permissible substituents include acyclic and cyclic, branched and unbranched branched, carbocyclic and heterocyclic, aromatic and nonaromatic substituent organic compounds. The substituent may be one or more. The term "substituted" means that when the above groups are substituted at one or more positions, the substituents include acyl, amino (including simple amino, monoalkylamino and dialkylamino, monoaryl and diarylamino, and alkylarylamino), acylamino (carbamoyl and ureido), alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, alkoxycarbonyl, carboxy, aminocarbonyl, mono-and dialkylaminocarbonyl, cyano, azido, halogen, hydroxy, nitro, trifluoromethyl, thio, alkylthio, arylthio, alkylthiocarbonyl, thiocarboxylate, lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, lower alkoxy, aryloxy, aryloxycarbonyloxy, benzyloxy, benzyl, sulfinyl, alkylsulfinyl, sulfonyl, sulfate, salts of these groups, Sulfonates, sulfonamides, phosphates, phosphonates, imides, formamides, and the like. Any of the above substituents may be further substituted, if permitted, for example with alkyl, aryl or other groups.

The term "solvate" as used herein refers to a physical association of a compound with one or more solvent molecules, whether organic or inorganic. The physical association includes hydrogen bonding. In some cases, the solvate can be isolated, for example, when one or more solvent molecules are incorporated into the crystal lattice of the crystal. "solvate" includes solution phases and solvates which may be separated. Solvates include, but are not limited to, hydrates, ethanolates, methanolates, hemi-ethanolates, and the like.

"pharmaceutically acceptable salts" of a compound refers to salts of a pharmaceutically acceptable compound. Salts (basic, acidic, or charged functional groups) of the desired compounds can retain or improve the biological activity and properties of the parent compound as defined herein, and are not biologically undesirable. The pharmaceutically acceptable Salts may be those mentioned by Berge et al in "Pharmaceutical Salts", J.pharm.Sci.66,1-19 (1977). Including but not limited to:

(1) organic acids include acetic acid, propionic acid, lactic acid, oxalic acid, glycolic acid, pivalic acid, tert-butylacetic acid, β -hydroxybutyric acid, valeric acid, caproic acid, cyclopentanepropionic acid, pyruvic acid, malonic acid, succinic acid, malic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, cyclohexylsulfamic acid, benzenesulfonic acid, sulfanilic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 3-phenylpropionic acid, lauryl sulfonic acid, lauryl sulfuric acid, oleic acid, palmitic acid, stearic acid, lauric acid, pamoic acid (pamoic acid), pamoic acid, pantothenic acid, lactobionic acid, alginic acid, diacidic acid, gluconic acid, glucoheptonic acid, hydroxynaphthoic acid, salicylic acid, muconic acid, and the like.

(2) When an acidic proton is present in the parent compound or it is substituted with a metal ion, a base may be added to give a salt. The metal ions include basic metal ions (e.g., lithium, sodium, potassium), alkaline earth metal ions (magnesium, calcium, barium) or other metal ions such as aluminum, zinc, iron, and the like. Organic bases include, but are not limited to, N' -dibenzylethylenediamine, ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, piperazine, chloroprocaine, procaine, choline, lysine, and the like.

Pharmaceutically acceptable salts can be synthesized from the parent compound, which contains a basic or acidic moiety, by conventional chemical methods. Typically, such salts are prepared by reacting the compound (free acid or base) with a stoichiometric amount of a base or acid in water or an organic solvent, or a mixture of the two. Salts may be prepared in situ during the final isolation or purification of the agent or by separately reacting the purified compound of the invention in free acid or base form with the desired corresponding base or acid and isolating the salt thus formed. The term "pharmaceutically acceptable salts" also includes zwitterionic compounds that contain a cationic group covalently bonded to an anionic group, which are referred to as "inner salts". The compounds of the present invention include all acid, salt, base and other ionic and nonionic forms. For example, if a compound of the present invention is an acid, the salt form of the compound is also included. Likewise, if a compound of the present invention is a salt, the acid and/or base form of the compound is also included.

The compounds provided herein may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. Unnatural proportions of isotopes can be defined from amounts found in nature to amounts consisting of 100% of the atom in question. For example, the compounds may incorporate a radioisotope, such as tritium (A), (B), (C), (D), (³H) Iodine-125 (¹²⁵I) Or carbon-14 (¹⁴C) Or a non-radioactive isotope such as deuterium (²H) Or carbon-13 (¹³C) In that respect Such isotopic variations may provide additional applications to those described elsewhere in this application. For example, isotopic variants of the compounds of the present invention may find additional utility, including but not limited to, as diagnostic and/or imaging agents, or as cytotoxic/radiotoxic therapeutic agents. In addition, isotopic variants can have altered pharmacokinetic and pharmacodynamic profiles, which can help to enhance safety, tolerability, or efficacy during treatment. All isotopic variations of the compounds provided herein, whether radioactive or not, are encompassed by the present invention.

Isotopic enrichment is the process by which one particular isotope is enriched (i.e., increased) and the corresponding other isotope is reduced or depleted by changing the relative abundance of the isotopes of a given element. The term "isotopically enriched" compound or derivative as used herein means that one or more specific isotopes are increased (i.e., one or more specific isotopic elements are enriched or augmented) in the compound. Typically, in an isotopically enriched compound or derivative, the specific isotopic element at a specific position of the compound is enriched or increased. It is to be understood, however, that the compound may be enriched or augmented with two or more isotopic elements, including different isotopes of the same element as well as respective isotopes of different elements. Further, isotopically enriched compounds can be isotopically enriched mixed forms, i.e., containing a plurality of specific isotopes or elements or both. The "isotopically enriched" compounds or derivatives used in the present invention have isotopic levels above this natural abundance. The level of isotopic enrichment depends on the natural abundance of the particular isotope itself. In some embodiments, the isotopic enrichment level of the compound or elements in the compound can be from about 2 to about 100 mole percent (%), e.g., about 2%, about 5%, about 17%, about 30%, about 51%, about 83%, about 90%, about 95%, about 96%, about 97%, about 98%, and greater than about 98%, about 99%, or 100%.

The term "naturally abundant element" or "naturally abundant atom" as used herein refers to an element or atom, respectively, of atomic mass that is most abundant in nature. For example, the natural abundance element of hydrogen is¹H, the natural abundance element of nitrogen is¹⁴N, the natural abundance element of oxygen is¹⁶O, the natural abundance element of carbon is¹²C and the like. A "non-isotopically enriched" compound is one in which all atoms or elements in the compound are isotopes of natural abundance, i.e., the atomic masses of all atoms or elements are the most abundant in nature.

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

The terms "administration," "applicator," or the like, when applied to, for example, a subject, cell, tissue, organ, or biological fluid, refer to contacting, for example, a CD73 inhibitor, a pharmaceutical composition or diagnostic agent comprising the CD73 inhibitor, with the subject, cell, tissue, organ, or biological fluid. In the case of cells, administration includes contacting the agent with the cell (e.g., in vitro or ex vivo), and contacting the agent with a fluid, wherein the fluid is in contact with the cell.

The terms "treat," "treating," or the like, refer to initiating action (e.g., administration of a CD73 inhibitor or a pharmaceutical composition comprising the same) after a disease, disorder, or condition, or symptom thereof, has been diagnosed, observed, so as to temporarily or permanently eliminate, alleviate, inhibit, slow down, or ameliorate at least one underlying cause of the disease, disorder, or condition afflicting the subject, or a symptom thereof associated with the disease, disorder, or condition. Thus, treatment includes inhibiting (e.g., arresting the development or further development of the disease, disorder or condition or clinical symptoms associated therewith) the active disease.

The term "in need of treatment" as used herein refers to the judgment made by a physician or other caregiver that a subject needs or will benefit from treatment. Such a determination is made based on various factors in the professional field of a doctor or caregiver.

The terms "prevent," "preventing," "prevention," or the like, refer to beginning to act (e.g., administering a CD73 inhibitor or a pharmaceutical composition comprising the same) in a manner (e.g., prior to onset of a disease, disorder, condition, or symptom thereof) so as to temporarily or permanently prevent, inhibit, suppress, or reduce the risk of a subject suffering from a disease, disorder, or condition, etc. (as determined, for example, by lack of clinical symptoms) or delay onset of a particular disease, disorder, or condition in the context of a subject predisposed to such a disease, disorder, or condition. In certain instances, the term also refers to slowing the progression of a disease, disorder or condition or inhibiting its development into a harmful or other undesirable state.

The term "in need of prophylaxis" as used herein refers to the judgment made by a physician or other caregiver that a subject needs or will benefit from prophylactic care. Such determinations are made based on various factors in the field of expertise of the physician or caregiver.

The terms "therapeutically effective amount" and "effective amount" are used interchangeably herein and refer to an amount of an agent, alone or as part of a pharmaceutical composition, that when administered to a subject, is administered to the subject in a single dose or as part of a series of doses, in an amount that is capable of having any detectable positive effect on any symptom, aspect or feature of administration to the disease, disorder or condition. The therapeutically effective amount can be determined by measuring the relevant physiological effects, and can be adjusted according to the dosing regimen and diagnostic analysis of the subject's condition, among others. For example, measuring the serum level of the CD73 inhibitor (or e.g., a metabolite thereof) at a particular time after administration can indicate whether a therapeutically effective amount has been used. In some embodiments, the terms "therapeutically effective amount" and "effective amount" refer to an amount or dose of a therapeutic agent, e.g., a compound, that provides a desired therapeutic, diagnostic, or prognostic effect in a subject following administration to the subject in a single or multiple doses. The effective amount can be readily determined by the attending physician or diagnostician by known techniques and by observing the results obtained under analogous circumstances. In determining the effective amount or dose of the compound to be administered, a number of factors are considered, including but not limited to: the size, age, and general health of the subject; the specific diseases involved; the degree of involvement or severity of the disease or disorder to be treated; responses of the subject individuals; the particular compound administered; a mode of administration; the bioavailability characteristics of the administered formulation; selecting a dosage regimen; the use of concomitant medication; and other related considerations.

The term "substantially pure" is used herein to indicate that the component comprises greater than about 50% of the total composition, and typically greater than about 60% of the total composition. More typically, "substantially pure" means that the composition has a target component that is at least 75%, at least 85%, at least 90% or more of the total composition. In some cases, the component of interest will comprise greater than about 90%, or greater than about 95% of the total content of the composition.

As used herein, the terms "CD 73-associated disease, disorder or condition" and "disease, disorder or condition mediated by CD 73" are used interchangeably to refer to any disease, disorder or condition that can benefit from treatment with a CD73 inhibitor. In general, diseases, disorders, and conditions related to or mediated by CD73 are those in which CD73 activity plays a biological, mechanistic, or pathological role. Such diseases, disorders and conditions may also be associated with the activity of one or more adenosine receptors. Non-limiting examples of CD 73-associated diseases, disorders, and conditions include tumor-associated disorders (cancer, tumors, etc.), immune-related disorders, inflammatory component conditions, microbial-associated conditions, CNS-related conditions and neurological conditions, and other diseases (such as, but not limited to, cardiovascular diseases, gastrointestinal diseases, metabolic diseases, liver diseases, lung diseases, ophthalmic diseases, and kidney diseases).

For example, inhibitors of CD73 may be useful in the prevention or treatment of proliferative disorders, cancer or tumors; increasing or enhancing an immune response; improved vaccination, including improved vaccine efficacy; and increase inflammation. The CD73 inhibitor disclosed by the invention can be used for treating immunodeficiency diseases related to immunodeficiency, immunosuppressive drug treatment, acute and/or chronic infection and aging. CD73 inhibitors may also be used to stimulate the immune system in patients with iatrogenically induced immunosuppression, including those who have received bone marrow transplantation, chemotherapy, or radiation therapy. In other embodiments, the CD73 inhibitor may be used to treat or prevent any viral, bacterial, fungal, parasitic or other infectious disease, disorder or condition, including but not limited to HIV and AIDS.

The pharmaceutical compositions provided herein can be formulated to be compatible with the intended method or route of administration; exemplary routes of administration are set forth herein. In addition, the pharmaceutical compositions may be used in combination with other therapeutically active agents or compounds described herein to treat or prevent the CD 73-related diseases, disorders, and conditions discussed herein.

Pharmaceutical compositions containing the active ingredient (e.g., CD73 inhibitor) may be in a form suitable for oral use, such as tablets, capsules, lozenges, troches, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups, solutions, microbeads, or elixirs. Pharmaceutical compositions for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents, for example sweetening agents, flavouring agents, colouring agents and preserving agents in order to provide pharmaceutically acceptable formulations. Tablets, capsules and the like typically contain the active ingredient in admixture with non-toxic pharmaceutically acceptable carriers or excipients which are suitable for the manufacture of tablets. These carriers or excipients may be, 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, for example starch, gelatin or acacia, and lubricating agents, for example magnesium stearate, stearic acid or talc.

Tablets, capsules and the like suitable for oral administration may be uncoated or coated using known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action. For example, a time delay material such as glycerol monostearate or glycerol distearate may be employed. They may also be coated by techniques known in the art to form osmotic therapeutic tablets for controlled release. Other agents include biodegradable or biocompatible particulate or polymeric materials such as polyesters, polyanilines, hydrogels, polyvinylpyrrolidone, polyanhydrides, polyglycolic acid, ethylene vinyl acetate, methylcellulose, carboxymethylcellulose, protamine sulfate or lactide/glycolide copolymers, polylactide/glycolide copolymers or ethylene vinyl acetate copolymers to control delivery of the administered composition. For example, oral formulations may be embedded in colloidal drug delivery systems or microcapsules prepared by coacervation techniques or by interfacial polymerization, by using hydroxymethylcellulose or gelatin microcapsules or poly (methylmethacylate) microcapsules, respectively. Colloidal dispersion systems include macromolecular complexes, nanocapsules, microspheres, microbeads and lipid-based systems including oil-in-water emulsions, micelles, mixed micelles and liposomes. Methods for preparing the above formulations will be apparent to those skilled in the art.

Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent (for example, calcium carbonate, calcium phosphate, kaolin or microcrystalline cellulose); or in the form of soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin, or olive oil. Aqueous suspensions contain the active material in admixture with excipients suitable for the manufacture thereof. Such excipients may be suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents, for example naturally-occurring phosphatides (for example lecithin), or condensation products of an alkylene oxide with fatty acids (for example polyoxyethylene stearate), or condensation products of ethylene oxide with long chain aliphatic alcohols (for example heptadecaethyleneoxycetanol), or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol (for example polyoxyethylene sorbitol monooleate), or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides polyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives.

Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Such as those described above, such as sweetening agents, may be added, and flavoring agents may also be added to provide a palatable oral preparation.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are known in the art.

The pharmaceutical compositions of the present invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil (for example, olive oil or arachis oil) or a mineral oil (for example, liquid paraffin), or mixtures thereof. Suitable emulsifying agents may be naturally-occurring gums, for example gum acacia and gum tragacanth; naturally occurring phospholipids, such as soy, lecithin and esters or partial esters derived from fatty acids; hexitols, such as sorbitan monooleate; condensation products of partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate.

The pharmaceutical compositions generally comprise a therapeutically effective amount of a CD73 inhibitor compound provided herein and one or more pharmaceutically and physiologically acceptable formulating agents. Suitable pharmaceutically or physiologically acceptable diluents, carriers or excipients include, but are not limited to, antioxidants (e.g., ascorbic acid and sodium disulfide), preservatives (e.g., benzyl alcohol, methyl paraben, ethyl or n-propyl paraben), emulsifiers, suspending agents, dispersants, solvents, fillers, detergents, buffers, carriers, diluents and/or adjuvants. For example, a suitable carrier may be a physiological saline solution or citrate buffered saline, possibly supplemented with other substances common in pharmaceutical compositions for parenteral administration. Neutral buffered saline or saline mixed with serum albumin are further exemplary carriers. One of skill in the art will readily know of various buffers that may be used in the pharmaceutical compositions and dosage forms contemplated herein. Typical buffering agents include, but are not limited to, pharmaceutically acceptable weak acids, weak bases, or mixtures thereof. For example, the buffer component may be a water soluble substance such as phosphoric acid, tartaric acid, lactic acid, succinic acid, citric acid, acetic acid, ascorbic acid, aspartic acid, glutamic acid, and salts thereof. Acceptable buffers include, for example, Tris buffer, N- (2-hydroxyethyl) piperazine-N' - (2-ethanesulfonic acid) (HEPES), 2- (N-morpholine) ethanesulfonic acid (MES), 2- (N-morpholine) ethanesulfonic acid sodium salt (MES), 3- (N-morpholine) propanesulfonic acid (MOPS), and Ntris [ hydroxymethyl ] methyl-3-aminopropanesulfonic acid (TAPS). After the pharmaceutical composition is formulated, it may be stored in sterile vials in the form of a solution, suspension, gel, emulsion, solid, dehydrated or lyophilized powder. Such formulations may be stored in a ready-to-use form, a lyophilized form requiring reconstitution prior to use, a liquid form requiring dilution prior to use, or other acceptable forms.

In some embodiments, the pharmaceutical composition is contained in a single-use container (e.g., a single-use vial, ampoule, syringe, or auto-injector), while in other embodiments, is contained in a multiple-use container (e.g., a multiple-use vial).

The formulation may also include a carrier to protect the composition from rapid degradation or disappearance from the body, such as a controlled release formulation, including liposomes, hydrogels, and microencapsulated delivery systems. For example, a time delay material such as glyceryl monostearate or glyceryl stearate alone, or in combination with a wax, may be used. Any drug delivery device may be used to deliver the CD73 inhibitor, including implants (e.g., implantable pumps) and catheter systems, slow syringe pumps and devices, all of which are well known to those skilled in the art.

The pharmaceutical compositions may also be in the form of sterile injectable aqueous or oleaginous suspensions. The suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which are mentioned herein. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol. Acceptable diluents, solvents and dispersion media which can be employed include water, ringer's solution, isotonic sodium chloride solution, Cremophor ELTM (BASF, Parsippany, N.J.) or Phosphate Buffered Saline (PBS), ethanol polyol, polyhydric alcohols (e.g., glycerol, propylene glycol and liquid polyethylene glycol) and suitable mixtures thereof. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono-or diglycerides. Furthermore, fatty acids (such as oleic acid) may be used in the preparation of injectables. Prolonged absorption of a particular injectable formulation can be brought about by the inclusion of agents that delay absorption (e.g., aluminum monostearate or gelatin).

The CD73 inhibitor compounds and compositions provided herein can be administered to a subject in any suitable manner known in the art. Suitable routes of administration include, but are not limited to, oral; parenteral, e.g., intramuscular, intravenous, subcutaneous (e.g., injection or implant), intraperitoneal, intracisternal, intraarticular, intracerebral (intraparenchymal and intracerebroventricular; nasal; vaginal; sublingual; intraocular; rectal; topical (e.g., transdermal); oral and inhalation.

The CD73 inhibitor compounds and compositions provided herein can be administered to a subject in such amounts: the amount depends on, for example, the administration target (e.g., desired resolution); the age, weight, sex, health and physical condition of the subject to which the formulation is administered; the route of administration; and the condition of a disease, disorder, condition, or symptom thereof. The dosing regimen also takes into account the presence, nature and extent of any adverse effects associated with the administered agent. Effective dosages and dosage regimens can be readily determined by, for example, safety and dose escalation assays, in vivo studies (e.g., animal models), and other methods known to those of skill in the art. Generally, the dosage parameters dictate that the dosage be less than the amount that is likely to have irreversible toxicity to the subject (maximum tolerated dose, MTD) and not less than the amount required to produce a measurable effect on the subject. These amounts are determined, for example, by pharmacokinetic and pharmacodynamic parameters associated with ADME, taking into account route of administration and other factors.

In some embodiments, the CD73 inhibitor may be administered at a dosage level of 0.01mg/kg to about 50mg/kg, or about 1mg/kg to about 25mg/kg, of the subject's body weight once a day, or multiple times a day, to achieve the desired therapeutic effect. For use in oral formulations, the compositions may be provided in the form of tablets or capsules containing from 1.0 mg to 1000 mg of the active ingredient, particularly 1mg, 3mg, 5mg, 10mg, 15mg, 20 mg, 25mg, 50mg, 75 mg, 100mg, 150mg, 200mg, 250mg, 300 mg, 400 mg, 500 mg, 600 mg, 750 mg, 800 mg, 900 mg or 1000 mg of the active ingredient.

In some embodiments, the desired dose of CD73 inhibitor is contained in a "unit dosage form". The phrase "unit dosage form" refers to physically discrete units, each unit containing a predetermined amount of the CD73 inhibitor alone, or in combination with one or more additional pharmaceutical agents, sufficient to produce the desired effect. It will be understood that the parameters of the unit dosage form will depend upon the particular agent and the effect to be achieved.

The invention also provides kits comprising CD73 inhibitor compounds or compositions. Kits are generally in the form of physical structures that contain various components and can be used, for example, to perform the methods provided herein. For example, a kit may include one or more CD73 inhibitors disclosed herein (e.g., provided in a sterile container), which may be in the form of a pharmaceutical composition suitable for administration to a subject. The CD73 inhibitor may be provided in a ready-to-use form (e.g., a tablet or capsule) or in a form that requires reconstitution or dilution (e.g., a powder) prior to administration, for example. When the CD73 inhibitor is in a form that requires reconstitution or dilution by the user, the kit may further include diluents (e.g., sterile water), buffers, pharmaceutically acceptable excipients, and the like, packaged with or separately from the CD73 inhibitor. When a combination therapy is employed, the kit may independently contain several therapeutic agents, or they may already be combined in the kit. Each component of the kit may be enclosed in a separate container, and all of the various containers may be in a single package. The kits of the invention can be designed to suitably maintain the conditions required for the components contained therein (e.g., refrigeration or freezing).

The kit may also contain a label or package insert containing information identifying the components therein and instructions for use (e.g., dosage parameters, clinical pharmacology of the active ingredient, including mechanism of action, pharmacokinetics and pharmacodynamics, adverse effects, contraindications, etc.). The label or insert may contain manufacturer information such as lot number and expiration date. The label or package insert may, for example, be integrated into the physical structure containing the components, contained separately within the physical structure, or attached to a component of the kit (e.g., an ampoule, tube, or vial).

Examples

The invention will be more readily understood by reference to the following examples, which are intended to illustrate the invention and are not to be construed as limiting the scope of the invention in any way.

Unless defined otherwise or clear from context to be otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It should be understood that any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention.

Synthesis of the Compounds:

the 2-chloropurine nucleoside derivative S-x (1.0mmol,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Separating by C column chromatography to obtain white solid.

Amino Compounds RNH used in the experiments₂There are two sources, depending on the direct availability of the particular compound: one is commercial source and the other is synthesized according to literature methods.

The compounds provided herein can be prepared by the above-described conventional methods, and the NMR and MS data of the corresponding compounds are shown in the examples below.

Example 1: preparation of compound 1:

2, 6-dichloro-9- (2',3',5' -tri-O-acetyl- β -D-ribofuranosyl) purine (5.0mmol,2236mg,1.0eq.) and benzylamine (5.0mmol,536mg,1.0eq.) were dissolved in 1, 4-dioxane (25 mL). DIEA (7.5mmol,969mg,1.5eq.) was added at room temperature, heated and stirred overnight, concentrated under reduced pressure and dissolved in dichloromethane (1000mL), extracted with water for 2 times, the organic phase was concentrated under reduced pressure and subjected to column chromatography to give an intermediate, which was dissolved in methanolic ammonia (1000mL) and stirred overnight at 35 deg.C, concentrated under reduced pressure and the crude product was subjected to column chromatography to give the 2-chloropurine nucleoside derivative S-II-11818 mg.

2-chloropurine nucleoside derivative S-1(1.0mmol,392mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸C columnChromatography gave 369mg of an off-white solid.

¹H NMR(500MHz,CD₃OD-d₄)δppm 2.46(t，2H)，4.23-4.74(m，7H)，6.01(d，1H)，7.19-7.38(m，5H)，8.59(s，1H)；¹³C NMR(125MHz,CD₃Cl-d₃)δppm 40.12，43.84，63.92，69.87，74.64，83.71，88.50，115.22，126.92，127.47，128.13，149.27，154.08，154.96，160.55；³¹P NMR(200MHz,CD₃Cl-d₃)δppm 12.94，18.11；m/z(ESI⁺)550.1.

Example 2: preparation of compound 6:

2, 6-dichloro-9- (2',3',5' -tri-O-acetyl- β -D-ribofuranosyl) purine (5.0mmol,2.2g,1.0eq.) and 1-naphthylmethylamine (5.0mmol,786mg,1.0eq.) were dissolved in 1, 4-dioxane (25mL), DIEA (7.5mmol,969mg,1.5eq.) was added at room temperature, the mixture was heated with stirring overnight, the mixture was concentrated under reduced pressure, dissolved in dichloromethane (1000mL), extracted with water for 2 times, the organic phase was concentrated under reduced pressure and subjected to column chromatography to give an intermediate, which was dissolved in methanolic ammonia (1000mL), and subjected to column chromatography at 35 ℃ overnight, and the crude product was subjected to column chromatography to give S-II-61.3g of a 2-chloropurine nucleoside derivative.

The 2-chloropurine nucleoside derivative S-6(1.0mmol,442mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 110mg of an off-white solid.

¹H NMR(500MHz,D₂O)δppm 0.88-0.91(m,3H),1.34-1.43(m,4H),1.67-1.68(m,2H),2.15-2.26(m,2H),4.16-4.22(m,2H),4.25-4.31(m,2H),4.38-4.41(m,1H),4.53-4.57(m,1H),4.74-4.76(m,H),6.13-1.15(m,1H),8.70-8.75(m,1H)；¹³C NMR(125MHz,D₂O)δppm13.20,21.60,27.24,27.55,63.46,67.12,70.15,74.34,84.08,84.14,87.38,120.98,142.83,150.24,152.49,153.22；³¹P NMR(200MHz,D₂O)δ16.15,18.97；m/z(ES^-)571.8.

Example 3: preparation of compound 7:

2, 6-dichloro-9- (2',3',5' -tri-O-acetyl- β -D-ribofuranosyl) purine (5.0mmol,2.2g,1.0eq.) and 2-naphthylmethylamine (5.0mmol,786mg,1.0eq.) were dissolved in 1, 4-dioxane (25mL), DIEA (7.5mmol,969mg,1.5eq.) was added at room temperature, the mixture was heated with stirring overnight, the mixture was concentrated under reduced pressure, dissolved in dichloromethane (1000mL), extracted with water for 2 times, the organic phase was concentrated under reduced pressure and subjected to column chromatography to give an intermediate, which was dissolved in methanolic ammonia (1000mL), and subjected to column chromatography at 35 ℃ overnight, and concentrated under reduced pressure to give the crude product 2-chloropurine nucleoside derivative S-II-71.15 g.

The 2-chloropurine nucleoside derivative S-7(1.0mmol,442mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 105mg of an off-white solid.

¹H NMR(500MHz,D₂O)δppm 2.11(t,J＝19.7Hz,2H),4.10(s,2H),4.29(s,1H),4.42(s,1H),4.55(s,1H),4.88(s,2H),5.79(s,1H),7.36-7.44(m,4H),7.71(d,J＝7.9Hz,1H),7.76(d,J＝7.2Hz,1H),7.87(d,J＝7.8Hz,1H),8.13(s,1H)；¹³C NMR(125MHz,D₂O)δppm27.57,42.23,63.39,70.05,74.16,83.59,86.92,117.78,122.74,125.48,125.88,126.05,126.26,128.24,128.43,130.48,132.19,133.08,139.02,148.62,153.86,154.44.³¹P NMR(202MHz,D₂O)δppm15.17,19.58；m/z(ES^-)598.2.

Example 4: preparation of compound 8:

the 2-chloropurine nucleoside derivative S-8(1.0mmol,415mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 58mg of an off-white solid.

¹H NMR(500MHz,D₂O)δppm 2.12(t,J＝19.7Hz,2H),4.08(s,2H),4.27(s,1H),4.40(s,1H),4.53(s,1H),4.65(s,2H),5.75(s,1H),7.36(s,3H),7.65(s,2H),7.69(d,J＝8.0Hz,2H),8.28(s,1H)；¹³C NMR(125MHz,D₂O)δppm 44.04,63.41,70.06,74.19,83.73,86.90,125.50,125.60,126.00,126.34,127.38,128.15,132.13,132.71,135.18,139.21,153.96,154.73；³¹P NMR(202MHz,D₂O)δppm 15.86,19.01；m/z(ES^-)598.4.

Example 5: preparation of compound 9:

2, 6-dichloro-9- (2',3',5' -tri-O-acetyl- β -D-ribofuranosyl) purine (5.0mmol,2.2g,1.0eq.) and memantine hydrochloride (5.0mmol,1.0g,1.0eq.) were dissolved in 1, 4-dioxane (25 mL). DiEA (12.5mmol,1.6g,2.5eq.) was added at room temperature, heated with stirring overnight, concentrated under reduced pressure in dichloromethane (1000mL), extracted 2 times with water, the organic phase was concentrated under reduced pressure and subjected to column chromatography to give an intermediate which was dissolved in methanolic ammonia (1000mL), stirred overnight at 35 deg.C, concentrated under reduced pressure and the crude product was subjected to column chromatography to give S-II-91.1 g of a 2-chloropurine nucleoside derivative.

The 2-chloropurine nucleoside derivative S-9(1.0mmol,463mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 200mg of off-white solid.

¹H NMR(500MHz,D₂O)δppm 0.76(s,6H),0.98-1.12(m,2H),1.22(s,2H),1.31(d,J＝11.3Hz,2H),1.70(dd,J＝29.4,11.9Hz,4H),1.90(s,2H),2.11(t,J＝19.9Hz,3H),4.07(s,2H),4.28(s,1H),4.44(dd,J＝6.5,2.4Hz,1H),4.67–4.63(m,1H),5.91(d,J＝5.7Hz,1H),8.33(s,1H)；¹³C NMR(125MHz,D₂O)δppm 25.96,26.95,29.97,32.08,39.23,42.32,50.32,55.08,63.73,70.04,74.33,83.78,87.45,116.10,138.34,148.62,153.57,154.16；³¹P NMR(200MHz,D₂O)δppm 18.12；m/z(ES^-)620.2。

Example 6: preparation of compound 10:

2-Chloropurine nucleoside derivative S-10(1.0mmol,451mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 60mg of off-white solid.

¹H NMR(500MHz,D₂O)δppm 2.27(t,J＝19.4Hz,2H),4.17(s,2H),4.35(s,1H),4.46(s,1H),6.02(s,1H),7.27(s,4H),7.54(s,2H),7.97(s,2H),8.33(s,1H)；¹³C NMR(125MHz,D₂O)δppm 16.73,25.47,26.48,27.50,57.39,63.69,70.09,74.31,83.84,87.70,113.16,120.00,122.84,124.66,126.42,138.03,143.08,148.86,152.70,154.12；³¹P NMR(200MHz,D₂O)δppm17.43,19.35–19.97；m/z(ES^-)608.0。

Example 7: preparation of compound 11:

2-Chloropurine nucleoside derivative S-11(1.0mmol,564mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 100mg of off-white solid.

¹H NMR(500MHz,D₂O)δppm 0.77(d,J＝6.5Hz,3H),1.22(s,14H),1.55(s,2H),1.92(s,4H),2.12(t,J＝19.8Hz,2H),2.67(d,J＝38.2Hz,4H),4.03(s,2H),4.19(s,1H),4.40(s,1H),4.60(s,1H),5.20(d,J＝5.6Hz,4H),5.88(d,J＝4.4Hz,1H),8.51(s,1H)；¹³C NMR(125MHz,D₂O)δppm 13.90,22.48,24.71,25.52,27.10,29.26,29.64,31.41,36.86,37.49,63.89,70.44,74.37,84.15,86.83,119.95,127.82,129.70,149.34,152.45,152.86,164.88,174.66；³¹P NMR(200MHz,D₂O)δppm 16.04,18.65；m/z(ES^-)720.4。

Example 8: preparation of compound 12:

2, 6-dichloro-9- (2',3',5' -tri-O-acetyl- β -D-ribofuranosyl) purine (5.0mmol,2230mg,1.0eq.) and 2-naphthylamine (5.0mmol,715mg,1.0eq.) were dissolved in 1, 4-dioxane (25 mL). DIEA (7.5mmol,645mg,1.5eq.) was added at room temperature, heated and stirred overnight, concentrated under reduced pressure to dichloromethane (1000mL), extracted with water for 2 times, the organic phase was concentrated under reduced pressure to give an intermediate for column chromatography, which was dissolved in methanolic ammonia (1000mL) and stirred at 35 ℃ overnight, concentrated under reduced pressure to give the crude product S-II-12670 mg of the nucleoside derivative of 2-chloropurine.

The 2-chloropurine nucleoside derivative S-12(1.0mmol,427mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 100mg of off-white solid.

¹H NMR(D₂O,500MHz)δppm 2.21-2.29(m,2H),4.20-4.22(m,2H),4.40-4.44(m,1H),4.56-4.60(m,1H),4.78-4.80(m,1H),6.09-6.10(m,1H),7.58-7.64(m,3H),7.71-7.73(m,1H),7.97-7.99(m,1H)8.02-8.04(m,2H),8.55(s,1H)；¹³C NMR(D₂O,125MHz)δppm26.38,27.37,28.36,63.61,70.33,84.01,86.76,118.06,121.57,122.25,125.52,126.23,126.41,139.82,149.74,153.39,153.47；³¹P NMR(D₂O,200MHz)δ16.14,18.96ppm；m/z(ES^-)583.9.

Example 9: preparation of compound 15:

2-Chloropurine nucleoside derivative S-15(1.0mmol,513mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 30mg of off-white solid.

¹H NMR(500MHz,D₂O)δppm 0.70(t,J＝6.3Hz,3H),1.06(s,16H),1.27(s,2H),1.58(s,2H),2.10(t,J＝19.7Hz,2H),4.12(d,J＝32.1Hz,4H),4.27(s,1H),4.46(s,1H),4.66(s,1H),6.00(d,J＝4.7Hz,1H),8.60(s,1H)；¹³C NMR(125MHz,D₂O)δppm 13.73,22.40,25.51,27.55,28.40,29.07,29.36,31.66,63.64,66.70,70.27,74.36,83.95,87.18,120.57,142.65,150.05,152.38,153.21；³¹P NMR(200MHz,D₂O)δppm 15.64,18.89；m/z(ES^-)670.1.

Example 10: preparation of compound 16:

2-Chloropurine nucleoside derivative S-16(1.0mmol,478mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 250mg of off-white solid.

¹H NMR(500MHz,D₂O)δppm 1.64(s,11H),1.91(s,4H),2.16(t,J＝19.8Hz,2H),2.29(s,2H),4.14(dddd,J＝6.8,5.9,4.1,1.6Hz,3H),4.30–4.37(m,1H),4.56–4.46(m,1H),6.11(d,J＝5.1Hz,1H),8.72(s,1H)；¹³C NMR(125MHz,D₂O)δppm 16.75,27.23,28.38,33.41,36.07,42.03,51.44,63.47,70.17,74.37,84.19,87.49,115.91,121.90,143.36,149.28,153.15,173.66；³¹P NMR(200MHz,D₂O)δppm 16.37,18.85；m/z(ES^-)634.1。

Example 11: preparation of compound 17:

2, 6-dichloro-9- (2',3',5' -tri-O-acetyl- β -D-ribofuranosyl) purine (5.0mmol,2.2g,1.0eq.) and 2-amantadine hydrochloride (5.0mmol,0.94g,1.0eq.) were dissolved in 1, 4-dioxane (25 mL). DIEA (12.5mmol,1.6g,2.5eq.) was added at room temperature, heated with stirring overnight, concentrated under reduced pressure to dichloromethane (1000mL), extracted with water for 2 times, and the organic phase was concentrated under reduced pressure to give an intermediate which was dissolved in methanolic ammonia (1000mL), and column chromatography was carried out under 35 ℃ overnight with stirring, under reduced pressure to give the 2-chloropurine nucleoside derivative S-II-17880 mg.

2-Chloropurine nucleoside derivative S-17(1.0mmol,435mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 30mg of off-white solid.

¹H NMR(500MHz,D₂O)δppm 1.60(d,J＝12.7Hz,2H),1.71(s,2H),1.82(d,J＝20.8Hz,7H),1.96(d,J＝19.6Hz,4H),2.14(t,J＝19.8Hz,2H),4.10(s,2H),4.20(s,1H),4.31(s,1H),4.47(t,J＝4.2Hz,1H),5.96(d,J＝5.6Hz,1H),8.41(s,1H)；¹³C NMR(125MHz,D₂O)δppm16.70,26.77,30.81,31.44,36.48,36.86,57.36,63.51,70.23,74.20,83.97,86.73,139.05,154.39；³¹P NMR(200MHz,D₂O)δppm 15.85,19.04；m/z(ES^-)592.0。

Example 12: preparation of compound 18:

2-Chloropurine nucleoside derivative S-18(1.0mmol,512mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 70mg of off-white solid.

¹H NMR(500MHz,CD₃OD-d₄)δppm 0.93(t,J＝6.8Hz,3H),1.52(s,18H),1.60–1.74(m,2H),2.33(t,J＝19.8Hz,2H),3.43(t,J＝6.6Hz,2H),4.27(d,J＝21.4Hz,3H),4.47–4.61(m,1H),4.71(t,J＝5.3Hz,1H),6.13(d,J＝5.4Hz,1H),8.76(s,1H)；¹³C NMR(125MHz,CD₃OD-d₄)δppm 13.01,22.30,26.62,28.98,29.28,31.63,39.56,63.95,70.57,74.91,84.44,87.86,118.78,142.32,150.79,151.78,151.97,154.17；³¹P NMR(200MHz,CD₃OD-d₄)δppm 16.03,20.25；m/z(ES^-):669.2.

Example 13: preparation of compound 19:

2, 6-dichloro-9- (2',3',5' -tri-O-acetyl- β -D-ribofuranosyl) purine (5.0mmol,2.2g,1.0eq.) and didodecylamine (5.0mmol,1.8g,1.0eq.) were dissolved in 1, 4-dioxane (25mL), DIEA (7.5mmol,1.0g,1.5eq.) was added at room temperature, the mixture was heated with stirring overnight, the mixture was concentrated under reduced pressure, dissolved in dichloromethane (1000mL), extracted with water for 2 times, the organic phase was concentrated under reduced pressure and subjected to column chromatography to give an intermediate, which was dissolved in methanolic ammonia (1000mL), and was stirred overnight at 35 ℃ and concentrated under reduced pressure, and the crude product was subjected to column chromatography to give S-II-191.3 g of a 2-chloropurine nucleoside derivative.

2-Chloropurine nucleoside derivative S-19(1.0mmol,637mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 150mg of an off-white solid.

¹H NMR(500MHz,CD₃OD-d₄)δppm 0.93(t,J＝6.6Hz,6H),1.32–1.46(m,36H),1.72(s,4H),2.36(t,J＝20.0Hz,2H),3.70(s,2H),4.11–4.32(m,5H),4.47(s,1H),4.66(t,J＝5.2Hz,1H),6.04(d,J＝5.4Hz,1H),8.37(s,1H)；¹³C NMR(125MHz,CD₃OD-d₄)δppm 13.08,22.34,26.44,29.08,29.34,31.68,64.24,70.51,74.51,83.84,87.46,118.27,137.84,151.64,153.46,154.25；³¹P NMR(200MHz,CD₃OD-d₄)δppm 16.20,19.99；m/z(ES^-):794.6.

Example 14: preparation of compound 20:

2, 6-dichloro-9- (2',3',5' -tri-O-acetyl- β -D-ribofuranosyl) purine (5.0mmol,2.2g,1.0eq.) and 2-aminoanthracene (5.0mmol,1.0g,1.0eq.) were dissolved in 1, 4-dioxane (25mL), DIEA (7.5mmol,1.0g,1.5eq.) was added at room temperature, the mixture was heated with stirring overnight, the mixture was concentrated under reduced pressure, dissolved in dichloromethane (1000mL), extracted with water for 2 times, the organic phase was concentrated under reduced pressure and subjected to column chromatography to give an intermediate, which was dissolved in methanolic ammonia (1000mL), and the mixture was stirred overnight at 35 ℃ and concentrated under reduced pressure, and the crude product was subjected to column chromatography to give S-II-20770mg of the 2-chloropurine nucleoside derivative.

2-Chloropurine nucleoside derivative S-20(1.0mmol,477mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 80mg of an off-white solid.

¹H NMR(500MHz,D₂O)δppm 2.25(t,J＝19.4Hz,2H),4.28(d,J＝58.1Hz,3H),4.46(s,2H),5.56(s,1H),7.19(s,3H),7.57(d,J＝73.7Hz,4H),7.85(s,2H),8.21(s,1H)；³¹PNMR(200MHz,D₂O)δppm 18.42,19.15；m/z(ES^-):633.9.

Example 15: preparation of compound 22:

2, 6-dichloro-9- (2',3',5' -tri-O-acetyl- β -D-ribofuranosyl) purine (5.0mmol,2.2g,1.0eq.) and 1-amantadine (5.0mmol,0.76g,1.0eq.) were dissolved in 1, 4-dioxane (25 mL). DIEA (7.5mmol,1.0g,1.5eq.) was added at room temperature, the mixture was heated with stirring overnight, the mixture was concentrated under reduced pressure, methylene chloride (1000mL) was dissolved in methylene chloride, extracted with water for 2 times, the organic phase was concentrated under reduced pressure and column chromatography was performed to give an intermediate, which was dissolved in methanolic ammonia (1000mL), and column chromatography was performed under 35 ℃ overnight with stirring, and under reduced pressure, the crude product was subjected to column chromatography to give S-II-221.3 g of a nucleoside derivative of 2-chloropurine.

2-Chloropurine nucleoside derivative S-22(1.0mmol,435mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 100mg of off-white solid.

¹H NMR(500MHz,D₂O)δppm 1.64(s,6H),2.06(d,J＝33.5Hz,9H),2.17(d,J＝19.9Hz,2H),4.09(s,2H),4.30(d,J＝0.9Hz,1H),4.45(ddd,J＝5.9,3.0,2.0Hz,1H),4.68–4.65(m,1H),5.94(d,J＝5.2Hz,1H),8.38(s,1H)；¹³C NMR(125MHz,D₂O)δppm 26.22,27.22,29.28,35.72,40.80,53.50,63.57,70.19,74.20,83.95,86.81,138.60,148.74,153.76,154.43；³¹P NMR(200MHz,D₂O)δppm 16.38,18.81；m/z(ES-)592.2.

Example 16: preparation of compound 23:

2-Chloropurine nucleoside derivative S-23(1.0mmol,481mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 141mg of an off-white solid.

¹H NMR(D₂O,500MHz)δppm 0.90-0.97(m,9H),1.18-1.20(m,1H),1.36-1.40(m,1H),1.75-1.80(m,1H),1.90-1.97(m,1H),2.00-2.30(m,2H),2.40-2.45(m,1H),4.20-4.25(m,2H),4.40-4.44(m,1H),4.44-4.57(m,1H),4.72-4.73(m,1H),4.91-5.05(m,1H),6.43-6.45(m,1H),9.02(m,1H)；¹³C NMR(D₂O,125MHz)δ12.74,18.03,18.90,35.91,44.47,47.41,48.45,63.50,70.16,74.34,83.22,84.06,84.13,87.37,120.84,142.72,150.23,152.41,153.24,153.45ppm；³¹P NMR(D₂O,200MHz)δ16.49,18.79ppm；m/z(ES^-)638.0.

Example 17: preparation of compound 31:

2, 6-dichloro-9- (2',3',5' -tri-O-acetyl- β -D-ribofuranosyl) purine (5.0mmol,2.2g,1.0eq.) and 2-azaspiro [4.5] decane (5.0mmol,0.70g,1.0eq.) were dissolved in 1, 4-dioxane (25mL), DIEA (7.5mmol,1.0g,1.5eq.) was added at room temperature, the mixture was heated with stirring overnight, the mixture was concentrated under reduced pressure in dichloromethane (1000mL), extracted with water for 2 times, the organic phase was concentrated under reduced pressure and subjected to column chromatography to give an intermediate, which was dissolved in methanolic ammonia (1000mL), and the mixture was stirred overnight at 35 ℃ and concentrated under reduced pressure to give crude 2-chloropurine nucleoside derivative S-II-311.1 g.

The 2-chloropurine nucleoside derivative S-31(1.0mmol,423mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis (phosphorus/phosphorus) chloride was added under ice-bath conditionsTrimethyl acid solution (4.0eq.) is stirred for 2-4h at 0 ℃ and the reaction is detected by TCL. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 140mg of an off-white solid.

¹H NMR(500MHz,D₂O)δppm 1.43(t,J＝22.8Hz,10H),1.83(d,J＝52.0Hz,2H),2.15(t,J＝19.7Hz,2H),3.38(s,1H),3.58(s,1H),3.79(s,1H),40.1(s,1H),4.12(s,2H),4.32(s,1H),4.49(t,J＝4.4Hz,1H),4.67–4.70(m,1H),5.97-5.98(d,J＝5.4Hz,1H),8.36-8.37(d,J＝6.8Hz,1H)；¹³C NMR(125MHz,D₂O)δppm 7.67,8.70,9.72,22.92,27.41,34.49,45.49,47.78,69.61,73.58,74.79,86.08,87.43,118.15,139.22,150.08,153.06,153.84；³¹P NMR(200MHz,D₂O)δppm 15.71,19.11；m/z(ES^-):580.0.

Example 18: preparation of compound 51:

2, 6-dichloro-9- (2',3',5' -tri-O-acetyl- β -D-ribofuranosyl) purine (5.0mmol,2.2g,1.0eq.) and nortropin (5.0mmol,0.86g,1.0eq.) were dissolved in 1, 4-dioxane (25mL), DIEA (7.5mmol,1.0g,1.5eq.) was added at room temperature, the mixture was heated with stirring overnight, the mixture was concentrated under reduced pressure, dissolved in dichloromethane (1000mL), extracted with water for 2 times, the organic phase was concentrated under reduced pressure and subjected to column chromatography to give an intermediate, which was dissolved in methanolic ammonia (1000mL), and the mixture was stirred overnight at 35 ℃ and concentrated under reduced pressure, and the crude product was subjected to column chromatography to give S-II-511.2 g of a 2-chloropurine nucleoside derivative.

2-Chloropurine nucleoside derivative S-51(1.0mmol,411mg,1.0eq.) was dissolved in trimethyl phosphate (10 mL). 5mL of methylene bis phosphorus chloride/trimethyl phosphate solution (4.0eq.) is added under the ice bath condition, the mixture is stirred for 2 to 4 hours under the condition of 0 ℃ and TCL detects the reaction. The reaction was quenched by the addition of triethylamine bicarbonate solution and the pH of the reaction was adjusted to 7 to 8. Extracting with dichloromethane, passing the aqueous phase through the reverse phase¹⁸Column chromatography gave 160mg of an off-white solid.

¹H NMR(500MHz,D₂O)δppm 1.78–2.42(m,11H),4.06(s,1H),4.12(s,2H),4.28–4.37(m,1H),4.45–4.55(m,1H),4.82–4.88(m,1H),5.38(dd,J＝3.4,2.4Hz,1H),5.99(d,J＝5.6Hz,1H),8.40(s,1H)；¹³C NMR(125MHz,D₂O)δppm 26.05,27.10,27.68,37.08,38.20,53.79,54.40,64.38,70.20,74.09,83.99,86.71,117.78,138.54,151.12,154.22；³¹P NMR(200MHz,D₂O)δppm 16.90,18.63；m/z(ES^-):568.0。

Biological assay

Screening assays for CD73 inhibitors

To evaluate the inhibitory effect of compounds on CD73, the Malachite Green phosphate assay kit (R & D, Cat # DY996) was used. Briefly, compounds were dissolved and diluted to the desired concentration with phosphate-free buffer (Tris-HCl, pH 7.3). mu.L of the compound solution was added to an equal volume of CD73 protein solution (2 Xconcentration, 0.5. mu.g/mL, Novoprotein, Cat # C446) followed by incubation at room temperature for 5 min. To each well 10 μ L of malachite green reagent a was added, mixed well and incubated at room temperature for 10 minutes. Then, 10 μ L of malachite green reagent B was added to each well, mixed well and incubated at room temperature for 20 minutes. Finally, the optical density per well was determined using a microplate reader at 620 nm. The activity of the compounds is shown in Table 2. The results of the experiments with compound 9 and compound 22 obtained from the CD73 inhibitor screening experiments are shown in fig. 1 and 2.

TABLE 2 Experimental test for the Activity of Compounds

Compound numbering	Inhibition of CD73 Performance
		1	+++
6	+++
		7	+++
8	++
		9	+++
10	++
		11	++
12	++
		15	+++
16	++
		17	+++
18	++
		19	+
20	++
		22	+++
23	++
		31	+++
51	+++

Inhibition of CD73, IC₅₀:+,>100nM；++,10-100nM；+++,<10nM

Cellular level analysis of CD73 Activity

The 5 × compound solution was prepared by dissolving and diluting the compound to the desired concentration in serum-free RMPI-1640 medium with 1 μ M AMP. A375 cells were collected and washed twice with PBS, then allowed to stand at 1.125X 10⁵The density of/mL was suspended in serum-free RMPI-1640 medium. Each aliquot of 80. mu.L of the cell suspension was placed in a 96-well plate, followed by addition of 20. mu.L of a 5X compound solution, gentle mixing, and 5% CO at 37 ℃₂The cells were incubated for 16 hours. After incubation, 50 μ Ι _ of supernatant was transferred from each well to a new 96-well plate. Subsequently, 2. mu.L of 2.5. mu.M ATP and 50. mu.L of Celltiter Glo reagent were added to each well in sequence. Fluorescence was measured using pherastar (bmg).

3. Method for evaluating pharmacokinetics of compound

After a single intravenous (1 mg/kg)/intragastric (1mg/kg) administration of the compound to fasted SD male rats, blood samples were collected at 0.08h, 0.25h, 0.5h, 1h, 2h, 4h, 6h, 8h and 24h after administration. Plasma was separated by centrifugation (8000rpm) and frozen (-20 ℃) until used for analysis. The concentration of the compound in rat plasma was determined by HPLC-MS/MS. Plasma was dispensed into appropriate tubes containing internal standard and methanol or acetonitrile. The tubes were shaken vigorously for 3 minutes to effect deproteinization, followed by centrifugation at 8000rmp for 5 minutes. The supernatant was transferred to an autosampler vial and injected into the chromatography system. Pharmacokinetic parameters, such as AUC, were calculated using WinMonlin 6.3 software_0-tCmax, tmax, t1/2, MRT, C1 and Vd. The absolute bioavailability was calculated as follows:f ═ AUC (i.g.) x dose (i.v.)]/[ AUC (i.v.) x dose (i.g.)]×100％。

While the invention has been described in detail with reference to the embodiments thereof, the embodiments are provided for the purpose of illustration and not for the purpose of limitation. Other embodiments that can be derived from the principles of the invention are intended to be within the scope of the invention as defined by the claims.

Claims

1. A compound of formula I, or a pharmaceutically acceptable ester or salt thereof

Wherein:

w is oxygen, sulfur or methylene;

x is selected from phosphoryl (-P (═ O) (OR) -), sulfonyl (-S (═ O)₂-) and a carbonyl (-C (═ O) -) moiety, wherein R is hydrogen, an ester-forming group, or a protecting group;

y is selected from phosphonate (-PO)₃R₂) Sulfonate (-SO)₃R) and a carboxylate (-CO)₂R), wherein R is hydrogen, an ester-forming group, or a protecting group;

R¹is hydroxy or hydrogen;

R²is chlorine or hydrogen; and

R³and R⁴Independently selected from hydrogen, alkyl, alkenyl and alkynyl, wherein R³And R⁴Has 11 to 30 carbon atoms; or

R³And R⁴Independently selected from hydrogen, alkyl, alkenyl and alkynyl, wherein R³And R⁴has-C (═ O) R⁵OR-C (═ O) OR⁵Wherein R is⁵Is alkyl, alkenyl or alkynyl having 11 to 30 carbon atoms; or

R³And R⁴Independently selected from hydrogen and a ring system containing a bicyclic, tricyclic, spirocyclic, fused or bridged ring carbocyclic or heterocyclic ring system, said carbocyclic system being aromatic or non-aromatic, the formerIs R³And R⁴Not hydrogen at the same time; or

R³Is hydrogen or lower alkyl, and R⁴Is an oligoethylene glycol residue or a derivative thereof. Such as, but not limited to, — (CH)₂CH₂O)_n─R⁶Where n is equal to 3 to 10, R⁶Selected from hydrogen or lower alkyl (e.g., methyl, ethyl, propyl, butyl, etc.); or

R³Is hydrogen or lower alkyl, and R⁴Is a substituted or unsubstituted arylalkyl, alkylaryl, or a group consisting of an aromatic hydrocarbon group (a carbon aryl or heteroaryl group) and an aliphatic hydrocarbon group.

2. The compound of claim 1, wherein R³Is hydrogen or lower alkyl; and R⁴is-C (═ O) R⁵OR-C (═ O) OR⁵Wherein R is⁵Is C₁₁To C₃₀Alkyl, alkenyl or alkynyl.

3. The compound of claim 1, wherein R³Is hydrogen or lower alkyl; and R⁴is-C (═ O) R⁵OR-C (═ O) OR⁵Wherein R is⁵Is a ring system having a carbocyclic or heterocyclic ring system containing bicyclic, tricyclic, spirocyclic, fused, or bridged rings, the carbocyclic ring system being aromatic or non-aromatic, and the heterocyclic ring system being substituted or unsubstituted.

4. The compound of any one of claims 1 to 3, wherein the compound is a compound of formula II or formula III, or a pharmaceutically acceptable salt or ester thereof:

5. a compound according to any one of claims 1 to 3, wherein the compound is of formula IV:

6. a compound according to any one of claims 1 to 3, wherein the compound is of formula V or formula VI:

7. a compound according to any one of claims 1 to 3, wherein the compound is of formula VII or VIII:

wherein:

r is hydrogen, an ester-forming group, or a protecting group.

8. A compound according to any one of claims 1 to 3, wherein the compound is of formula IX, or a pharmaceutically acceptable salt or ester thereof:

wherein:

r is hydrogen, an ester-forming group, or a protecting group;

R¹is hydroxy or hydrogen;

R²is hydrogen or chlorine; and

R³is hydrogen or lower alkyl, and R⁴Is alkyl, alkenyl or alkynyl having 11 to 30 carbon atoms; or

R³Is hydrogen or lower alkyl, andR⁴is a substituent comprising a bicyclic, tricyclic, or polycyclic ring system, wherein the ring system is a fused ring system, a spiro ring system, a bridged ring system, or a parallel ring system, and the ring system is a carbocyclic ring, an aliphatic ring, an aromatic ring, a heterocyclic ring, or a combination thereof; or

R³Is hydrogen or lower alkyl, and R⁴is-C (═ O) R⁵OR-C (═ O) OR⁵Wherein R is⁵Is alkyl, alkenyl or alkynyl having 11 to 30 carbon atoms; or

R³Is hydrogen or lower alkyl, and R⁴is-C (═ O) R⁵OR-C (═ O) OR⁵Wherein R is⁵Is a substituent containing a carbon or heterocyclic ring system containing a bicyclic, tricyclic, spiro, fused or bridged ring, said carbocyclic or heterocyclic ring system being aromatic or non-aromatic, said heterocyclic ring system being substituted or unsubstituted, wherein said ring is a carbocyclic ring, an aliphatic ring, an aromatic ring, a heterocyclic ring, or combinations thereof; or R³Is hydrogen or lower alkyl, and R⁴Is unsubstituted or substituted 1-adamantyl, α -naphthylmethyl or β -naphthylmethyl, or

R³、R⁴And the nitrogen to which they are attached form a fused three-membered ring structure.

9. The compound of claim 8, wherein R⁴Is a group containing a adamantyl moiety; alternatively, R⁴Is substituted or unsubstituted 1-adamantyl or 2-adamantyl; alternatively, R⁴Is 1-adamantylethyl, 1-adamantylpropyl or 1-adamantylbutyl, wherein the adamantyl moiety can be substituted or unsubstituted.

10. A compound according to claim 8, R⁴Is a group containing a naphthyl moiety; alternatively, R⁴Is a substituted or unsubstituted α -naphthyl or β -naphthyl group, alternatively, R⁴α -naphthylmethyl or β -naphthylmethyl, in the absence or presence of other substitutions, and optionally R⁴Selected from naphthyl ethylNaphthylpropyl, wherein the naphthyl moiety can be unsubstituted or substituted.

11. The compound of claim 8, wherein the fused three-membered ring structure is a substituted or unsubstituted carbazolyl moiety.

12. The compound of any one of claims 1 to 11, wherein the compound is a compound listed in table 1, or a pharmaceutically acceptable salt or ester thereof.

13. The compound of any one of claims 1 to 12, wherein C, H, O and the N atom in the compound are each independently selected from a naturally abundant atom and an isotopically enriched atom.

14. The compound of claim 13, wherein said isotopically enriched atoms are selected from those of carbon¹²C、¹³C and¹⁴c; selected from hydrogen¹H、²H and³h; selected from oxygen¹⁶O、¹⁷O and¹⁸o; and is selected from nitrogen¹⁴N and¹⁵N。

15. a pharmaceutical composition comprising a compound of any one of claims 1 to 14, or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable carrier.

16. The pharmaceutical composition of claim 15, wherein the pharmaceutically acceptable carrier comprises a cream, an emulsion, a gel, a liposome, or a nanoparticle.

17. The pharmaceutical composition of claim 15 or 16, wherein the composition is suitable for oral administration.

18. The pharmaceutical composition of claim 17, wherein the composition is a hard-shelled gelatin capsule, a soft-shelled gelatin capsule, a cachet, a pill, a tablet, a lozenge, a powder, a granule, a pellet, or a dragee.

19. The pharmaceutical composition of claim 17, wherein the composition is in the form of a solution, an aqueous liquid suspension, a non-aqueous liquid suspension, an oil-in-water liquid emulsion, a water-in-oil liquid emulsion, an elixir, or a syrup.

20. The pharmaceutical composition of any one of claims 17 to 19, wherein the composition has an enteric coating.

21. The pharmaceutical composition of any one of claims 17 to 19, wherein the composition is formulated for controlled release.

22. The pharmaceutical composition of claim 15, wherein the composition is injectable.

23. A method of treating or preventing a CD 73-related disease, disorder or condition in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the compound of any one of claims 1-14 or the pharmaceutical composition of any one of claims 15-22, thereby treating or preventing a CD 73-related disease, disorder or condition in the subject.

24. The method of claim 23, wherein the compound is administered in an amount effective to reverse, slow or stop the progression of CD 73-mediated immunosuppression in the subject.

25. The method of claim 23 or 24, wherein the CD 73-associated disease, disorder or condition is cancer.

26. The method of claim 25, wherein the cancer is a cancer of the prostate, colon, rectum, pancreas, cervix, stomach, endometrium, brain, liver, bladder, ovary, testis, head, neck, skin, mesothelial membrane, white blood cells, esophagus, breast, muscle, connective tissue, lung, adrenal gland, thyroid, kidney, or bone.

27. The method of claim 25, wherein the cancer is glioblastoma, mesothelioma, renal cell carcinoma, gastric cancer, sarcoma, choriocarcinoma, basal cell carcinoma of the skin, or testicular seminoma.

28. The method of claim 25, wherein the cancer is selected from or includes melanoma, colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, leukemia, brain tumors, lymphoma, ovarian cancer, kaposi's sarcoma, and the like.

29. The method of claim 23 or 24, wherein the disease, disorder or condition associated with CD73 is selected from or includes rheumatoid arthritis, renal failure, lupus, asthma, psoriasis, colitis, pancreatitis, allergy, fibrosis, anemic fibromyalgia, alzheimer's disease, congestive heart failure, stroke, aortic valve stenosis, arteriosclerosis, osteoporosis, parkinson's disease, infection, crohn's disease, ulcerative colitis, allergic contact dermatitis, eczema, systemic sclerosis, and multiple sclerosis, and the like.

30. The pharmaceutical composition of any one of claims 15 to 22, further comprising at least one additional therapeutic agent.

31. The pharmaceutical composition of claim 30, wherein the at least one additional therapeutic agent is a chemotherapeutic agent, an immune and/or inflammation modulator, an anti-hypercholesterolemic agent, or an anti-infective agent.

32. The pharmaceutical composition of claim 30, wherein the at least one additional therapeutic agent is an immune checkpoint inhibitor.

33. The method of any of claims 23 to 29, further comprising: administering at least one additional therapeutic agent to the subject.

34. The method of claim 33, wherein the at least one additional therapeutic agent is administered simultaneously or sequentially with the compound or composition.

35. The method of claim 33 or 34, wherein the at least one additional therapeutic agent is a chemotherapeutic agent, an immune and/or inflammation modulator, an anti-hypercholesterolemic agent, or an anti-infective agent.

36. A kit comprising a compound or pharmaceutically acceptable salt or ester according to any one of claims 1 to 14, and at least one additional therapeutic agent.

37. The kit of claim 36, wherein the at least one additional therapeutic agent is a chemotherapeutic agent, an immune and/or inflammation modulator, an anti-hypercholesterolemic agent, an anti-infective agent, or an immune checkpoint inhibitor.

38. The kit of claim 36 or 37, further comprising a buffer or excipient, and/or instructions for use thereof.

39. A method of treating cancer in a subject, the method comprising: administering to the subject a therapeutically effective amount of a compound of any one of claims 1 to 14, or a pharmaceutically acceptable salt or ester thereof, and an immune checkpoint inhibitor, thereby treating cancer in the subject.

40. The method of claim 39, wherein said administering is prior to, concurrent with, or subsequent to radiation therapy.

41. The method of claim 39, wherein the compound and the immune checkpoint inhibitor are administered in combination.

42. The method of claim 39, wherein the compound and the immune checkpoint inhibitor are administered sequentially.

43. The method of claim 42, wherein the compound is administered after the immune checkpoint inhibitor.

44. The method of claim 43, wherein the compound is administered prior to the immune checkpoint inhibitor.

45. The pharmaceutical composition of claim 32, the kit of claim 37, or the method of any one of claims 39 to 44, wherein the immune checkpoint inhibitor is selected from or comprises Yiprioman, Navolumab, and lambrolizumab, among others.