WO2020106625A1

WO2020106625A1 - Compounds and uses thereof

Info

Publication number: WO2020106625A1
Application number: PCT/US2019/062025
Authority: WO
Inventors: Andrew J. Mcriner; Junyi Zhang; Yanbin Liu; Neil E. Westlund; Ying Zhang; Dawn M. TROAST
Original assignee: Xios Therapeutics Inc
Current assignee: Xios Therapeutics Inc
Priority date: 2018-11-19
Filing date: 2019-11-18
Publication date: 2020-05-28
Anticipated expiration: 2021-05-19

Abstract

The present invention features compounds useful in the treatment of adenosine or adenosine receptor related disorders.

Description

COMPOUNDS AND USES THEREOF

Background

Current cancer treatments include surgery, chemotherapy, radiation therapy, and photodynamic therapy. However, none of these treatments is completely effective, and each has its own associated side effects.

The inflammatory response helps eliminate harmful agents from the body, including tumors. However, inflammation is also a non-specific response that can harm healthy tissue. The cells involved in inflammation include immune cells (e.g., T cells, B cells, dendritic cells, macrophages, or granulocytes), the vascular endothelium, vascular smooth muscle cells, and fibroblasts. Inflammation is normally a localized action that results in isolation of the damaging agent and injured tissue. There exists a need to attenuate active immune responses through inhibitory and negative feedback pathways to prevent collateral damage in the healthy tissue. Collectively known as immune checkpoint pathways, inhibitory signaling networks provide a negative feedback mechanism that is important for immunomodulation and protection of healthy tissues from the damage of inflammatory responses.

The adenosine receptors are important immune checkpoints in cancer therapy. The present disclosure relates to compositions and methods for the treatment of disorders related to adenosine or adenosine receptors, such as cancer.

Summary of the Invention

The present invention features compounds useful to modulate levels of adenosine, e.g., in a subject in need thereof. In some embodiments, the compounds described herein modulate adenosine levels via antagonism of the adenosine A2A receptor. In some embodiments, the compounds described herein modulate adenosine levels via antagonism of the adenosine A_åB receptor. In some embodiments, the compounds described herein modulate adenosine levels via antagonism of both the adenosine A2_A and A2B receptors. In some embodiments, the compounds described herein are useful in the treatment of disorders associated with an alteration in the activity level of the adenosine A2A receptor. In some embodiments, the compounds described herein are useful in the treatment of disorders associated with an alteration in the activity level of the adenosine A2B receptor. The compounds of the invention, alone or in combination with other pharmaceutically active agents, can be used for treating such disorders, which include cancer.

In one aspect, the invention provides a compound, or a pharmaceutically acceptable salt thereof, having the structure of Formula I:

where each of R¹, R², and R³ is, independently, H or optionally substituted C1-C6 alkyl;

each of R⁵ and R⁶ is, independently, H or optionally substituted C1-C6 alkyl, or R⁵ and R⁶, together with the atom to which each is attached, combine to form an optionally substituted C3-C10 carbocyclyl;

B is optionally substituted C₃-Cio carbocyclylene or optionally substituted C3-C9 heterocyclylene;

R5 _R6 0 h

L² is V " ^_r n \

orr ^ / ' ^*

ml is 1 , 2, or 3;

m2 is 0, 1 , 2, or 3; and

R⁴ is optionally substituted C6-C10 aryl, optionally substituted C3-C9 heterocyclyl, optionally substituted C1-C9 heteroaryl, or optionally substituted C₁-C₆ heteroalkyl,

or a pharmaceutically acceptable salt thereof.

In some embodiments, if R¹ is H, R² is CH3, R³ is H, and L¹ is

, then R⁴ is substituted C6-C10 aryl, optionally substituted C3-C9 heterocyclyl, optionally substituted C1 -C9 heteroaryl, or optionally substituted C1-C6 heteroalkyl.

In some embodiments, R¹ is H. In some embodiments, R¹ is optionally substituted C1-C6 alkyl. In some embodiments,

In some embodiments, R² is H. In some embodiments, R² is optionally substituted C1 -C6 alkyl. In

/CH₃

some embodiments, R² is A.

In some embodiments, R³ is H. In some embodiments, R³ is optionally substituted C1 -C6 alkyl. In some embodiments,

R5 R6 A

In some embodiments, L¹ is /

In some embodiments, ml is 1 or 2. In some embodiments, ml is 1 . In some embodiments, ml is 2.

In some embodiments, R⁵ is H. In some embodiments, R⁵ is optionally substituted C1-C6 alkyl. In

C^H3

some embodiments, R⁵ is A.

In some embodiments, R⁶ is H. In some embodiments, R⁶ is optionally substituted C1-C6 alkyl. In

/CH3 _S /CH₃

some embodiments, R⁶ is A . In some embodiments, R⁶ is H or A

In some embodiments, R⁵ and R⁶, together with the atom to which each is attached, combine to form an optionally substituted C3-C10 carbocyclyl.

In some embodiments, R⁵ and R⁶, together with the atom to which each is attached, combine to form an optionally substituted C3-C6 carbocyclyl. In some embodiments, R⁵ and R⁶, together with the atom to which each is attached, combine to form an optionally substituted C3-C5 carbocyclyl.

In some embodiments, B is optionally substituted C3-C10 carbocyclylene. In some embodiments,

B is optionally substituted C3-C9 heterocyclylene.

. ,

0 R⁵ R⁶

In some embodiments, L² is

. In some embodiments, L² is

In some embodiments, m2 is 1 , 2, or 3. In some embodiments, m2 is 1 or 2. In some embodiments, m2 is 1 . In some embodiments, m2 is 2.

In some embodiments, R⁴ is optionally substituted C6-C10 aryl.

In some embodiments,

where n is 0, 1 , 2, 3, 4, or 5; and

each R^a is, independently, C1-C6 perfluoroalkyl or C3-C8 cycloalkyl.

In some embodiments, each R^a is, independently,

, or

In some embodiments, each R^a is

.

In some embodiments, n is 1 or 2.

In some embodiments, R⁴ is optionally substituted C3-C9 heterocyclyl.

where o is 0, 1 , 2, or 3;

each R^b is, independently, halo or optionally substituted C1-C6 alkyl; and R⁷ is H or optionally substituted C1-C6 alkyl.

,

In some embodiments,

some embodiments,

In some embodiments, R⁴ is

. , In some

embodiments,

In some embodiments, each R^b is, independently, halo or C1-C6 alkyl.

In some embodiments, o is 0 or 1 . In some embodiments, o is 0. In some embodiments, o is 1 .

In some embodiments, R⁷ is H. In some embodiments, R⁷ is optionally substituted C1-C6 alkyl. In some embodiments,

In some embodiments,

In some embodiments, R⁴ is optionally substituted C1 -C9 heteroaryl.

In some embodiments,

where

p is 0, 1 , 2, or 3;

q is 0, 1 , 2, or 3;

each of X^a, X^b, X^c, and X^d is, independently, N, CH, or CR^C;

X^e is O, S, NR⁸, or CR^cR^d;

each R^c is, independently, halo or optionally substituted C1-C6 alkyl; and

R⁸ is H or optionally substituted C1-C6 alkyl.

in some embodiments, R⁴ is

. ,

In some embodiments, q is 0 or 1 . In some embodiments, q is 0. In some embodiments, q is 1 .

In some embodiments, each of X^a and X^b is, independently, N or CH. In some embodiments, each of X^c and X^d is, independently, N or CH. In some embodiments, X^e is O or S. In some

embodiments, X^e is NH or CH2.

In some embodiments, X^a is N. In some embodiments, X^a is CH.

In some embodiments, X^b is N. In some embodiments, X^b is CH.

In some embodiments, X^c is N. In some embodiments, X^c is CH.

In some embodiments, X^d is N. In some embodiments, X^d is CH.

In some embodiments, X^e is NH. In some embodiments, X^e is CH2.

In some embodiments, p is 0 or 1 . In some embodiments, p is 0. In some embodiments, p is 1 .

In some embodiments, each R^c is, independently, halo or C1-C6 alkyl.

In some embodiments, R⁸ is H. In some embodiments, R⁸ is optionally substituted C1-C6 alkyl. In some embodiments,

In some embodiments, R⁴ is optionally substituted Ci-Ce heteroalkyl. In some embodiments,

In some embodiments,

In some embodiments,

In one aspect, the invention provides a compound, or a pharmaceutically acceptable salt thereof, selected from any one of compounds 1 -80 in Table 1 .

Table 1. Compounds of the Invention

In one aspect, the invention provides a pharmaceutical composition comprising any of the foregoing compounds (e.g. a compound of Formula I, such as any of compounds 1-86 in Table 1 ) and a pharmaceutically acceptable excipient.

In another aspect, the invention provides a method for modulating the level of adenosine in a cell.

This method includes contacting a cell with an effective amount of any of the foregoing compounds or pharmaceutical compositions.

In yet another aspect, the invention provides a method for modulating the level of adenosine in a subject, the method comprising administering to the subject an effective amount of a compound of any of the foregoing compounds or pharmaceutical compositions.

In a further aspect, the invention provides a method for the treatment of a disorder associated with adenosine (e.g., cancer) in a subject in need thereof, the method comprising administering an effective amount of any of the foregoing compounds or pharmaceutical compositions.

In another aspect, the invention provides a method for modulating the activity of the adenosine A2_A receptor in a cell. This method includes contacting a cell with an effective amount of any of the foregoing compounds or pharmaceutical compositions.

In yet another aspect, the invention provides a method for modulating the activity of the adenosine A2_A receptor in a subject, the method comprising administering to the subject an effective amount of a compound of any of the foregoing compounds or pharmaceutical compositions.

In a further aspect, the invention provides a method for the treatment of a disorder associated with the adenosine A2_A receptor (e.g., cancer) in a subject in need thereof, the method comprising administering an effective amount of any of the foregoing compounds or pharmaceutical compositions.

In another aspect, the invention provides a method for modulating the activity of the adenosine A2_B receptor in a cell. This method includes contacting a cell with an effective amount of any of the foregoing compounds or pharmaceutical compositions.

In yet another aspect, the invention provides a method for modulating the activity of the adenosine A2_B receptor in a subject, the method comprising administering to the subject an effective amount of a compound of any of the foregoing compounds or pharmaceutical compositions. In a further aspect, the invention provides a method for the treatment of a disorder associated with the adenosine A2B receptor (e.g., cancer) in a subject in need thereof, the method comprising administering an effective amount of any of the foregoing compounds or pharmaceutical compositions.

In another aspect, the invention provides a method for modulating the activity of the adenosine AZA and the adenosine ASB receptor in a cell. This method includes contacting a cell with an effective amount of any of the foregoing compounds or pharmaceutical compositions.

In yet another aspect, the invention provides a method for modulating the activity of the adenosine A2A and the adenosine A2B receptor in a subject, the method comprising administering to the subject an effective amount of a compound of any of the foregoing compounds or pharmaceutical compositions.

In a further aspect, the invention provides a method for the treatment of a disorder associated with the adenosine A2A and the adenosine A_åB receptor (e.g., cancer) in a subject in need thereof, the method comprising administering an effective amount of any of the foregoing compounds or

pharmaceutical compositions.

In another aspect, the invention provides a method of modulating an immune response in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of any of the foregoing compounds or pharmaceutical compositions.

In yet another aspect, the invention provides a method for modulating the activity of the dopamine D2 receptor in a cell, the method comprising contacting a cell with an effective amount of any of the foregoing compounds or pharmaceutical compositions.

In a further aspect, the invention provides a method for modulating the activity of the dopamine D2 receptor in a subject, the method comprising administering to the subject an effective amount of a compound of any of the foregoing compounds or pharmaceutical compositions.

In a further aspect, the invention provides a method of inhibiting inflammation in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the foregoing compounds or pharmaceutical compositions.

In another aspect, the invention provides a method of treating cancer in a subject, the method comprising administering to the subject an effective amount of any of the foregoing compounds or pharmaceutical compositions.

In some embodiments, the method further comprises administering to the subject an additional anticancer therapy (e.g., an immunotherapy such as a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, and/or adoptive T-cell transfer therapy, a chemotherapeutic or cytotoxic agent, and/or or radiotherapy). In some embodiments, the additional anticancer therapy and any of the foregoing compounds or pharmaceutical compositions are administered within 28 days of each other each in an amount that together are effective to treat the subject.

In some embodiments of any of the foregoing methods, the subject has a compromised immune system. In some embodiments, the cancer has failed to respond to a previously administered an immunotherapy and/or the cancer is resistant to an immunotherapy.

In some embodiments of any of the foregoing methods, the cancer is breast cancer such as triple negative breast cancer, colon cancer, renal cell cancer, non-small cell lung cancer, hepatocellular carcinoma, gastric cancer, ovarian cancer, pancreatic cancer, esophageal cancer, prostate cancer, sarcoma, glioblastoma, diffuse large B-cell lymphoma, leukemia (e.g., acute myeloid leukemia), or melanoma. In some embodiments of any of the foregoing methods, the cancer is melanoma. In some embodiments of any of the foregoing methods, the cancer is breast cancer. In some embodiments of any of the foregoing methods, the cancer is renal cell cancer. In some embodiments of any of the foregoing methods, the cancer is pancreatic cancer. In some embodiments of any of the foregoing methods, the cancer is non-small cell lung cancer. In some embodiments of any of the foregoing methods, the cancer is colon cancer. In some embodiments of any of the foregoing methods, the cancer is ovarian cancer. In some embodiments of any of the foregoing methods, the cancer is glioblastoma. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is prostate cancer. In some

embodiments, the cancer is diffuse large B-cell lymphoma. In some embodiments, the cancer is leukemia (e.g., acute myeloid leukemia).

In some embodiments, the cancer is a metastatic cancer, a migrating cancer, or a non-metastatic cell migration cancer.

In certain embodiments of any of the foregoing methods, the cancer is a drug resistant cancer or has failed to respond to a prior therapy (e.g., a cancer resistant to, or a cancer that has failed to respond to prior treatment with vemurafenib, dacarbazine, a CTLA4 inhibitor, a PD1 inhibitor, interferon therapy, a BRAF inhibitor, a MEK inhibitor, radiotherapy, temozolimide, irinotecan, a CAR-T therapy, herceptin, pertuzumab, tamoxifen, capecitabine, docetaxol, platinum agents (e.g., carboplatin), a taxane (e.g., paclitaxel and/or docetaxel), ALK inhibitors, MET inihibitors, pemetrexed, protein-bound paclitaxel (ABRAXANE®), doxorubicin, gemcitabine, bevacizumab, eribulin mesylate (HALAVEN®), neratinib, a PARP inhibitor, ARN810, an mTOR inhibitor, topotecan, gemcitabine, a VEGFR2 inhibitor, a folate receptor antagonist, demcizumab, fosbretabulin, or a PDL1 inhibitor).

In particular embodiments, the cancer is melanoma (e.g., metastatic melanoma) that is resistant to, or has failed to respond to prior treatment with, vemurafenib, dacarbazine, interferon therapy, a CTLA- 4 inhibitor, a BRAF inhibitor, a MEK inhibitor, a PD1 inhibitor, a PDL-1 inhibitor, and/or a CAR-T therapy.

In some embodiments, the cancer is glioblastoma that is resistant to, or has failed to respond to prior treatment with, temozolimide, radiotherapy, bevacizumab, irinotecan, a VEGFR2 inhibitor, a CAR-T therapy, and/or an mTOR inhibitor. In some embodiments, the cancer is non-small cell lung cancer such as metastatic non-small cell lung cancer (e.g., EGFR-wild type non-small cell lung cancer and/or squamous non-small cell lung cancer) that is resistant to, or has failed to respond to prior treatment with, an EGFR inhibitor, platinum agents (e.g., carboplatin), bevacizumab, an ALK inhibitor, a MET inhibitor, a taxane (e.g., paclitaxel and/or doceltaxel), gemcitabine, pemetrexed, radiotherapy, a PD1 inhibitor, a PDL1 ihibitor, and/or a CAR-T therapy. In some embodiments, the cancer is a breast cancer (e.g., triple negative breast cancer) that is resistant to, or has failed to respond to prior treatment with, herceptin, pertuzumab, tamoxifen, capecitabine, docetaxel, carboplatin, paclitaxel, protein-bound paclitazel (ABRAXANE®), doxorubicin, gemcitabine, bevacizumab, eribuline mesylate (HALAVEN®), neratinib, a PARP inhibitor, a PD1 inhibitor, a PDL1 inhibitor, a CAR-T therapy, ARN810, and/or an mTOR inhibitor.

In some embodiments, the cancer is ovarian cancer (e.g., metastatic ovarian cancer) that is resistant to, or has failed to respond to prior treatment with, a PARP inhibitor, bevacizumab, platinum agents such as carboplatin, paclitaxel, docetaxel, topotecan, gemcitabine, a VEGR2 inhibitor, a folate receptor antagonist, a PD1 inhibitor, a PDL1 inhibitor, a CAR-T therapy, demcizumab, and/or fosbretabulin. Chemical Terms

It is to be understood that the terminology employed herein is for the purpose of describing particular embodiments and is not intended to be limiting.

Compounds described herein also include isotopically-labeled compounds wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds described herein include and are not limited to ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ³⁶CI, ¹⁸F, ¹²³|, ¹²⁵l, ¹³N, ¹⁵N, ¹⁵0, ¹⁷0, ¹⁸0, ³²P, and ³⁵S. In certain embodiments, isotopically-labeled compounds are useful in drug and/or substrate tissue distribution studies. In another embodiment, substitution with heavier isotopes such as deuterium affords greater metabolic stability (for example, increased in vivo half- life or reduced dosage requirements). In yet another embodiment, substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵0 and ¹³N, is useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. Isotopically-labeled compounds are prepared by any suitable method or by processes using an appropriate isotopically-labeled reagent in place of the non-labeled reagent otherwise employed.

The term“acyl,” as used herein, represents a FI or an alkyl group, as defined herein, that is attached to a parent molecular group through a carbonyl group, as defined herein, and is exemplified by formyl (i.e., a carboxyaldehyde group), acetyl, trifluoroacetyl, propionyl, and butanoyl. Exemplary unsubstituted acyl groups include from 1 to 6, from 1 to 1 1 , or from 1 to 21 carbons.

The term“alkyl,” as used herein, refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon radical of 1 to 20 carbon atoms (e.g., 1 to 16 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms). An alkylene is a divalent alkyl group.

The term“alkenyl,” as used herein, alone or in combination with other groups, refers to a straight- chain or branched hydrocarbon residue having a carbon-carbon double bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms).

The term“alkynyl,” as used herein, alone or in combination with other groups, refers to a straight- chain or branched hydrocarbon residue having a carbon-carbon triple bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms).

The term“amino,” as used herein, represents -N(R^N1)2, wherein each R^N1 is, independently, FI, OH, NO2, N(R^N2)2, S0₂0R^N2, S02R^N2, SOR^N2, an /V-protecting group, alkyl, alkoxy, aryl, arylalkyl, cycloalkyl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), wherein each of these recited R^N1 groups can be optionally substituted; or two R^N1 combine to form an alkylene or heteroalkylene, and wherein each R^N2 is, independently, H, alkyl, or aryl. The amino groups of the invention can be an unsubstituted amino (i.e., - NH2) or a substituted amino (i.e., -N(R^N1 )2).

The term“aryl,” as used herein, refers to an aromatic mono- or polycarbocyclic radical of 6 to 12 carbon atoms having at least one aromatic ring. Examples of such groups include, but are not limited to, phenyl, naphthyl, 1 ,2,3,4-tetrahydronaphthyl, 1 ,2-dihydronaphthyl, indanyl, and 1 H-indenyl.

The term“arylalkyl,” as used herein, represents an alkyl group substituted with an aryl group. Exemplary unsubstituted arylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C1-6 alkyl C6-10 aryl, C1-10 alkyl C6-10 aryl, or C1-20 alkyl Ce-io aryl), such as, benzyl and phenethyl. In some embodiments, the akyl and the aryl each can be further substituted with 1 , 2, 3, or 4 substituent groups as defined herein for the respective groups.

The term“azido,” as used herein, represents a -N3 group.

The term“cyano,” as used herein, represents a -CN group.

The terms“carbocyclyl," as used herein, refer to a non-aromatic C3-12 monocyclic, bicyclic, or tricyclic structure in which the rings are formed by carbon atoms. Carbocyclyl structures include cycloalkyl groups and unsaturated carbocyclyl radicals.

The term“cycloalkyl,” as used herein, refers to a saturated, non-aromatic, monovalent mono- or polycarbocyclic radical of three to ten, preferably three to six carbon atoms. This term is further exemplified by radicals such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and adamantyl.

The term“halogen,” as used herein, means a fluorine (fluoro), chlorine (chloro), bromine (bromo), or iodine (iodo) radical.

The term“heteroalkyl,” as used herein, refers to an alkyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkyl group can be further substituted with 1 , 2, 3, or 4 substituent groups as described herein for alkyl groups. Examples of heteroalkyl groups are an“alkoxy” which, as used herein, refers alkyl-O- (e.g., methoxy and ethoxy). A heteroalkylene is a divalent heteroalkyl group.

The term“heteroalkenyl,” as used herein, refers to an alkenyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkenyl group can be further substituted with 1 , 2, 3, or 4 substituent groups as described herein for alkenyl groups. Examples of heteroalkenyl groups are an“alkenoxy” which, as used herein, refers alkenyl-O-. A heteroalkenylene is a divalent heteroalkenyl group.

The term“heteroalkynyl,” as used herein, refers to an alkynyl group, as defined herein, in which one or more of the constituent carbon atoms have been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkynyl group can be further substituted with 1 , 2, 3, or 4 substituent groups as described herein for alkynyl groups. Examples of heteroalkynyl groups are an“alkynoxy” which, as used herein, refers alkynyl-O-. A heteroalkynylene is a divalent heteroalkynyl group.

The term“heteroaryl,” as used herein, refers to an aromatic mono- or polycyclic radical of 5 to 12 atoms having at least one aromatic ring containing one, two, or three ring heteroatoms selected from N,

O, and S, with the remaining ring atoms being C. One or two ring carbon atoms of the heteroaryl group may be replaced with a carbonyl group. Examples of heteroaryl groups are pyridyl, pyrazoyl, benzooxazolyl, benzoimidazolyl, benzothiazolyl, imidazolyl, oxaxolyl, and thiazolyl.

The term“heteroarylalkyl,” as used herein, represents an alkyl group substituted with a heteroaryl group. Exemplary unsubstituted heteroarylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as C1-6 alkyl C2-9 heteroaryl, C1-10 alkyl C2-9 heteroaryl, or C1-20 alkyl C2-9 heteroaryl). In some embodiments, the akyl and the heteroaryl each can be further substituted with 1 , 2, 3, or 4 substituent groups as defined herein for the respective groups.

The term“heterocycly!,” as used herein, denotes a mono- or polycyclic radical having 3 to 12 atoms having at least one ring containing one, two, three, or four ring heteroatoms selected from N, O or S, wherein no ring is aromatic. Examples of heterocyclyl groups include, but are not limited to, morpholinyl, thiomorpholinyl, furyl, piperazinyl, piperidinyl, pyranyl, pyrrol id inyl, tetrahydropyranyl, tetrahydrofuranyl, and 1 ,3-dioxanyl.

The term“heterocyclylalkyl,” as used herein, represents an alkyl group substituted with a heterocyclyl group. Exemplary unsubstituted heterocyclylalkyl groups are from 7 to 30 carbons (e.g., from 7 to 16 or from 7 to 20 carbons, such as Ci-6 alkyl C2-9 heterocyclyl, C1-10 alkyl C2-9 heterocyclyl, or C1 -20 alkyl C2-9 heterocyclyl). In some embodiments, the akyl and the heterocyclyl each can be further substituted with 1 , 2, 3, or 4 substituent groups as defined herein for the respective groups.

The term“hydroxyl,” as used herein, represents an -OH group.

The term“/V-protecting group,” as used herein, represents those groups intended to protect an amino group against undesirable reactions during synthetic procedures. Commonly used /V-protecting groups are disclosed in Greene,“Protective Groups in Organic Synthesis,” 3^rd Edition (John Wiley &

Sons, New York, 1999). /V-protecting groups include acyl, aryloyl, or carbamyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, a-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4

nitrobenzoyl, and chiral auxiliaries such as protected or unprotected D, L or D, L-amino acids such as alanine, leucine, and phenylalanine; sulfonyl-containing groups such as benzenesulfonyl, and p- toluenesulfonyl; carbamate forming groups such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl,

p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4- dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl,

3.4.5-trimethoxybenzyloxycarbonyl, 1-(p-biphenylyl)-1 -methylethoxycarbonyl, a,a-dimethyl-

3.5-dimethoxybenzyloxycarbonyl, benzhydryloxy carbonyl, t-butyloxycarbonyl,

diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2,-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxy carbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, and phenylthiocarbonyl, arylalkyl groups such as benzyl, triphenylmethyl, and benzyloxymethyl, and silyl groups, such as trimethylsilyl. Preferred /V-protecting groups are alloc, formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, alanyl, phenylsulfonyl, benzyl, t-butyloxycarbonyl (Boc), and benzyloxycarbonyl (Cbz).

The term“nitro,” as used herein, represents an -NO2 group.

The term“thiol,” as used herein, represents an -SH group.

The alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl (e.g., cycloalkyl), aryl, heteroaryl, and heterocyclyl groups may be substituted or unsubstituted. When substituted, there will generally be 1 to 4 substituents present, unless otherwise specified. Substituents include, for example: aryl (e.g., substituted and unsubstituted phenyl), carbocyclyl (e.g., substituted and unsubstituted cycloalkyl), halogen (e.g., fluoro), hydroxyl, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy, or thioalkoxy), heteroaryl, heterocyclyl, amino (e.g., NH2 or mono- or dialkyl amino), azido, cyano, nitro, or thiol. Aryl, carbocyclyl (e.g., cycloalkyl), heteroaryl, and heterocyclyl groups may also be substituted with alkyl (unsubstituted and substituted such as arylalkyl (e.g., substituted and unsubstituted benzyl)).

Compounds of the invention can have one or more asymmetric carbon atoms and can exist in the form of optically pure enantiomers, mixtures of enantiomers such as, for example, racemates, optically pure diastereoisomers, mixtures of diastereoisomers, diastereoisomeric racemates or mixtures of diastereoisomeric racemates. The optically active forms can be obtained for example by resolution of the racemates, by asymmetric synthesis or asymmetric chromatography (chromatography with a chiral adsorbents or eluant). That is, certain of the disclosed compounds may exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Enantiomers are pairs of stereoisomers whose mirror images are not superimposable, most commonly because they contain an asymmetrically substituted carbon atom that acts as a chiral center. "Enantiomer" means one of a pair of molecules that are mirror images of each other and are not superimposable. Diastereomers are stereoisomers that are not related as mirror images, most commonly because they contain two or more asymmetrically substituted carbon atoms and represent the configuration of substituents around one or more chiral carbon atoms. Enantiomers of a compound can be prepared, for example, by separating an enantiomer from a racemate using one or more well-known techniques and methods, such as, for example, chiral chromatography and separation methods based thereon. The appropriate technique and/or method for separating an enantiomer of a compound described herein from a racemic mixture can be readily determined by those of skill in the art. "Racemate" or "racemic mixture" means a compound containing two enantiomers, wherein such mixtures exhibit no optical activity; i.e., they do not rotate the plane of polarized light.“Geometric isomer" means isomers that differ in the orientation of substituent atoms in relationship to a carbon-carbon double bond, to a cycloalkyl ring, or to a bridged bicyclic system. Atoms (other than H) on each side of a carbon- carbon double bond may be in an E (substituents are on opposite sides of the carbon- carbon double bond) or Z (substituents are oriented on the same side) configuration. "R," "S," "S*," "R*," "E," "Z," "cis," and "trans," indicate configurations relative to the core molecule. Certain of the disclosed compounds may exist in atropisomeric forms. Atropisomers are stereoisomers resulting from hindered rotation about single bonds where the steric strain barrier to rotation is high enough to allow for the isolation of the conformers. The compounds of the invention may be prepared as individual isomers by either isomer-specific synthesis or resolved from an isomeric mixture. Conventional resolution techniques include forming the salt of a free base of each isomer of an isomeric pair using an optically active acid (followed by fractional crystallization and regeneration of the free base), forming the salt of the acid form of each isomer of an isomeric pair using an optically active amine (followed by fractional crystallization and regeneration of the free acid), forming an ester or amide of each of the isomers of an isomeric pair using an optically pure acid, amine or alcohol (followed by chromatographic separation and removal of the chiral auxiliary), or resolving an isomeric mixture of either a starting material or a final product using various well known chromatographic methods. When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99% or 99.9%) by weight relative to the other

stereoisomers. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by weight optically pure. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by weight pure. Percent optical purity is the ratio of the weight of the enantiomer or over the weight of the enantiomer plus the weight of its optical isomer. Diastereomeric purity by weight is the ratio of the weight of one diastereomer or over the weight of all the diastereomers. When the stereochemistry of a disclosed compound is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by mole fraction pure relative to the other stereoisomers. When a single enantiomer is named or depicted by structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by mole fraction pure. When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is at least 60%,

70%, 80%, 90%, 99% or 99.9% by mole fraction pure. Percent purity by mole fraction is the ratio of the moles of the enantiomer or over the moles of the enantiomer plus the moles of its optical isomer.

Similarly, percent purity by moles fraction is the ratio of the moles of the diastereomer or over the moles of the diastereomer plus the moles of its isomer. When a disclosed compound is named or depicted by structure without indicating the stereochemistry, and the compound has at least one chiral center, it is to be understood that the name or structure encompasses either enantiomer of the compound free from the corresponding optical isomer, a racemic mixture of the compound or mixtures enriched in one enantiomer relative to its corresponding optical isomer. When a disclosed compound is named or depicted by structure without indicating the stereochemistry and has two or more chiral centers, it is to be understood that the name or structure encompasses a diastereomer free of other diastereomers, a number of diastereomers free from other diastereomeric pairs, mixtures of diastereomers, mixtures of diastereomeric pairs, mixtures of diastereomers in which one diastereomer is enriched relative to the other

diastereomer(s) or mixtures of diastereomers in which one or more diastereomer is enriched relative to the other diastereomers. The invention embraces all of these forms.

Definitions

In this application, unless otherwise clear from context, (i) the term“a” may be understood to mean“at least one”; (ii) the term“or” may be understood to mean“and/or”; (iii) the terms“comprising” and “including” may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps; and (iv) the terms“about” and “approximately" may be understood to permit standard variation as would be understood by those of ordinary skill in the art; and (v) where ranges are provided, endpoints are included.

As used herein, the term“adenosine AZA receptor” refers to the protein encoded by the

ADORA2A gene in mammals, e.g., humans. For example, the adenosine A_åA receptor refers to a peptide having the sequence of SEQ ID NO: 1 and/or a polypeptide with at least 90% identity (e.g., at least 95% identity, at least 98% identity, at least 99% identity) to SEQ ID NO: 1.

SEQ ID NO: 1

MPIMGSSVYITVELAIAVLAILGNVLVCWAVWLNSNLQNVVWLNSNLQNVADIAVGVLAIPFAITISTGFCAA

CHGCLFIACFVLVLTQSSIFSLLAIAIDRYIAIRIPLRYNGLVTGTRAKGIIAICWVLSFAIGLTPMLGWNNCGQ

PKEGKNHSQGCGEGQVACLFEDVVPMNYMVYFNFFACVLVPLLLMLGVYLRIFLAARRQLKQMESQPLP

GERARSTLQKEVHAAKSLAIIVGLFALCWLPLHIINCFTFFCPDCSHAPLWLMYLAIVLSHTNSVVNPFIYAY

RIREFRQTFRKIIRSHVLRQQEPFKAAGTSARVLAAHGSDGEQVSLRLNGHPPGVWANGSAPHPERRPN

GYALGLVSGGSAQESQGNTGLPDVELLSHELKGVCPEPPGLDDPLAQDGAGVS

As used herein, the term“adenosine A2A receptor activity” is measured by any method known in the art. For example, human adenosine A2_A receptor activity may be measured by a radioligand competition assay. A exemplary radioligand competition assay protocol may include: 6 nM of [³FI]-labeled CGS 21680 ligand (K_<j = 27 nM) bound to HEK293 cells expressing the human A2_A receptor. The ability of each test compound to compete for the radioligand binding at eight concentration points may be measured by scintillation counting. Data may be analyzed by non-linear regression to generate IC50 values and the inhibition constants (Ki) were calculated using the Cheng Prusoff equation.

As used herein, the term“adenosine A2B receptor” refers to the protein encoded by the

ADORA2B gene in mammals, e.g., humans. For example, the adenosine AåB receptor refers to a peptide having the sequence of SEQ ID NO: 2 and/or a polypeptide with at least 90% identity (e.g., at least 95% identity, at least 98% identity, at least 99% identity) to SEQ ID NO: 2.

SEQ ID NO: 2

MLLETQDALYVALELVIAALSVAGNVLVCAAVGTANTLQTPTNYFLVSLAAADVAVGLFAIPFAITISLGFCT

DFYGCLFLACFVLVLTQSSIFSLLAVAVDRYLAICVPLRYKSLVTGTRARGVIAVLWVLAFGIGLTPFLGWN

SKDSATNNCTEPWDGTTNESCCLVKCLFENVVPMSYMVYFNFFGCVLPPLLIMLVIYIKIFLVACRQLQRT

ELMDHSRTTLQREIHAAKSLAMIVGIFALCWLPVHAVNCVTLFQPAQGKNKPKWAMNMAILLSHANSVVN

PIVYAYRNRDFRYTFHKIISRYLLCQADVKSGNGQAGVQPALGVGL

As used herein, the term“adenosine AIB receptor activity” is measured by any method known in the art. For example, human adenosine A2B receptor activity may be measured by a radioligand competition assay. A exemplary radioligand competition assay protocol may include: 6 nM of [³H]-labeled CGS 21680 ligand (K_d = 27 nM) bound to FIEK293 cells expressing the human A2B receptor. The ability of each test compound to compete for the radioligand binding at eight concentration points may be measured by scintillation counting. Data may be analyzed by non-linear regression to generate IC50 values and the inhibition constants (Ki) were calculated using the Cheng Prusoff equation.

As used herein, the term“administration” refers to the administration of a composition (e.g., a compound or a preparation that includes a compound as described herein) to a subject or system.

Administration to an animal subject (e.g., to a human) may be by any appropriate route. For example, in some embodiments, administration may be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal,

intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal and vitreal.

The term“cancer” refers to a condition caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, and lymphomas.

As used herein, a“combination therapy” or“administered in combination” means that two (or more) different agents or treatments are administered to a subject as part of a defined treatment regimen for a particular disease or condition. The treatment regimen defines the doses and periodicity of administration of each agent such that the effects of the separate agents on the subject overlap. In some embodiments, the delivery of the two or more agents is simultaneous or concurrent and the agents may be co-form u I ated. In other embodiments, the two or more agents are not co-formulated and are administered together or in a sequential manner as part of a prescribed regimen. In some embodiments, administration of two or more agents or treatments in combination is such that the reduction in a symptom, or other parameter related to the disorder is greater than what would be observed with one agent or treatment delivered alone or in the absence of the other. The effect of the two treatments can be partially additive, wholly additive, or greater than additive (e.g., synergistic). Sequential or substantially simultaneous administration of each therapeutic agent can be effected by any appropriate route including, but not limited to, oral routes, intravenous routes, intramuscular routes, and direct absorption through mucous membrane tissues. The therapeutic agents can be administered by the same route or by different routes. For example, a first therapeutic agent of the combination may be administered by intravenous injection while a second therapeutic agent of the combination may be administered orally.

A cancer“determined to be drug resistant,” as used herein, refers to a cancer that is drug resistant based on unresponsiveness or decreased responsiveness to a chemotherapeutic agent, or is predicted to be drug resistant based on a prognostic assay (e.g., a gene expression assay). By a“drug resistant” cancer is meant a cancer that does not respond, or exhibits a decreased response to, one or more chemotherapeutic agents (e.g., any agent described herein).

The term "CTLA-4 inhibitor,” as used herein, refers to a compound such as an antibody capable of inhibiting the activity of the protein that in humans is encoded by the CTLA4 gene. Known CTLA-4 inhibitors include ipilimumab.

By“determining the level of a protein” is meant the detection of a protein, or an mRNA encoding the protein, by methods known in the art either directly or indirectly. “Directly determining” means performing a process (e.g., performing an assay or test on a sample or“analyzing a sample” as that term is defined herein) to obtain the physical entity or value. “Indirectly determining” refers to receiving the physical entity or value from another party or source (e.g., a third party laboratory that directly acquired the physical entity or value). Methods to measure protein level generally include, but are not limited to, western blotting, immunoblotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), immunoprecipitation, immunofluorescence, surface plasmon resonance, chemiluminescence, fluorescent polarization, phosphorescence, immunohistochemical analysis, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, liquid chromatography (LC)-mass spectrometry, microcytometry, microscopy, fluorescence activated cell sorting (FACS), and flow cytometry, as well as assays based on a property of a protein including, but not limited to, enzymatic activity or interaction with other protein partners. Methods to measure mRNA levels are known in the art.

As used herein, the term“disorder related to the adenosine A2_A receptor,” refers to any disease or disorder that may derive a therapeutic benefit from modulation (e.g., inhibition) of the activity of the adenosine AåA receptor, e.g., cancer.

An“effective amount” of a compound may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the compound to elicit the desired response. A therapeutically effective amount encompasses an amount in which any toxic or detrimental effects of the compound are outweighed by the therapeutically beneficial effects. A therapeutically effective amount also encompasses an amount sufficient to confer benefit, e.g., clinical benefit.

As used herein, the term“failed to respond to a prior therapy” or“refractory to a prior therapy,” refers to a cancer that progressed despite treatment with the therapy.

As used herein, the term“inhibiting inflammation” refers to preventing inflammation or decreasing inflammation.

By“modulating an immune response,” is meant altering (e.g., stimulating or suppressing) an immune response. The level of immune response may be measured using any method known in the art. By“modulating the activity of a protein," is meant increasing or decreasing the level of an activity related to the protein, or a related downstream effect. The activity level a protein may be measured using any method known in the art.

By“modulating the activity of the adenosine receptor,” is meant increasing or decreasing the level of an activity related to the adenosine receptor (e.g., the adenosine AIA receptor, the adenosine AåB receptor, the adenosine Ai receptor, or the adenosine A3 receptor) , or a related downstream effect. A non-limiting example of modulation of adenosine receptor includes increasing the level of immune cells in a subject by inhibiting the adenosine A2A receptor.

By“modulating the activity of the dopamine receptor,” is meant increasing or decreasing the level of an activity related to the dopamine receptor (e.g., the dopamine Di receptor or the dopamine D2 receptor), or a related downstream effect. A non-limiting example of modulation of the dopamine receptor includes increasing the level of immune cells in a subject by inhibiting the dopamine D2 receptor.

By“level” is meant a level of a protein, or mRNA encoding the protein, as compared to a reference. The reference can be any useful reference, as defined herein. By a“decreased level” or an “increased level” of a protein is meant a decrease or increase in protein level, as compared to a reference (e.g., a decrease or an increase by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, or more; a decrease or an increase of more than about 10%, about 15%, about 20%, about 50%, about 75%, about 100%, or about 200%, as compared to a reference; a decrease or an increase by less than about 0.01 -fold, about 0.02-fold, about 0.1 -fold, about 0.3-fold, about 0.5-fold, about 0.8-fold, or less; or an increase by more than about 1 .2-fold, about 1 .4-fold, about 1 .5-fold, about 1 .8-fold, about 2.0-fold, about 3.0-fold, about 3.5-fold, about 4.5-fold, about 5.0-fold, about 10-fold, about 15-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 100-fold, about 1000-fold, or more). A level of a protein may be expressed in mass/volume (e.g., g/dL, mg/ml_, pg/mL, ng/mL) or percentage relative to total protein or mRNA in a sample.

As used herein,“metastatic tumor” refers to a tumor or cancer in which the cancer cells forming the tumor have a high potential to or have begun to, metastasize, or spread from one location to another location or locations within a subject, via the lymphatic system or via haematogenous spread, for example, creating secondary tumors within the subject. Such metastatic behavior may be indicative of malignant tumors. In some cases, metastatic behavior may be associated with an increase in cell migration and/or invasion behavior of the tumor cells.

Examples of cancers that can be defined as metastatic include but are not limited to non-small cell lung cancer, breast cancer, ovarian cancer, colorectal cancer, biliary tract cancer, bladder cancer, brain cancer including glioblastomas and medullablastomas, cervical cancer, choriocarcinoma, endometrial cancer, esophageal cancer, gastric cancer, hematological neoplasms, multiple myeloma, leukemia, intraepithelial neoplasms, livercancer, lymphomas, neuroblastomas, oral cancer, pancreatic cancer, prostate cancer, sarcoma, skin cancer including melanoma, basocellular cancer, squamous cell cancer, testicular cancer, stromal tumors, germ cel! tumors, thyroid cancer, and renal cancer.

As used herein,“migrating cancer” refers to a cancer in which the cancer cells forming the tumor migrate and subsequently grow as malignant implants at a site other than the site of the original tumor. The cancer cells migrate via seeding the surface of the peritoneal, pleural, pericardial, or subarachnoid spaces to spread into the body cavities; via invasion of the lymphatic system through invasion of lymphatic cells and transport to regional and distant lymph nodes and then to other parts of the body; via haematogenous spread through invasion of blood cells; or via invasion of the surrounding tissue.

Migrating cancers include metastatic tumors and cell migration cancers, such as ovarian cancer, mesothelioma, and primary lung cancer, each of which is characterized by cellular migration.

“Non-metastatic cell migration cancer” as used herein refers to cancers that do not migrate via the lymphatic system or via haematogenous spread.

The term“PD-1 inhibitor,” as used herein, refers to a compound such as an antibody capable of inhibiting the activity of the protein that in humans is encoded by the PDCD1 gene. Known PD-1 inhibitors include nivolumab, pembrolizumab, pidilizumab, and BMS 936559.

The term“PD-L1 inhibitor,” as used herein, refers to a compound such as an antibody capable of inhibiting the activity of the protein that in humans is encoded by the CD274 gene. Known PD-L1 inhibitors include atezolizumab and durvalumab.

The term“pharmaceutical composition,” as used herein, represents a composition containing a compound described herein formulated with a pharmaceutically acceptable excipient, and manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gelcap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or in any other pharmaceutically acceptable formulation.

A“pharmaceutically acceptable excipient,” as used herein, refers any ingredient other than the compounds described herein (for example, a vehicle capable of suspending or dissolving the active compound) and having the properties of being substantially nontoxic and non-inflammatory in a patient. Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspensing or dispersing agents, sweeteners, and waters of hydration. Exemplary excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

As used herein, the term“pharmaceutically acceptable salt” means any pharmaceutically acceptable salt of the compound of formula (I). For example pharmaceutically acceptable salts of any of the compounds described herein include those that are within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al.,

J. Pharmaceutical Sciences 66:1 -19, 1977 and in Pharmaceutical Salts: Properties , Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting a free base group with a suitable organic acid.

The compounds of the invention may have ionizable groups so as to be capable of preparation as pharmaceutically acceptable salts. These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds of the invention be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases and methods for preparation of the appropriate salts are well-known in the art. Salts may be prepared from pharmaceutically acceptable non-toxic acids and bases including inorganic and organic acids and bases.

Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, 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, toluenesulfonate, undecanoate, and valerate salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine.

By a“reference” is meant any useful reference used to compare protein or mRNA levels. The reference can be any sample, standard, standard curve, or level that is used for comparison purposes. The reference can be a normal reference sample or a reference standard or level. A“reference sample” can be, for example, a control, e.g., a predetermined negative control value such as a“normal control" or a prior sample taken from the same subject; a sample from a normal healthy subject, such as a normal cell or normal tissue; a sample (e.g., a cell or tissue) from a subject not having a disease; a sample from a subject that is diagnosed with a disease, but not yet treated with a compound of the invention; a sample from a subject that has been treated by a compound of the invention; or a sample of a purified protein (e.g., any described herein) at a known normal concentration. By“reference standard or level” is meant a value or number derived from a reference sample. A“normal control value” is a pre-determ ined value indicative of non-disease state, e.g., a value expected in a healthy control subject. Typically, a normal control value is expressed as a range (“between X and Y”), a high threshold (“no higher than X”), or a low threshold (“no lower than X”). A subject having a measured value within the normal control value for a particular biomarker is typical!'/ referred to as“within normal limits” for that biomarker. A norma! reference standard or level can be a value or number derived from a normal subject not having a disease or disorder (e.g., cancer); a subject that has been treated with a compound of the invention. In preferred embodiments, the reference sample, standard, or level is matched to the sample subject sample by at least one of the following criteria: age, weight, sex, disease stage, and overall health. A standard curve of levels of a purified protein, e.g., any described herein, within the normal reference range can also be used as a reference.

As used herein, the term“subject” refers to any organism to which a composition in accordance with the invention may be administered, e.g., for experimental, diagnostic, prophylactic, and/or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans). A subject may seek or be in need of treatment, require treatment, be receiving treatment, be receiving treatment in the future, or be a human or animal who is under care by a trained professional for a particular disease or condition.

As used herein, the terms "treat," "treated," or "treating" mean both therapeutic treatment and prophylactic or preventative measures wherein the object is to prevent or slow down (lessen) an undesired physiological condition, disorder, or disease, or obtain beneficial or desired clinical results. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of a condition, disorder, or disease; stabilized (i.e., not worsening) state of condition, disorder, or disease; delay in onset or slowing of condition, disorder, or disease progression; amelioration of the condition, disorder, or disease state or remission (whether partial or total), whether detectable or undetectable; an amelioration of at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present disclosure; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

The details of one or more embodiments of the invention are set forth in the description below. Other features, objects, and advantages of the invention will be apparent from the description and from the claims.

Brief Description of the Drawings

FIG 1. is a graph of the quantitation of IL-12 cytokine secreted from dendritic cells that had been differentiated and activated in the presence or absence of 5’-(N-ethylcarboxamido)adenosine (NECA) and compounds of the invention.

FIG. 2 is a graph of the quantitation of IL-6 cytokine secreted from dendritic cells that had been differentiated and activated in the presence or absence of NECA and compounds of the invention. Detailed Description of the Invention

Recent immunotherapy approaches based on inhibiting immune checkpoint receptors offer a promising avenue to achieve robust, durable therapeutic responses in a number of solid tumors.

Effective immunotherapy, whether by checkpoint blockade or adoptive cell therapy, is limited in most patients by a key barrier: the immunosuppressive tumor microenvironment. A considerable proportion of patients remain unresponsive and demonstrate an innate resistance to anti-programmed cell death protein 1 (PD1 ), anti-PD1 ligand 1 (PDL1 ) and anti-cytotoxic T lymphocyte antigen 4 (CTLA4) therapies in advanced cancer. Unresponsive patients are often characterized by an immunosuppressed state of non- T-cell-inflamed tumors, which is contrary to T-cell-inflamed tumors where tumor-infiltrating lymphocytes can recognize tumor antigens and promote T-cell mediated killing and tumor regression.

The suppression of tumor-specific T cells is orchestrated by both intrinsic and extrinsic factors including the production of immunosuppressive oncometabolites (e.g., tryptophan and adenosine) and by the activity of a variety of stromal myeloid and lymphoid cells. The myeloid cells comprise various cell types, including neutrophils, macrophages, myeloid-derived suppressor cells (MDSCs) and dendritic cells (DCs) that regulate inflammation and immune state to maintain tissue homeostasis.

Adenosine Receptors

The action of adenosine is mediated by a family of G-protein coupled receptors (GPCR). Based on biochemical and pharmacological criteria, four subtypes of adenosine receptors have been described: A2A, A2B, Ai, and A3. The Ai and A3 receptors inhibit adenylyl cyclase, while the A2A and A2A receptors stimulate adenylyl cyclase.

The adenosine A2_A receptor has dynamic expression pattern on a subset of immune cells and is an important checkpoint in cancer therapy. Most of the immunomodulatory effects of adenosine signaling are thought to be mediated through the high-affinity adenosine A2_A receptor, which is expressed primarily on T-cells, and through the low-affinity adenosine A2B receptor, which is expressed primarily on myeloid cells.

The regulation of adenosine metabolism and signaling contributes to the resolution of tissue inflammation and modulation of adaptive T cell immunity in a healthy individual. With cancer, however, adenosine production is induced in response to hypoxia and its presence and downstream functions create an immunosuppressive state that is protective of a tumor rather than the patient. In order to achieve an effective anti-tumor immune response, blocking the adenosine signaling-mediated immune checkpoint is important.

The concentration of adenosine, typically sub-micromolar in the extracellular space, can reach levels as high as 100 pM within the tumor microenvironment. The tumor microenvironment is thereby primed for immunosuppression by this metabolite exerting its functions through binding to A2_A receptors and A2B receptors expressed on primarily lymphoid and myeloid cells, respectively. Adenosine binds to A2_A receptors with a high affinity, and low concentrations of adenosine are able to trigger a cAMP- dependent signaling cascade into T cells and NK cells that suppresses their immunosurveillance capabilities. Small molecule antagonists of A2_A receptor rescue lymphocytes from the

immunosuppressive effects of exogenous adenosine in T cell activation and cytotoxicity in vitro assays. A2_A receptor antagonists also act in synergy with checkpoint inhibitors to block tumor progression and metastases in animal models of cancer, demonstrating the immunosuppressive impact of endogenous adenosine in vivo.

Compounds

The invention features compounds useful for modulating the activity of the adenosine A2_A receptor, e.g., for the treatment of cancer. Exemplary compounds described herein include compounds having a structure according to Formula I:

or pharmaceutically acceptable salts thereof.

In some embodiments, the compound has the structure of any one of compounds 1 -80 in Table 1.

Other embodiments, as well as exemplary methods for the synthesis or production of these compounds, are described herein.

Pharmaceutical Uses

The compounds described herein are useful in the methods of the invention and, while not bound by theory, are believed to exert their desirable effects through modulation of the levels of adenosine, for example, through their ability to modulate the level, status, and/or activity of an adenosine receptor (e.g., the adenosine A2A receptor and/or the adenosine A2B receptor) in mammals, e.g., humans.

An aspect of the present invention relates to methods of modulating an immune response in a subject in need thereof. In some embodiments, the compound is administered in an amount and for a time effective to stimulate an immune response in a subject in need thereof.

A further aspect of the present invention relates to inhibiting inflammation in a subject in need thereof. In some embodiments, the compound is administered in an amount and for a time effective to prevent inflammation in a subject in need thereof.

Another aspect of the present invention relates to methods of treating disorders related to the adenosine receptors (e.g., the adenosine A2_A receptors) such as cancer, in a subject in need thereof. In some embodiments, the compound is administered in an amount and for a time effective to result in one of (or more, e.g., 2 or more, 3 or more, 4 or more of): (a) reduced tumor size, (b) reduced rate of tumor growth, (c) increased tumor cell death (d) reduced tumor progression, (e) reduced number of metastases, (f) reduced rate of metastasis, (g) decreased tumor recurrence (h) increased survival of subject, or (i) increased progression free survival of subject.

Treating cancer can result in a reduction in size or volume of a tumor. For example, after treatment, tumor size is reduced by 5% or greater (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%,

90% or greater) relative to its size prior to treatment. Size of a tumor may be measured by any reproducible means of measurement. For example, the size of a tumor may be measured as a diameter of the tumor. Treating cancer may further result in a decrease in number of tumors. For example, after treatment, tumor number is reduced by 5% or greater (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) relative to number prior to treatment. Number of tumors may be measured by any reproducible means of measurement, e.g., the number of tumors may be measured by counting tumors visible to the naked eye or at a specified magnification (e.g., 2x, 3x, 4x, 5x, 10x, or 50x).

Treating cancer can result in a decrease in number of metastatic nodules in other tissues or organs distant from the primary tumor site. For example, after treatment, the number of metastatic nodules is reduced by 5% or greater (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or greater) relative to number prior to treatment. The number of metastatic nodules may be measured by any reproducible means of measurement. For example, the number of metastatic nodules may be measured by counting metastatic nodules visible to the naked eye or at a specified magnification (e.g., 2x, 10x, or 50x).

Treating cancer can result in an increase in average survival time of a population of subjects treated according to the present invention in comparison to a population of untreated subjects. For example, the average survival time is increased by more than 30 days (more than 60 days, 90 days, or 120 days). An increase in average survival time of a population may be measured by any reproducible means. An increase in average survival time of a population may be measured, for example, by calculating for a population the average length of survival following initiation of treatment with the compound of the invention. An increase in average survival time of a population may also be measured, for example, by calculating for a population the average length of survival following completion of a first round of treatment with a pharmaceutically acceptable salt of the invention.

Treating cancer can also result in a decrease in the mortality rate of a population of treated subjects in comparison to an untreated population. For example, the mortality rate is decreased by more than 2% (e.g., more than 5%, 10%, or 25%). A decrease in the mortality rate of a population of treated subjects may be measured by any reproducible means, for example, by calculating for a population the average number of disease-related deaths per unit time following initiation of treatment with a pharmaceutically acceptable salt of the invention. A decrease in the mortality rate of a population may also be measured, for example, by calculating for a population the average number of disease-related deaths per unit time following completion of a first round of treatment with a pharmaceutically acceptable salt of the invention.

Combination Formulations and Uses Thereof

The compounds of the invention can be combined with one or more therapeutic agents. In particular, the therapeutic agent can be one that treats or prophylactically treats any cancer described herein.

A compound of the invention can be used alone or in combination with an additional therapeutic agent, e.g., other agents that treat cancer or symptoms associated therewith, or in combination with other types of treatment to treat cancer. In combination treatments, the dosages of one or more of the therapeutic compounds may be reduced from standard dosages when administered alone. For example, doses may be determined empirically from drug combinations and permutations or may be deduced by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6, 2005). In this case, dosages of the compounds when combined should provide a therapeutic effect.

In some embodiments, the second agent may be a checkpoint inhibitor. In one embodiment, the inhibitor of checkpoint is an inhibitory antibody (e.g., a monospecific antibody such as a monoclonal antibody). The antibody may be, e.g., humanized or fully human. In other embodiments, the inhibitor of checkpoint is a fusion protein, e.g., an Fc-receptor fusion protein. In some embodiments, the inhibitor of checkpoint is an agent, such as an antibody, that interacts with a checkpoint protein. In other embodiments, the inhibitor of checkpoint is an agent, such as an antibody, that interacts with the ligand of a checkpoint protein. In one embodiment, the inhibitor of checkpoint is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of CTLA-4 (e.g., an anti-CTLA4 antibody such as ipilimumab/Yervoy or tremelimumab). In on embodiment, the inhibitor of checkpoint is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of PD-1 (e.g., nivolumab/Opdivo®; pembrolizumab/Keytruda®;

pidilizumab/CT-011 ). In one embodiment, the inhibitor of checkpoint is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of PDL1 (e.g., MPDL3280A/RG7446; MEDI4736; MSB0010718C; BMS 936559). In one embodiment, the inhibitor of checkpoint is an inhibitor (e.g., an inhibitory antibody or Fc fusion or small molecule inhibitor) of PDL2 (e.g., a PDL2/lg fusion protein such as AMP 224). In one embodiment, the inhibitor of checkpoint is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of B7-H3 (e.g., MGA271 ), B7-H4, BTLA, HVEM, TIM3, GAL9, LAG 3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK 1 , CHK2, A2_A receptor, A2B receptor, B-7 family ligands, or a combination thereof.

In any of the combination embodiments described herein, the first and second therapeutic agent are administered simultaneously or sequentially, in either order. The first therapeutic agent may be administered immediately, up to 1 hour, up to 2 hours, up to 3 hours, up to 4 hours, up to 5 hours, up to 6 hours, up to 7 hours, up to, 8 hours, up to 9 hours, up to 10 hours, up to 11 hours, up to 12 hours, up to 13 hours, 14 hours, up to hours 16, up to 17 hours, up 18 hours, up to 19 hours up to 20 hours, up to 21 hours, up to 22 hours, up to 23 hours up to 24 hours or up to 1-7, 1 -14, 1-21 or 1 -30 days before or after the second therapeutic agent.

Pharmaceutical Compositions

The compounds of the invention are preferably formulated into pharmaceutical compositions for administration to human subjects in a biologically compatible form suitable for administration in vivo. Accordingly, in another aspect, the present invention provides a pharmaceutical composition comprising a compound of the invention in admixture with a suitable diluent, carrier, or excipient.

The compounds of the invention may be used in the form of the free base, in the form of salts, solvates, and as prodrugs. All forms are within the scope of the invention. In accordance with the methods of the invention, the described compounds or salts, solvates, or prodrugs thereof may be administered to a patient in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. The compounds of the invention may be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pump, or transdermal administration and the pharmaceutical compositions formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, rectal, and topical modes of administration. Parenteral administration may be by continuous infusion over a selected period of time.

A compound of the invention may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard or soft shell gelatin capsules, or it may be compressed into tablets, or it may be incorporated directly with the food of the diet. For oral therapeutic administration, a compound of the invention may be incorporated with an excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, and wafers.

A compound of the invention may also be administered parenterally. Solutions of a compound of the invention can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington’s Pharmaceutical Sciences (2003, 20^th ed.) and in The United States Pharmacopeia: The National Formulary (USP 24 NF19), published in 1999.

The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be fluid to the extent that may be easily administered via syringe.

Compositions for nasal administration may conveniently be formulated as aerosols, drops, gels, and powders. Aerosol formulations typically include a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container, which can take the form of a cartridge or refill for use with an atomizing device. Alternatively, the sealed container may be a unitary dispensing device, such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve which is intended for disposal after use. Where the dosage form comprises an aerosol dispenser, it will contain a propellant, which can be a compressed gas, such as compressed air or an organic propellant, such as fluorochlorohydrocarbon. The aerosol dosage forms can also take the form of a pump-atomizer.

Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, where the active ingredient is formulated with a carrier, such as sugar, acacia, tragacanth, gelatin, and glycerine. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base, such as cocoa butter.

The compounds of the invention may be administered to an animal, e.g., a human, alone or in combination with pharmaceutically acceptable carriers, as noted herein, the proportion of which is determined by the solubility and chemical nature of the compound, chosen route of administration, and standard pharmaceutical practice.

Dosages

The dosage of the compounds of the invention, and/or compositions comprising a compound of the invention, can vary depending on many factors, such as the pharmacodynamic properties of the compound; the mode of administration; the age, health, and weight of the recipient; the nature and extent of the symptoms; the frequency of the treatment, and the type of concurrent treatment, if any; and the clearance rate of the compound in the animal to be treated. One of skill in the art can determine the appropriate dosage based on the above factors. The compounds of the invention may be administered initially in a suitable dosage that may be adjusted as required, depending on the clinical response. In general, satisfactory results may be obtained when the compounds of the invention are administered to a human at a daily dosage of, for example, between 0.05 mg and 3000 mg (measured as the solid form). Dose ranges include, for example, between 10-1000 mg (e.g., 50-800 mg). In some embodiments, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg of the compound is administered.

Alternatively, the dosage amount can be calculated using the body weight of the patient. For example, the dose of a compound, or pharmaceutical composition thereof, administered to a patient may range from 0.1 -50 mg/kg (e.g., 0.25-25 mg/kg). In exemplary, non-limiting embodiments, the dose may range from 0.5-5.0 mg/kg (e.g., 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or 5.0 mg/kg) or from 5.0-20 mg/kg (e.g., 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg/kg).

EXAMPLES

Synthesis of Compounds of the Invention

The compounds of this invention can be synthesized according to one or more of the examples shown below. The variables recited in the general schemes below are as defined for Formula I.

Example 1. Synthesis of Compound 6

N-( 2-chlorobenzyl)-6~methyl-4-oxo-3, 4~dihydrofuro[2, 3-d]pyrimidine-5~carboxamide

6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (100 mg, 0.515 mmol) and (2- chlorophenyl)methanamine (85 mg, 0.6 mmol) were dissolved in DMF (3 mL), then FIATU (294 mg, 0.773 mmol) was added followed by DIEA (269pL, 1.545 mmol). The reaction mixture was stirred at room temperature overnight. DMF was removed under reduced pressure, then to the residue was added EtOAc and H2O. Organics were extracted with EtOAc (3 x 50mL), washed with water (3 x 50mL), dried (Na₂S0₄) and concentrated in vacuo. The title compound was purified by flash column chromatography on silica gel (DCM/MeOH) to give 80 mg product (49% yield)

m/z (ES⁺) [M+H]⁺ = 318

¹H NMR (300 MHz, DMSO-d6) 13.176 (s, 1 H), 10.617 (t, 1 H), 8.238 (s, 1 H), 7.458 (m, 2H), 7.338 (m, 2H), 4.599 (d, 2H), 2.739 (s, 3H). Example 2. Synthesis of Compound 7

N-( 3-chlorobenzyl)-6-methyl-4-oxo-3, 4-dihydrofuro[2, 3-d]pyrimidine-5-carboxamide

6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (100 mg, 0.515 mmol) and (3- chlorophenyl)methanamine (85 mg, 0.6 mmol) were dissolved in DMF (3 ml_), then HATU (294 mg, 0.773 mmol) was added followed by DIEA (269pL, 1 .545 mmol). The reaction mixture was stirred at room temperature overnight. DMF was removed under reduced pressure, then to the residue was added EtOAc and H2O. Organics were extracted with EtOAc (3 x 50mL), washed with water (3 x 50mL), dried (Na2S04) and concentrated in vacuo. The title compound was purified by flash column chromatography on silica gel (DCM/MeOH) to give 45 mg product (28% yield)

m/z (ES⁺) [M+H]⁺ = 318

¹H NMR (300 MHz, DMSO-d6) 13.198 (s, 1 H), 10.625 (s, 1 H), 8.264 (s, 1 H), 7.40 (m, 2H), 7.330 (m, 2H), 4.546 (d, 2H), 2.731 (s, 3H).

Example 3. Synthesis of Compound 8

N-(4-chlorobenzyl)-6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxamide

6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (100 mg, 0.515 mmol) and (4- chlorophenyl)methanamine (85 mg, 0.6 mmol) were dissolved in DMF (3 ml_), then HATU (294 mg, 0.773 mmol) was added followed by DIEA (269pL, 1 .545 mmol). The reaction mixture was stirred at room temperature overnight. DMF was removed under reduced pressure, then to the residue was added EtOAc and H2O. Organics were extracted with EtOAc (3 x 50mL), washed with water (3 x 50mL), dried (Na2S04) and concentrated in vacuo. The title compound was purified by flash column chromatography on silica gel (DCM/MeOH) to give 15 mg product (9 % yield)

m/z (ES⁺) [M+H]⁺ = 318

¹ H NMR (300 MHz, DMSO-d6) 1 1 .031 (broad s, 1 H), 8.168 (s, 1 H), 7.370 (m, 4H), 4.475 (d, 2H), 2.707 (s, 3H). Example 4. Synthesis of Compound 13

N-( 3, 4-dichlorobenzyl)-6~methyl-4-oxo-3, 4-dihydrofuro[2, 3-d]pyrimidine-5-carboxamide

6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (100 mg, 0.515 mmol) and (3,4-dichlorophenyl)methanamine (106 mg, 0.6 mmol) were dissolved in DMF (3 mL), then HATU (294 mg, 0.773 mmol) was added followed by DIEA (269pL, 1.545 mmol). The reaction mixture was stirred at room temperature overnight. DMF was removed under reduced pressure, then to the residue was added EtOAc and FbO. Organics were extracted with EtOAc (3 x 50mL), washed with water (3 x 50ml_), dried (Na2S04) and concentrated in vacuo. The title compound was purified by flash column chromatography on silica gel (DCM/MeOFI) to give 25 mg product (14% yield)

m/z (ES⁺) [M+H]⁺ = 352

¹H NMR (300 MHz, DMSO-d6) 13.202 (s, 1 H), 10.688 (s, 1 H), 8.257 (s, 1 H), 7.608 (d, 1 H), 7.358 (d, 1 H), 4.544 (d, 2H), 2.714 (s, 3H).

Example 5. Synthesis of Compound 10

6-methyl-4-oxo-N-(4-(trifluoromethyl)benzyl)-3,4-dihydrofuro[2,3~d]pyrimidine-5-carboxamide

6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (100 mg, 0.515 mmol) and (4- (trifluoromethyl)phenyl)methanamine (105 mg, 0.6 mmol) were dissolved in DMF (3 mL), then HATU (294 mg, 0.773 mmol) was added followed by DIEA (269pL, 1.545 mmol). The reaction mixture was stirred at room temperature overnight. DMF was removed under reduced pressure, then to the residue was added EtOAc and H2O. Organics were extracted with EtOAc (3 x 50mL), washed with water (3 x 50mL), dried (Na2S04) and concentrated in vacuo. The title compound was purified by flash column chromatography on silica gel (DCM/MeOH) to give 60 mg product (33% yield)

m/z (ES⁺) [M+H]⁺ = 352

¹H NMR (300 MHz, DMSO-d6) 10.672 (s, 1 H), 8.263 (s, 1 H), 7.700 (d, 2H), 7.58 (d, 2H), 4.623 (d, 2H), 2.724 (s, 3H). Example 6. Synthesis of Compound 5

A solution of 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (203mg, 1 .05mmol), (3-(trifluoromethyl)phenyl)methanamine (189mg, 1 .08mmol), triethylamine (438ul, 3.21 mmol) and dichloromethane (6mL) was added a solution of 2-chloro-1 ,3-dimethylimidazolinium chloride (224mg, 1 33mmol) in dichloromethane (4mL) and stirred at room temperature overnight. The mixture was loaded directly on to silica (12g) and purified by flash column chromatography using a gradient of

dichloromethane-methanol as eluent. The product was repurified on silica (12g) using a gradient of dichloromethane-methanol as eluent. The product was triturated with dichloromethane (4mL) overnight then filtered to afford 6-methyl-4-oxo-N-(3-(trifluoromethyl)benzyl)-3,4-dihydrofuro[2,3-d]pyrimidine-5- carboxamide (197mg) as a white solid.

¹ H NMR (300 MHz, DMSO-d6) d 13.19 (s, 1 H), 10.62 (t, 1 H), 8.26 (s, 1 H), 7.63 (m, 4H), 4.62 (d, 2H), 2.73 (s, 3H). m/z (ES⁺) [M+H]⁺ = 352.

Example 7. Synthesis of Compound 30

A mixture of 4-bromo-2,3-dihydro-1 H-inden-1 -one (1 .078g, 5.13mmol), methanesulfonic acid (15mL) and dichloromethane (30mL) at 0 °C was treated slowly with sodium azide (500mg, 7.69mmol). The mixture was allowed to warm to room temperature and stirred overnight. The mixture was diluted with 1 N sodium hydroxide (50ml) and extracted with dichloromethane (3 x 50mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica (40g) using a gradient of dichloromethane-ethyl acetate to afford 5-bromo-3,4-dihydroisoquinolin-1 (2H)-one (800mg).

A solution of 5-bromo-3,4-dihydroisoquinolin-1 (2H)-one (800mg, 3.56mmol) in DMF (20mL) at 0 °C was treated with 60% sodium hydride (350mg, 8.75mmol) and stirred for 5 minutes. Methyl iodide (0.5mL, 8.03mmol) was added and the mixture was allowed to warm to room temperature and the stirring was continued for 1 hour. The mixture was diluted with ethyl acetate (70mL) and washed with water (3 x 60mL), dried with anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford 5~bromo-2-methyl-3,4-dihydroisoquinolin-1 (2H)-one (690mg) that was used without further purification.

A mixture of 5-bromo-2-methyl-3,4~dihydroisoquinolin-1 (2H)-one (690mg, 2.88mmol), zinc cyanide ( 1.01 g, 8.60mmol), palladium tetrakis(triphenylphosphine) (140mg, 0.09mmol) and DMF (7.5mL) was heated under microwave irradiation at 150 °C for 40 minutes. The mixture was diluted with brine (50ml) and extracted with dichloromethane (3 x 50mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude product product was purified by flash column chromatography on silica (24g) using a gradient of dichloromethane-ethyl acetate to afford 2-methyi-1-oxo-1 ,2,3,4-tetrahydroisoquinoline-5-carbonitrile (440mg).

A solution of 2-methyl-1 -oxo-1 ,2,3,4-tetrahydroisoquinoline-5-carbonitrile (440mg, 2.37mmol) in methanol (25mL) was treated with Raney nickel (ca. 1 mL, aqueous slurry), to which was applied a balloon of hydrogen gas. The reaction was stirred overnight then filtered through celite. The filtrate was concentrated under reduced pressure then purified by flash column chromatography on silica (12g) using a gradient of dichloromethane-methanol to afford 5-(aminomethyl)-2-methyl-3,4-dihydroisoquinolin--1 (2H)- one (183mg) as a pale yellow oil.

A solution of 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (187mg, 0.96mmol), afford 5-(aminomethyl)-2-methyl-3,4-dihydroisoquinolin-1(2H)-one (183mg, 0.96mmol), triethylamine (393pL, 2.88mmol) and dichloromethane (5ml) was added a solution of 2-chioro-1 ,3- dimethylimidazolinium chloride (211 mg, 1 25mmol) in dichloromethane (4mL) and stirred at room temperature overnight. The mixture was concentrated under reduced pressure and purified by reverse phase HPLC using a gradient of water-acetonitrile-0.1 % formic acid to afford 6-methyl-N-((2-methyl-1 - oxo-1 ,2,3,4-tetrahydroisoquinolin-5-yl)methyl)-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxamide (76mg) as a white solid.

¹H NMR (300 MHz, DMSO-d6) d 13.10 (s, 1 H), 10.44 (t, 1 H), 8.23 (s, 1 H), 7.24 (t, 1 H), 7.05 (m, 2H), 4.52 (d, 2H), 3.26 (s, 3H), 2.92 (m, 2H), 2.72 (s, 3H), 2.53 (m, 2H). m/z (ES⁺) [M+H]⁺ = 367 .

Example 8. Synthesis of Compound 9

Synthesis of N-cyclopentyl-4-oxo-3, 4-dihydrofuro[2, 3-d]pyrimidine-5-carboxamide ( 9)

To a solution of 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxy!ic acid (150 mg, 0.77 mmol) in DMF (4 mL) was added HATU (293 mg, 1.93 mmol). To this mixture was added

cyclopentanamine (75 pL mg, 0.77 mmol) followed by DIEA (300 pL, 2.31 mmol). The reaction was allowed to stir at room temperature overnight. The mixture was diluted with water and extracted with ethyl acetate. Organics were washed with water and brine. Organics were dried over sodium sulfate, filtered and concentrated to give the desired product. Material was purified using a silica gel column with a gradient of 0-100% Ethyl Acetate in Hexanes. Obtained 3.8 mg of pure N-cyclopentyl-4-oxo-3,4- dihydrofuro[2,3-d]pyrimidine-5-carboxamide m/z (ES⁺) [M+H]⁺ = 262.05.

HNMR (300 MHz, CDCI3) 10.21 (d, 1 H), 8.26 (s, 1 H), 4.20 (m, 1 H), 2.72 (S, 3H), 1 .87 (m, 2H), 1.69-1.50 (m, 6H).

Example 9. Synthesis of Compound 31

Synthesis of N-(3-((dimethylamino)methyl)benzyl)-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxamide

To a solution of 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (100 mg, 0.52 mmol) in DMF (4 ml.) was added HATU (259 mg, 0.68 mmol). To this mixture was added 1-(3- (aminomethyl)phenyl)-N,N-dimethylmethanamine (82 mg, 0.52 mmol) followed by DIEA (272 mI_, 1.56 mmol). The reaction was allowed to stir at room temperature overnight, resulting in the precipitation of a solid. The solid was collected and washed with DCM and MeOH, resulting in the isolation of 23 mg of pure N-(3-((dimethylamino)methyl)benzyl)-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxamide. m/z (ES⁺) [M+H]⁺ = 341.12.

HNMR (300 MHz, CDCI3) 10.58 (m, 1 H), 8.24 (s, 1 H), 8.16 (s, 1 H), 7.28 (m, 4H), 4.51 (m, 2H), 3.56 (s, 2H), 2.73 (s, 3H), 2.26 (s, 6H).

Example 10. Synthesis of Compound 27

Synthesis of N-(isoquinolin-5-ylmethyl)~4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxamide (27)

To a solution of 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (50 mg, 0.26 mmol) in DCM (4 ml_) was added isoquinolin-5-ylmethanamine (43 mg, 0.26 mmol) and TEA (72 pL,

0.52) mmol. To this mixture was added 2-chloro-1 ,3-dimethylimidazolinium chloride (77 mg, 0.34 mmol).

After 30 minutes the reaction showed poor conversion, so additional 2-chloro-1 ,3-dimethylimidazolinium chloride (77 mg, 0.34 mmol) was added. Additional 2-chloro-1 ,3-dimethy!imidazolinium chloride was added two more times and the reaction was allowed to stir for 2 hours at room temperature before good conversion was observed. The reaction mixture was concentrated, and purified by prep-HPLC using a 5- 80% Acetonitrile in water gradient, resulting in the isolation of 47 mg of pure N-(isoquinolin-5-ylmethyl)-4- oxo-3, 4-dihydrofuro[2,3-d]pyrimidine-5-carboxamide. m/z (ES⁺) [M+H]⁺ - 335.14.

HNMR (300 MHz, CDCI3) 13.16 (s, 1 H), 10.70 (s, 1 H), 9.33 (s, 1 H), 8.57 (d, 1 H), 8.25 (s, 1 H), 8.08 (m, 2H), 7.83 (m, 1 H), 7.70 (m, 1 H), 4.99 (m, 2H), 2.77 (s, 3H).

Example 11. Synthesis of Compound 14

Synthesis of 5-bromo-2-methyl-1 ,4-dihydroisoquinolin-3(2H)-one (i-11a)

To methanesulfonic acid (24.7 mL) in a flask at 0 °C degree was added phosphorus pentoxide (2.95 g) in batches. After addition, the mixture was stirred at room temperature overnight. To this mixture was added 2-(2-bromophenyl)-N-methylacetamide (5.0 g, 22.12 mmol) and paraformaldehyde (1.0 g).

The resulting mixture was heated at 90 °C for 4 hours, cooled to room temperature. Ice-water was added to the mixture to three times of the original volume. Resulting mixture was basified using 4 N NaOH aqueous solution to pH = 10. Solid was collected by filtration and purified by flash column

chromatography on silica gel eluting with 0-100% ethyl acetate in dichloromethane to afford 2.6 g of a yellow solid m/z (ES⁺) [M+H]⁺ = 240

Synthesis of 2-methyl-3-oxo- 1,2,3, 4-tetrahydroisoquinoline-5-carbonitrile (i-11b)

A mixture of 5-bromo-2-methyl-1 ,4-dihydroisoquinolin-3(2H)-one (i-11a) (600 mg, 2.50 mmol), Zn(CN)2 (880 mg, 7.5 mmol) and palladium tetrakis triphenylphosphine (120 mg) in DMF (6 mL) was heated in a microwave oven at 150 °C for 3 hours. After being cooled to room temperature, the mixture was diluted with dichloromethane (10 mL), filtered through celite and concentrated. Product was purified by column chromatography on silica gei eluting with 0-10% methanol in dichloromethane. Fractions containing the desired product were combined and concentrated. Product was further purified on silica gel eluting with 0-100% ethyl acetate in dichloromethane to provide the title compound (185 mg) m/z (ES⁺) [M+H]⁺ = 187. Synthesis of 5-(aminomethyl)-2-methyl-1,4-dihydroisoquinolin-3(2H)-one (i-11 c)

A mixture of 2-methyl-3-oxo-1 ,2,3,4-tetrahydroisoquinoline-5-carbonitrile (i-1 1 b) (95 mg, 0.51 mmol) and 10% Pd/C (100 mg) in methanol (5 ml_) was stirred under hydrogen balloon over weekend and filtered through celite. Filtrate was concentrated to provide 74 mg of the amine. It was used without further purification m/z (ES⁺) [M+H]⁺ = 191 .

Synthesis of 6-methyl-N-( ( 2-methyl-3-oxo- 1,2,3, 4-tetrahydroisoquinolin-5-yl)methyl)-4-oxo-3, 4- dihydrofuro[2,3-d]pyrimidine-5-carboxamide (compound 14)

To a solution of 5-(aminomethyl)-2-methyl-1 ,4-dihydroisoquinolin-3(2H)-one (i-1 1 c) (50 mg, 0.26 mmol), 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (51 mg, 0.26 mmol) and triethylamine (147 pL, 1 .0 mmol) in dichloromethane (2.5 mL) was added a solution of 2-chloro-1 ,3- dimethylimidazolinium chloride (49 mg, 0.26 mmol) in 0.55 ml. of dichloromethane. The resulting mixture was stirred at room temperature for 2 hours. Product was purified by column chromatography on silica gel eluting with 0-10% methanol in dichloromethane. Fractions containing the desired product were collected and concentrated. The residue was treated with methanol (1 mL). Product was collected by filtration to provide the title compound as a white solid (47 mg) m/z (ES⁺) [M+H]⁺ = 367.

¹H NMR (300 MHz, DMSO-c/₆) d 13.10 (1 , 1 H), 10.44 (s, 1 H), 8.26 (s, 1 H), 7.24-7.18 (m, 3H), 4.53-4.48 (m, 4H), 3.58 (s, 2H), 2.96 (s, 3H), 2.72 (s, 3H).

Example 12. Synthesis of Compound 11

N-( 2, 3-dichlorobenzyl)-6-methyl-4-oxo-3, 4-dihydrofuro[2, 3-d]pyrimidine-5-carboxamide

To a solution of 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (95 mg, 0.5 mmol) in DMF (3 mL), (2,3-dichlorophenyl)methanamine (88 mg, 0.5 mmol), EDC (1 15 mg, 0.6 mmol), HOBT(81 mg, 0.6 mmol) and TEA (125 mg, 1 .25 mmol) were added sequentially. The mixture was stirred at RT for 12 hours, diluted with ethyl acetate(10 mL), washed with water (10 mL), saturated aqueous sodium bicarbonate solution (10 mL) and brine (5 mL), dried over anhydrous NaåS04 and concentrated under vacuum. This crude product was then purified on silica gel column chromatography eluting with dichloromethane/methanol (95:5) to give the desired product as a solid (55 mg)

m/z (ES⁺) [M+H]⁺ = 352.2

¹H NMR (300 MHz, DMSO): d 10.67 (m, 1 H), 8.26 (s, 1 H), 7.59 (m, 1 H), 7.42 (m, 2H), 4.61 (d, 2H), 2.71 (s, 3H). Example 13. Synthesis of Compound 15

6-methyl-4-oxo-N-( 2-( trifluoromethyl)benzyl)-3, 4-dihydrofuro[2, 3-d]pyrimidine-5-carboxamide

The title compound was prepared following the same procedure for the preparation of

(Compound 13) except using (2-(trifluoromethyl)phenyl)methanamine as the starting material m/z (ES⁺) [M+H]⁺ = 352.1

¹H NMR (300 MHz, DMSO): d 10.63 (m, 1 H), 8.26 (s, 1 H), 7.97 (d, 1 H), 7.55 (m, 2H), 7.39 (m, 2H), 4.68 (d, 2H), 2.73 (s, 3H).

Example 14. Synthesis of Compound 2

tert-butyl 2-( 2-( dimethylamino)-2-oxoethyl)benzylcarbamate ( i- 14a)

A solution of 2-(2-(((tert-butoxycarbonyl)amino)methyl)phenyl)acetic acid (265 mg, 1 .0 mmol), HATU (380 mg, 1.0 mmol), DIEA (387 mg, 3.0 mmol) and dimethylamine hydrochloride ( 123 mg, 1 .5 mmol) in 10 mL DCM was stirred at room temperature for 3 hours. The resulting solution was concentrated under vacuum to afford crude product which was purified by flash column to give the product (263 mg, 90 %) as a yellow solid m/z (ES⁺) [M+H]⁺ = 293.

2-(2-(aminomethyl)phenyl)-N,N-dimethylacetamide (i-14b)

To a solution of i-14a (263 mg, 0.90 mmol) in dichloromethane (8 mL) was added CF3COOH (2 mL) at r.t and the reaction mixture was stirred at room temperature for 2 hours. The resulting solution was concentrated under vacuum to afford a crude product (225 mg, 130 %) as yellow oil. m/z (ES⁺) [M+H]⁺ = 193. N-(2-(2-(dimethylamino)-2-oxoethyl)benzyl)-6-methyl-4-oxo-3,4-dihydrofuro[2, 3-d]pyrimidine-5~ carboxamide (Compound 2)

A solution of 2-(2-(aminomethyl)phenyl)-N,N-dimethylacetamide (150 mg, 0.78 mmol), 6-methyl- 4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (152 mg, 0.78 mmol), HATU(380 mg, 1.0 mmol) and K2C03 (414 mg, 3.0 mmol) in 10 mL DMF was stirred at room temperature for 6 hours. The resulting solution was concentrated under vacuum to afford crude product which was purified by prep-HPLC to give the product (57 mg, 20 %) as a yellow solid m/z (ES⁺) [M+H]⁺ = 369.

¹H NMR (400 MHz, DMSO) d 13.14 (d, 1 H), 10.42 (t, 1 H), 8.24 (d, 1 H), 7.37 - 7.29 (m, 1 H), 7.25 - 7.18 (m, 2H), 7.12 - 7.09 (m, 1 H), 4.46 (d, 2H), 3.80 (s, 2H), 3.03 (s, 3H), 2.81 (s, 3H), 2.73 (s, 3H).

Example 15. Synthesis of Compound 32

Synthesis of 1 -methyl-1 H-indazole-4-carboxamide

A solution of 1 -methyl-1 H-indazole-4-carboxylic acid (148mg, 0.84mmol), triethylamine (285ul, 2.04mmol) and dioxane (6mL) at room temperature was treated with CDI (199mg, 1 23mmol) and heated to 75 °C for 1 hour. The reaction mixture was cooled to room temperature and treated with concentrated ammonium hydroxide (1.5mL) and stirred for 10 minutes. The mixture was concentrated under reduced pressure then purified by reverse phase chromatography (C18) using a gradient of water-acetonitrile as eluent to afford 1 -methyl-1 H-indazole-4-carboxamide (161 mg) as a yellow solid.

Synthesis of (1 -methyl- 1 H-indazol-4-yl)methanamine

Lithium aluminum hydride (76mg, 2.00mmol) was added to a solution of 1 -methyl-1 H-indazole-4- carboxamide (161 mg, 0.92mmol) in THF (6mL) and stirred for 5 minutes. The mixture was then heated at reflux for 1 hour before being cooled to 0 °C. Water (76pL), 15%aq. NaOH (76pL) and water (220pL) were added in succession and the mixture was diluted with DCM-methanol (9:1 ) and filtered through celite. The filtrate was concentrated under reduced pressure to afford (1 -methyl-1 H-indazol-4- yl)methanamine as a sticky brown solid (276mg) that was used without further purification.

Synthesis of 6-methyl-N-( ( 1 -methyl- 1 H-indazol-4-yl)methyl)-4-oxo-3, 4-dihydrofuro[2, 3-d]pyrimidine-5- carboxamide

A mixture of (1 -methyl-1 H-indazol-4-yl)methanamine (276mg, crude from previous step), 6- methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (75mg, 0.39mmol) and triethylamine (360pL, 2.58mmol) in DCM (6ml) was treated with 2-chloro-1 ,3-dimethylimidazolinium chloride (200mg,

1 18mmol) in DCM (4mL) and stirred for 3 hours. The mixture was diluted was purified directly by flash chromatography on silica gel using a gradient of DCM-methanol as eluent to afford 6-methyl-N-((1- methyl-1 H-indazol-4-yl)methyl)-4~oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxamide (43mg).

¹H NMR (300 MHz, DMSO-d6) d 13.15 (s, 1 H), 10.68 (t, 1 H), 8.24 (s, 1 H), 8.15 (s, 1 H), 7.55 (d, 1 H), 7.34 (m, 1 H), 7.09 (d, 1 H), 4.82 (d, 2H), 4.04 (s, 3H), 2.74 (s, 3H). m/z (ES⁺) [M+H]⁺ = 338 .

Example 16. Synthesis of Compound 44

Synthesis of 7-bromo-2-methylisoindolin-1-one

A solution of 7-bromoisoindolin-1 -one (885 mg, 4.21 mmol) in DMF (19 mL) at 0 °C was treated with 60% sodium hydride (167 mg, 4.18mmol) and stirred for 10 minutes. Methyl iodide (0.60mL, 9.460mmol) was added and stirring continued for 2 hours at 0 °C. The mixture was diluted with ethyl acetate (50ml_) and washed with water (3 x 50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure and purified by flash chromatography on silica gel using a gradient of hexanes-ethyl acetate as eluent to afford 7-bromo-2-methylisoindolin-1-one (593 mg) as a pale yellow solid.

Synthesis of 2-methyl~3~oxoisoindoline-4-carbonitrile

A mixture of 7-bromo-2-methylisoindolin-1-one (593 mg, 2.62 mmol), zinc cyanide (1.125 g, 9.58 mmol) and Pd(PPh3)4 (140 mg, 0.12 mmol) in DMF (7 ml) was heated to 140 °C for 3 hours under microwave irradiation. The mixture was diluted with brine and extracted with DCM (x3). The combined organic extracts were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure and purified by flash chromatography on silica gel using a gradient of DCM-ethyl acetate as eluent to afford 2-methyl-3-oxoisoindoline-4-carbonitrile (395 mg).

Synthesis of tert-butyl ( ( 2-methyl-3~oxoisoindolin-4-yl)methyl)carbamate

A solution of 2-methyi-3-oxoisoindoline-4-carbonitrile (395 mg, 2.30 mmol) in methanol (25mL) was treated with Raney nickel (1 mL aqueous slurry) then 4N HCI-dioxane (2 mL). A balloon of hydrogen was applied overnight with stirring. Upon completion of the reaction the flask was purged with nitrogen then the contents filtered through celite. The filtrate was concentrated under reduced pressure then treated with dioxane (20 mL) and 1.2N sodium hydroxide (5.0 mL, 4.17 mmol) then di-tert- butyldicarbonate (980 mg, 4.54 mmol). The mixture was stirred at room temperature for 1 hour then diluted with brine and extracted with DCM (x3). The combined organic extracts were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure and purified by flash chromatography on silica gel using a gradient of DCM-ethyl acetate as eluent to afford tert-butyl ((2-methyl-3- oxoisoindolin-4-yl)methyl)carbamate (191 mg).

Synthesis of 7-(aminomethyl)-2-methylisoindolin-1-one hydrochloride

A solution of tert-butyl ((2-methyl-3-oxoisoindolin-4-yl)methyl)carbamate (191 mg) in DCM (3 ml_) was treated with 4N HCI-dioxane (2 ml_, 8.0 mmol) and stirred at room temperature for 3 days then concentrated under reduced pressure to afford 7-(aminomethyl)-2-methylisoindolin-1 -one hydrochloride (145 mg).

Synthesis of 6-methyl-N-( ( 2-methyl-3-oxoisoindolin-4-yl)methyl)-4-oxo-3, 4-dihydrofuro[2, 3-d]pyrimidine-5- carboxamide

A mixture of 7-(aminomethyl)-2-methylisoindolin-1 -one hydrochloride (83mg, 0.39mmol), 6- methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (76mg, 0.39mmol) and triethylamine (360pL, 2.58mmol) in DCM (6ml_) was treated with 2-chloro-1 ,3-dimethylimidazolinium chloride (92mg, 0.54mmol) in DCM (3mL) and stirred overnight. The mixture was diluted was purified directly by flash chromatography on silica gel using a gradient of DCM-methanol as eluent to afford 6-methyl-N-((2- methyl-3-oxoisoindolin-4-yl)methyl)-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxamide (24mg).

¹H NMR (300 MHz, DMSO-d6) d 8.23 (s, 1 H), 7.50-7.37 (m, 3H), 5.03 (d, 2H), 4.45 (s, 2H), 3.07 (s, 3H), 2.72 (s, 3H). m/z (ES⁺) [M+H]⁺ = 353.

Example 17. Synthesis of Compound 35

Synthesis of tert-butyl (quinazolin-8-ylmethyl)carbamate (i-17a)

A mixture of quinazoline-8-carbaldehyde (0.20 g, 1.2 mmol), NaB(CN)H3 (0.22 g, 3.72 mmol) in methanol (8 ml_) containing 7 N ammonia was heated in a microwave oven at 110 °C for 20 minutes. Solvent was removed and residue was taken into a mixed solution of dioxane (3 mL) and water (1 mL), sodium hydroxide (0.14 g, 3.6 mmol) was added followed by addition of BOC anhydride (0.30 g, 1.4 mmol). The resulting mixture was stirred at room temperature overnight and dichloromethane ( 20 mL) was added. Organic layer was separated, dried over sodium sulfate and concentrated. Product was purified by flash column chromatography on silica gel eluting with 0-100% ethyl acetate in hexanes to afford 0.070 g of the title compound m/z (ES⁺) [M+H]⁺ = 260.

Synthesis of 6-methyl-4-oxo-N-(quinazolin-8-ylmethyl)-3, 4-dihydro†uro[2, 3-d]pyrimidine-5-carboxamide To a solution of compound i-17b (0.070 g, 0.27 mmol) in dichloromethane (2 mL) was added 4 N HCI in dioxane (1 mL). The resulting mixture was stirred at room temperature for 1 .5 hours. Solvent was removed to dryness to provide the HCI salt i-17c.

To a solution of the HCI salt i-17c (0.27 mmol), 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine~ 5-carboxylic acid (5) (0.072 g, 0.34 mmol) and TEA (0.27 mL, 1 .94 mmol) in dichloromethane (2 mL) was added a solution of 2-Chloro-1 ,3-dimethylimidazolinium chloride (0.058 g, 0.34 mmol) in 1 .0 mL of dichloromethane. The resulting mixture was stirred at room temperature for 2 hours. Product was purified by column chromatography on silica gel eluting with 0-10% methanol in dichloromethane to provide the title compound (0.028 g) as a white solid m/z (ES⁺) [M+H]⁺ = 336.

¹H NMR (300 MHz, DMSO-d₆) d 10.75 (m, 1 H), 9.63 (s, 1 H), 9.38 (s 1 H), 8.22 (s, 1 H), 8.16 (s,

1 H), 8.10 (d, J = 7.8 Hz, 1 H), 8.00 (d, 0 = 6.9 Hz, 1 H), 7.75 (m, 1 H), 5.10 (d, J = 5.7 Hz, 2H), 2.72 (s,

3H).

Example 18: Synthesis of Compound 81

To a solution of 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (100 mg, 0.52 mmol) and phenylmethylamine (64 mg, 0.60 mmol) in DMF (3 mL) was added HATU (294 mg, 0.77 mmol) followed by DIEA (0.27 mL, 1 .55 mmol). The reaction was stirred at rt for 12 hs. The solvent was concentrated under reduced pressure and the residue was distributed between EtOAc (10 mL) and H2O (10 mL). The organic layer was washed with H2O (5 mL ^c 3), dried over anhydrous Na₂S0₄, filtered and concentrated. The crude compound was purified by flash column chromatography on silica gel

(MeOH/DCM) to afford the title compound (70 mg, 48%). ¹H NMR (300 MHz, DMSO-de) d 10.62 (t, 1 H), 8.25 (s, 1 H), 7.35 (d, 4H), 7.27 (m, 1 H), 4.52 (d, 2H), 2.72 (s, 3H); m/z (ES⁺) [M+H]⁺ = 284.0.

Example 19. Synthesis of Compound 9

Compound 9 was prepared using the procedure in Example 18 with the corresponding carboxylic acid and amine. Yield: 2%. ¹H NMR (300 MHz, CDCb) 610.21 (d, 1 H), 8.26 (s, 1 H), 4.20 (m, 1 H), 2.72 (S, 3H), 1 .87 (m, 2H), 1 .69-1 .50 (m, 6H); m/z (ES⁺) [M+H]⁺ = 262.1 . Example 20. Synthesis of Compound 82

Compound 82 was prepared using the procedure in Example 18 with the corresponding carboxylic acid and amine. Yield: 1 1 %. ¹H NMR (300 MHz, DMSO-de) d 12.59 (s, 1 H), 8.30 (s, 1 H), 7.73 (d, 2H), 7.39 (d, 2H), 7.1 1 (t, 1 H), 2.80 (s, 3H); m/z (ES⁺) [M+H]⁺ = 269.9.

Example 21. Synthesis of Compound 84

Compound 84 was prepared using the procedure in Example 18 with the corresponding carboxylic acid and amine. Yield: 20%. ¹H NMR (300 MHz, CDCI3) d 13.14 (s, 1 H), 10.65 (d, 1 H), 8.26 (s, 1 H), 7.37-7.22 (m, 5H), 5.13 (q, 1 H), 2.69 (s, 3H), 1 .47 (d, 3H); m/z (ES⁺) [M+H]⁺ = 297.9.

Example 22. Synthesis of Compound 20

Compound 20 was prepared using the procedure in Example 18 with the corresponding carboxylic acid and amine. Yield: 12%. ¹H NMR (300 MHz, CDCI3) 13.14 (s, 1 H), 10.14 (d, 1 H), 3.14 (m, 2H), 2.88 (s, 1 H), 2.731 (s, 3H), 1 .72 (m, 5H), 1 .51 (m, 1 H), 1 .19 (m, 3H), 0.98 (m, 2H); m/z (ES⁺) [M+H]⁺ = 290.2.

Example 23. Synthesis of Compound 85

Compound 85 was prepared using the procedure in Example 18 with the corresponding carboxylic acid and amine. Yield: 85%. ¹H NMR (300 MHz, DMSO-de) d 13.17 (brs, 1 H), 10.69 (d, 1 H), 8.28 (s, 1 H), 7.32 (d, 4H), 7.24 (m, 1 H), 4.92 (q, 1 H), 2.70 (s, 3H), 1 .77 (m, 2H), 0.88 (m, 3H); m/z (ES⁺) [M+H]⁺ = 31 1 .9.

Example 24. Synthesis of Compound 26

Compound 26 was prepared using the procedure in Example 18 with the corresponding carboxylic acid and amine. Yield: 30%. ¹H NMR (400 MHz, DMSO-de) d 13.15 (brs, 1 H), 10.64 (d, 1 H), 8.21 (s, 1 H), 7.28 (m, 1 H), 7.17 (m, 2H), 7.03 (d, 1 H), 4.68 (s, 2H), 2.72 (s, 3H), 2.03 (m, 1 H), 0.92 (q, 2H), 0.65 (m, 2H); m/z (ES⁺) [M+H]⁺ = 324.0.

Example 25. Synthesis of Compound 38

Compound 38 was prepared using the procedure in Example 18 with the corresponding carboxylic acid and amine. Yield: 30%. ¹H NMR (300 MHz, DMSO-de) d 13.30 (brs, 1 H), 11 .97 (s, 1 H), 8.32 (s, 1 H), 7.93 (d, 1 H), 7.53 (d, 1 H), 7.37 (m, 1 H), 7.23 (m, 1 H), 2.79 (s, 3H); m/z (ES⁺) [M+H]⁺ = 304.1 .

Example 26. Synthesis of Compound 41

Compound 41 was prepared using the procedure in Example 18 with the corresponding carboxylic acid and amine. Yield: 8%. ¹H NMR (400 MHz, DMSO-de) d 10.97 (brs, 1 H), 8.16 (s, 1 H), 6.78 (m, 1 H), 6.76 (s, 2H), 4.43 (d, 2H), 4.30 (m, 2H), 4.28 (m, 2H), 2.69 (s, 3H); m/z (ES⁺) [M+H]⁺ = 342.0.

Example 27. Synthesis of Compound 69

Compound 69 was prepared using the procedure in Example 18 with the corresponding carboxylic acid and amine. Yield: 7%. ¹H NMR (300 MHz, DMSO-de) d 13.18 (s, 1 H), 10.63 (s, 1 H), 8.27 (s, 1 H), 7.36 (m, 1 H), 7.24 (d, 2H), 4.59 (s, 2H), 2.75 (s, 3H); m/z (ES⁺) [M+H]⁺ = 320.0 . Example 28. Synthesis of Compound 76

Compound 76 was prepared using the procedure in Example 18 with the corresponding carboxylic acid and amine. Yield: 21 %. ¹H NMR (300 MHz, DMSO-de) d 13.12 (s, 1 H), 10.14 (t, 1 H), 8.23 (s, 1 H), 3.24-3.17 (m, 2H), 2.70 (s, 3H), 2.10-2.00 (m, 1 H)1.78-1.68 (m, 2H), 1 .62-1 .40 (m, 4H), 1 .30-1 .16

(m, 2H); m/z (ES⁺) [M+H]⁺ = 276.0.

Example 29. Synthesis of Compound 21

Compound 21 was prepared using the procedure in Example 35 with the corresponding carboxylic acid and amine. Yield: 22%. ¹H NMR (300 MHz, DMSO-de) d 13.08 (brs, 1 H), 10.22 (s, 1 H), 8.23 (s, 1 H), 7.26 (s, 4H), 7.20 (m, 1 H), 3.53 (m, 2H), 2.82 (m, 2H), 2.70 (s, 3H); m/z (ES⁺) [M+H]⁺ = 298.1 .

Example 30. Synthesis of Compound 36

Compound 36 was prepared using the procedure in Example 35 with the corresponding carboxylic acid and amine. Yield: 14%. ¹H NMR (300 MHz, CDCI3) d 13.207 (s, 1 H), 10.666 (s, 1 H), 8.250 (s, 1 H), 7.749 (m, 2H), 7.391 (m, 1 H), 4.635 (s, 2H), 2.720 (s, 3H); m/z (ES⁺) [M+H]⁺ = 370.1 .

Example 31. Synthesis of Compound 47

Compound 47 was prepared using the procedure in Example 35 with the corresponding carboxylic acid and amine. Yield: 36%. ¹ H NMR (400 MHz, DMSO-de) d 10.72 (d, 1 H), 8.28 (s, 1 H), 7.70 (d, 2H), 7.61 (d, 2H), 5.21 (m, 1 H), 2.67 (s, 3H), 1 .50 (s, 3H); m/z (ES⁺) [M+H]⁺ = 365.9 .

Example 32. Synthesis of Compound 65

Compound 65 was prepared using the procedure in Example 35 with the corresponding carboxylic acid and amine. Yield: 19%. ¹H NMR (400 MHz, DMSO-de) d 13.10 (s, 1 H), 10.62 (s, 1 H), 8.96 (s, 1 H), 8.39 (d, 1 H), 8.22 (s, 1 H), 7.91 (d, 1 H), 7.72 (s, 1 H), 7.59 (d, 2H), 5.16 (s, 2H), 2.74 (s, 3H); m/z (ES-) [M+H]⁺ = 335.1 .

Example 33. Synthesis of Compound 68

Compound 41 was prepared using the procedure in Example 35 with the corresponding carboxylic acid and amine. Yield: 1 1 %. ¹H NMR (400 MHz, DMSO-de) d 13.16 (s, 1 H), 10.59 (d, 1 H), 8.27 (s, 1 H), 7.42 (m, 2H), 7.20 (d, 2H), 4.57 (s, 2H), 2.73 (s, 3H); m/z (ES⁺) [M+H]⁺ = 302.1 .

Example 34. Synthesis of Compound 70

Compound 70 was prepared using the procedure in Example 35 with the corresponding carboxylic acid and amine. Yield: 27%. ¹H NMR (400 MHz, DMSO-de) d 13.15 (brs, 1 H), 10.44 (t, 1 H), 8.28 (s, 1 H), 7.22 (m, 3H), 4.51 (d, 2H), 4.50 (s, 2H), 3.57 (s, 2H), 3.40 (q, 2H), 2.72 (s, 3H),1 .08 (t, 3H); m/z (ES⁺) [M+H]⁺ = 381 .1 . Example 35. Synthesis of Compound 71

Compound 71 was prepared using the procedure in Example 35 with the corresponding carboxylic acid and amine. Yield: 21 %. ¹H NMR (400 MHz, DMSO-de) d 13.19 (brs, 1 H), 10.76 (d, 1 H), 8.26 (s, 1 H), 7.33 (m, 1 H), 7.23 (m, 2H), 5.36 (m, 1 H), 2.65 (s, 3H), 1.50 (d, 3H); m/z (ES⁺) [M+H]⁺ = 333.9. Example 36. Synthesis of Compound 72

Compound 72 was prepared using the procedure in Example 35 with the corresponding carboxylic acid and amine. Yield: 19%. ¹H NMR (400 MHz, DMSO-de) d 13.17 (brs, 1 H), 10.74 (d, 1 H), 8.28 (s, 1 H), 7.42 (t, 1 H), 7.33 (m, 1 H), 7.19 (m, 2H), 5.32 (m, 1 H), 2.67 (s, 3H), 1 .47 (d, 3H); m/z (ES⁺) [M+H]⁺ = 315.9.

Example 37. Synthesis of Compound 73

Compound 73 was prepared using the procedure in Example 35 with the corresponding carboxylic acid and amine. Yield: 41 %. ¹ H NMR (400 MHz, DMSO-de) d 13.20 (brs, 1 H), 10.44 (t, 1 H), 8.24 (s, 1 H), 7.20 (m, 3H), 4.51 (m, 4H), 3.58 (s, 2H), 3.38 (m, 2H), 2.73 (s, 3H), 1 .52 (q, 2H), 0.84 (t, 3H); m/z (ES⁺) [M+H]⁺ = 395.1 .

Example 38. Synthesis of Compound 79

Compound 79 was prepared using the procedure in Example 35 with the corresponding carboxylic acid and amine. Yield: 96%. ¹ H NMR (400 MHz, DMSO-de) d 13.20 (brs, 1 H), 10.01 (d, 1 H), 8.24 (s, 1 H), 3.86 (m, 1 H), 2.68 (s, 3H), 1 .67 (m, 5H), 1 .39 (m, 1 H), 1 .03-1.17 (m, 5H), 1 .08 (d, 3H); m/z (ES⁺) [M+H]⁺ = 304.2.

Example 39. Synthesis of Compound 12

Compound 12 was prepared using the procedure in Example 35 with the corresponding carboxylic acid and amine. Yield: 32%. ¹H NMR (300 MHz, DMSO-de) 13.15 (s, 1 H), 10.09 (s, 1 H), 8.24 (s, 1 H), 3.3-2.9 (m, 2H), 2.70 (s, 3H), 2.50 (m, 1 H), 2.10-1 .90 (m, 2H), 1 .81 -1 .73 (m, 4H); m/z (ES⁺)

[M+H]⁺ = 262.0.

Example 40. Synthesis of Compound 25

Compound 25 was prepared using the procedure in Example 35 with the corresponding carboxylic acid and amine. Yield: 23%. ¹H NMR (400 MHz, DMSO) d 13.196 (brs, 1 H), 10.657 (d, 1 H), 8.278 (s, 1 H), 7.378 (m, 1 H), 7.218 (t, 2H), 7.078 (t, 1 H), 5.138 (m, 1 H), 2.679 (s, 3H), 1 .480 (d, 3H); m/z (ES⁺) [M+H]⁺ = 316.0.

Example 41. Synthesis of Compound 62

To a solution of 6-methyl-4-oxo-3,4~dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (150 mg, 0.77 mmol) and isoquinolin-8-ylmelhaiiamine (179 mg, 0.77 rrimoi) in DMF (5 ml) was added (i -cyano-2~ ethoxy-2-oxoethylidenaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate (COMU, 396 mg, 0.89 mmol) and TEA (0.372, 2.7 mmol). The reaction was stirred at rt for 2 h before the solvent was removed. The resulting residue was diluted with EtOAc (10 mL), washed with H2O (10 mL><3). The organic phase was dried over anhydrous Na₂SC>₄, filtered and concentrated. The crude product was purified on silica (DCM/MeOH/TEA) followed by HPLCs (H2O/ACN 5-100%) to afford the titie product (8 mg, 3.2%). ¹H NMR (400 MHz, MeOD-de) d 9.62 (d, 1 H), 8.49 (d, 1 H), 8.09 (d, 1 H), 7.91 (m, 2H), 7.77 (m, 2H), 5.18 (s, 2H), 2.80 (s, 3H); m/z (ES⁺) [M+Hf = 335.1 .

Example 42. Synthesis of Compound 52

To a solution of 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (150 mg, 0.77 mmol) and (R)-6,7,8,9-tetrahydro-5H-benzo[7]annulen-5-amine (124 mg, 0.77 mmol) in DMF (5 mL) was added COMU (396 mg, 0.89 mmol) and TEA (0.375 mL, 2.7 mmol). The reaction was stirred at rt for 2 hs before the solvent was removed. The residue was dissolved in DMSO then purified by HPLC (H2O/ACN 5-100%) to afford the title product (1 14 mg, 44%). ¹ H NMR (400 MHz, DMSO-de) d 13.21 (s, 1 H), 10.63 (d, 1 H), 8.27 (s, 1 H), 7.28 (s, 1 H), 7.09 (s, 3H), 5.26 (m, 1 H), 3.02 (m, 1 H), 2.84 (m, 1 H), 2.73 (s, 3H), 2.10 (m, 1 H), 1 .82 (m, 3H), 1 .63 (m, 2H); m/z (ES⁺) [M+H]⁺ = 337.9. Example 43. Synthesis of Compound 29

To a solution of 2-(2-(aminomethyl)phenyl)acetic acid (220 mg, 1 .33 mmol), potassium carbonate (404 mg, 2.93 mmol) in dioxane (6 mL) and hteO (2 mL) was added Fmoc chloride (380 mg, 1.47 mmol). The reaction was stirred at rt for 2 h before it was acidified to pH 2 and diluted with water (50 mL). The resulting mixture was extracted with EtOAc (50 mLx3). The combined organic extracts were dried over anhydrous Na₂SC>₄, filtered and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica (EtOAc/DCM) to afford the title product (277 mg, 25%). m/z (ES⁺)

[M+H]⁺ = 388.0.

To a solution of 2-(2-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)phenyl)acetic acid (277 mg, 0.72 mmol), ethanolamine (0.053 mL, 0.88 mmol), and TEA (0.2 mL, 1.47 mmol) in DMF (6 mL) was added COMU (369 mg, 0.86 mmol). The reaction was stirred at rt for 1 h before it was diluted with sat. NaHCOs (50 mL) and extracted with EtOAc (50 mLx3). The combined organic phase was dried over anhydrous Na₂S0₄, filtered and concentrated. The crude product was purified by flash column chromatography on silica (MeOH/DCM) to afford the title product (192 mg, 62%). m/z (ES⁺) [M+H]⁺ = 431.0.

To a solution of (9H-fluoren-9-yl)methyl (2-(2-((2-hydroxyethyl)amino)-2- oxoethyl)benzyl)carbamate (1 .65 g, 3.84 mmol) in DCM (50 mL) was added Dess-Martin reagent (1 .63 g, 3.84 mmol). The reaction was stirred at rt for 2 hs before the solution was concentration under reduced pressure. The crude product was purified by flash column chromatography (EtOAc/DCM) to afford (9H- fluoren-9-yl)methyl (2-(2-oxo~2-((2-oxoethyl)amino)ethyl)benzyl)carbamate (1 .22 g, 74%) which was redissolved in THF (22 mL). To the resulting solution was added Burgess reagent (1 .3 g, 5.59 mmol) and the mixture was heated at 65°C for 12 h. The mixture was diluted with methanol (10 mL), filtered through celite and concentrated. The crude product was purified by flash column chromatography on silica (EtOAc/Hexanes) to afford the title product (178 mg, 15%). m/z (ES⁺) [M+H]⁺ = 41 1 .0.

Step d. Synthesis of xazol-2~ylmethyl)phenyl)methanamine

To a solution of (9H-fluoren-9-yl)methyl (2-(oxazol-2-ylmethyl)benzyl)carbamate (178 mg, 0.43 mmol) in dioxane (3 mL) was added piperidine (2 mL) . The reaction was heated at 50 °C for 1 h. The solution was concentrated under reduced pressure and the crude was purified by reverse phase HPLC to afford the title product (38mg, 47%). m/z (ES⁺) [M+H]⁺ = 189.0.

Step e. Synthesis of 6-methyl-N-(2-(oxazol-2-ylmethyl)benzyl)-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-· carboxamide (compound 29)

To a solution of 6-methyl-4~oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (39 mg, 0.20 mmol), (2-(oxazol~2-ylmethyl)phenyl)methanamine (38 mg, 0.20 mmol), TEA (0.084 mL, 0.61 mmol) in DCM (2 mL) was added a solution of 2-chloro-1 ,3-dimethylimidazolinium chloride (44 mg, 0.26 mmol) in DCM (2 mL). The reaction was stirred at rt for 12 h. The reaction solution was concentrated under reduced pressure and the crude was purified by reverse phase HPLC to afford the title product (12 mg, 16%) as a white solid. ¹H NMR (300 MHz, DMSO-de) d 13.10 (s, 1 H), 10.60 (s, 1 H), 8.23 (s, 1 H), 7.98 (d, 1 H), 7.33 (m, 1 H), 7.30-7.15 (m, 3H), 7.10 (d, 1 H), 4.56 (d, 2H), 4.26 (s, 2H), 2.71 (s, 3H). m/z (ES⁺) [M+H]⁺ = 365.0.

Example 44. Synthesis of Compound 45

Step c step d

Step a. Synthesis of 4-bromo-1 -methylindolin-2-one

A solution of 4-bromo-1-methylindoline-2,3-dione (970 mg, 4.05 mmol) in hydrazine hydrate (30 mL) was heated in a microwave at 130° C for 1 h. During the reaction a solid crashed out of solution. The mixture was then cooled to 0°C and ice water (5 mL) was added to quench the reaction. The resulting mixture was stirred for 30 minutes at 0 °C. Filtration afforded the title product (300 mg, 33%). m/z (ES⁺) [M+H]⁺ = 226.0.

Step b. Synthesis of 1-methyl-2-oxoindoline-4-carbonitrile

To a solution of 4-bromo-1-methylindolin-2-one (250 mg, 1.11 mmol) in DMF (3 mL) was added Zn(CN)2 (385 mg, 3.33 mmol) followed by Pd(PPh3)4 (52 mg, 0.04 mmol). The reaction was stirred at 150°C for 40 mins. The reaction solution was extracted with DCM (20 mL><3). The combined organic phase was washed with brine (20 mL), dried over anhydrous Na2SC>₄, filtered and concentrated. The crude product was purified by silica gel column chromatography (EtOAc/DCM) to afford the title product (160 mg, 84% yield), m/z (ES⁺) [M+H]⁺ = 173.0.

Step c. Synthesis of 4-(aminomethyl)~ 1 -methylindolin-2-one

To a solution of 1 -methyl-2-oxoindoline-4-carbonitrile (80 mg, 0.46 mmol) in MeOH (15 mL) was added Pd/C (25 mg) followed by cone. HCI (0.003 mL). This mixture was then stirred under an atmosphere of H2(g) for 48 hs. The solution was filtered through celite and used for the next step without further purification m/z (ES⁺) [M+H]⁺ = 176.9.

To a solution of 4-(aminomethyl)~1 -methylindolin-2-one (80 mg, 0.45 mmol) in DCM (7 mL) was added 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (0.087 mL, 0.45 mmol), TEA (0.125 mL, 0.90 mmol), and 2-chloro-1 ,3-dimethylimidazolinium chloride (100 mg, 0.59 mmol). The reaction was stirred at rt for 1 h. The reaction mixture was concentrated, purified on the waters prep- HPLC using 5-100% Acetonitrile in Water to afford the title product (30 mg, 19%). ¹ H NMR (300 MHz, CDCI3) 6 13.15 (s, 1 H), 10.52 (s, 1 H), 8.26 (s, 1 H), 7.26 (t, 1 H), 6.99 (d, 1 H), 6.90 (d, 1 H), 4.45 (d, 2H), 3.61 (s, 2H), 3.13 (s, 3H), 2.72 (s, 3H); m/z (ES⁺) [M+H]⁺ = 353.1.

Example 45. Synthesis of Compound 66

Step a. Synthesis of 1 -methyl-1 H-benzo[d]imidazole-4-carbonitrile

To a solution of 4-bromo-1 -methyl-1 H-benzo[d]imidazole (150 mg, 0.71 mmol) in DMF (5 mL) was added Zn(CN)2 (250, 2.13 mmol) followed by Pd(PPfi3)4 (33 mg, 0.03 mmol). The reaction was stirred at 150°C for 12 hs and then heated to 160°C in a microwave for another 24 h. The reaction was diluted with H2O (5 mL) and extracted with DCM (20 mL). The organic layer was washed with brine (10 mL), dried over anhydrous Na2S04, filter and concentrate. The crude product was used for the next step without further purification m/z (ES⁺) [M+H]⁺ = 180.0.

To a solution of 1 -methyl-2-oxoindoline-4-carbonitrile (100 mg, 0.64 mmol) in methanol (15 mL) was added Pd/C 10% (30 mg) followed by cone. HCI (0.03 mL). This mixture was then stirred under H2 for 2 h. The solution was filtered through celite and used for the next step without further purification m/z (ES⁺) [M+H]⁺ = 158.0.

To a solution of 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (120 mg, 0.62 mmol) in DMF (5 ml_) was added HATU (306 mg, 0.81 mmol), (1 -methyl-1 H-benzo[d]imidazol-4- yl)methanamine (0.10 mL, 0.62 mmol), and DIEA (0.324, 1 .86 mmol). The reaction was stirred at rt for 12 hs. The mixture was diluted with H2O (5 mL) and extracted with EtOAc (15 mL). The organic layer was washed with H2O (5 mL) and brine (5 mL), dried over anhydrous Na2SC>4, filtered and concentrated. The crude product was purified by prep-HPLC to afford the title product (5 mg, 3%). ¹H NMR (300 MHz, CDCI3) d 13.02 (s, 1 H), 10.56 (d, 1 H), 8.18 (d, 2H), 7.47 (d, 1 H), 7.17 (m, 2H), 4.90 (m, 2H), 3.85 (d, 3H), 2.72 (s, 3H); m/z (ES⁺) [M+H]⁺ = 338.0.

Example 46. Synthesis of Compound 67

To a solution of 3-bromo-2-nitrobenzaldehyde (723 mg, 3.14 mmol), iron powder (828 mg, 14.79 mmol) in ethanol (9 mL) and water (1.5 mL) was added cone. HCI (2 drops). The reaction was heated to 80°C for 3 h. The reaction was cooled to rt, diluted with EtOAc (50 mL), filtered through celite, and concentrated. The product was dissolved in MeOH/DCM (1 mL/9 mL), dried over anhydrous Na2S04, filtered and concentrated under reduced pressure to afford 2-amino-3-bromobenzaldehyde (584mg) which was used without further purification m/z (ES⁺) [M+H]⁺ = 200.0.

Step b. Synthesis of 8-bromo-2-methylquinazoline

To a solution of 2-amino-3-bromobenzaldehyde (584 mg, 2.92 mmol) in EtOH (10 mL) was added acetimidamide hydrochloride (1.90 g, 20.16 mmol) and sodium carbonate (2.20 g, 20.75 mmol). The reaction was heated at 120°C for 4 days. The mixture was then cooled to rt, diluted with brine (10 mL), and extracted with DCM (10 mLx3). The combined organic extracts were dried over anhydrous NazSCTi, filtered and concentrated. The crude product was purified by flash column chromatography on silica gel (EtOAc/Hexanes) to afford the title product (153 mg, 24%). m/z (ES⁺) [M+H]⁺ = 223.0 .

Step c. Synthesis of 2~methylquinazoline~8-carbonitrile

To a solution of 8-bromo-2-methylqnina7oline (1 53 mg_, 0 69 mmol) in DMF (4.5 mL) was added ZnCN (304 mg, 2.59 mmol) and PdCbdppf.CLECb (78 mg, 0.10 mmol). The mixture was heated to 160°C under microwave irradiation for 2.5 h. The mixture was diluted with brine (5 mL) and extracted with DCM (10 mLx3). The combined organic extracts were dried over anhydrous Na₂S0₄, filtered and concentrated. The crude product was purified by flash column chromatography on silica gel

(EtOAc/DCM) to afford the title product (58 mg, 50%). m/z (ES⁺) [M+H]⁺ = 170.0.

To a solution of 2-methylquinazoline-8-carbonitrile (58 mg, 0.34 mmol) in 7 N ammonia-methanol (15 mL) was added Raney Ni (1 mL, aqueous slurry). The suspension was purged with nitrogen and stirred under an atmosphere of H2(g) at rt for 2 h. The mixture was filtered through celite and

concentrated to afford the title product used without further purification m/z (ES⁺) [M+H]⁺ = 176.0.

To a solution of crude (2-methyl-3,4-dihydroquinazolin-8-yl)methanamine in dioxane (4 ml_) was added 2N NaOH (0.70 ml_, 0.35 mmol) and di-tert-butyl dicarbonate (134 mg, 0.62 mmol). The reaction was stirred at rt for 72 h. The mixture was diluted with H2O (10 ml_) and extracted with DCM (10 ml_x3). The combined organic extracts were dried over anhydrous Na2SC>4, filtered and concentrated. The residue was re-dissolved in MeOH (2 ml_) and 2N NaOH (0.5 ml_). The resulting mixture was heated to 60°C for 3 h. The mixture was diluted with H2O (10 ml_) and extracted with DCM (10 ml_x3). The combined organic extracts were dried over anhydrous a2S04, filtered and concentrated to afford the title product used without further purification, m/z (ES⁺) [M+H]⁺ = 276.0.

To a solution of above crude tert-butyl ((2-methyl-3,4-dihydroquinazolin-8-yl)methyl)carbamate in chloroform (4 ml_) was added manganese(IV) oxide (452 mg). The mixture was heated to 50 °C for 1 h. before it was filtered through celite and concentrated. The crude product was purified by flash column chromatography on silica gel (EtOAc/DCM) to afford the title product (63 mg, overall 67%, step d-f). m/z (ES⁺) [M+H]⁺ = 274.0.

Step g. Synthesis of (2-methylquinazolin-8-yl)methanamine hydrochloride

A solution of tert- butyl ((2-methylquinazolin-8-yl)methyl)carbamate (63 mg, 0.23 mmol) in 4 N HCI-dioxane (1 ml_, 4.0 mmol) and methanol (0.5 mL) was heated to 50°C for 2 h. The mixture was concentrated to afford the title product (63 mg) m/z (ES⁺) [M+H]⁺ = 174.0.

To a solution of (2-methylquinazolin-8-yl)methanamine hydrochloride (63 mg, 0.30 mmol) in DCM (5 mL) was added 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxyiic acid (58 mg, 0.30 mmol), TEA (0.20 mL, 1 .43 mmol), and a solution of 2-chloro-1 ,3-dimethylimidazolinium chloride (74 mg, 0.44 mmol) in DCM (2 mL). The reaction was stirred at rt for 12 hs. The mixture was diluted with sat. NaHCC (10 mL) and extracted with DCM (10 mLx3). The combined organic extracts were dried over anhydrous Na2SC>4, filtered and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel (MeOH/DCM) to afford the title product (12 mg, 1 1 %). ¹H NMR (300 MHz, DMSO-d6) d 10.98 (1 H), 9.52 (1 H), 8.19 (2H), 8.00 (1 H), 7.91 (1 H), 7.64 (1 H), 5.06 (2H), 2.84 (3H), 2.71 (3H). m/z (ES⁺) [M+H]⁺ = 350.0.

Example 47. Synthesis of Compound 74

Step d

Step a. Synthesis of 8-Bromo-1, 4-dihydroisoquinolin-3( 2H)-one

To a solution of 4-Bromo-1 ,3-dihydro-2H-inden-2-one (650 mg, 3.08 mmol) in cone. HCI (20 ml) was added NaN3 (400 mg, 6.16 mmol) potionwise at 0 °C. The reaction was stirred at 0°C for 20 mins and then at rt for 12 hs. The pH of the reactio solution was adjusted to 9 at 0 °C using potasium carbonate. The resulting mixture was extracted with DCM (20 m x3). The combined organic phase was collected and allowed to stand at rt until the solid crashed out. The filtration afforded the title product (225 mg, 33%). m/z (ES⁺) [M+H]⁺ = 21 1 .0.

To a solution of 8-bromo-1 ,4-dihydroisoquinolin-3(2H)-one (225 mg, 1 mmol) in DMF (5 ml) was added Zn(CN)2 (342 mg, 2.88 mmol) and Pd(PPfi3)₄ (59 mg, 0.051 mmol). The reaction solution was purged with N2 for 1 minute and then heated to 150 °C for 6 hs. After cooled down to rt, the mixture was filtered through celite. The filtrate was concentrated and the crude was purified by silica gel column chromatography (EtOAc/Hexanes) to afford the title compound (130 mg, 76%). m/z (ES⁺) [M+H]⁺ = 173.1 .

A solution of 3-oxo-l ,2,3,4-tetrahydroisoquinoline-8-carbonitrile(130 mg, 0.76 mmol) in 7N Nhb in MeOH(5 mL) was purged with N2 and added Ra-Ni (100 mg, slury in water). The mixture was then stirred under H2 bloon for 12 hs. The mixture was filtered through celite and the filtrate was concentrated to afford the title product without further purification (95 mg, 71 %). m/z (ES⁺) [M+H]⁺ = 177.0.

To a solution of 6-methyl-4-oxo-3,4-dihydrofuro[2,3-d]pyrimidine-5-carboxylic acid (105 mg, 0.54 mmol) and 8-(aminomethyl)-1 ,4-dihydroisoquinolin-3(2H)-one (95 mg, 0.54 mmol) in DCM (5 mL) was added TEA (0.5 mL, 3.58 mmol) and 2-chloro-1 ,3-dimethylimidazolinium chloride (109 mg, 0.65 mmol). The reaction was stirred at rt for 1 h before the reaction solution was diluted with EtOAc (10 mL), washed with H2O (10 mLx3) and brine (5ml). The organic phase was dried over anhydrous Na2SC>4, filtered and concentrated. The crude product was purified on silica gel column chromatography (DCM/MeOH) to afford the title product (31 mg, 17%). ¹H NMR (400 MHz, DMSO-de) d 13.15 (brs, 1 H), 10.45 (t, 1 H), 8.25 (s, 1 H), 8.02 (t, 1 H), 7.22 (m, 2H), 7.13 (m, 1 H), 4.50 (d, 2H), 4.41 (s, 2H), 3.49 (s, 2H), 2.72 (s, 3H); m/z (ES⁺) [M+H]⁺ = 353.0.

Example 48. Human AåA Receptor Binding by Compounds of the Invention

Protocol: Compound binding to the human A2A receptor was assessed in a radioligand competition assay. 6 nM of [³H]-labeled CGS 21680 ligand (Kd = 27 nM) was bound to HEK293 cells expressing the human A2_A receptor. The ability of each test compound to compete for the radioligand binding at eight concentration points was measured by scintillation counting. Data was analyzed by nonlinear regression to generate IC50 values and the inhibition constants (K,) were calculated using the Cheng Prusoff equation.

Results: As shown in Table 2, compounds of the invention bind to human A2_A receptor with inhibition constants less than 10 nM.

Table 2. Human A2A Receptor Binding Data

'+++' = <10 nM; '++' = 10-100 nM,‘+’ = 100-1000 nM

Example 49. Rat Functional A2_A Receptor Assay for Compounds of the Invention

Protocol: An A2A receptor agonist, NECA, was added at 100 nM to rat PC12 cells that endogenously express the A2A receptor. After incubation for 10 min at RT, cAMP, an intracellular signaling molecule, was measured in an HTRF assay. The ability of each compound to inhibit this cAMP production was evaluated at eight concentrations. Data was analyzed by non-linear regression to generate IC50 values for each compound. The apparent dissociation constants (KB) were calculated using a modified Cheng Prusoff equation KB= IC50/I +(A/ECSOA) where A is the concentration of reference agonist in the assay, and ECSOA is the EC50 value of this reference agonist.

Results: As shown in Table 3, compounds of the invention are able to compete with NECA and inhibit cAMP production as shown by the dissociation constant of the reference agonist.

Table 3. Rat Functional A2_A Receptor Data

‘+++’ = <10 nM; '++' = 10-100 nM,‘+’ = 100-1000 nM

Example 50. TNFa and IFNy Recovery Assay for Compounds of the Invention

Protocol: PBMCs were prepared from fresh human blood using a Lymphoprep™ kit and the purified PBMCs were resuspended in RPMI1640 media with 10% FBS. PBMCs were then stimulated to produce cytokines with the addition of anti-CD3/CD28 coated bead at a 1 :6 bead/cell ratio. A total of

200,000 PBMC were added to each well of a 96-well plate along with either 150 nM or 300 nM NEC A and various concentrations of test compounds. The PBMC plate was incubated for 24 hours at 37 °C and then an aliquot of the media was taken to read TNFa levels using a standard ELISA assay. IFNy levels were read using a standard ELISA assay after 48 hours of incubation. Percent recovery for each cytokine was calculated by comparing the cytokine concentration in the presence of compound to the

concentration in the presence of DMSO without NECA.

Results: As shown in Tables 4-7, compounds of the invention are able to compete with NECA to increase recovery of TNFa and IFNy.

Table 4. TNFa Recovery Data

Table 5. IFNy Recovery Data

Table 6. TNFa Recovery Data

Table 7. IFNY Recovery Data

Example 51. Quantitation of IL-12 and IL-6 Cytokine Secreted from Dendritic Cells Treated with Compounds of the Invention

The ability of an AIA receptor and an AZAIAZB receptor antagonist to reverse the effect of NECA, a nonhydrolyzable analog of adenosine, on the maturation and activation of dendritic cells was studied, starting with the differentiation of human immature DC.

Protocol: Fifty thousand human monocytes that had been negatively selected from PBMCs (Astarte Biologies, #1008) were cultured in a 200 pL final volume containing serum free media (X-Vivo 15; Lonza #04-418Q), 10 ng/mL of human GM-CSF (Peprotech #300-03) and IL-4 (Peprotech #200-04). NECA (Sigma #E2387) was solubilized in DMSO at 20 mM and serially diluted in serum free media to working stocks containing either 400 mM or 40 pM in 2% DMSO. An equivalent serial dilution was employed to give a working stock of just 2% DMSO in serum free media. With the addition of 5 pl· of one of these working stocks to a given well, the final concentration of NECA was either 0, 1 or 10 pM.

Similarly, the A2_A and A_åA/b antagonists were solubilized at 10 mM in DMSO and serially diluted to 400 pM in 2% DMSO. With the addition of 5 pl of these or the 2% DMSO working stock, the final

concentrations in a given well was either 0 or 10 pM of an antagonist. The final concentration of DMSO was 0.1 % in all wells. Cell were incubated for 5 days at 37°C in 6% CO2. The immature DC were fully matured/activated by the addition of poly IC in 20 pl of media (Sigma #1530) to a final concentration of 20 pg/ml. After a further 24 h incubation, conditioned media was collected and frozen. A final 20x dilution of the conditioned media was evaluated for cytokines with the V-plex Proinflammatory Panel 1 (MSD #R15056).

IL-12 (FIG. 1 ) and IL-6 (FIG. 2) levels were measured in the conditioned media 24 h after immature DC had been activated with 20 pg/mL poly IC. Immature DC had been differentiated from monocytes, and subsequently activated, in the presence or absence of 1 pM and 10 pM NECA, a nonhydrolyzable analog of adenosine, 10 pM of an A2A antagonist (compound 42), or 10 pM of a dual A2_A/A2_B antagonist (compound 84).

Result: As shown in FIG. 1 and FIG. 2, an AZA antagonist (compound 47) is not able to reverse the effect of NECA on the IL-12 and IL-6 levels, while a dual AZAIAZB antagonist (compound 84) is effective at reversing the effect of NECA on the IL-12 and IL-6 levels.

Other Embodiments

While the present invention has been described with reference to what are presently considered to be the preferred examples, it is to be understood that the invention is not limited to the disclosed examples. To the contrary, the invention is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. Where a term in the present application is found to be defined differently in a document incorporated herein by reference, the definition provided herein is to serve as the definition for the term.

Other embodiments are in the claims.

Claims

What is claimed is:

1 . A compound having the structure:

wherein X is O or S;

each of R¹ , R², and R³ is, independently, H or optionally substituted C1-C6 alkyl;

B is optionally substituted C3-C10 carbocyclylene or optionally substituted C3-C9 heterocyclylene;

m2 is 0, 1 , 2, or 3;

R⁴ is optionally substituted C6-C10 aryl, optionally substituted C3-C9 heterocyclyl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C6 heteroalkyl; and

wherein if R¹ is H, R² is CH3, R³ is H, and L¹ is

, then R⁴ is substituted Ce-Cio aryl, optionally substituted C3-C9 heterocyclyl, optionally substituted C1-C9 heteroaryl, or optionally substituted C1-C6 heteroalkyl,

or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1 , wherein R¹ is optionally substituted C1-C6 alkyl.

3. The compound of claim 2, wherein R¹ is

4. The compound of claim 1 , wherein R¹ is H.

5. The compound of any one of claims 1 to 4, wherein R² is optionally substituted C1-C6 alkyl.

CH₃

6. The compound of claim 5, wherein R² is X

7. The compound of any one of claims 1 to 6, wherein R³ is H.

R5 _r6

8. The compound of any one of claims 1 to 7, wherein U is

9. The compound of claim 8, wherein ml is 1.

10. The compound of claim 8 or 9, wherein R⁵ is H.

11. The compound of claim 8 or 9, wherein R⁵ is optionally substituted C1-C6 alkyl.

CH₃

12. The compound of claim 11 , wherein R⁵ is X

13. The compound of any one of claims 8 to 12, wherein R⁶ is H.

14. The compound of any one of claims 8 to 12, wherein R⁶ is optionally substituted Ci~C6 alkyl.

CH₃

15. The compound of claim 14, wherein R⁶ is

16. The compound of claim 8 or 9, wherein R⁵ and R⁶, together with the atom to which each is attached, combine to form an optionally substituted C3-C10 carbocyclyl.

17. The compound of claim 16, wherein R⁵ and R⁶, together with the atom to which each is attached, combine to form an optionally substituted C3-C₅ carbocyclyl.

18. The compound of claim 17, wherein U is

19. The compound of any one of claims 1 to 7, wherein U is

20. The compound of claim 19, wherein B is optionally substituted C3-C10 carbocyclylene. 21 . The compound of claim 20, wherein

22. The compound of claim 19, wherein B is optionally substituted C3-C9 heterocyclylene.

23. The compound of claim 22, wherein

24. The compound of claim 19, wherein

0

25. The compound of any one of claims 19 to 24, wherein L²

R⁵ R⁶

26. The compound of any one of claims 19 to 24, wherein L² is

27. The compound of claim 26, wherein m2 is 1 , 2, or 3.

28. The compound of claim 26 or 7, wherein m2 is 1 or 2.

29. The compound of any one of claims 1 to 28, wherein R⁴ is optionally substituted C6-C10 aryl.

30. The compound of claim 29, wherein R⁴ is

wherein

n is 0, 1 , 2, 3, 4, or 5; and

each R^a is, independently, C1-C6 perfluoroalkyl or C3-C8 cycloalkyl.

31 . The compound of claim 30, wherein each R^a is, independently,

32. The compound of claim 30 or 31 , wherein each R^a is

33. The compound of any one of claims 30 to 32, wherein n is 1 or 2.

34. The compound of any one of claims 1 to 28, wherein R⁴ is optionally substituted C3-C9 heterocyclyl.

wherein

o is 0, 1 , 2, or 3;

each R^b is, independently, halo or optionally substituted C1-C6 alkyl; and

R⁷ is H or optionally substituted C1-C6 alkyl.

36. The compound of claim 35, wherein each R^b is, independently, halo or Ci~C₆ alkyl.

37. The compound of claim 35 or 36, wherein o is 0 or 1.

38. The compound of any one of claims 35 to 37, wherein R⁷ is H.

39. The compound of any one of claims 35 to 37, wherein R⁷ is optionally substituted C1-C6 alkyl.

40. The compound of claim 39, wherein R⁷ is

.

41. The compound of claim 34, wherein R⁴ is

42. The compound of any one of claims 1 to 28, wherein R⁴ is optionally substituted C1 -C9 heteroaryl.

wherein

p is 0, 1 , 2, or 3;

q is 0, 1 , 2, or 3;

each of X^a, X^b, X^c, and X^d is, independently, N, CH, or CR^C; and X^e is O, S, NR⁸, or CR^cR^d;

each R^c is, independently, halo or optionally substituted C1-C6 alkyl; and R⁸ is H or optionally substituted C1-C6 alkyl.

44. The compound of claim 43, wherein

45. The compound of claim 44, wherein

46. The compound of claim 44 or 45, wherein q is 0 or 1 .

47. The compound of claim 43, wherein

48. The compound of claim 47, wherein

49. The compound of claim 47 or 48, wherein each of X^a and X^b is, independently, N or CH.

50. The compound of claim 49, wherein X^a is N.

51 . The compound of claim 49, wherein X^a is CH.

52. The compound of claim 50 or 51 , wherein X^b is N.

53. The compound of claim 50 or 51 , wherein X^b is CH.

54. The compound of claim 43, wherein

56. The compound of claim 54 or 55, wherein each of X^c and X^d is, independently, N or CH.

57. The compound of claim 56, wherein X^c is N.

58. The compound of claim 56, wherein X° is CH.

59. The compound of claim 57 or 58, wherein X^d is N.

60. The compound of claim 57 or 58, wherein X^d is CH.

61. The compound of any one of claims 57 to 60, wherein X^® is NH or CH2.

62. The compound of claim 61 , wherein X^e is NH.

63. The compound of claim 61 , wherein X^e is CH2.

64. The compound of any one of claims 47 to 63, wherein p is 0 or 1.

65. The compound of any one of claims 47 to 64, wherein p is 0.

66. The compound of any one of claims 1 to 28, wherein R⁴ is optionally substituted C1-C6 heteroalkyl.

CH₃

L ^ch

67. The compound of claim 66, wherein R⁴ is ^ ^ⁿ3 .

68. A pharmaceutical composition comprising a compound of any one of claims 1 to 67 and a pharmaceutically acceptable excipient.

69. A method for modulating the level of adenosine in a cell, the method comprising contacting a cell with an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68.

70. A method for modulating the level of adenosine in a subject, the method comprising administering an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68 to the subject.

71 . A method for the treatment of a disorder related to adenosine in a subject in need thereof, the method comprising administering an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68 to the subject.

72. A method for modulating the activity of the adenosine A2_A receptor in a cell, the method comprising contacting a cell with an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68.

73. A method for modulating the activity of the adenosine A_åA receptor in a subject, the method comprising administering an effective amount of a compound of any one of claims 1 to 67 or a

pharmaceutical composition of claim 68 to the subject.

74. A method for the treatment of a disorder related to the adenosine A2A receptor in a subject in need thereof, the method comprising administering an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68 to the subject.

75. A method for modulating the activity of the adenosine A2B receptor in a cell, the method comprising contacting a cell with an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68.

76. A method for modulating the activity of the adenosine A2B receptor in a subject, the method comprising administering an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68 to the subject.

77. A method for the treatment of a disorder related to the adenosine A2B receptor in a subject in need thereof, the method comprising administering an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68 to the subject.

78. A method of modulating an immune response in a subject in need thereof, the method comprising administering an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68 to the subject.

79. A method for modulating the activity of the dopamine D2 receptor in a cell, the method comprising contacting a ceil with an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68.

80. A method for modulating the activity of the dopamine D2 receptor in a subject, the method comprising administering an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68 to the subject.

81. A method of inhibiting inflammation in a subject in need thereof, the method comprising administering an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68 to the subject.

82. A method of treating cancer in a subject, the method comprising administering an effective amount of a compound of any one of claims 1 to 67 or a pharmaceutical composition of claim 68 to the subject.

83. The method of claim 82, wherein the method further comprises administering to the subject an additional anticancer therapy.

84. The method of claim 83, wherein the additional anticancer therapy is an immunotherapy, chemotherapeutic or cytotoxic agent, or radiotherapy.

85. The method of claim 84, wherein the immunotherapy is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, or adoptive T-cell transfer therapy.

86. The method of any one of claims 82 to 85, wherein the subject has a compromised immune system.

87. The method of any one of claims 82 to 86, wherein the cancer has failed to respond to a previously administered an immunotherapy.

88. The method of any one of claims 82 to 87, wherein the cancer is resistant to an

immunotherapy.

89. The method of any one of claims 83 to 88, wherein the additional anticancer therapy and the compound of any one of claims 1 to 67 or pharmaceutical composition of claim 68 are administered within 28 days of each other each in an amount that together are effective to treat the subject.