WO2020139803A1 - Benzothiadiazine derivatives and compositions comprising the same for treating disorders mediated by adenosine - Google Patents

Abstract

Disclosed herein are novel inhibitors of CD73. More specifically, the novel CD73 inhibitors are derived from benzothiadiazine. Further, the present disclosure is related to the use of said benzothiadiazine derivatives in the treatment of diseases and/or disorders mediated by adenosine, such as AIDS, an autoimmune disease, atherosclerosis, a cancer, an infectious disease, ischemia-reperfusion injury and pre-cancerous syndrome. Also encompassed in the present disclosure are pharmaceutical compositions comprising said benzothiadiazine derivatives.

Description

BENZOTHIADIAZINE DERIVATIVES AND COMPOSITIONS COMPRISING THE SAME FOR TREATING DISORDERS MEDIATED BY ADENOSINE

CROSS-REFERENCE TO RELATED APPLICATION

This application claims the priority of US Provisional Application No.

62/784,557, filed December 24, 2018, the content of which is incorporated by reference in its entirety for all purposes.

BACKGROUND OF THE INVENTION

1. FIELD OF THE INVENTION

The present disclosure in general relates to the field of benzothiadiazines; more particularly to the derivatives of benzothiadiazines, and their use as CD73 inhibitors in the treatment of diseases and/or disorders mediated by adenosine.

2. DESCRIPTION OF RELATED ART

CD73 or ecto-5 '-nucleotidase (5 '-NT) is an enzyme that anchors to the cell membrane through a glycosylphosphatidylinositol (GPI) linkage, has ecto-enzyme activity. The primary function of CD73 is to convert extracellular nucleotides (e.g., 5-AMP) to their corresponding nucleosides (e.g., adenosine) in the extracellular space.

Adenosine is an endogenous modulator of diverse physiological functions, including the cardiovascular system, the central nervous system (CNS), and the immune system, where extracellular adenosine acts on a variety of immune cells and mediates anti-inflammatory effects. Adenosine also promotes fibrosis (excess matrix production) in a variety of tissues. As inhibition of CD73 results in decreased adenosine, thus CD73 inhibitors can be used for the treatment of diseases or disorders mediated by adenosine and its actions on adenosine receptors. Thus, CD73 inhibitors can be used for enhancing immune responses, as well as for treating a wide range of conditions, including neurological and CNS disorders, ischemic, immune and/or infectious diseases.

In light of the role that CD73 plays in disease pathogenesis, there is a need for new inhibitor of CD73. Unexpectedly, inventors of the present disclosure found several novel benzothiadiazine derivatives may antagonize the function of CD73.

Accordingly, these novel benzothiadiazine derivatives may be useful as lead compounds for the development of therapeutic agents for the treatment of diseases and/or disorders mediated by adenosine. SUMMARY

The present disclosure is based on unexpected discovery that certain

benothiadiazine derivatives are potent inhibitor of CD73, thus are useful as lead compounds for the development of medicaments for the treatment of diseases and/or disorders mediated by adenosine, such as cancer, autoimmune diseases, pre-cancerous syndrome, adult immune deficiency syndrome (AIDS), atherosclerosis,

ischemia-reperfusion injury, and/or infectious diseases.

Accordingly, one aspect of the present disclosure is to provide a novel compound having the structure of formula (I), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, and stereoisomer thereof:

R is phenyl or aryl;

R3 is H, Ci -20 alkyl, or -(ClUj_n-phenyl, in which n is an integral between 1 to 10; R4 is H, Ci -20 alkyl, phenyl, or heteroaryl; and

each Ci -20 alkyl, phenyl and heteroaryl is optionally substituted with at least one substituent selected from the group consisting of halogen, hydroxyl, C_M lower alkyl, haloalkyl, alkoxyl, haloalkoxy, -CN, and -COOH.

In certain embodiments, the compound of formula (I) can be the compound of formula (II), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, and stereoisomer thereof:

X, Y, and Z are independently C or N, provided that when one of X, Y, and Z is N, then the other two are independently C;

R₃ is H, Ci-₄ lower alkyl, or -Cth-phenyl;

R^a and R^b are independently H, -OR₅, phenyl, pyridinyl, or thiophenyl, in which R5 is Ci-4 lower alkyl or phenyl;

R^c is nil or at least one substituent selected from the group consisting of halogen, -OH, Ci-₄ lower alkyl, haloalkyl, alkoxyl, and haloalkoxy;

each phenyl and pyridinyl is optionally substituted with at least one substituent selected from the group consisting of halogen, -OH, -OR₆, -NH₂, -NR₇R₈ alkyl, haloalkyl, and -CN; and

R6, R₇, and Re are independently C_1-4 lower alkyl or phenyl optionally substituted with at least one halogen.

In one embodiment, in formula (II), X, Y, and Z are independently C; R₃ is H;

R^a is phenyl substituted with at least one fluoro; R^b is H; and R^c is methoxy.

In another embodiment, in Formula (II), X and Y are independently C, and Z is N; R₃ is H; R^a is phenyl substituted with two fluoro; R^b is H; and R^b is nil.

In certain embodiments, the compound of formula (I) can be the compound of formula (III), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, and stereoisomer thereof: wherein, R^a is phenyl optionally substituted with at least one halogen.

In certain embodiments, the compound of formula (I) can be the compound of formula (IV), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, and stereoisomer thereof:

Wherein:

X is C or -C=0; and

R^c is nil or at least one substituent selected from the group consisting of halogen, -OH, Ci-4 lower alkyl, haloalkyl, alkoxyl, and haloalkoxy.

In one embodiment, in formula (IV), X is C; and R^c is methoxy.

In another embodiment, in formula (IV), X is -C=0; and R^c is methoxy.

A further aspect of the present disclosure is to provide a pharmaceutical composition for the treatment of a subject having or suspected of having a disease and/or disorder mediated by adenosine. The disease and/or disorder mediated by adenosine may be AIDS, an autoimmune disease, atherosclerosis, a cancer, an infectious disease, ischemia-reperfusion injury or pre-cancerous syndrome. The pharmacological composition comprises a therapeutically effective amount of the compound of formula (I) to (IV); and a pharmaceutically acceptable carrier.

The compound of formula (I) to (IV) is present at a level of about 0.1 % to 99% by weight, based on the total weight of the pharmaceutical composition. In some embodiments, the compound of formula (I) to (IV) is present at a level of at least 1% by weight, based on the total weight of the pharmaceutical composition. In certain embodiments, the compound of formula (I) to (IV) is present at a level of at least 5% by weight, based on the total weight of the pharmaceutical composition. In still other embodiments, the compound of formula (I) to (IV) is present at a level of at least 10% by weight, based on the total weight of the pharmaceutical composition. In still yet other embodiments, the compound of formula (I) to (IV) is present at a level of at least 25% by weight, based on the total weight of the pharmaceutical composition.

According to some preferred embodiments, the pharmaceutical composition further comprises an anti-neoplastic agent, an immunosuppressant, or an anti-infectious agent.

In some examples, the anti-neoplastic agent may be an anti-microtubule agent, a platinum coordination complex, an alkylating agent, an antibiotic, a topoisomerase II inhibitor, a topoisomerase I inhibitor, an anti-metabolite, a hormone, a signal transduction pathway inhibitor, a non-receptor tyrosine kinase, an angiogenesis inhibitor, an immunotherapeutic agent, a pro-apoptotic agent, cell cycle signaling inhibitor, a proteasome inhibitor, an inhibitor of cancer metabolism, an anti-PD-Ll agent, a PD-1 antagonist, an immune-modulator, Stimulator of Interferon Genes (STING) modulating compound, CD39 inhibitor, A2a and A2a adenosine antagonists, a Toll-like receptor 4 (TLR4) inhibitor, an anti-ICOS antibody or an anti-OX40 antibody. In some examples the immunosuppressant may be a corticosteroid, a calcineurin inhibitor, a mammalian target of rapamycin (mTOR) inhibitor, an Inosine monophosphate dehydrogenase (IMPDH) inhibitor, a therapeutic protein, or a monoclonal antibody. In some examples, the anti-infectious agent may be an anti-biotic that inhibits the growth of Gram-negative or Gram-positive bacteria.

The present disclosure also encompasses a method for the treatment of a subject having a disease and/or disorder mediated by adenosine. The method comprises the step of administering the present pharmaceutical composition to the subject, so as to ameliorate, mitigate and/or prevent the symptoms of the disease and/or disorder mediated by adenosine. The disease and/or disorder mediated by adenosine may be AIDS, an autoimmune disease, atherosclerosis, a cancer, an infectious disease, ischemia-reperfusion injury or pre-cancerous syndrome.

Examples of the autoimmune disease treatable by the present method include, but are not limited to, multiple sclerosis, psoriasis, systemic lupus erythemotosis (SLE), Type I diabetes mellitus, and Wegener’s granulomatosis.

Examples of the infectious disease treatable by the present method include, but are not limited to, bacterial, fungal and viral infections.

Examples of cancer treatable by the present method include, but are not limited to, acinar cell carcinoma, acute lymphoblastic leukemia, acute myelogenous leukemia, adenocarcinoma, adenosquamous carcinoma, Bannayan-Zonana syndrome, bladder cancer, breast cancer, buccal cancer, Burkitt's lymphoma, cervical cancer, colon cancer, chronic myelogenous leukemia, chronic lymphocytic leukemia, chronic neutrophilic leukemia, Cowden disease, ductal adenocarcinoma, endometrial cancer, Edwing’s sarcoma, ependymoma, erythroleukemia, esophageal cancer, follicular lymphoma, gastric cancer, gastrointestinal stromal tumor, giant cell tumor of bone, glioma, glioblastoma, astrocytoma, glioblastoma multiforme, glucagonoma, hairy-cell leukemia, head and neck cancer, hodgkins lymphoma, insulinoma, immunoblastic large cell leukemia, kidney cancer, Lhermitte-Duclos disease, lobular carcinoma, liver cancer, lymphoblastic T cell leukemia, malignant lymphoma, medulloblastoma, melanoma, mantle cell leukemia, mesothelioma, multiple myeloma, megakaryoblastic leukemia, nasopharangeal cancer, non-hodgkins lymphoma, non-small cell lung cancer, osteosarcoma, ovarian cancer, pancreatic cancer, plasmacytoma, promyelocytic leukemia, prostate cancer, Rhabdomyosarcoma, sarcoma, salivary gland cancer, skin cancer, small cell lung cancer, testicular cancer, thyroid cancer, urothelial cancer, vulval cancer, and Wilm’s tumor.

According to some preferred embodiments, the method further includes the step of administering to the subject another agent that is known to improve the symptoms of the disease and/or disorder mediated by adenosine, such as an anti-neoplastic agent, an immunosuppressant, or an anti-infectious agent before, together with, or after the administration of the present pharmaceutical composition. The details of one or more embodiments of this disclosure are set forth in the accompanying description below. Other features and advantages of the invention will be apparent from the detail descriptions, and from claims.

It is to be understood that both the foregoing general description and the following detailed description are by examples, and are intended to provide further explanation of the invention as claimed.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The detailed description provided below in connection with the appended drawings is intended as a description of the present disclosure and is not intended to represent the only forms in which the present disclosure may be constructed or utilized.

1. Definitions

When a range of values is listed, it is intended to encompass each value and sub range within the range. For example“Ci^” is intended to encompass, Ci, Ci, C3, C4, Ci_^t, Ci-3, Ci-2, C2^, C2-3, and C3^.

Unless otherwise indicated, the term“alkyl” means a straight chain, branched and/or cyclic (“cycloalkyl”) hydrocarbon having from 1 to 20 (e.g., 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, or 1) carbon atoms. Alkyl moieties having from 1 to 4 carbons (Ci^ alkyl) are referred to as“lower alkyl.” Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, isobutyl,

2-isopropyl-3 -methyl butyl, pentyl, pentan-2-yl, hexyl, isohexyl, heptyl, heptan-2-yl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl and dodecyl. Cycloalkyl moieties may be monocyclic or multicyclic, and examples include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Unless otherwise specified, each instance of an alkyl group is independently optionally substituted, /._<?.,

unsubstituted (an“unsubstituted alkyl”) or substituted (a“substituted alkyl”) with one or more substituents. In certain embodiments, the alkyl group is substituted C2-10 alkyl. In some embodiments, cycloalkyl is a monocyclic, saturated carbocyclyl group having from 3 to 6 ring carbon atoms (“C3-6 cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C5-6 cycloalkyl”). Examples of C5-6 cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (Ce). Unless otherwise specified, each instance of a cycloalkyl group is independently unsubstituted (an“unsubstituted cycloalkyl”) or substituted (a“substituted cycloalkyl”) with one or more substituents. In certain embodiments, the cycloalkyl group is unsubstituted C3-10 cycloalkyl. In certain embodiments, the cycloalkyl group is substituted C3-10 cycloalkyl.

Unless otherwise indicated, the term“aryl” means an aromatic ring or a partially aromatic ring system composed of carbon and hydrogen atoms. An aryl moiety may comprise multiple rings bound or fused together. Examples of aryl moieties include phenyl, naphthyl, fluorenyl, and fluorenone. In further examples, the aryl refers to fluorenon. Unless otherwise specified, each instance of an aryl group is independently optionally substituted, /._<?., unsubstituted (an“unsubstituted aryl”) or substituted (a “substituted aryl”) with one or more substituents. In certain embodiments, the aryl group is a substituted phenyl. In other embodiments, the aryl group is an unsubstituted fluorenyl. In further embodiments, the aryl is an unsubstituted fluorenone.

Unless otherwise indicated, the term“heteroaryl” means an aryl moiety wherein at least one of its carbon atoms has been replaced with a heteroatom (e.g., N, O or S).

In some embodiments, a heteroaryl group is a 5-10 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”). In some embodiments, a heteroaryl group is a 5-8 membered aromatic ring system having ring carbon atoms and 1—4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In some embodiments, a heteroaryl group is a 5-6 membered aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In some embodiments, the 5-6 membered heteroaryl has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, each instance of a heteroaryl group is independently optionally substituted, /._<?., unsubstituted (an “unsubstituted heteroaryl”) or substituted (a“substituted heteroaryl”) with one or more substituents. Exemplary 5-membered heteroaryl groups containing one heteroatom include, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, without limitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, without limitation, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, without limitation, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, without limitation, pyridinyl.

Exemplary 6-membered heteroaryl groups containing two heteroatoms include, without limitation, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, without limitation, triazinyl and tetrazinyl, respectively.

Unless otherwise indicated, the term“alkylaryl” or“alkyl-aryl” means an alkyl moiety bound to an aryl moiety.

Unless otherwise indicated, the term“alkoxy” means an -O-alkyl group.

Examples of alkoxy groups include, but are not limited

to, -OCH₃, -OCH2CH3, -0(CH₂)₂CH₃, -0(CH₂)₃CH₃, -0(CH₂)₄CH₃, and -0(CH₂)₅CH₃. The term“lower alkoxy” refers to -0-(lower alkyl), such as -OCH₃ and -OCH₂CH₃.

Unless otherwise indicated, the terms“halogen” and“halo” encompass fluoro, chloro, bromo, and iodo.

The term“amino” refers to a moiety of the formula: -N(R)₂, wherein each instance of R is independently a substituent described herein, or two instances of R are connected to form substituted or unsubstituted heterocyclyl. In certain embodiments, the amino is unsubstituted amino (i.e., -NH₂). In certain embodiments, the amino is a substituted amino group, wherein at least one instance of R is not hydrogen.

Unless otherwise indicated, the term“substituted,” when used to describe a chemical structure or moiety, refers to a derivative of that structure or moiety wherein one or more of its hydrogen atoms is substituted with an atom, chemical moiety or functional group such as, but not limited to, -OH, -CHO, alkoxy, alkanoyloxy (e.g., -OAc), alkenyl, alkyl (e.g., methyl, ethyl, propyl, t-butyl), aryl, aryloxy, halo, or haloalkyl (e.g., -CCh, -CP₃, -C(CF₃)₃).

In a particular embodiment, the term“substituted,” when used to describe a chemical structure or moiety, refers to a derivative of that structure or moiety wherein one or more of its hydrogen atoms is substituted with one or more of: alkoxy, alkanoyloxy, alkyl, aryl, halo, haloalkyl, or hydroxyl. Unless otherwise indicated, one or more adjectives immediately preceeding a series of nouns is to be construed as applying to each of the nouns. For example, the phrase“optionally substituted alky, pnehyl, aryl, or heteroaryl” has the same meaning as “optionally substituted alky, optionally substituted alkyl, optionally substituted phenyl, optionally substituted aryl, or optionally substituted heteroaryl.”

The present disclosure is not intended to be limited in any manner by the above exemplary listing of substituents.

The term“solvate” refers to forms of the compound that are associated with a solvent, usually by a solvolysis reaction. This physical association may include hydrogen bonding. Conventional solvents include water, methanol, ethanol, acetic acid, dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), diethyl ether, and the like. The compounds described herein may be prepared, e.g., in crystalline form, and may be solvated. Suitable solvates include pharmaceutically acceptable solvates and further include both stoichiometric solvates and non-stoichiometric solvates. In certain instances, the solvate will be capable of isolation, for example, when one or more solvent molecules are incorporated in the crystal lattice of a crystalline solid. “Solvate” encompasses both solution-phase and isolatable solvates. Representative solvates include hydrates, ethanolates, and methanolates.

The term“hydrate” refers to a compound which is associated with water.

Typically, the number of the water molecules contained in a hydrate of a compound is in a definite ratio to the number of the compound molecules in the hydrate. Therefore, a hydrate of a compound may be represented, for example, by the general formula R x H2O, wherein R is the compound, and x is a number greater than 0. A given compound may form more than one type of hydrate, including, e.g., monohydrates (x is 1), lower hydrates (x is a number greater than 0 and smaller than 1, e.g., hemihydrates (R-0.5 H2O)), and polyhydrates (x is a number greater than 1, e.g., dihydrates (R-2 H2O) and hexahydrates (R-6 H2O)).

It is also to be understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed“isomers”. Isomers that differ in the arrangement of their atoms in space are termed“stereoisomers”.

Stereoisomers that are not mirror images of one another are termed

“diastereomers” and those that are non-superimposable mirror images of each other are termed“enantiomers”. When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (/._<?., as (+) or (-)-isomers respectively). A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a“racemic mixture”.

It should also be noted that if the stereochemistry of a structure or a portion of a structure is not indicated with, for example, bold or dashed lines, the structure or the portion of the structure is to be interpreted as encompassing all stereoisomers of it.

Similarly, names of compounds having one or more chiral centers that do not specify the stereochemistry of those centers encompass pure stereoisomers and mixtures thereof. Moreover, any atom shown in a drawing with unsatisfied valences is assumed to be attached to enough hydrogen atoms to satisfy the valences.

Unless otherwise indicated,“an effective amount” of a compound is an amount sufficient to provide a therapeutic benefit in the treatment or management of a disease or condition, or to delay or minimize one or more symptoms associated with the disease or condition. A therapeutically effective amount of a compound is an amount of therapeutic agent, alone or in combination with other therapies, which provides a therapeutic benefit in the treatment or management of the disease or condition. The term “effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms or causes of a disease or condition, or enhances the therapeutic efficacy of another therapeutic agent.

Unless otherwise indicated, the terms“treat,”“treating” and“treatment” contemplate an action that occurs while a patient is suffering from the specified disease or disorder, which reduces the severity of the disease or disorder, or one or more of its symptoms, or retards or slows the progression of the disease or disorder.

The term“pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid or by using other methods known in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3- phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N⁺(C,-₄ alky 1)₄ salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate, and aryl sulfonate.

The term“pharmaceutically acceptable carrier” refers to a carrier, whether diluent or excipient, that is compatible with the other ingredients of a formulation and not deleterious to the recipient thereof.

The terms "administration of a composition" or "administering a composition" is defined to include an act of providing a compound or pharmaceutical composition of the present invention to the subject in need of treatment.

Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in the respective testing measurements. Also, as used herein, the term“about” generally means within 10%, 5%, 1%, or 0.5% of a given value or range. Alternatively, the term“about” means within an acceptable standard error of the mean when considered by one of ordinary skill in the art. Other than in the

operating/working examples, or unless otherwise expressly specified, all of the numerical ranges, amounts, values and percentages such as those for quantities of materials, durations of times, temperatures, operating conditions, ratios of amounts, and the likes thereof disclosed herein should be understood as modified in all instances by the term“about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth in the present disclosure and attached claims are approximations that can vary as desired. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

The singular forms“a”,“and”, and“the” are used herein to include plural referents unless the context clearly dictates otherwise.

2. Novel Compounds

The compounds as described herein can have the structure of formula (I), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, and stereoisomer thereof:

In the formul (I), R may be phenyl or aryl; R3 may be H, Ci-20 alkyl, or -(CH2)- _n-phenyl, in which n is an integral between 1 to 10; and R4 may be H, Ci-20 alkyl, phenyl, or heteroaryl. Optionally, each Ci-20 alkyl, phenyl and heteroaryl is substituted with at least one substituent selected from the group consisting of halogen, hydroxyl, C_M lower alkyl, haloalkyl, alkoxyl, haloalkoxy, aryloxy, -CN, and -COOH.

In certain embodiments, R is phenyl optionally substituted with hydroxyl. In other embodiments, R is unsubstituted aryl, such as fluorenyl and fluorenone.

In certain embodiments, R3 is H, while R4 is phenyl or pyridinyl optionally substituted with at least one substituent selected from the group consisting of halogen, Ci-4 lower alkyl, alkoxy, haloalkoxy, -CN, and -COOH.

Exemplary compounds of formula (I) may be, 3-(3-methoxyphenyl)-5-phenylthieno[2,3-d]pyrimidine-2,4(lH,3H)-dione

(compound 1); or

5-(3-hydroxyphenyl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)- dione (compound 3).

In certain embodiments, the compound of formula (I) can be of formula (II), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, and stereoisomer thereof:

X, Y, and Z are independently C or N, provided that when one of X, Y, and Z is

N, then the other two are independently C;

R3 is H, Ci-4 lower alkyl, or -Cth-phenyl;

R^a and R^b are independently H, -OR₅, phenyl, pyridinyl, or thiophenyl, in which R5 is alkyl or phenyl;

R^c is nil or at least one substituent selected from the group consisting of halogen,

-OH, Ci-4 lower alkyl, haloalkyl, alkoxyl, and haloalkoxy;

each phenyl and pyridinyl is optionally substituted with at least one substituent selected from the group consisting of halogen, -OH, -OR6, -NH2, -NR7R8, alkyl, haloalkyl, and -CN; and

R₆, R7, and Rx are independently C1-4 lower alkyl or phenyl optionally substituted with at least one halogen.

In some embodiments, X, Y, and Z are independently C; R₃ is H; R^a is phenyl optionally substituted with at least one substituent selected from the group consisting of halogen, hydroxyl, alkoxy, haloalkoxy, -CN, and haloalkyl; R^b is H; and R^c is at least one substituent selected from the group consisting of halogen, alkoxy, haloalkyl, and haloalkoxy.

In certain embodiments, X, Y, and Z are independently C; R3 is H; R^ais pyridinyl optionally substituted with at least one substituent that is halogen or haloalkyl; R^b is H; and R^c is methoxy.

In other embodiments, X, Y, and Z are independently C; R3 is H; R^ais thiophenyl; R^b is H; and R^c is methoxy.

In further embodiments, X and Z are independently C, Y is N; R3 is H; R^ais phenyl optionally substituted with at least one substituent selected from the group consisting of halogen and haloalkoxy; R^b is H; and R^c is nil, alkoxy, or a halogen.

In still further embodiments, Y and Z are independently C, X is N; R3 is H; R^a is phenyl substituted with at least one halogen; R^b is H; and R^c is nil.

In still further embodiments, X and Y are independently C, Z is N; R3 is H; R^a is phenyl substituted with at least one halogen; R^b is H; and R^c is nil.

Exemplary compounds of formula (II) may be selected from the group consisting of:

5-(2'-hydroxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione (compound 2);

5-(4'-hydroxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione (compound 4);

5-([l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3 H)-dione (compound 5);

5-(3'-methoxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione (compound 6);

5-(3'-hydroxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione (compound 7);

5-(2'-hydroxy-3'-methoxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3- d]pyrimidine-2,4(lH,3H)-dione (compound 8);

5-(4'-methoxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione (compound 9);

3-(3-methoxyphenyl)-5-(4'-(trifluoromethoxy)-[l,r-biphenyl]-4-yl)thieno[2,3-d] pyrimidine-2,4(lH,3H)-dione (compound 10); 5-([l,r-biphenyl]-4-yl)-3-(4-chlorophenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)- dione (compound 11);

5-([l,r-biphenyl]-4-yl)-3-(4-chloro-3-(trifluoromethyl)plienyl)tliieno[2,3-d]pyri midine-2,4(lH,3H)-dione (compound 12);

3-(3-methoxyphenyl)-5-(4'-(trifluoromethyl)-[l,r-biphenyl]-4-yl)thieno[2,3-d]p yrimidine-2,4(lH,3H)-dione (compound 13);

5-([l,r-biphenyl]-4-yl)-3-(2,4-difluorophenyl)thieno[2,3-d]pyrimidine-2,4(lH,3 H)-dione (compound 14);

5-(4'-fluoro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine- 2,4(lH,3H)-dione (compound 15);

5-(3',4'-dilluoro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimid ine-2,4(lH,3H)-dione (compound 16);

5-([l,r-biphenyl]-4-yl)-3-(2,4-dimethoxyphenyl)thieno[2,3-d]pyrimidine-2,4(l H,3H)-dione (compound 17);

5-(3'-fluoro-4'-methoxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d] pyrimidine-2, 4(lH,3H)-dione (compound 18);

5-(2',4'-dichloro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimid ine-2,4(lH,3H)-dione (compound 19);

5-(3',5'-dichloro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimid ine-2,4(lH,3H)-dione (compound 21);

5-([l,r-biphenyl]-4-yl)-3-(3-fluorophenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)- dione (compound 22);

5-(4'-chloro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine- 2,4(lH,3H)-dione (compound 23);

5-(4-(5-fluoropyridin-3-yl)phenyl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine -2,4(lH,3H)-dione (compound 24);

5-([l,r-biphenyl]-4-yl)-3-(6-chloropyridin-3-yl)thieno[2,3-d]pyrimidine-2,4(lH ,3H)-dione (compound 25);

3-(4-chloro-3-(trifluoromethyl)phenyl)-5-(4'-(triiluoromethoxy)-[l,r-biphenyl]-

4-yl)thieno[2,3-d]pyrimidine-2,4(lH,3H)-dione (compound 26);

5-(4-(6-fluoropyridin-2-yl)phenyl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine -2,4(lH,3H)-dione (compound 27); 4'-(3-(4-methoxyphenyl)-2,4-dioxo-l,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-5- yl)-[l,l'-biphenyl]-4-carbonitrile (compound 28);

3-(4-methoxyphenyl)-5-(4-(6-methoxypyridin-2-yl)phenyl)thieno[2,3-d]pyrimid ine-2,4(lH,3H)-dione (compound 29);

5-([l,r-biphenyl]-3-yl)-3-(4-methoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3 H)-dione (compound 30);

3-(3-methoxyphenyl)-5-(4-(thiophen-2-yl)phenyl)thieno[2,3-d]pyrimidine-2,4(l H,3H)-dione (compound 31);

3-(3-methoxyphenyl)-5-(4-(5-(trifluoromethyl)pyridin-3-yl)phenyl)thieno[2,3-d] pyrimidine-2, 4(lH,3H)-dione (compound 32);

3-(3-methoxyphenyl)-5-(3-phenoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)- dione (compound 33);

3-(3-methoxyphenyl)-5-(4-phenoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)- dione (compound 34);

5-(4'-fluoro-[l,r-biphenyl]-3-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine- 2,4(lH,3H)-dione (compound 35);

3-(4-chloro-3-(trifluoromethyl)phenyl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thie no[2,3-d]pyrimidine-2,4(lH,3H)-dione (compound 38);

3-(2-chloro-5-(trifluoromethyl)phenyl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thie no[2,3-d]pyrimidine-2,4(lH,3H)-dione (compound 39);

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(4-(trifluoromethoxy)phenyl)thieno[2,3- d]pyrimidine-2,4(lH,3H)-dione (compound 40);

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(pyridin-3-yl)thieno[2,3-d]pyrimidine-2, 4(lH,3H)-dione (compound 41);

3-(3-chloro-4-methylphenyl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thieno[2,3-d] pyrimidine-2, 4(lH,3H)-dione (compound 42);

3-(4-chloro-3-(trifluoromethyl)phenyl)-5-(2',4'-difluoro-[l,r-biphenyl]-4-yl)thie no[2,3-d]pyrimidine-2,4(lH,3H)-dione (compound 43);

3-(2-chloro-5-(trifluoromethyl)phenyl)-5-(4'-(triiluoromethoxy)-[l,r-biphenyl]-

4-yl)thieno[2,3-d]pyrimidine-2,4(lH,3H)-dione (compound 44);

5-(2',4'-dilluoro-[l,r-biphenyl]-4-yl)-3-(4-(triiluoromethoxy)phenyl)thieno[2,3- d]pyrimidine-2,4(lH,3H)-dione (compound 45); 5-(2',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(pyridin-3-yl)thieno[2,3-d]pyrimidine-2, 4(lH,3H)-dione (compound 46);

5-(4'-(trifluoromethoxy)- [1,1 '-biphenyl]-4-yl)-3-(4-(trifluoromethoxy)plienyl)tlii eno[2,3-d]pyrimidine-2,4(lH,3H)-dione (compound 47);

3-(6-chloropyridin-3-yl)-5-(4'-(trifluoromethoxy)-[l,r-biphenyl]-4-yl)thieno[2,3 -d]pyrimidine-2,4(lH,3H)-dione (compound 48);

3-(3,5-dichlorophenyl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thieno[2,3-d]pyrimi dine-2,4(lH,3H)-dione (compound 49);

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(pyridin-4-yl)thieno[2,3-d]pyrimidine-2, 4(lH,3H)-dione (compound 50);

3-(6-chloropyridin-3-yl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thieno[2,3-d]pyri midine-2,4(lH,3H)-dione (compound 51);

3-(pyridin-3-yl)-5-(4'-(trilluoromethoxy)-[l,r-biphenyl]-4-yl)thieno[2,3-d]pyri midine-2,4(lH,3H)-dione (compound 52);

5-(3',4'-dilluoro-[l,r-biphenyl]-4-yl)-3-(pyridin-2-yl)thieno[2,3-d]pyrimidine-2, 4(lH,3H)-dione (compound 53);

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(2-methoxypyridin-4-yl)thieno[2,3-d]py rimidine-2,4(lH,3H)-dione (compound 54);

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)-l-methylthieno[2,3- d]pyrimidine-2,4(lH,3H)-dione; and

l-benzyl-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3- d]pyrimidine-2,4(lH,3H)-dione.

In certain embodiments, the compound of formula (I) can be of formula (III), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, and stereoisomer thereof:

Wherein:

R^a is phenyl optionally substituted with at least one halogen.

According to one preferred embodiment, in the formula (III), R_a is phenyl. Exemplary compound of formula (III) is

5-([l,l'-biphenyl]-4-yl)-3-ethylthieno[2,3-d]pyrimidine-2,4(lH,3H)-dione (compound

20).

In further embodiments, the compound of formula (I) can be of formula (IV), or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, and stereoisomer thereof:

X is C or -C=0; and

R^c is nil or at least one substituent selected from the group consisting of halogen, -OH, Ci-4 lower alkyl, haloalkyl, alkoxyl, and haloalkoxy.

In one embodiment, in formula (IV), X is C; and R^c is methoxy.

In another embodiment, in formula (IV), X is -C=0, and Rc is methoxy.

Exemplary compounds of formula (IV) may be any of:

5-(7-fluoro-9H-fluoren-2-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-2,4(l H,3H)-dione (compound 36); or

5-(7-fluoro-9-oxo-9H-fluoren-2-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione (compound 37).

Each compounds of the invention contain one or more stereocenters, thus can exist as racemic mixtures of enantiomers or mixtures of diastereomers. This invention thus encompasses stereomerically pure forms of such compounds, as well as mixtures of those forms. Stereoisomers may be asymmetrically synthesized or resolved using standard techniques such as crystallization, chromatography, and the use of a resolving agent. One preferred way of separating enantiomers from a racemic mixture is by use of preparative high performance liquid chromatography (HPLC). Alternatively, the racemic may be separated into its enantiomers by reacting with an optically active form of a resolving agent in the presence of a solvent. Depending on the optical form of the resolving agent, one of the two enantiomers is separated out as an insoluble salt with high yield and high optical purity, while the opposite enantiomer remains in the solution. The present invention thus further encompasses stereoisomeric mixtures of compounds disclosed herein. It also encompasses configurational isomers of compounds disclosed herein (e.g., cis and trans isomers, whether or not involving double bonds), either in admixture or in pure or substantially pure form.

In certain embodiments, compounds of the invention are the compounds described herein, and pharmaceutically acceptable salts, solvates, hydrates, co-crystals, and stereoisomers. In certain embodiments, compounds of the invention are the compounds of any one of formulae (I) to (IV), and pharmaceutically acceptable salts, solvates, hydrates, co-crystals, and stereoisomers. In certain embodiments, compounds of the invention are the compounds of any one of Formulae (I) to (IV), and

pharmaceutically acceptable salts thereof.

According to preferred embodiments of the present disclosure, the compound of formula (I) may suppress the activity of CD73. Accordingly, the compound of formula (I) acts as an antagonist of CD73, thus may be useful as a lead compound for the development of a medicament suitable for the treatment of diseases and/or disorders mediated by adenosine.

Any of the compounds described herein can be prepared by routine methods known in the art. In some embodiments, a compound as described herein (e.g., a compound of formula (I)) is synthesized by procedures described in working examples, i.e., schemes I or II. The structure of each compound is verified by mass and NMR analysis; while its function is evaluated by the inhibition on CD73 enzyme activity.

3. Pharmaceutical Formulation

This invention encompasses pharmaceutical compositions for the treatment of a disease and/or a disorder mediated by adenosine. The pharmaceutical composition comprises a therapeutically effective amount of a compound of formula (I) to (IV) of the present invention. The compound of formula (I) to (IV) is present at a level of about 0.1 % to 99% by weight, based on the total weight of the pharmaceutical composition. In some embodiments, the compound of formula (I) to (IV) is present at a level of at least 1% by weight, based on the total weight of the pharmaceutical composition. In certain embodiments, the compound of formula (I) to (IV) is present at a level of at least 5% by weight, based on the total weight of the pharmaceutical composition. In still other embodiments, the compound of formula (I) to (IV) is present at a level of at least 10% by weight, based on the total weight of the pharmaceutical composition. In still yet other embodiments, the compound of formula (I) to (IV) is present at a level of at least 25% by weight, based on the total weight of the pharmaceutical composition.

According to embodiments of the present disclosure, the disease and/or disorder mediated by adenosine may be AIDS, an autoimmune disease, atherosclerosis, a cancer, an infectious disease, ischemia-reperfusion injury or pre-cancerous syndrome.

Examples of the autoimmune disease treatable by the present pharmaceutical composition include, but are not limited to, multiple sclerosis, psoriasis, systemic lupus erythemotosis (SLE), Type I diabetes mellitus, and Wegener’s granulomatosis.

Examples of the infectious disease treatable by the present pharmaceutical composition include, but are not limited to, bacterial, fungal and viral infections.

In some preferred embodiments, the pharmaceutical composition may further comprise an anti-neoplastic agent, an immunosuppressant, or an anti-infectious agent.

The anti-neoplastic agent that may be used with the present pharmaceutical composition is an anti-microtubule agent, a platinum coordination complex, an alkylating agent, an antibiotic, a topoisomerase II inhibitor, a topoisomerase I inhibitor, an anti-metabolite, a hormone, a signal transduction pathway inhibitor, a non-receptor tyrosine kinase, an angiogenesis inhibitor, an immunotherapeutic agent, a pro-apoptotic agent, cell cycle signaling inhibitor, a proteasome inhibitor, an inhibitor of cancer metabolism, an anti-PD-Ll agent, a PD-1 antagonist, an immune-modulator, Stimulator of Interferon Genes (STING) modulating compound, CD39 inhibitor, A2a and A2a adenosine antagonists, Toll-like receptor 4 (TLR4) inhibitor, anti-ICOS antibody or anti- 0X40 antibody.

The immunosuppressant that may be used with the present pharmaceutical composition is a corticosteroid, a calcineurin inhibitor, a mammalian target of rapamycin (mTOR) inhibitor, an Inosine monophosphate dehydrogenase (IMPDH) inhibitor, a therapeutic protein, or a monoclonal antibody. Examples of corticosteroid suitable as an immunosuppressant include, but are not limited to, prednisone, budesonide, and prednisolone. Examples of calcineurin inhibitor suitable as an immunosuppressant include, but are not limited to, cyclosporine, and tacrolimus.

Examples of mTOR inhibitor suitable as an immunosuppressant include, but are not limited to, sirolimus, and everolimus. Examples of IMDH inhibitor suitable as an immunosuppressant include, but are not limited to, azathioprine, leflunomide, and mycophenolate. Examples of therapeutic protein suitable as an immunosuppressant include, but are not limited to, abatacept, adalimumab, anakinra, certolizumab, etanercept, golimumab, infliximab, ixekizumab, natalizumab, rituximab, secukinumab, tocilizumab, ustekinumab, and vedolizumab. Examples of monoclonal antibody suitable as an immunosupressant include, but are not limited to, basiliximab, daclizumab, and muromonab.

The anti-infectious agent suitable for use in the present pharmaceutical composition may be an anti-biotic that inhibits the growth of Gram-negative or

Gram-positive bacteria. Examples of antibiotic suitable as an anti-infectious agent include, but are not limited to, acumycin, ampicillin, amoxycillin, amphotericins, antimycins, anglomycin, avermectins, azithromycin, boromycin, carbomycins, carbapenem, ceftazidime, cethromycin, chloramphenicol, chalcomycin, ciprofloxacin, concanamycins, cirramycin, clarithromycin, colistin, cycloxacillin, daptomycin, desmethyl azithromycin, desertomycins, dihydropikromycin, dirithromycin, doxycycline, enramycin, erythromycin, flurithromycin, flumequin gentamycin, juvenimicins, kujimycins, lankamycins, lincomycin, litorin, leucomycins,

megalomicins, meropenem, methymycin, midecamycins, mycinamicin I, mycinamicin II, mycinamicin III, mycinamicin IV, mycinamicin V, mycinamicin VI, mycinamicin VII, mycinamicin VIII, narbomycin, neoantimycin, neomethymycin, netilmicin, neutromycin, niddamycins, norfioxacin, oleandomycins, oligomycins, ossamycin, oxacillin, oxolinic acid, penicillin, pikromycin, piperacillin, platenomycins, rapamycins, relomycin, rifamycins, rosaramicin, roxithromycin, virginiamycin, spiramycin, sporeamycin, staphococcomycin, streptomycin, sulfamethoxazole, swalpamycin, telithromycin, teicoplanin, timentin, tobramycin, ticarcillin, trimethoprim, tetracyclin, zlocillin, and/or a combination thereof. Certain pharmaceutical compositions are single unit dosage forms suitable for oral, mucosal (e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (e.g., subcutaneous, intravenous, bolus injection, intramuscular, or intra-arterial), or transdermal administration to a patient. Examples of dosage forms include, but are not limited to: tablets; caplets; capsules, such as soft elastic gelatin capsules; cachets; troches; lozenges; dispersions; suppositories; ointments; cataplasms (poultices); pastes; powders; dressings; creams; plasters; solutions; patches; aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage forms suitable for oral or mucosal administration to a patient, including suspensions (e.g., aqueous or non-aqueous liquid suspensions, oil-in-water emulsions, or a water-in-oil liquid emulsions), solutions, and elixirs; liquid dosage forms suitable for parenteral administration to a patient; and sterile solids (e.g., crystalline or amorphous solids) that can be reconstituted to provide liquid dosage forms suitable for parenteral administration to a patient.

The formulation should suit the mode of administration. For example, oral administration requires enteric coatings to protect the compounds of this invention from degradation within the gastrointestinal tract. Similarly, a formulation may contain ingredients that facilitate delivery of the active ingredient(s) to the site of action. For example, compounds may be administered in liposomal formulations, in order to protect them from degradative enzymes, facilitate transport in circulatory system, and effect delivery across cell membranes to intracellular sites.

Similarly, poorly soluble compounds may be incorporated into liquid dosage forms (and dosage forms suitable for reconstitution) with the aid of solubilizing agents, emulsifiers and surfactants such as, but not limited to, cyclodextrins (e.g.,

a-cyclodextrin or b-cyclodextrin), and non-aqueous solvents, such as, but not limited to, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethyl formamide, dimethyl sulfoxide (DMSO), biocompatible oils (e.g. , cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols, fatty acid esters of sorbitan, and mixtures thereof (e.g., DMSO:com oil).

The composition, shape, and type of a dosage form will vary depending on its use. For example, a dosage form used in the acute treatment of a disease may contain larger amounts of one or more of the active ingredients it comprises than a dosage form used in the chronic treatment of the same disease. Similarly, a parenteral dosage form may contain smaller amounts of one or more of the active ingredients it comprises than an oral dosage form used to treat the same disease. These and other ways in which specific dosage forms encompassed by this invention will vary from one another will be readily apparent to those skilled in the art.

3.1 Oral Dosage Forms

Pharmaceutical compositions of the present invention suitable for oral administration can be presented as discrete dosage forms, such as, but are not limited to, tablets (e.g., chewable tablets), caplets, capsules, and liquids (e.g., flavored syrups). Such dosage forms contain predetermined amounts of active ingredients, and may be prepared by methods of pharmacy well known to those skilled in the art.

Typical oral dosage forms are prepared by combining the active ingredient(s) in an intimate admixture with at least one excipient according to conventional

pharmaceutical compounding techniques. Excipients can take a wide variety of forms depending on the form of preparation desired for administration.

Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit forms. If desired, tablets can be coated by standard aqueous or non-aqueous techniques. Such dosage forms can be prepared by conventional methods of pharmacy. In general, pharmaceutical compositions and dosage forms are prepared by uniformly and intimately admixing the active ingredients with liquid carriers, finely divided solid carriers, or both, and then shaping the product into the desired presentation if necessary. Disintegrants may be incorporated in solid dosage forms to facility rapid dissolution. Lubricants may also be incorporated to facilitate the manufacture of dosage forms (e.g., tablets).

3.2 Parenteral Dosage Forms

Parenteral dosage forms can be administered to patients by various routes including, but not limited to, subcutaneous, intravenous (including bolus injection), intramuscular, and intra-arterial. Because their administration typically bypasses patients’ natural defenses against contaminants, parenteral dosage forms are specifically sterile or capable of being sterilized prior to administration to a patient. Examples of parenteral dosage forms include, but are not limited to, solutions ready for injection, dry products ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, suspensions ready for injection, and emulsions. Suitable vehicles that can be used to provide parenteral dosage forms of the invention are well known to those skilled in the art. Examples include, but are not limited to: water; aqueous vehicles such as, but not limited to, sodium chloride solution, Ringer’s solution, and Dextrose; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

3.3 Transdermal, Topical and Mucosal Dosage Forms

Transdermal, topical, and mucosal dosage forms include, but are not limited to, ophthalmic solutions, sprays, aerosols, creams, lotions, ointments, gels, solutions, emulsions, suspensions, or other forms known to one of skill in the art. Transdermal dosage forms include“reservoir type” or“matrix type” patches, which can be applied to the skin and worn for a specific period of time to permit the penetration of a desired amount of active ingredients.

Suitable excipients (e.g. , carriers and diluents) and other materials that can be used to provide transdermal, topical, and mucosal dosage forms are well known to those skilled in the pharmaceutical arts, and depend on the particular tissue to which a given pharmaceutical composition or dosage form will be applied.

Depending on the specific tissue to be treated, additional components may be used prior to, in conjunction with, or subsequent to treatment with active ingredients of the invention. For example, penetration enhancers may be used to assist in delivering active ingredients to the tissue.

The pH of a pharmaceutical composition or dosage form, or of the tissue to which the pharmaceutical composition or dosage form is applied, may also be adjusted to improve delivery of one or more active ingredients. Similarly, the polarity of a solvent carrier, its ionic strength, or tonicity can be adjusted to improve delivery.

Compounds such as stearates may also be added to pharmaceutical compositions or dosage forms to advantageously alter the hydrophilicity or lipophilicity of one or more active ingredients so as to improve delivery. In this regard, stearates can serve as a lipid vehicle for the formulation, as an emulsifying agent or surfactant, and as a delivery-enhancing or penetration-enhancing agent. Different salts, hydrates or solvates of the active ingredients can be used to further adjust the properties of the resulting composition 4. Kits

Also encompasses within the present disclosure is an article of manufacture or “kit,” containing materials useful for the treatment or prophylaxis of a disease and/or disorder mediated by adenosine in a subject.

In one embodiment, the kit comprises a container comprising the compound of the present disclosure. The kit is suitable for the treatment or prophylaxis of a disease and/or disorder mediated by adenosine, such as AIDS, an autoimmune disease, atherosclerosis, a cancer, an infectious disease, ischemia-reperfusion injury or pre-cancerous syndrome. Suitable containers include, for example, bottles, vials, syringes, blister pack, and etc. The container may be formed from a variety of materials such as glass, or plastic. The contain may hold a compound of the present disclosure or a pharmaceutical formulation thereof, in an amount effective for the treatment or prophylaxis of the disease and/or disorder mediated by adenosine, and may have a sterile access port, for example, the container may be an intravenous solution bag or a vail having a stopper pierceable by a hypodermic injection needle). The kit may further comprise a label or package insert on or associated with the container. The label or package insert indicates that the composition is used for treating condition of choice. Alternatively or additionally, the kit may further comprise a second container comprising a pharmaceutically acceptable buffer, such as a phosphate-buffered saline, Ringer’s solution or dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

The kit may further include directions for the administration of the compound of the present invention and, if present, the second formulation for treating or preventing the disease and/or disorder mediated by adenosine. For example, if the kit comprises a first composition comprising the compound of the present disclosure, and a second pharmaceutical formulation, the kit may further include directions for the simultaneous, sequential, or separate administration of the first and second pharmaceutical

compositions to a patient in need thereof.

In another embodiment, the kits are suitable for the delivery of solid oral forms of a compound of the present disclosure, such a kit includes, for example, a number of unit dosages. Such kits include card having the dosages oriented in the order of their intended use. An example of such kit is a“blister pack.” Blister packs are well known in the packaging industry and are widely used for packaging pharmaceutical unit dosage forms. If desired, an aid may be provided, for example, in the form of numbers, letters, or other markings or with a calendar insert, designating the days in the treatment schedule in which the dosage can be administered.

According to one embodiment, the Kit may include, at least, (a) a first container containing any of the present compound of formula (I) to (IV); and optionally, (b) a second container containing a second therapeutic agent that is any of a known CD73 antagonist, an anti-neoplastic agent, an anti-biotic, or an immunosuppressant; and (c) a legend associated with the kit for instructing a user how to use the kit. The legend may be in a form of pamphlet, tape, CD, VCD or DVD.

5. Method of Use

The present invention encompasses a method for the treatment of a subject having a disease and/or a disorder mediated by adenosine. The method comprises the step of administering the present pharmaceutical composition, which comprises a therapeutically effective amount of any of the compound of formula (I) to (IV) of the present disclosure, to the subject, so as to ameliorate, mitigate and/or prevent the symptoms of the disease and/or disorder mediated by adenosine.

According to embodiments of the present disclosure, the disease and/or disorder mediated by adenosine may be AIDS, an autoimmune disease, atherosclerosis, a cancer, an infectious disease, ischemia-reperfusion injury or pre-cancerous syndrome.

Examples of cancer treatable by the present method include, but are not limited to, acinar cell carcinoma, acute lymphoblastic leukemia, acute myelogenous leukemia, adenocarcinoma, adenosquamous carcinoma, Bannayan-Zonana syndrome, bladder cancer, breast cancer, buccal cancer, Burkitt's lymphoma, cervical cancer, colon cancer, chronic myelogenous leukemia, chronic lymphocytic leukemia, chronic neutrophilic leukemia, Cowden disease, ductal adenocarcinoma, endometrial cancer, Edwing’s sarcoma, ependymoma, erythroleukemia, esophageal cancer, follicular lymphoma, gastric cancer, gastrointestinal stromal tumor, giant cell tumor of bone, glioma, glioblastoma, astrocytoma, glioblastoma multiforme, glucagonoma, hairy-cell leukemia, head and neck cancer, hodgkins lymphoma, insulinoma, immunoblastic large cell leukemia, kidney cancer, Lhermitte-Duclos disease, lobular carcinoma, liver cancer, lymphoblastic T cell leukemia, malignant lymphoma, medulloblastoma, melanoma, mantle cell leukemia, mesothelioma, multiple myeloma, megakaryoblastic leukemia, nasopharangeal cancer, non-hodgkins lymphoma, non-small cell lung cancer, osteosarcoma, ovarian cancer, pancreatic cancer, plasmacytoma, promyelocytic leukemia, prostate cancer, Rhabdomyosarcoma, sarcoma, salivary gland cancer, skin cancer, small cell lung cancer, testicular cancer, thyroid cancer, urothelial cancer, vulval cancer, or Wilm’ s tumor.

Examples of the autoimmune disease treatable by the present method include, but are not limited to, multiple sclerosis, psoriasis, systemic lupus erythemotosis (SLE), Type I diabetes mellitus, and Wegener’s granulomatosis.

Examples of the infectious disease treatable by the present method include, but are not limited to, bacterial, fungal and viral infections.

Accordingly, in some embodiments, the method further includes the step of administering to the subject, an anti-neoplastic agent, an immunosuppressant, or an anti-infectious agent before, together with, or after the administration of the present pharmaceutical composition.

The anti-neoplastic agent suitable for use in the present method is an anti-microtubule agent, a platinum coordination complex, an alkylating agent, an antibiotic, a topoisomerase II inhibitor, a topoisomerase I inhibitor, an anti-metabolite, a hormone, a signal transduction pathway inhibitor, a non-receptor tyrosine kinase, an angiogenesis inhibitor, an immuno therapeutic agent, a pro-apoptotic agent, cell cycle signaling inhibitor, a proteasome inhibitor, an inhibitor of cancer metabolism, an anti-PD-Ll agent, a PD-1 antagonist, an immune-modulator, Stimulator of Interferon Genes (STING) modulating compound, CD39 inhibitor, A2a and A2a adenosine antagonists, Toll-like receptor 4 (TLR4) inhibitor, anti-ICOS antibody or anti-OX40 antibody.

The immunosuppressant suitable for use in the present method may be a corticosteroid, a calcineurin inhibitor, a mammalian target of rapamycin (mTOR) inhibitor, an Inosine monophosphate dehydrogenase (IMPDH) inhibitor, a therapeutic protein, or a monoclonal antibody. Examples of corticosteroid suitable as an immunosuppressant include, but are not limited to, prednisone, budesonide, and prednisolone. Examples of calcineurin inhibitor suitable as an immunosuppressant include, but are not limited to, cyclosporine, and tacrolimus. Examples of mTOR inhibitor suitable as an immunosuppressant include, but are not limited to, sirolimus, and everolimus. Examples of IMDH inhibitor suitable as an immunosuppressant include, but are not limited to, azathioprine, leflunomide, and mycophenolate.

Examples of therapeutic protein suitable as an immunosuppressant include, but are not limited to, abatacept, adalimumab, anakinra, certolizumab, etanercept, golimumab, infliximab, ixekizumab, natalizumab, rituximab, secukinumab, tocilizumab,

ustekinumab, and vedolizumab. Examples of monoclonal antibody suitable as an immunosupressant include, but are not limited to, basiliximab, daclizumab, and muromonab.

The anti-infectious agent suitable for use in the present method may be an anti-biotic that inhibits the growth of Gram-negative or Gram-positive bacteria.

Examples of antibiotic suitable for use as an anti-infectious agent include, but are not limited to, acumycin, ampicillin, amoxycillin, amphotericins, antimycins, anglomycin, avermectins, azithromycin, boromycin, carbomycins, carbapenem, ceftazidime, cethromycin, chloramphenicol, chalcomycin, ciprofloxacin, concanamycins, cirramycin, clarithromycin, colistin, cycloxacillin, daptomycin, desmethyl azithromycin, desertomycins, dihydropikromycin, dirithromycin, doxycycline, enramycin,

erythromycin, flurithromycin, flumequin gentamycin, juvenimicins, kujimycins, lankamycins, lincomycin, litorin, leucomycins, megalomicins, meropenem, methymycin, midecamycins, mycinamicin I, mycinamicin II, mycinamicin III, mycinamicin IV, mycinamicin V, mycinamicin VI, mycinamicin VII, mycinamicin VIII, narbomycin, neoantimycin, neomethymycin, netilmicin, neutromycin, niddamycins, norfloxacin, oleandomycins, oligomycins, ossamycin, oxacillin, oxolinic acid, penicillin, pikromycin, piperacillin, platenomycins, rapamycins, relomycin, rifamycins, rosaramicin, roxithromycin, virginiamycin, spiramycin, sporeamycin, staphococcomycin,

streptomycin, sulfamethoxazole, swalpamycin, telithromycin, teicoplanin, timentin, tobramycin, ticarcillin, trimethoprim, tetracyclin, zlocillin, and/or a combination thereof.

The amount, route of administration and dosing schedule of the present compounds will depend upon factors such as the specific indication to be treated, prevented, or managed, and the age, sex and condition of the patient. The roles played by such factors are well known in the art, and may be accommodated by routine experimentation.

The present invention will now be described more specifically with reference to the following embodiments, which are provided for the purpose of demonstration rather than limitation. While they are typically of those that might be used, other procedures, methodologies, or techniques known to those skilled in the art may alternatively be used.

EXAMPLES

Materials and Methods

CD73 activity assay.

Human CD73 catalyzes the conversion of AMP to adenosine and generate inorganic phosphate (Pi). Inhibition of CD73 enzyme activity by the test compound was determined by measuring the amount of Pi released using Malachite Green assay kit (SensoLyte® MG Phosphate Assay Kit, AnaSpec, USA) and compare to the DMSO vehicle control. Stock solutions of each compound were prepared in DMSO. The final concentration of DMSO in all samples/assays/experiments was 2.5% (v/v). The enzymatic reaction was started by adding purified C-terminal His-tagged CD73 (BPS#71184) protein (3.2ng per reaction) into lx assay buffer (BPS#74000), AMP (100 mM), and test compound in total volume of 80 pL in 96-well plate (costar #3370) for 25 min at 23 _°C. Then Malachite green reagent (20 pL) was added and incubated for 10 min at room temperature, and the plate was read at 620 nm using a SpectraMax i3x microplate reader. All samples were run in triplicate. Test compound inhibition was expressed as percent inhibition of DMSO controls.

EXAMPLE 1 Preparation of the compound of formula (I)

The compound of Formula (I) were prepared by general reaction schemes described bellowed.

Scheme 1-1. Synthesis of pyrimidinedione derivative. (Method A)

To a mixture of PdC Cdppf) (0.1 eq), substituted l-(bromophenyl)ethanone (1 eq) (compounds 1-1, 1-2) and boric acid was added DMF and K2CO3 _(aq) (4 eq) under inert atmosphere. The mixture was stirred overnight at 110°C. After the reaction completed, the reaction mixture was filtered with celite and then extracted with EA.

The organic layer was dried over with MgSCri and concentrated under reduced pressure. The substituted 1-phenylethanone 1-3 was purified by column chromatography.

To a solution of substituted 1-phenylethanone 1-3 (1 eq) in ethanol was added ethyl 2-cyanoacetate (1 eq), morpholine (1 eq) and sulfur (1 eq). The reaction mixture was stirred and refluxed overnight. After the reaction completed, the solvent was removed under reduced pressure. The residue was washed with cold ethanol and then dried under vacuum. The crude product was dissolved in dichloromethane and washed with brine. The organic layer was collected and concentrated under reduced pressure. The ethyl 2-amino-4-phenylthiophene-3-carboxylate derivative 1-4 was purified by column chromatography.

The ethyl 2-amino-4-phenylthiophene-3-carboxylate derivative 1-4 (1 eq) and isocyante (3 eq) were added to acetic acid (2.5 mL / mole) and stirred at ambient temperature for 18 hours. The reaction mixture was poured into water (6 mL / mmole). The precipitate was filtered and washed with water. The solid was suspended in water (12 ml / mmole) and added the concentrated NaOH _(aq) until pH 12-14. The suspension was stirred at 100 °C for 2h then cooled to room temperature. The mixture was acidified by glacial acetic acid. The pyrimidinedione derivative 1-5 was collected, washed with water and then dried under vacuum at 40 °C.

Scheme 1-2. Synthesis of pyrimidinedione derivative. (Method B)

To a mixture of amine (3eq) and DMAP (1.5eq) was added acetonitrile and stirred under N2 at 0°C. Phenyl chloroformate was slowly added to the reaction mixture at 0°C. Then triethylamine was added into the reaction mixture. The mixture was stirred at room temperature for 1 hour and then heated to 50-60°C for 1 hour. After the reaction mixture returned to the room temperature, the carbamate derivative 1-6 was collected without further purification.

The mixture of 2-amino-4-phenylthiophene-3-carboxylate derivative 1-4 (3eq) and carbamate 1-6 was added acetonitrile and heated to 80-90°C in seal for 2-3 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give thiophenyl urea derivate 1-7.

The thiophenyl urea derivate 1-7 (1 eq) was added 30% NaOMe (5 mL / mmole) and MeOH (15 mL / mmole) and heated to 50~60°C in the sealed tube. After the reaction completed, the reaction mixture was neutralized with Dowex H⁺ (Dowex 50WX8H⁺). The precipitate was filtered and washed with methanol. The solvent was removed under reduced pressure and pyrimidinedione derivative 1-5 was collected without further purification. Scheme 1-3. Synthesis of pyrimidinedione derivative. (Method C)

The substituted 2-acetylfluorene or 2-acetylflourenone 1-8 was prepared as described by Berkovic, S. ( Isr . J. Chem. 1963, 1, 1-11.) and Zhang, Xin et al. ( Green Chem. 2011, 13, 1891-1896).

To a solution of substituted 2-acetylfluorene or 2-acetylflourenone 1-8 (1 eq) in ethanol was added ethyl 2-cyanoacetate (1 eq), morpholine (1 eq) and sulfur (1 eq). The reaction mixture was stirred and refluxed overnight. After the reaction completed, the solvent was removed under reduced pressure. The residue was washed with cold ethanol and then dried under vacuum. The crude product was dissolved in dichloromethane and washed with brine. The organic layer was collected and concentrated under reduced pressure. The ethyl 2-amino-4-fluorenylthiophene-3-carboxylate derivative 1-9 was purified by column chromatography.

The ethyl 2-amino-4-phenylthiophene-3-carboxylate derivative 1-9 (1 eq) and isocyante (3 eq) were added to acetic acid (2.5 mL / mole) and stirred at ambient temperature for 18 hours. The reaction mixture was poured into water (6 mL / mmole). The precipitate was filtered and washed with water. The solid was suspended in water (12 ml / mmole) and added the concentrated NaOH _(aq) until pH 12-14. The suspension was stirred at 100 °C for 2h then cooled to room temperature. The mixture was acidified by glacial acetic acid. The pyrimidinedione derivative 1-10 was collected, washed with water and then dried under vacuum at 40 °C.

Compound 1

3-(3-methoxyphenyl)-5-phenylthieno[2,3-d]pyrimidine-2,4(lH,3H)-dione Method A, Yield: 52.5%, ¾ NMR (500 MHz, DMSO): d 3.73 (s, 3H), 56.82-6.83 (d, J = 7.80Hz, 1H), 56.87 (s, 1H), 56.94-6.96 (d, J = 8.75Hz, 1H), 57.04 (s, 1H), 57.30-7.35 (m, 4H), 57.42-7.44 (d, J = 7.09Hz, 2H). LC-MS (ESI): Calcd for [C14H19N2O3S] 351.07 [M +1]⁺, found 351.0[M +1]⁺.

Compound 2

5-(2'-hydroxy-[l,l'-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione

Method A, Yield: 40.9%, ¾ NMR (500 MHz, CDC13): d 3.80 (s, 3H),

56.75-6.89 (m, 3H), 56.94-6.99 (t, J = 8.37Hz, 3H), 57.20-7.25 (m, 1H), 57.37-7.42 (t, J = 7.18Hz, 2H), 57.43-7.48 (d, J = 8.37Hz, 2H), 57.59-7.63 (d, J = 7.18, 2H). LC-MS (ESI): Calcd for [C₂₅H₁₈N₂O₄S] 443.1 [M +1]⁺, found 443.2 [M +1]⁺.

Compound 3

5-(3-hydroxyphenyl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)- dione

Method A, Yield: 57.9%, ¾ NMR (500 MHz, CDCb): 5 3.78 (s, 3H), 55.16 (s, 1H), 56.71 (s, 1H), 56.79 (m, 2H), 56.80-6.84 (d, J = 7.82Hz, 1H), 56.95 (m, 1H), 57.02-7.04 (m, 1H), 57.18-7.21 (t, J = 8.00Hz, 1H), 57.36-7.39 ( t, J = 8.02, 1H), 59.49 ( s, 1H). LC-MS (ESI): Calcd for [C19H14N2O4S] 367.07 [M+l]⁺, found 367.2[M +1]⁺.

Compound 4

5-(4'-hydroxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione

Method A, Yield: 21.4%, ¾ NMR (500 MHz, DMSO): 5 3.74 (s, 3H), 56.83-6.88 (m, 2H), 56.96 (m, 1H), 57.08 (s, 1H), 57.32-7.36 ( m, 2H), 57.42-7.54 ( m, 4H), 57.61-7.62 (d, J = 8.59Hz, 2H), 57.65-7.67 (d, J = 8.30Hz, 2H). LC-MS (ESI): Calcd for [C25H18N2O4S] 427.1 [M +1]⁺, found 426.9[M +1]⁺.

Compound 5

5-([l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3

H)-dione

Method A, Yield: 39.3%, ¾ NMR (500 MHz, DMSO): 5 3.74 (s, 3H),

56.83-6.84 (d, J = 7.62Hz, 2H), 56.88 (s, 1H), 56.94-6.96 (m, 1H), 57.08 (s, 1H), 57.32-7.37 (q, J = 8.76Hz, 2H), 57.45-7.48 (t, J = 8.30Hz, 2H), 57.52-7.54 (d, J = 8.59Hz, 2H), 57.61-7.62 (d, J = 8.59Hz, 2H), 57.65-7.67 (d, J = 7.29Hz, 2H).

LC-MS(ESI): m/z Calcd for [C25H18N2O3S] 427.1 [M +1]⁺, found 426.9[M +1]⁺.

Compound 6

5-(3'-methoxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione Method A, Yield: 24.7%, ¾ NMR (500 MHz, DMSO): d 3.73 (s, 3H), 53.81 (s, 1H), 56.77 (m, 2H), 56.91-6.62 (m, 3H), 57.16 (s, 1H), 57.21-7.22 (d, J = 8.10Hz, 1H), 57.29-7.32 (t, J = 8.78Hz, 1H), 57.35-7.38 (t, J = 8.78Hz, 1H), 57.51-7.52 (d, J = 8.78Hz, 2H), 57.58-7.59 (d, J = 8.10Hz, 2H). LC-MS(ESI): m/z Calcd for [C25H18N2O4S]

457.11[M +1]⁺, found 457.01[M +1]⁺.

Compound 7

5-(3'-hydroxy-[l,l'-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione

Method A, Yield: 43.6%, ¾ NMR (500 MHz, DMSO): d 3.74 (s, 3H),

56.74-6.76 (m, 1H), 56.82-6.84 (d, J =8.37Hz, 1H), 56.87 (s, 1H), 56.94-6.96 (m, 1H), 57.00 (s, 1H), 57.05-7.07 (d, J = 8.37Hz, 2H), 57.23-7.24 (t, J = 8.37Hz, 1H),

57.32-7.34 (t, J = 8.93Hz, 1H), 57.50-7.54 (m ,4H), 59.53 (s,lH). LC-MS(ESI): m/z Calcd for [C25H18N2O4S] 457.11[M +1]⁺, found 457.01[M +1]⁺.

Compound 8

5-(2'-hydroxy-3'-methoxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3- d]pyrimidine-2,4(lH,3H)-dione

Method A, Yield: 51.9%, ¾ NMR (500 MHz, DMSO): d 3.74 (m, 3H), 53.83-3.84 (m, 3H), 56.83-6.88 (m, 4H), 56.95-6.96 (m, 2H), 57.07 (s, 1H), 57.32-7.35 (t, J = 7.61Hz, 1H), 57.44-7.49 (m, 4H), 58.62 (s, 1H). LC-MS(ESI): m/z Calcd for [C26H20N2O5S] 473.11[M +1]⁺, found 473.22[M +1]⁺.

Compound 9

5-(4'-methoxy-[l,l'-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione

Method A, Yield: 60.5%, ¾ NMR (500 MHz, DMSO): d 3.74 (s, 3H), 53.78 (s, 3H), 56.83-6.85 (d, J = 8.24Hz, 1H), 56.88 (s, 1H), 56.94-6.96 (m, 1H), 57.01-7.03 (d, J

= 8.30Hz, 2H), 57.07 (s, 1H), 57.32-7.35 (t, J = 7.84Hz, 1H), 57.48-7.50 (d, J = 8.76, 2H), 57.55-7.57 (d, J = 7.84, 2H), 57.59-7.61 (d, J = 8.37Hz, 2H). LC-MS(ESI): m/z Calcd for [C26H20N2O4S] 457.11[M +1]⁺, found 456.9[M +1]⁺.

Compound 10

3-(3-methoxyphenyl)-5-(4'-(trifluoromethoxy)-[l,r-biphenyl]-4-yl)thieno[2,3-d] pyrimidine-2,4(lH,3H)-dione

Method A, Yield: 58.8%, ¾ NMR (500 MHz, DMSO): d 3.74 (s, 3H),

56.81-6.85 (d, J = 5.93Hz, 1H), 56.87 (s, 1H), 56.93-6.97 (m, 1H), 57.09 (s, 1H), 57.30-7.36 (t, J = 7.91Hz, 1H), 57.43-7.47 (d, J = 8.9Hz, 1H), 57.53-7.57 (d, J =

6.92Hz, 2H), 57.61-7.65 (d, J = 6.92, 2H), 57.77-7.81 (d, J = 8.9, 2H). LC-MS(ESI): m/z Calcd for [C26H17F3N2O4S] 511.09[M +1]⁺, found 511.0[M +1]⁺. Compound 11

5-([l,r-biphenyl]-4-yl)-3-(4-chlorophenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)- dione

Method A, Yield: 8.3%, ¾ NMR (500 MHz, DMSO): d 7.04 (s, 1H),

57.29-7.31 (d, J = 8.38Hz, 2H), 57.34-7.37 (t, J = 7.54Hz, 1H), 57.44-7.54 (m, 6H), 57.60-7.62 (d, J = 8.38Hz, 2H), 7.65-7.67 (d, J = 7.54Hz, 2H). LC-MS(ESI): m/z Calcd for[C₂₄Hi₅ClN₂O₂S]431.05[M +1]⁺, found 430.8[M +1]⁺.

Compound 12

5-([l,l'-biphenyl]-4-yl)-3-(4-chloro-3-(trifluoromethyl)phenyl)thieno[2,3-d]pyri midine-2,4(lH,3H)-dione

Method A, Yield: 30.8%, ¾ NMR (500 MHz, DMSO): d 7.08 (s, 1H),

57.36-7.37 (t, J = 7.38Hz, 1H), 57.45-7.48 (t, J = 7.84Hz, 2H), 57.52-7.54 (d, J = 7.84Hz, 2H), 57.61-7.63 (d, J = 7.84Hz, 2H), 57.65-7.67 (d, J = 7.84Hz, 2H), 57.82-7.84 (d, J =

7.84Hz, 1H), 57.90 (s, 1H). LC-MS(ESI): m/z Calcd for[C₂₄Hi₅ClN₂0₂S] 499.04[M +1]⁺, found 498.6[M +1]⁺.

Compound 13

3-(3-methoxyphenyl)-5-(4'-(trifluoromethyl)-[l,r-biphenyl]-4-yl)thieno[2,3-d]p yrimidine-2,4(lH,3H)-dione Method A, Yield: 74.5%, ¾ NMR (500 MHz, DMSO): d 3.74 (s, 3H),

56.83-6.85 (d, J = 7.76Hz, 1H), 56.88 (s, 1H), 56.94-6.97 (m, 1H), 57.13 (s, 1H), 57.32-7.36 (t, J = 8.07Hz, 1H), 57.57-7.58 (d, J = 8.27Hz, 2H), 57.69-7.13 (d, J = 8.3Hz, 2H), 57.80-7.82 (d, J = 8.34Hz, 2H) 57.89-7.90 (d, J = 8.19Hz, 2H). LC-MS(ESI): m/z

Calcd for[C₂₆Hi₇F₃N₂0₃S] 495.09[M +1]⁺, found 495.0[M +1]⁺.

Compound 14

5-([l,l'-biphenyl]-4-yl)-3-(2,4-difluorophenyl)thieno[2,3-d]pyrimidine-2,4(lH,3

H)-dione

Method A, Yield: 52%, ¾ NMR (500 MHz, DMSO): d 7.14 (s, 1H), 57.17-7.22 (t, J = 9.65Hz, 1H), 57.34-7.39 (t, J = 8.17Hz, 1H), 57.40-7.49 (m, 3H), 57.51-7.59 (d, J = 8.91Hz, 1H), 57.60-7.71 (m, 4H). LC-MS(ESI): m/z Calcd for [C24H14F2N2O2S] 433.07[M +1]⁺, found 433[M +1]⁺.

Compound 15

5-(4'-fluoro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-

2,4(lH,3H)-dione

Method A, Yield: 67.4%, ¾ NMR (500 MHz, DMSO): d 3.73-3.75 (s, 3H), 56.81-6.85 (d, J = 7.97Hz, 1H), 56.88 (s, 1H), 56.93-6.97 (m, 1H), 57.08 (s, 1H), 57.25-7.36 (m, 3H), 57.50-7.55(d, J = 7.97Hz, 2H), 57.58-7.61(d, J = 8.31, 2H), 57.67-7.78(m, 3H). LC-MS(ESI): m/z Calcd for [C25H17FN2O3S] 445.09[M +1]⁺, found 445.1[M +1]⁺.

Compound 16

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimid ine-2,4(lH,3H)-dione

Method A, Yield: 35.7%, ¾ NMR (500 MHz, DMSO): d 3.74 (s, 3H),

56.82-6.85 (d, J = 6.39Hz, 1H), 56.89 (s, 1H), 56.94-6.97 (m, 1H), 57.14 (s, 1H), 57.28-7.37 (m, 3H), 57.41-7 47(m, 1H), 57.51-7.55(d, J = 7.76, 2H), 57.55-7.58(d, J = 8.22Hz, 2H). LC-MS(ESI): m/z Calcd for [C25H17FN2O3S] 445.09[M +1]⁺, found 445.1[M +1]⁺.

Compound 17

5-([l,r-biphenyl]-4-yl)-3-(2,4-dimethoxyphenyl)thieno[2,3-d]pyrimidine-2,4(l

H,3H)-dione

Method A, Yield: 18.9%, ¾ NMR (500 MHz, DMSO): d 3.69 (s, 3H), 53.79 (s, 3H), 56.54-6.57 (m, 1H), 56.64-6.66 (d, J = 2.76Hz, 1H), 57.08-7.11 (t, J = 4.22Hz, 1H),

57.34-7.38 (t, J = 7.56Hz, 1H), 57.44-7 48(t, J = 8.24Hz, 2H), 57.50-7.53(d, J = 8.76, 2H), 57.60-7.63(d, J = 8.76Hz, 2H), 57.65-7.67(d, J = 8.24Hz, 2H). LC-MS(ESI): m/z Calcd for [C26H20N2O3S] 457.11[M +1]⁺, found 456.9[M +1]⁺. Compound 18

5-(3'-fluoro-4'-methoxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d] pyrimidine-2, 4(lH,3H)-dione

Method A, Yield: 64.3%, ¾ NMR (500 MHz, DMSO): d 3.74 (s, 3H), 53.87 (s,

3H), 56.82-6.85 (d, J = 7.76Hz, 1H), 56.87-6.89 (t, J = 2.19Hz, 1H), 56.94-6.97 (m, 1H), 57.08 (s, 1H), 57.22-7.26(t, J = 8.96Hz, 1H), 57.32-7.35(t, J = 8.09, 1H), 57.46-7 48(d, J = 8.41Hz, 1H), 57.48-7.5 l(d, J = 8.30Hz, 2H), 57.53-7.57 (m, 1H), 57.59-7.53(d, J = 8.63Hz, 2H). LC-MS(ESI): m/z Calcd for [C26H19FN2O4S] 475.1[M +1]⁺, found 475.2[M +1]⁺.

Compound 19

5-(2',4'-dichloro-[l,r-biphenyl]-4-yl)-3-(3-methoxyplienyl)tliieno[2,3-d]pyrimid ine-2,4(lH,3H)-dione

Method A, Yield: 66.8%, ¾ NMR (500 MHz, DMSO): d 3.74 (s, 3H),

56.80-6.84 (d, J = 6.86Hz, 1H), 56.86 (s, 1H), 56.91-6.97 (d, J = 6.86Hz, 1H), 57.08 (s, 1H), 57.08 (s, 1H), 57.30-7.35(t, J = 8.42Hz, 1H), 57.36-7.40(d, J = 8.11, 2H),

57.41-7.45(d, J = 8.11Hz, 1H), 57.49-7.56(m, 3H), 57.72-7.74 (d, J = 2.08Hz, 1H). LC-MS(ESI): m/z Calcd for [C25H16CI2N2O3S] 495.03[M +1]⁺, found 496.7[M +1]⁺.

Compound 20

5-([l,l'-biphenyl]-4-yl)-3-ethylthieno[2,3-d]pyrimidine-2,4(lH,3H)-dione Method A, Yield: 19.3%, ¾ NMR (500 MHz, DMSO): 51.07-1.11 (t, J = 7.05Hz, 3H), 53.83-3.88(m, 2H), 57.05 (s, 1H), 57.36-7.40 (t, J = 7.05z, 1H), 57.46-7.51 (t, J = 8.71Hz, 2H), 57.53-7.56 (d, J = 8.30, 2H), 57.64-7.68(d, J = 8.71Hz, 2H), 57.68-7.73(d, J = 7.88, 2H). LC-MS(ESI): m/z Calcd for [C20H16N2O2S] 348.09[M +1]⁺, found 348.7[M +1]⁺.

Compound 21

5-(3',5'-dichloro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimid ine-2,4(lH,3H)-dione

Method A, Yield: 38.7%, ¾ NMR (500 MHz, DMSO): 53.74 (s, 3H),

56.82-6.86(d, J = 8.17Hz, 1H), 56.87-6.90 (t, J = 2.40Hz, 1H), 56.93-6.97 (m, 1H), 57.13 (s, 1H), 57.32-7.36 (t, J = 8.05, 1H), 57.52-7.57(d, J = 8.41Hz, 2H), 57.72-7 75(d, J = 1.89, 2H). LC-MS(ESI): m/z Calcd for [C25H16CI2N2O3S] 495.03[M +1]⁺, found 495.1[M +1]⁺.

Compound 22

5-([l,r-biphenyl]-4-yl)-3-(3-fluorophenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)- dione

Method A, Yield: 19.4%, ¾ NMR (500 MHz, DMSO): 57.08-7.11 (s, 1H),

57.13-7.17(d, J = 8.50Hz, 1H), 57.21-7.26 (m, 2H), 57.34-7.38 (t, J = 7.94Hz, 1H), 57.44-7.50 (m, 3H), 57.51-7.55 (d, J = 7.94, 2H), 57.60-7.64(d, J = 8.22Hz, 2H), 57.64-7.68(d, J = 6.80, 2H). LC-MS(ESI): m/z Calcd for [C24H15FN2O2S] 415.08[M +1]⁺, found 415.1[M +1]⁺.

Compound 23

5-(4'-chloro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-

2,4(lH,3H)-dione

Method A, Yield: 34.1%, ¾ NMR (500 MHz, DMSO): 53.73 (s, 1H),

56.81-6.82(d, J = 7.72Hz, 1H), 56.85 (s, 1H), 56.93-6.95 (m, 1H), 57.05 (s, 1H), 57.31-7.34 (t, J = 8.30, 2H), 57.50-7.54(m, 4H), 57.60-7.62(d, J = 8.60, 2H),

57.68-7.70(d, J = 8.60, 2H). LC-MS(ESI): m/z Calcd for [C25H17CIN2O3S] 461.06[M +1]⁺, found 461.3[M +1]⁺.

Compound 24

5-(4-(5-fluoropyridin-3-yl)phenyl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine -2,4(lH,3H)-dione

Method A, Yield: 20.6%, ¾ NMR (600 MHz, DMSO): 5 3.74 (s, 3H), 56.82-6.84 (d, J = 7.97Hz, 1H), 56.87 (s, 1H), 56.94-6.95 (d, J = 8.29Hz, 1H), 57.11 (s, 1H), 57.32-7.35 (t, J = 9.21Hz, 1H), 57.57-7.58 (d, J = 7.83Hz, 2H) , 57.74-7.76 (d, J = 6.91Hz, 2H) , 58.05-8.07 (d, J = 10.14Hz, 1H), 5 8.56 (s, 1H) , 5 8.81 (s, 1H).

LC-MS(ESI): m/z Calcd for [C24H16FN3O3S] 446.1 [M +1]⁺, found 446.3[M +1]⁺.

Compound 25

5-([l,r-biphenyl]-4-yl)-3-(6-chloropyridin-3-yl)thieno[2,3-d]pyrimidine-2,4(lH

,3H)-dione Method A, Yield: 98%, ¾ NMR (600 MHz, DMSO): d 7.16 (s, 1H), 7.35-7.38 (m, 1H), 57.45-7.48 (t, J = 8.37Hz, 3H), 57.20-7.25 (m, 1H), 57.37-7.42 (t, J = 7.18Hz, 2H), 57.43-7.48 (d, J = 8.50Hz, 2H), 57.53-7.54 (d, J = 8.76, 2H) , 57.62-7.64 (d, J = 8.50, 2H) , 57.66-7.68 (m, 3H) , 57.88-7.90 (m, 1H) 58.39-8.40 (m, 1H), d 12.60 (s, 1H).

LC-MS(ESI): m/z Calcd for [C23H14CIN3O2S] 432.05[M +1]⁺, found 432.15[M +1]⁺.

Compound 26

3-(4-chloro-3-(trifluoromethyl)phenyl)-5-(4'-(trifluoromethoxy)-[l,r-biphenyl]-

4-yl)thieno [2,3 -d]pyrimidine-2,4( 1 H,3H)-dione

Method A, Yield: 52.5%, ¾ NMR (600 MHz, DMSO): d 7.13 (s, 1H),

57.44-7.45 (d, J = 8.29Hz, 2H), 57.54-7.55 (d, J = 8.29Hz, 2H), 57.63-7.65 (d, J = 6.91Hz, 2H), 57.67-7.69 (d, J = 8.75Hz, 1H), 57.78-7.79 (d, J = 8.75Hz, 2H), 57.83-7.84 (d, J = 9.75Hz, 1H), 57.92 (s, 1H). LC-MS(ESI): m/z Calcd for [C26H13CIF6N2O3S] 582.0 [M +1]⁺, found 583.4[M +1]⁺.

Compound 27

5-(4-(6-fluoropyridin-2-yl)phenyl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine

-2,4(lH,3H)-dione

Method A, Yield: 43.9%, ¾ NMR (600 MHz, DMSO): d 3.73 (s, 3H),

56.82-6.84 (d, J = 7.83Hz, 1H), 56.87 (s, 1H), 56.94-6.95 (d, J = 8.29Hz, 1H),

57.11-7.12 (d, J = 7.37Hz, 2H), 57.32-7.35 (t, J = 7.83Hz, 1H), 57.57-7.58 (d, J = 7.83Hz, 2H), 57.92-7.93 (d, J = 7.37Hz, 1H), 58.00-8.01 (d, J = 7.37Hz, 2H) ,

58.04-8.08 (m, 1H). LC-MS(ESI): m/z Calcd for [C24H16FN3O3S] 446.1 [M +1]⁺, found

446.4[M +1]⁺.

Compound 28

4'-(3-(4-methoxyphenyl)-2,4-dioxo-l,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-5- yl)-[l,l’-biphenyl] -4-carbonitrile

Method A, Yield: 8.6%, ¾ NMR (600 MHz, DMSO): d 3.73 (s, 3H), 56.77 (s, 2H), 56.91 (s, 1H), 57.29(s, 1H), 57.57-7.58 (d, J = 8.32Hz, 2H), 57.68-7.70 (d, J = 8.32Hz, 2H), 57.78-7.93 (m, 5H). LC-MS(ESI): m/z Calcd for [C26H17N3O3S]

452.10[M +1]⁺, found 452.24[M +1]⁺.

Compound 29

3-(4-methoxyphenyl)-5-(4-(6-methoxypyridin-2-yl)phenyl)thieno[2,3-d]pyrimid ine-2,4(lH,3H)-dione

Method A, Yield: 66.0%, ¾ NMR (600 MHz, DMSO): d 3.74 (s, 3H), 53.88 (s, 3H), 56.83-6.85 (m, 1H), 56.88-6.89 (t, J = 2.33Hz, 1H), 56.90-6.91 (d, J = 8.58Hz, 1H),

56.94-6.96 (m, 1H), 57.10 (s, 1H), 57.33-7.35 (t, J = 7.89Hz, 1H), 57.51-7.53 (d, J = 8.57Hz, 2H), 57.60-7.62 (d, J = 8.32Hz, 2H), 58.00-8.02 (m,lH) , 58.48-8.49 (d, J = 2.51Hz, 1H). LC-MS(ESI): m/z Calcd for [C25H19N3O4S] 458.11[M +1]⁺, found 457.91[M +1]⁺.

Compound 30

5-([l,r-biphenyl]-3-yl)-3-(4-methoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3

H)-dione

Method A, Yield: 70.7%, ¾ NMR (600 MHz, DMSO): d 3.73 (s, 3H), 56.83-6.84 (d, J = 7.65Hz, 1H), 56.88-6.89 (t, J = 2.27Hz, 1H), 56.93-6.95 (m, 1H), 57.16 (s, 1H), 57.32-7.36 (m, 2H), 57.41-7.46 (m, 4H), 57.58-7.59 (m, 1H) , 57.63-7.65 (d, J = 7.37Hz, 2H), 57.71 (s, 3H). LC-MS(ESI): m/z Calcd for [C25H18N2O3S] 427.10[M +1]⁺, found 427.02[M +1]⁺.

Compound 31

3-(3-methoxyphenyl)-5-(4-(thiophen-2-yl)phenyl)thieno[2,3-d]pyrimidine-2,4(l

H,3H)-dione

Method A, Yield: 43.1%, ¾ NMR (600 MHz, DMSO): d 3.73 (s, 3H), 56.83-6.85 (d, J = 6.52Hz, 1H), 56.88-6.89 (t, J = 2.40Hz, 1H), 56.94-6.96 (m, 1H), 57.08 (s, 1H), 57.32-7.35 (t, J = 8.57Hz, 1H), 57.46-7.48 (d, J = 8.76Hz, 2H),

57.55-7.56 (d, J = 5.47Hz, 1H) , 57.62-7.64 (m, 1H), 57.65-7.67 (d, J = 8.76Hz, 2H). LC-MS(ESI): m z Calcd for [C23H16N2O3S2] 433.06[M +1]⁺, found 432.97[M +1]⁺ Compound 32

3-(3-methoxyphenyl)-5-(4-(5-(trifluoromethyl)pyridin-3-yl)phenyl)thieno[2,3-d] pyrimidine-2,4(lH,3H)-dione Method A, Yield: 67.4%, ¾ NMR (600 MHz, DMSO): d 3.73 (s, 3H),

56.83-6.85 (d, J = 7.71Hz, 1H), 56.89-6.90 (t, J = 2.24Hz, 1H), 56.94-6.96 (m, 1H), 57.15 (s, 1H), 57.33-7.35 (t, J = 8.08Hz, 1H), 57.59-7.61 (d, J = 8.58Hz, 2H),

57.81-7.82 (d, J = 8.73Hz, 2H) , 58.45 (s, 1H), 59.21 (s, 1H). LC-MS(ESI): m/z Calcd for [C25H16 F3N3O3S] 496.09 [M +1]⁺, found 496.04[M +1]⁺.

Compound 33

3-(3-methoxyphenyl)-5-(3-phenoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)- dione

Method A, Yield: 5.1%, ¾ NMR (600 MHz, DMSO): d 3.74 (s, 3H),

56.80-6.82 (d, J = 8.37Hz, 1H), 56.85 (s, 1H), 56.92-6.96 (m, 2H), 57.00-7.01 (d, J = 7.08Hz, 2H), 57.05 (s, 1H), 57.10-7.12 (t, J = 6.14Hz, 2H), 57.21-7.22 (d, J = 7.56, 1H), 57.31-7.35 (m, 4H). LC-MS(ESI): m/z Calcd for [C25H18N2O4S] 443.10[M +1]⁺, found 442.86[M +1]⁺.

Compound 34

3-(3-methoxyphenyl)-5-(4-phenoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)- dione

Method A, Yield: 32%, ¾ NMR (600 MHz, DMSO): 53.73 (s, 3H), 56.82-6.83 (d, J = 8.58Hz, 1H), 56.93-6.95 (m, 3H), 57.00-7.03 (m, 3H), 57.12-7.15 (t, J = 7.08Hz, 1H), 57.32-7.35 (t, J = 8.03Hz, 1H), 57.37-7.40 (t, J = 8.03Hz, 2H), 57.44-7.45 (d, J = 8.97Hz, 2H). LC-MS(ESI): m/z Calcd for [C25H18N2O4S] 443.10[M +1]⁺, found 443.09[M +1]⁺.

Compound 35

5-(4'-fluoro-[l, -biphenyl]-3-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-

2,4(lH,3H)-dione

Method A, Yield: 30.2%, ¾ NMR (600 MHz, DMSO): 5 3.73 (s, 3H),

56.82-6.84 (d, J = 6.91Hz, 1H), 56.88 (s, 1H), 56.93-6.95 (d, J = 7.54Hz, 1H), 57.16 (s, 1H), 57.25-7.28 (t, J = 8.80Hz, 2H), 57.31-7.34 (t, J = 8.80Hz, 1H), 57.39-7.44 (m, 2H), 57.56-7.57 (d, J = 6.91Hz, 1H), 57.67-7.69 (m, 3H). LC-MS(ESI): m/z Calcd for

[C25H17_FN2O3S] 445.09[M +1]⁺, found 445.15[M +1]⁺.

Compound 36

5-(7-fluoro-9H-fluoren-2-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-2,4(l

H,3H)-dione

Method C, Yield: 56.2%, ¾ NMR (600 MHz, DMSO): 53.73 (s, 3H), 53.92 (s, 2H), 56.83 (d, J = 8.39Hz, 1H), 56.88 (s, 1H), 56.94-6.96 (d, J = 8.80Hz, 1H), 57.08 (s,

1H), 57.19-7.22 (t, J = 8.37Hz, 1H), 57.32-7.35 (t, J = 8.00Hz, 1H), 57.41-7.43 (d, J = 9.22Hz, 1H), 57.41-7.45 (d, J = 8.16Hz, 1H), 57.63 (s, 1H), 57.81-7.82 (d, J = 8.22Hz, 1H), 57.90-7.92 (q, J = 5.02Hz, 1H). LC-MS(ESI): m/z Calcd for [C₂₆H₁₇FN₂O₃S] 457.09 [M +1]⁺, found 456.93[M +1]⁺.

Compound 37

5-(7-fluoro-9-oxo-9H-fluoren-2-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione

Method C, Yield: 47.3%, ¾ NMR (600 MHz, DMSO): 53.73 (s, 3H), 56.84-6.86 (d, J = 7.87Hz, 1H), 56.90 (s, 1H), 56.95-6.97 (d, J = 8.66Hz, 1H), 57.23 (s, 1H), 57.33-7.35 (t, J = 8.66Hz, 1H), 57.43-7.47 (m, 2H), 57.66-7.68 (t, J = 5.85Hz, 2H), 57.75-7.77 (d, J = 8.45Hz, 1H), 57.84-7.86 (q, J = 4.55Hz, 1H). LC-MS(ESI): m/z Calcd for[C₂₆Hi₅FN₂O₄S]471.07[M +1]⁺, found 471.36[M +1]⁺.

Compound 38

3-(4-chloro-3-(trifluoromethyl)phenyl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thie no[2,3-d]pyrimidine-2,4(lH,3H)-dione

Method A, Yield: 48.8%, ¾ NMR (600 MHz, DMSO): 57.14 (s, 1H) 57.52 (s, 2H), 57.54 (s, 2H), 57.65-7.66 (d, J = 8.21Hz, 2H), 57.68-7.69 (d, J = 8.89Hz, 1H), 57.76-7.79 (m, 1H), 57.84-7.85 (d, J = 8.68Hz, 1H), 57.93 (s, 1H). LC-MS(ESI): m/z Calcd for [C25H12CIF5N2O2S] 535.02 [M +1]⁺, found 535.35[M +1]⁺

Compound 39

3-(2-chloro-5-(trifluoromethyl)phenyl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thie no[2,3-d]pyrimidine-2,4(lH,3H)-dione Method A, Yield: 56.5%, ¾ NMR (600 MHz, DMSO): 57.19 (s, 1H)

57.52-7.54 (m, 4H), 57.66-7.68 (d, J = 8.22Hz, 2H), 57.76-7.79 (m, 1H), 57.85-7.89 (m, 2H), 58.04 (s, 1H). LC-MS(ESI): m/z Calcd for[C₂₅Hi₂ClF₅N₂O₂S]535.02[M +1]⁺, found 535.36[M +1]⁺.

Compound 40

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(4-(trifluoromethoxy)phenyl)thieno[2,3- d]pyrimidine-2,4(lH,3H)-dione

Method A, Yield: 54.0%, ¾ NMR (600 MHz, DMSO): 57.12 (s, 1H) 57.45 (s,

4H), 57.52-7.54 (d, J = 8.35Hz, 4H), 57.64-7.65 (d, J = 8.21Hz, 2H), 57.76-7.79 (m, 1H). LC-MS(ESI): m/z Calcd for[C₂₅Hi₃F₅N₂O₃S]517.06[M +1]⁺, found 517.73[M +1]⁺.

Compound 41

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(pyridin-3-yl)thieno[2,3-d]pyrimidine-2, 4(lH,3H)-dione

Method A, Yield: 19.4%, ¾ NMR (600 MHz, DMSO): 57.16 (s, 1H)

57.49-7.54 (m, 5H), 57.65-7.66 (d, J = 8.34Hz, 2H), 57.76-7.79 (m, 2H), 58.50-8.51 (d, J = 2.43Hz, 1H), 58.56-8.57 (m, 1H). LC-MS(ESI): m/z Calcd

for[C₂₃Hi₃F₂N₃O₂S]434.07[M +1]⁺, found 434.55[M +1]⁺. Compound 42

3-(3-chloro-4-methylphenyl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thieno[2,3-d] pyrimidine-2, 4(lH,3H)-dione

Method A, Yield: 18.4%, ¾ NMR (600 MHz, DMSO): 52.35 (s, 3H), 57.12 (s,

1H), 57.16-7.18 (d, J = 8.88Hz, 1H), 57.41-7.42 (m, 2H), 57.52-7.54 (m, 4H),

57.64-7.66 (d, J =9.29Hz, 2H), 57.76-7.79 (m, 1H). LC-MS(ESI): m/z Calcd

for[C₂₅Hi₅ClF₂N₂O₂S]481.05[M +1]⁺, found 481.44[M +1]⁺.

Compound 43

3-(4-chloro-3-(trifluorometliyl)plienyl)-5-(2',4'-difluoro-[l,r-biplienyl]-4-yl)tliie no[2,3-d]pyrimidine-2,4(lH,3H)-dione

Method A, Yield: 56.2%, ¾ NMR (600 MHz, DMSO): 57.15 (s, 1H),

57.18-7.21 (t, J = 8.28Hz, 1H), 57.35-7.39 (t, J = 8.87Hz, 1H), 57.47-7.49 (d, J = 6.50Hz, 2H), 57.53-7.54 (d, J = 8.68Hz, 2H), 57.57-7.60 (m, 1H), 57.68-7.69 (d, J = 8.01Hz, 1H),

57.84-7.85 (d, J = 8.34Hz, 1H), 57.93 (s, 1H). LC-MS(ESI): m/z Calcd

for[C₂₅Hi₂ClF₅N₂O₂S]535.02[M +1]⁺, found 535.36[M +1]⁺

Compound 44

3-(2-chloro-5-(trifluoromethyl)phenyl)-5-(4'-(trifluoromethoxy)-[l,r-biphenyl]- 4-yl)thieno [2,3 -d]pyrimidine-2,4( 1 H,3H)-dione Method A, Yield: 33%, ¾ NMR (600 MHz, CDCb): d 6.78 (s, 1H), 7.27 (s, 1H), 7.53-7.60 (m, 6H), 7.63-7.72 (m, 3H), 10.98 (s, 1H). LCMS (ESI): m/z Calcd for [C₂₆H_I3C1F₆N₂0₃S+H]⁺ 583.02, found 583.65 [M+H]⁺.

Compound 45

5-(2',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(4-(trifluoromethoxy)phenyl)thieno[2,3- d]pyrimidine-2,4(lH,3H)-dione

Method A, Yield: 50.4%, ¾ NMR (600 MHz, DMSO): 57.13 (s, 1H),

57.17-7.21 (t, J = 8.11Hz, 1H), 57.28-7.30 (d, J = 8.11Hz, 1H), 57.34-7.38 (t, J =

10.28Hz, 1H), 57.45 (s, 4H), 57.47 (s, 1H), 57.48 (s, 1H), 57.53 (s, 1H), 57.55 (s, 1H), 57.57-7.60 (t, J = 6.85Hz, 1H), 57.66-7.67 (d, J = 8.76Hz, 1H). LC-MS(ESI): m/z Calcd for [C25H13CIF5N2O3S] 517.06 [M +1]⁺, found 517.67[M +1]⁺.

Compound 46

5-(2',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(pyridin-3-yl)thieno[2,3-d]pyrimidine-2,

4(lH,3H)-dione

Method A, Yield: <10%, ¾ NMR (600 MHz, DMSO): 57.17 (s, 1H),

57.18-7.21(t, J = 8.54, 1H), 57.35-7.39 (t, J = 10.39, 1H), 57.47-7.49 (d, J = 8.28Hz, 2H), 57.50-7.52 (m, 1H), 57.54-7.56 (d, J = 8.63, 2H), 57.57-7.60(t, J = 9.34Hz, 1H), 57.78-7.80(m, 1H). LC-MS(ESI): m/z Calcd for [C23H13F2N3O2S] 434.07 [M +1]⁺, found 434.43[M +1]⁺.

Compound 47

5-(4'-(trifluoromethoxy)- [1,1 '-biphenyl]-4-yl)-3-(4-(trifluorometlioxy)plienyl)tlii eno[2,3-d]pyrimidine-2,4(lH,3H)-dione

Method A, Yield: 79%, ¾ NMR (600 MHz, CDCI3): 5 6.77 (s, 1H), 7.27 (s,

1H), 7.30-7.36 (m, 4H), 7.52-7.58 (m, 6H), 10.24 (s, 1H). LCMS (ESI): m/z Calcd for [C₂₆H_I4F₆N₂0₄S+H]⁺ 565.06, found 565.38 [M+H]⁺.

Compound 48

3-(6-chloropyridin-3-yl)-5-(4'-(trifluoromethoxy)-[l,r-biphenyl]-4-yl)thieno[2,3

-d]pyrimidine-2,4(lH,3H)-dione

Method A, Yield: 23%, ¾ NMR (600 MHz, CDCb): 5 6.82 (s, 1H), 7.27 (d, 2H), 7.46 (d, 1H), 7.54-7.62 (m, 7H), 8.37 (d, 1H) 10.23 (s, 1H). LCMS (ESI): m/z

Calcd for [C₂₄Hi₃ClF N₃0₃S+H]⁺ 516.03, found 516.52 [M+H]⁺.

Compound 49

3-(3,5-dichlorophenyl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thieno[2,3-d]pyrimi dine-2,4(lH,3H)-dione

Method A, Yield: 67.5%, ¾ NMR (600 MHz, DMSO): 57.12 (s, 1H), 57.50(s, 2H), 57.52-7.56 (d, J = 8.44Hz, 4H), 57.65 (s, 1H), 57.67-7.68 (m, 2H), 57.76.7.79 (m, 1H). LC-MS(ESI): m/z Calcd for [C24H12CI2F2N2O2S] 501.00[M +1]⁺, found 501.40[M +1]⁺.

Compound 50

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(pyridin-4-yl)thieno[2,3-d]pyrimidine-2,

4(lH,3H)-dione

Method A, Yield: 5.9%, ¾ NMR (600 MHz, DMSO): 57.16 (s, 1H),

57.42-7.43(d, J = 6.03Hz, 2H), 57.53-7.54 (d, J = 8.04Hz, 5H), 57.65-7.66 (d, J =

8.54Hz, 3H), 58.68-8.69 (d, J = 6.04Hz, 2H). LC-MS(ESI): m/z Calcd for

[C23H13F2N3O2S] 434.07 [M +1]⁺, found 434.32[M +1]⁺.

Compound 51

3-(6-chloropyridin-3-yl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thieno[2,3-d]pyri midine-2,4(lH,3H)-dione

Method A, Yield: 2.1%, ¾ NMR (600 MHz, DMSO): 57.16 (s, 1H),

57.53-7.54(d, J = 8.70Hz, 4H), 57.65-7.67 (d, J = 9.37Hz, 3H), 57.78-7.80 (m, 1H), 57.88-7.90 (m, 1H), 58.39 (s, 1H). LC-MS(ESI): m/z Calcd for [C23H12CI F2N3O2S] 468.03[M +1]⁺, found 468.30[M +1]⁺.

Compound 52

3-(pyridin-3-yl)-5-(4'-(trifluoromethoxy)-[l,r-biphenyl]-4-yl)thieno[2,3-d]pyri midine-2,4(lH,3H)-dione Method A, Yield: 16%, ¾ NMR (600 MHz, DMSO): d 7.17 (s, 1H), 7.43-7.48 (m, 2H), 7.51-7.58 (m, 3H), 7.63-7.68 (m, 2H), 7.76-7.84 (m, 3H), 8.52 (d, 1H), 8.58 (d, 1H), 12.57 (s, 1H). LCMS (ESI): m/z Calcd for ^HwFsNsOsS+Hr 482.07, found 482.25 [M+H]⁺.

Compound 53

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(pyridin-2-yl)thieno[2,3-d]pyrimidine-2,

4(lH,3H)-dione

Method B, Yield: 82.1%, ¾ NMR (600 MHz, DMSO): d 7.16 (s, 1H),

57.45-7.47 (m, 2H), d7.52 (s, 2H), d7.53 (s, 2H), d 7.65-7.66 (d, J = 8.73Hz, 2H), d 7.76-7.79 (m,lH), d 7.94-7.97 (m, 1H), d 8.56-8.58 (m, 1H). LCMS (ESI): m/z Calcd for [C23H13F2N3O2S] 434.07 [M +1] ⁺, found 434.15[M +1] ⁺.

Compound 54

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(2-methoxypyridin-4-yl)thieno[2,3-d]py rimidine-2,4(lH,3H)-dione

Method B, Yield: 60.8%, ¾ NMR (600 MHz, DMSO): d 3.87 (s, 3H), d 6.87 (s, 1H), d6.98-6.99 (d, J = 5.44Hz, 1H), d7.15 (s, 1H), d 7.53-7.54 (d, J = 7.76Hz, 4H), d 7.62-7.66 (d, J = 7.76Hz, 2H), d 7.76-7.79 (m, 1H), d 8.23-8.24 (d, J = 5.53Hz, 1H).

LCMS (ESI): m/z Calcd for [C24H15F2N3O2S] 464.08 [M +1] ⁺, found 464.19[M +1] ⁺. Compound 55

5-(3',4'-difluoro-[l, -biphenyl]-4-yl)-3-(3-methoxyphenyl)-l-methylthieno[2,3- d]pyrimidine-2,4(lH,3H)-dione

To a solution of compound 16 (0.043 mmol) in DMF (2.0 mL) was added NaH

(0.065 mmol). After 5 minutes, Mel (0.13 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hr. The mixture was poured into H2O and extracted with dichloromethane (DCM). The organic phase was dried (MgSCL), filtered, and concentrated in vacuo. The residue was purified by MPLC (EA/Hex = 1/3) to furnish the desire adduct. Yield: 99%, 1H NMR (500 MHz, DMSO-dd): d 3.54 (s, 3H), 3.74 (s, 3H), 6.84 (d, J = 7.8 Hz, 1H), 6.87 (t, 7 = 2.2 Hz, 1H), 6.97 (dd, J = 8.5; 2.5 Hz, 1H), 7.27-7.38 (m, 4H), 7.40-7.48 (m, 1H), 7.52-7.60 (m, 4H). LCMS (ESI): m/z Calcd for [C₂₆H_I8F₂N₂0₃S+H]⁺ 477.10, found 477.45 [M+H]⁺.

Compound 56

l-benzyl-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3- d]pyrimidine-2,4(lH,3H)-dione

To a solution of Compound 16 (0.043 mmol) in DMF (2.0 mL) was added NaH (0.065 mmol). After 5 minutes, benzyl bromide (BnBr) (0.13 mmol) was added, and the reaction mixture was stirred at room temperature for 1 Hr. The mixture was poured into H2O and extracted with DCM. The organic phase was dried (MgSCL), filtered, and concentrated in vacuo. The residue was purified by medium pressure liquid chromatography (MPLC) (EA/Hex = 3/7) to furnish the desire adduct. Yield : 46%, 1H NMR (500 MHz, DMSO-i 6): d 3.76 (s, 3H), 5.26 (s, 2H), 6.91-6.94 (m, 1H), 6.97-7.01 (m, 2H), 7.22 (s, 1H), 7.28-7.33 (m, 1H), 7.34-7.45 (m, 6H), 7.46-7.49 (m, 2H), 7.52-7.58 (m, 4H). LCMS (ESI): m/z Calcd for [C₃₂H₂₂F₂N₂0₃S+H]⁺ 553.13, found 553.27 [M+H]⁺.

EXAMPLE 2 Characterization of compound of formula (I)

In this example, the ompounds of Example 1 were test for their respective activities against CD73 in accordance with procedures described in the“Materials and Methods” section. Results are summarized in Table 1.

Table 1. Inhibition of CD73 activity

Potency : CD73 IC50 : + means >1 mM, ++ means 500 nM to 1 mM, +++ means < 500 nM.

It was found that compounds 15 and 16 were most potent, while the rest of the compounds exhibited fair to moderate inhibitory activity toward CD73. It will be understood that the above description of embodiments is given by way of example only and that various modifications may be made by those with ordinary skill in the art. The above specification, examples and data provide a complete description of the structure and use of exemplary embodiments of the invention.

Although various embodiments of the invention have been described above with a certain degree of particularity, or with reference to one or more individual embodiments, those with ordinary skill in the art could make numerous alterations to the disclosed embodiments without departing from the spirit or scope of the present disclosure.

Claims

WHAT IS CLAIMED IS:

1. A compound of Formula (I):

wherein:

R is phenyl or aryl;

R3 is H, Ci-20 alkyl, or -(CFFVphenyl, in which n is an integral between 1 to 10;

R4 is H, Ci-20 alkyl, phenyl, or heteroaryl; and

each Ci-20 alkyl, phenyl and heteroaryl is optionally substituted with at least one substituent selected from the group consisting of halogen, C1-4 lower alkyl, haloalkyl, alkoxyl, haloalkoxy, -CN, and -COOH;

or a pharmaceutically acceptable salt, solvate, hydrate, co-crystal, and stereoisomer thereof.

2. The compound of claim 1, wherein the compound is a CD73 inhibitor.

3. The compound of claim 2, wherein the compound of formula (I) has the structure of formula (II),

wherein: X, Y, and Z are independently C or N, provided that when one of X, Y, and Z is N, then the other two are independently C;

R3 is H, Ci-4alkyl, or -Cth-phenyl;

R^a and R^b are independently H, -OR₅, phenyl, pyridinyl, or thiophenyl, in which R5 is Ci-4 lower alkyl or phenyl;

R^c is nil or at least one substituent selected from the group consisting of halogen, -OH, Ci-4 lower alkyl, haloalkyl, alkoxyl, and haloalkoxy;

each phenyl and pyridinyl is optionally substituted with at least one substituent selected from the group consisting of halogen, -OH, -OR₆, -NH2, -NR7R8, alkyl, haloalkyl, and -CN; and

R6, R₇, and Re are independently C_1-4 lower alkyl or phenyl optionally substituted with at least one halogen.

4. The compound of claim 2, wherein the compound of formula (I) has the structure of formula (III),

wherein:

R^a is phenyl optionally substituted with at least one halogen.

5. The compound of claim 2, wherein the compound of formula (I) has the structure of formula (IY), X is C or -C=0; and

R^c is nil or at least one substituent selected from the group consisting of halogen, -OH, Ci-4 lower alkyl, haloalkyl, alkoxyl, and haloalkoxy.

6. The compound of claim 2, wherein the compound is selected from the group consisting of:

3-(3-methoxyphenyl)-5-phenylthieno[2,3-d]pyrimidine-2,4(lH,3H)-dione;

5-(2'-hydroxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-

2,4(lH,3H)-dione;

5-(3-hydroxyphenyl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)-di one;

5-(4'-hydroxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine- 2,4(lH,3H)-dione;

5-([l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)

-dione;

5-(3'-methoxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-

2,4(lH,3H)-dione;

5-(3'-hydroxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-

2,4(lH,3H)-dione;

5-(2'-hydroxy-3'-methoxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d] pyrimidine-2,4(lH,3H)-dione; 5-(4'-methoxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-

2,4(lH,3H)-dione;

3-(3-methoxyphenyl)-5-(4'-(trifluoromethoxy)-[l,l'-biphenyl]-4-yl)thieno[2,3-d]p yrimidine-2,4(lH,3H)-dione;

5-([l,r-biphenyl]-4-yl)-3-(4-chlorophenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)-di one;

5-([l,r-biphenyl]-4-yl)-3-(4-chloro-3-(trifluoromethyl)phenyl)thieno[2,3-d]pyrim idine-2,4(lH,3H)-dione;

3-(3-methoxyphenyl)-5-(4'-(trifluoromethyl)-[l,r-biphenyl]-4-yl)thieno[2,3-d]pyr imidine-2,4(lH,3H)-dione;

5-([l,r-biphenyl]-4-yl)-3-(2,4-difluorophenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H

)-dione;

5-(4'-fluoro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-2,

4(lH,3H)-dione;

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione;

5-([l,r-biphenyl]-4-yl)-3-(2,4-dimethoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,

3H)-dione;

5-(3'-fluoro-4'-methoxy-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]p yrimidine-2,4(lH,3H)-dione;

5-(2',4'-dichloro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione;

5-([l,r-biphenyl]-4-yl)-3-ethylthieno[2,3-d]pyrimidine-2,4(lH,3H)-dione;

5-(3',5'-dichloro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione;

5-([l,r-biphenyl]-4-yl)-3-(3-fluorophenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)-di one;

5-(4'-chloro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-2,

4(lH,3H)-dione;

5-(4-(5-fluoropyridin-3-yl)phenyl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-

2,4(lH,3H)-dione;

5-([l,r-biphenyl]-4-yl)-3-(6-chloropyridin-3-yl)thieno[2,3-d]pyrimidine-2,4(lH,3

H)-dione; 3-(4-chloro-3-(trifluoromethyl)phenyl)-5-(4'-(trifluoromethoxy)-[l,r-biphenyl]-4- yl)thieno [2 ,3 -d]pyrimidine-2 ,4( 1 H, 3H) -dione ;

5-(4-(6-fluoropyridin-2-yl)phenyl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-

2,4(lH,3H)-dione;

4'-(3-(4-methoxyphenyl)-2,4-dioxo-l,2,3,4-tetrahydrothieno[2,3-d]pyrimidin-5-yl )- [ 1 , 1’-biphenyl] -4-carbonitrile;

3-(4-methoxyphenyl)-5-(4-(6-methoxypyridin-2-yl)phenyl)thieno[2,3-d]pyrimidin e-2,4(lH,3H)-dione;

5-([l,r-biphenyl]-3-yl)-3-(4-methoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)

-dione;

3-(3-methoxyphenyl)-5-(4-(thiophen-2-yl)phenyl)thieno[2,3-d]pyrimidine-2,4(lH,

3H)-dione;

3-(3-methoxyphenyl)-5-(4-(5-(trifluoromethyl)pyridin-3-yl)phenyl)thieno[2,3-d]p yrimidine-2,4(lH,3H)-dione;

3-(3-methoxyphenyl)-5-(3-phenoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)-di one;

3-(3-methoxyphenyl)-5-(4-phenoxyphenyl)thieno[2,3-d]pyrimidine-2,4(lH,3H)-di one;

5-(4'-fluoro-[l,r-biphenyl]-3-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-2,

4(lH,3H)-dione;

5-(7-fluoro-9H-fluoren-2-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-2,4(l

H,3H)-dione;

5-(7-fluoro-9-oxo-9H-fluoren-2-yl)-3-(3-methoxyphenyl)thieno[2,3-d]pyrimidine-

2,4(lH,3H)-dione;

3-(4-chloro-3-(trifluoromethyl)phenyl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thien o [2, 3 -d]pyrimidine-2 ,4( 1 H, 3H) -dione ;

3-(2-chloro-5-(trifluoromethyl)phenyl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thien o [2, 3 -d]pyrimidine-2 ,4( 1 H, 3H) -dione ;

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(4-(trifluoromethoxy)phenyl)thieno[2,3-d] pyrimidine-2, 4(lH,3H)-dione;

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(pyridin-3-yl)thieno[2,3-d]pyrimidine-2,4

(lH,3H)-dione; 3-(3-chloro-4-methylphenyl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thieno[2,3-d]py rimidine-2,4(lH,3H)-dione;

3-(4-chloro-3-(trifluoromethyl)phenyl)-5-(2',4'-difluoro-[l,r-biphenyl]-4-yl)thien o [2, 3 -d]pyrimidine-2 ,4( 1 H, 3H) -dione ;

3-(2-chloro-5-(trifluoromethyl)phenyl)-5-(4'-(trifluoromethoxy)-[l,r-biphenyl]-4- yl)thieno [2 ,3 -d]pyrimidine-2 ,4( 1 H, 3H) -dione ;

5-(2',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(4-(trifluoromethoxy)phenyl)thieno[2,3-d] pyrimidine-2, 4(lH,3H)-dione;

5-(2',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(pyridin-3-yl)thieno[2,3-d]pyrimidine-2,4

(lH,3H)-dione;

5-(4'-(trifluoromethoxy)- [1,1 '-biphenyl]-4-yl)-3-(4-(trifluoromethoxy)phenyl)thie no[2,3-d]pyrimidine-2,4(lH,3H)-dione;

3-(6-chloropyridin-3-yl)-5-(4'-(trifluoromethoxy)-[l,r-biphenyl]-4-yl)thieno[2,3- d]pyrimidine-2,4(lH,3H)-dione;

3-(3,5-dichlorophenyl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thieno[2,3-d]pyrimidi ne-2,4(lH,3H)-dione;

5-(3',4'-dilluoro-[l,r-biphenyl]-4-yl)-3-(pyridin-4-yl)thieno[2,3-d]pyrimidine-2,4

(lH,3H)-dione;

3-(6-chloropyridin-3-yl)-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)thieno[2,3-d]pyrimi dine-2,4(lH,3H)-dione;

3-(pyridin-3-yl)-5-(4'-(trilluoromethoxy)-[l,r-biphenyl]-4-yl)thieno[2,3-d]pyrimi dine-2,4(lH,3H)-dione;

5-(3',4'-dilluoro-[l,r-biphenyl]-4-yl)-3-(pyridin-2-yl)thieno[2,3-d]pyrimidine-2,4

(lH,3H)-dione;

5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(2-methoxypyridin-4-yl)thieno[2,3-d]pyri midine-2,4(lH,3H)-dione;

5-(3',4'-dilluoro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)-l-methylthieno[2,3-d] pyrimidine-2, 4(lH,3H)-dione; and

l-benzyl-5-(3',4'-difluoro-[l,r-biphenyl]-4-yl)-3-(3-methoxyphenyl)thieno[2,3-d] pyrimidine-2, 4(lH,3H)-dione.

7. A pharmaceutical composition comprising the compound of claim 6, and a pharmaceutically acceptable carrier.

8. A method of treating a subject having or suspected of having a disease mediated by adenosine comprising administering to the subject an effective amount of the pharmaceutical composition of claim 7.

9. The method of claim 8, wherein the disease mediated by adenosine is AIDS, an autoimmune disease, atherosclerosis, a cancer, an infectious disease,

ischemia-reperfusion injury or pre-cancerous syndrome.

10. The method of claim 9, wherein the autoimmune disease is multiple sclerosis, psoriasis, systemic lupus erythemotosis (SLE), Type I diabetes mellitus, or Wegener’s granulomatosis.

11. The method of claim 10, wherein the cancer is acinar cell carcinoma, acute lymphoblastic leukemia, acute myelogenous leukemia, adenocarcinoma,

adenosquamous carcinoma, Bannayan-Zonana syndrome, bladder cancer, breast cancer, buccal cancer, Burkitt's lymphoma, cervical cancer, colon cancer, chronic myelogenous leukemia, chronic lymphocytic leukemia, chronic neutrophilic leukemia, Cowden disease, ductal adenocarcinoma, endometrial cancer, Edwing’s sarcoma, ependymoma, erythroleukemia, esophageal cancer, follicular lymphoma, gastric cancer,

gastrointestinal stromal tumor, giant cell tumor of bone, glioma, glioblastoma, astrocytoma, glioblastoma multiforme, glucagonoma, hairy-cell leukemia, head and neck cancer, hodgkins lymphoma, insulinoma, immunoblastic large cell leukemia, kidney cancer, Lhermitte-Duclos disease, lobular carcinoma, liver cancer, lymphoblastic T cell leukemia, malignant lymphoma, medulloblastoma, melanoma, mantle cell leukemia, mesothelioma, multiple myeloma, megakaryoblastic leukemia,

nasopharangeal cancer, non-hodgkins lymphoma, non-small cell lung cancer, osteosarcoma, ovarian cancer, pancreatic cancer, plasmacytoma, promyelocytic leukemia, prostate cancer, Rhabdomyosarcoma, sarcoma, salivary gland cancer, skin cancer, small cell lung cancer, testicular cancer, thyroid cancer, urothelial cancer, vulval cancer, or Wilm’ s tumor.

12. The method of claim 8, further comprising administering to the subject an anti-neoplastic agent, an immunosuppressant, or an anti-infectious agent.

13. The method of claim 12, wherein the anti-neoplastic agent is selected from the group consisting of an anti -microtubule agent, a platinum coordination complex, an alkylating agent, an antibiotic, a topoisomerase II inhibitor, a topoisomerase I inhibitor, an anti-metabolite, a hormone, a signal transduction pathway inhibitor, a non-receptor tyrosine kinase, an angiogenesis inhibitor, an immunotherapeutic agent, a pro-apoptotic agent, cell cycle signaling inhibitor, a proteasome inhibitor, an inhibitor of cancer metabolism, an anti-PD-Ll agent, a PD-1 antagonist, an immune-modulator, Stimulator of Interferon Genes (STING) modulating compound, CD39 inhibitor, A2a and A2a adenosine antagonists, Toll-like receptor 4 (TLR4) inhibitor, anti-ICOS antibody and anti- 0X40 antibody.

14. The method of claim 12, wherein the immunosuppressant is a corticosteroid, a calcineurin inhibitor, a mammalian target of rapamycin (mTOR) inhibitor, an inosine monophosphate dehydrogenase (IMPDH) inhibitor, a therapeutic protein, or a monoclonal antibody.

15. The method of claim 12, wherein the anti-infectious agent is an antibiotic selected from the group consisting of acumycin, ampicillin, amoxycillin, amphotericins, antimycins, anglomycin, avermectins, azithromycin, boromycin, carbomycins, carbapenem, ceftazidime, cethromycin, chloramphenicol, chalcomycin, ciprofloxacin, concanamycins, cirramycin, clarithromycin, colistin, cycloxacillin, daptomycin, desmethyl azithromycin, desertomycins, dihydropikromycin, dirithromycin, doxycycline, enramycin, erythromycin, flurithromycin, flumequin gentamycin, juvenimicins, kujimycins, lankamycins, lincomycin, litorin, leucomycins,

megalomicins, meropenem, methymycin, midecamycins, mycinamicin I, mycinamicin II, mycinamicin III, mycinamicin IV, mycinamicin V, mycinamicin VI, mycinamicin VII, mycinamicin VIII, narbomycin, neoantimycin, neomethymycin, netilmicin, neutromycin, niddamycins, norfloxacin, oleandomycins, oligomycins, ossamycin, oxacillin, oxolinic acid, penicillin, pikromycin, piperacillin, platenomycins, rapamycins, relomycin, rifamycins, rosaramicin, roxithromycin, virginiamycin, spiramycin, sporeamycin, staphococcomycin, streptomycin, sulfamethoxazole, swalpamycin, telithromycin, teicoplanin, timentin, tobramycin, ticarcillin, trimethoprim, tetracyclin, and zlocillin.