ENPP1 and / or ENPP3 inhibitors

Abstract

Compounds, compositions and methods are provided for the inhibition of ENPP1 and / or ENPP3.

Description

Attorney Docket No.: STAN-2201WO ENPP1 AND / OR ENPP3 INHIBITORS CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the priority benefit of United States Provisional Application No.63 / 646,591, filed May 13, 2024, the disclosure of which is incorporated herein by reference in its entirety. INTRODUCTION

[0002] Cyclic guanosine monophosphate-adenosine monophosphate (cGAMP) activates the Stimulator of Interferon Genes (STING) pathway, which is an important anti-cancer innate immune pathway. The cGAS-cGAMP-STING pathway gets activated in presence of cytoplasmic DNA either due to microbial infection or patho-physiological condition, including cancer and autoimmune disorder. Cyclic GMP-AMP synthase (cGAS) belongs to the nucleotidyltransferase family and is a universal DNA sensor that is activated upon binding to cytosolic dsDNA to produce the signaling molecule (2’-5’, 3’-5’) cyclic GMP- AMP (or 2′, 3′-cGAMP or cyclic guanosine monophosphate–adenosine monophosphate, cGAMP). Acting as a second messenger during microbial infection, 2′, 3′-cGAMP binds and activates STING, leading to production of type I interferon (IFN) and other co-stimulatory molecules that trigger the immune response. Besides its role in infectious disease, the STING pathway has emerged as a target for cancer immunotherapy and autoimmune diseases.

[0003] Ectonucleotide pyrophosphatase / phosphodiesterase 1 (ENPP1) is the dominant hydrolase of cGAMP that can degrade cGAMP. ENPP1 is a member of the ecto-nucleotide pyrophosphatase / phosphodiesterase (ENPP) family, and is a type II transmembrane glycoprotein comprising two identical disulfide-bonded subunits. ENPP1 has broad specificity to cleave a variety of substrates, including phosphodiester bonds of nucleotides and nucleotide sugars, and pyrophosphate bonds of nucleotides and nucleotide sugars. ENPP1 may function to hydrolyze nucleoside 5' triphosphates to their corresponding monophosphates and may also hydrolyze diadenosine polyphosphates. SUMMARY

[0004] Compounds, compositions and methods are provided for the inhibition of ENPP1. Aspects of the subject methods include contacting a sample with an ENPP1 inhibitor compound to inhibit the cGAMP hydrolysis activity of ENPP1. In some cases, the ENPP1 inhibitor compound is cell impermeable. ENPP1 inhibitor compounds can act extracellularlyAttorney Docket No.: STAN-2201WO to block the degradation of cGAMP. Also provided are pharmaceutical compositions and methods for treating cancer. Aspects of the methods include administering to a subject a therapeutically effective amount of an ENPP1 inhibitor to treat the subject for cancer. In certain cases, the cancer is a solid tumor cancer. Also provided are methods of administering radiation therapy to a subject in conjunction with administering an ENPP1 inhibitor to the subject. The radiation therapy can be administered in the subject methods at a dosage and / or frequency effective to reduce radiation damage to the subject, but still instigate an immune response.

[0005] These and other advantages and features of the disclosure will become apparent to those persons skilled in the art upon reading the details of the compositions and methods of use, which are more fully described below. DEFINITIONS

[0006] Before embodiments of the present disclosure are further described, it is to be understood that this disclosure is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.

[0007] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of embodiments of the present disclosure.

[0008] It must be noted that as used herein and in the appended claims, the singular forms “a”, “and”, and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a compound" includes not only a single compound but also a combination of two or more compounds, reference to "a substituent" includes a single substituent as well as two or more substituents, and the like.

[0009] In describing and claiming the present invention, certain terminology will be used in accordance with the definitions set out below. It will be appreciated that the definitions provided herein are not intended to be mutually exclusive. Accordingly, some chemical moieties may fall within the definition of more than one term.Attorney Docket No.: STAN-2201WO

[0010] The phrases “for example,” “for instance,” “such as,” or “including” are meant to introduce examples that further clarify more general subject matter. These examples are provided only as an aid for understanding the disclosure, and are not meant to be limiting in any fashion.

[0011] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

[0012] The terms "active agent," “antagonist”, "inhibitor", "drug" and "pharmacologically active agent" are used interchangeably herein to refer to a chemical material or compound which, when administered to an organism (human or animal) induces a desired pharmacologic and / or physiologic effect by local and / or systemic action.

[0013] The terms “treatment,” “treating,” and the like, refer to obtaining a desired pharmacologic and / or physiologic effect, such as reduction of tumor burden. The effect may be prophylactic in terms of completely or partially preventing a disease or symptom thereof and / or may be therapeutic in terms of a partial or complete cure for a disease and / or adverse effect attributable to the disease. “Treatment” is meant to cover any treatment of a disease in a mammal, particularly in a human, and includes: (a) preventing the disease or a symptom of a disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it (e.g., including diseases that may be associated with or caused by a primary disease (as in liver fibrosis that can result in the context of chronic HCV infection); (b) inhibiting the disease, i.e., arresting its development; and (c) relieving the disease, i.e., causing regression of the disease (e.g., reduction in of tumor burden).

[0014] The term “pharmaceutically acceptable salt” means a salt which is acceptable for administration to a patient, such as a mammal (salts with counterions having acceptable mammalian safety for a given dosage regime). Such salts can be derived from pharmaceutically acceptable inorganic or organic bases and from pharmaceutically acceptable inorganic or organic acids. “Pharmaceutically acceptable salt” refers to pharmaceutically acceptable salts of a compound, which salts are derived from a variety of organic and inorganic counter ions well known in the art and include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and whenAttorney Docket No.: STAN-2201WO the molecule contains a basic functionality, salts of organic or inorganic acids, such as hydrochloride, hydrobromide, formate, tartrate, besylate, mesylate, acetate, maleate, oxalate, and the like.

[0015] The terms “individual,” “host,” “subject,” and “patient” are used interchangeably herein, and refer to an animal, including, but not limited to, human and non-human primates, including simians and humans; rodents, including rats and mice; bovines; equines; ovines; felines; canines; and the like. "Mammal" means a member or members of any mammalian species, and includes, by way of example, canines; felines; equines; bovines; ovines; rodentia, etc. and primates, e.g., non-human primates, and humans. Non-human animal models, e.g., mammals, e.g. non-human primates, murines, lagomorpha, etc. may be used for experimental investigations.

[0016] The terms “determining,” “measuring,” “assessing,” and “assaying” are used interchangeably and include both quantitative and qualitative determinations.

[0017] The terms "polypeptide" and "protein", used interchangeably herein, refer to a polymeric form of amino acids of any length, which can include coded and non-coded amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides having modified peptide backbones. The term includes fusion proteins, including, but not limited to, fusion proteins with a heterologous amino acid sequence, fusions with heterologous and native leader sequences, with or without N-terminal methionine residues; immunologically tagged proteins; fusion proteins with detectable fusion partners, e.g., fusion proteins including as a fusion partner a fluorescent protein, β-galactosidase, luciferase, etc.; and the like.

[0018] The terms "nucleic acid molecule" and “polynucleotide" are used interchangeably and refer to a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or analogs thereof. Polynucleotides may have any three-dimensional structure, and may perform any function, known or unknown. Non-limiting examples of polynucleotides include a gene, a gene fragment, exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, control regions, isolated RNA of any sequence, nucleic acid probes, and primers. The nucleic acid molecule may be linear or circular.Attorney Docket No.: STAN-2201WO

[0019] A "therapeutically effective amount" or "efficacious amount" means the amount of a compound that, when administered to a mammal or other subject for treating a disease, condition, or disorder, is sufficient to effect such treatment for the disease, condition, or disorder. The "therapeutically effective amount" will vary depending on the compound, the disease and its severity and the age, weight, etc., of the subject to be treated.

[0020] The term “unit dosage form,” as used herein, refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of a compound (e.g., an aminopyrimidine compound, as described herein) calculated in an amount sufficient to produce the desired effect in association with a pharmaceutically acceptable diluent, carrier or vehicle. The specifications for unit dosage forms depend on the particular compound employed and the effect to be achieved, and the pharmacodynamics associated with each compound in the host.

[0021] The terms "pharmaceutically acceptable excipient," "pharmaceutically acceptable diluent," "pharmaceutically acceptable carrier," and "pharmaceutically acceptable adjuvant" refer to an excipient, diluent, carrier, or adjuvant that is useful in preparing a pharmaceutical composition that are generally safe, non-toxic and neither biologically nor otherwise undesirable, and include an excipient, diluent, carrier, and adjuvant that are acceptable for veterinary use as well as human pharmaceutical use. "A pharmaceutically acceptable excipient, diluent, carrier and adjuvant" as used in the specification and claims includes both one and more than one such excipient, diluent, carrier, and adjuvant.

[0022] The term "pharmaceutical composition" is meant to encompass a composition suitable for administration to a subject, such as a mammal, especially a human. In general a “pharmaceutical composition” is sterile, and preferably free of contaminants that are capable of eliciting an undesirable response within the subject (e.g., the compound(s) in the pharmaceutical composition is pharmaceutical grade). Pharmaceutical compositions can be designed for administration to subjects or patients in need thereof via a number of different routes of administration including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, intracheal, intramuscular, subcutaneous, and the like.

[0023] The phrase "having the formula" or "having the structure" is not intended to be limiting and is used in the same way that the term "comprising" is commonly used. The term "independently selected from" is used herein to indicate that the recited elements, e.g., R groups or the like, can be identical or different.Attorney Docket No.: STAN-2201WO

[0024] The terms “may,” "optional," "optionally," or “may optionally” mean that the subsequently described circumstance may or may not occur, so that the description includes instances where the circumstance occurs and instances where it does not. For example, the phrase "optionally substituted" means that a non-hydrogen substituent may or may not be present on a given atom, and, thus, the description includes structures wherein a non- hydrogen substituent is present and structures wherein a non-hydrogen substituent is not present.

[0025] “Acyl” refers to the groups H-C(O)-, alkyl-C(O)-, substituted alkyl-C(O)-, alkenyl- C(O)-, substituted alkenyl-C(O)-, alkynyl-C(O)-, substituted alkynyl-C(O)-, cycloalkyl-C(O)- , substituted cycloalkyl-C(O)-, cycloalkenyl-C(O)-, substituted cycloalkenyl-C(O)-, aryl- C(O)-, substituted aryl-C(O)-, heteroaryl-C(O)-, substituted heteroaryl-C(O)-, heterocyclyl- C(O)-, and substituted heterocyclyl-C(O)-, wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic are as defined herein. For example, acyl includes the “acetyl” group CH3C(O)-

[0026] The term "alkyl" refers to a branched or unbranched saturated hydrocarbon group (i.e., a mono-radical) typically although not necessarily containing 1 to about 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, octyl, decyl, and the like, as well as cycloalkyl groups such as cyclopentyl, cyclohexyl and the like. Generally, although not necessarily, alkyl groups herein may contain 1 to about 18 carbon atoms, and such groups may contain 1 to about 12 carbon atoms. The term "lower alkyl" intends an alkyl group of 1 to 6 carbon atoms. "Substituted alkyl" refers to alkyl substituted with one or more substituent groups, and this includes instances wherein two hydrogen atoms from the same carbon atom in an alkyl substituent are replaced, such as in a carbonyl group (i.e., a substituted alkyl group may include a -C(=O)- moiety). The terms "heteroatom-containing alkyl" and "heteroalkyl" refer to an alkyl substituent in which at least one carbon atom is replaced with a heteroatom, as described in further detail infra. If not otherwise indicated, the terms "alkyl" and "lower alkyl" include linear, branched, cyclic, unsubstituted, substituted, and / or heteroatom-containing alkyl or lower alkyl, respectively.

[0027] The term “substituted alkyl” is meant to include an alkyl group as defined herein wherein one or more carbon atoms in the alkyl chain have been optionally replaced with a heteroatom such as -O-, -N-, -S-, -S(O)n- (where n is 0 to 2), -NR- (where R is hydrogen orAttorney Docket No.: STAN-2201WO alkyl) and having from 1 to 5 substituents selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, - SO-aryl, -SO-heteroaryl, -SO2-alkyl, -SO2-aryl, -SO2-heteroaryl, and -NR’R”, wherein R’ and R” may be the same or different and are chosen from hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.

[0028] The term "alkenyl" refers to a linear, branched or cyclic hydrocarbon group of 2 to about 24 carbon atoms containing at least one double bond, such as ethenyl, n-propenyl, isopropenyl, n-butenyl, isobutenyl, octenyl, decenyl, tetradecenyl, hexadecenyl, eicosenyl, tetracosenyl, and the like. Generally, although again not necessarily, alkenyl groups herein may contain 2 to about 18 carbon atoms, and for example may contain 2 to 12 carbon atoms. The term "lower alkenyl" intends an alkenyl group of 2 to 6 carbon atoms. The term "substituted alkenyl" refers to alkenyl substituted with one or more substituent groups, and the terms "heteroatom-containing alkenyl" and "heteroalkenyl" refer to alkenyl in which at least one carbon atom is replaced with a heteroatom. If not otherwise indicated, the terms "alkenyl" and "lower alkenyl" include linear, branched, cyclic, unsubstituted, substituted, and / or heteroatom-containing alkenyl and lower alkenyl, respectively.

[0029] The term "alkynyl" refers to a linear or branched hydrocarbon group of 2 to 24 carbon atoms containing at least one triple bond, such as ethynyl, n-propynyl, and the like. Generally, although again not necessarily, alkynyl groups herein may contain 2 to about 18 carbon atoms, and such groups may further contain 2 to 12 carbon atoms. The term "lower alkynyl" intends an alkynyl group of 2 to 6 carbon atoms. The term "substituted alkynyl" refers to alkynyl substituted with one or more substituent groups, and the terms "heteroatom- containing alkynyl" and "heteroalkynyl" refer to alkynyl in which at least one carbon atom is replaced with a heteroatom. If not otherwise indicated, the terms "alkynyl" and "lower alkynyl" include linear, branched, unsubstituted, substituted, and / or heteroatom-containing alkynyl and lower alkynyl, respectively.

[0030] The term "alkoxy" refers to an alkyl group bound through a single, terminal ether linkage; that is, an "alkoxy" group may be represented as -O-alkyl where alkyl is as defined above. A "lower alkoxy" group refers to an alkoxy group containing 1 to 6 carbon atoms, andAttorney Docket No.: STAN-2201WO includes, for example, methoxy, ethoxy, n-propoxy, isopropoxy, t-butyloxy, etc. Substituents identified as "C1-C6 alkoxy" or "lower alkoxy" herein may, for example, may contain 1 to 3 carbon atoms, and as a further example, such substituents may contain 1 or 2 carbon atoms (i.e., methoxy and ethoxy). The designations “-OMe” and “MeO-” refer to a methoxy group.

[0031] The term “substituted alkoxy” refers to the groups substituted alkyl-O-, substituted alkenyl-O-, substituted cycloalkyl-O-, substituted cycloalkenyl-O-, and substituted alkynyl- O- where substituted alkyl, substituted alkenyl, substituted cycloalkyl, substituted cycloalkenyl and substituted alkynyl are as defined herein.

[0032] The term "aryl", unless otherwise specified, refers to an aromatic substituent generally, although not necessarily, containing 5 to 30 carbon atoms and containing a single aromatic ring or multiple aromatic rings that are fused together, directly linked, or indirectly linked (such that the different aromatic rings are bound to a common group such as a methylene or ethylene moiety). Aryl groups may, for example, contain 5 to 20 carbon atoms, and as a further example, aryl groups may contain 5 to 12 carbon atoms. For example, aryl groups may contain one aromatic ring or two or more fused or linked aromatic rings (i.e., biaryl, aryl-substituted aryl, etc.). Examples include phenyl, naphthyl, biphenyl, diphenylether, diphenylamine, benzophenone, and the like. "Substituted aryl" refers to an aryl moiety substituted with one or more substituent groups, and the terms "heteroatom- containing aryl" and "heteroaryl" refer to aryl substituent, in which at least one carbon atom is replaced with a heteroatom, as will be described in further detail infra. Aryl is intended to include stable cyclic, heterocyclic, polycyclic, and polyheterocyclic unsaturated C3-C14moieties, exemplified but not limited to phenyl, biphenyl, naphthyl, pyridyl, furyl, thiophenyl, imidazoyl, pyrimidinyl, and oxazoyl; which may further be substituted with one to five members selected from the group consisting of hydroxy, C1-C8 alkoxy, C1-C8 branched or straight-chain alkyl, acyloxy, carbamoyl, amino, N-acylamino, nitro, halogen, trifluoromethyl, cyano, and carboxyl (see e.g. Katritzky, Handbook of Heterocyclic Chemistry). If not otherwise indicated, the term "aryl" includes unsubstituted, substituted, and / or heteroatom-containing aromatic substituents.

[0033] The term "aralkyl" refers to an alkyl group with an aryl substituent, and the term "alkaryl" refers to an aryl group with an alkyl substituent, wherein "alkyl" and "aryl" are as defined above. In general, aralkyl and alkaryl groups herein contain 6 to 30 carbon atoms. Aralkyl and alkaryl groups may, for example, contain 6 to 20 carbon atoms, and as a further example, such groups may contain 6 to 12 carbon atoms.Attorney Docket No.: STAN-2201WO

[0034] The term "alkylene" refers to a di-radical alkyl group. Unless otherwise indicated, such groups include saturated hydrocarbon chains containing from 1 to 24 carbon atoms, which may be substituted or unsubstituted, may contain one or more alicyclic groups, and may be heteroatom-containing. "Lower alkylene" refers to alkylene linkages containing from 1 to 6 carbon atoms. Examples include, methylene (--CH2--), ethylene (--CH2CH2--), propylene (--CH2CH2CH2--), 2-methylpropylene (--CH2--CH(CH3)--CH2--), hexylene (-- (CH2)6--) and the like.

[0035] Similarly, the terms "alkenylene," "alkynylene," "arylene," "aralkylene," and "alkarylene" refer to di-radical alkenyl, alkynyl, aryl, aralkyl, and alkaryl groups, respectively.

[0036] The term "amino" refers to the group -NRR’ wherein R and R’ are independently hydrogen or nonhydrogen substituents, with nonhydrogen substituents including, for example, alkyl, aryl, alkenyl, aralkyl, and substituted and / or heteroatom-containing variants thereof.

[0037] The terms "halo" and "halogen" are used in the conventional sense to refer to a chloro, bromo, fluoro or iodo substituent.

[0038] “Carboxyl,” “carboxy” or “carboxylate” refers to –CO2H or salts thereof.

[0039] “Cycloalkyl” refers to cyclic alkyl groups of from 3 to 10 carbon atoms having single or multiple cyclic rings including fused, bridged, and spiro ring systems. Examples of suitable cycloalkyl groups include, for instance, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl and the like. Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantanyl, and the like.

[0040] The term “substituted cycloalkyl” refers to cycloalkyl groups having from 1 to 5 substituents, or from 1 to 3 substituents, selected from alkyl, substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, - SO2-substituted alkyl, -SO2-aryl and -SO2-heteroaryl.Attorney Docket No.: STAN-2201WO

[0041] The term "heteroatom-containing" as in a "heteroatom-containing alkyl group" (also termed a "heteroalkyl" group) or a "heteroatom-containing aryl group" (also termed a "heteroaryl" group) refers to a molecule, linkage or substituent in which one or more carbon atoms are replaced with an atom other than carbon, e.g., nitrogen, oxygen, sulfur, phosphorus or silicon, typically nitrogen, oxygen or sulfur. Similarly, the term "heteroalkyl" refers to an alkyl substituent that is heteroatom-containing, the term "heterocycloalkyl" refers to a cycloalkyl substituent that is heteroatom-containing, the terms "heterocyclic" or “heterocycle” refer to a cyclic substituent that is heteroatom-containing, the terms "heteroaryl" and "heteroaromatic" respectively refer to "aryl" and "aromatic" substituents that are heteroatom-containing, and the like. Examples of heteroalkyl groups include alkoxyaryl, alkylsulfanyl-substituted alkyl, N-alkylated amino alkyl, and the like. Examples of heteroaryl substituents include pyrrolyl, pyrrolidinyl, pyridinyl, quinolinyl, indolyl, furyl, pyrimidinyl, imidazolyl, 1,2,4-triazolyl, tetrazolyl, etc., and examples of heteroatom-containing alicyclic groups are pyrrolidino, morpholino, piperazino, piperidino, tetrahydrofuranyl, etc.

[0042] “Heteroaryl” refers to an aromatic group of from 1 to 15 carbon atoms, such as from 1 to 10 carbon atoms and 1 to 10 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur within the ring. Such heteroaryl groups can have a single ring (such as, pyridinyl, imidazolyl or furyl) or multiple condensed rings in a ring system (for example as in groups such as, indolizinyl, quinolinyl, benzofuran, benzimidazolyl or benzothienyl), wherein at least one ring within the ring system is aromatic, provided that the point of attachment is through an atom of an aromatic ring. In certain embodiments, the nitrogen and / or sulfur ring atom(s) of the heteroaryl group are optionally oxidized to provide for the N-oxide (N→O), sulfinyl, or sulfonyl moieties. This term includes, by way of example, pyridinyl, pyrrolyl, indolyl, thiophenyl, and furanyl. Unless otherwise constrained by the definition for the heteroaryl substituent, such heteroaryl groups can be optionally substituted with 1 to 5 substituents, or from 1 to 3 substituents, selected from acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substituted alkoxy, substituted alkenyl, substituted alkynyl, substituted cycloalkyl, substituted cycloalkenyl, amino, substituted amino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxyl, carboxylalkyl, cyano, halogen, nitro, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO2-alkyl, - SO2-substituted alkyl, -SO2-aryl and -SO2-heteroaryl, and trihalomethyl.Attorney Docket No.: STAN-2201WO

[0043] The terms “heterocycle,” “heterocyclic” and “heterocyclyl” refer to a saturated or unsaturated group having a single ring or multiple condensed rings, including fused bridged and spiro ring systems, and having from 3 to 15 ring atoms, including 1 to 4 hetero atoms. These ring heteroatoms are selected from nitrogen, sulfur and oxygen, wherein, in fused ring systems, one or more of the rings can be cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, provided that the point of attachment is through the non-aromatic ring. In certain embodiments, the nitrogen and / or sulfur atom(s) of the heterocyclic group are optionally oxidized to provide for the N-oxide, -S(O)-, or –SO2- moieties.

[0044] Examples of heterocycles and heteroaryls include, but are not limited to, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, dihydroindole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, phthalimide, 1,2,3,4-tetrahydroisoquinoline, 4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene, benzo[b]thiophene, morpholinyl, thiomorpholinyl (also referred to as thiamorpholinyl), 1,1-dioxothiomorpholinyl, piperidinyl, pyrrolidine, tetrahydrofuranyl, and the like.

[0045] Unless otherwise constrained by the definition for the heterocyclic substituent, such heterocyclic groups can be optionally substituted with 1 to 5, or from 1 to 3 substituents, selected from alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO-substituted alkyl, -SO-aryl, -SO- heteroaryl, -SO2-alkyl, -SO2-substituted alkyl, -SO2-aryl, -SO2-heteroaryl, and fused heterocycle.

[0046] "Hydrocarbyl" refers to univalent hydrocarbyl radicals containing 1 to about 30 carbon atoms, including 1 to about 24 carbon atoms, further including 1 to about 18 carbon atoms, and further including about 1 to 12 carbon atoms, including linear, branched, cyclic, saturated and unsaturated species, such as alkyl groups, alkenyl groups, aryl groups, and the like. A hydrocarbyl may be substituted with one or more substituent groups. The termAttorney Docket No.: STAN-2201WO "heteroatom-containing hydrocarbyl" refers to hydrocarbyl in which at least one carbon atom is replaced with a heteroatom. Unless otherwise indicated, the term "hydrocarbyl" is to be interpreted as including substituted and / or heteroatom-containing hydrocarbyl moieties.

[0047] By "substituted" as in "substituted hydrocarbyl," "substituted alkyl," "substituted aryl," and the like, as alluded to in some of the aforementioned definitions, is meant that in the hydrocarbyl, alkyl, aryl, or other moiety, at least one hydrogen atom bound to a carbon (or other) atom is replaced with one or more non-hydrogen substituents. Examples of such substituents include, without limitation, functional groups, and the hydrocarbyl moieties C1- C24 alkyl (including C1-C18 alkyl, further including C1-C12 alkyl, and further including C1-C6 alkyl), C2-C24alkenyl (including C2-C18alkenyl, further including C2-C12alkenyl, and further including C2-C6 alkenyl), C2-C24 alkynyl (including C2-C18 alkynyl, further including C2-C12 alkynyl, and further including C2-C6alkynyl), C5-C30aryl (including C5-C20aryl, and further including C5-C12 aryl), and C6-C30 aralkyl (including C6-C20 aralkyl, and further including C6- C12aralkyl). The above-mentioned hydrocarbyl moieties may be further substituted with one or more functional groups or additional hydrocarbyl moieties such as those specifically enumerated. Unless otherwise indicated, any of the groups described herein are to be interpreted as including substituted and / or heteroatom-containing moieties, in addition to unsubstituted groups.

[0048] “Sulfonyl” refers to the group SO2-alkyl, SO2-substituted alkyl, SO2-alkenyl, SO2- substituted alkenyl, SO2-cycloalkyl, SO2-substituted cylcoalkyl, SO2-cycloalkenyl, SO2- substituted cylcoalkenyl, SO2-aryl, SO2-substituted aryl, SO2-heteroaryl, SO2-substituted heteroaryl, SO2-heterocyclic, and SO2-substituted heterocyclic, wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, and substituted heterocyclic are as defined herein. Sulfonyl includes, by way of example, methyl-SO2-, phenyl-SO2-, and 4-methylphenyl-SO2-.

[0049] By the term “functional groups” is meant chemical groups such as halo, hydroxyl, sulfhydryl, C1-C24alkoxy, C2-C24alkenyloxy, C2-C24alkynyloxy, C5-C20aryloxy, acyl (including C2-C24 alkylcarbonyl (-CO-alkyl) and C6-C20 arylcarbonyl (-CO-aryl)), acyloxy (- O-acyl), C2-C24alkoxycarbonyl (-(CO)-O-alkyl), C6-C20aryloxycarbonyl (-(CO)-O-aryl), halocarbonyl (-CO)-X where X is halo), C2-C24 alkylcarbonato (-O-(CO)-O-alkyl), C6-C20 arylcarbonato (-O-(CO)-O-aryl), carboxy (-COOH), carboxylato (-COO-), carbamoyl (-(CO)- NH2), mono-substituted C1-C24 alkylcarbamoyl (-(CO)-NH(C1-C24 alkyl)), di-substitutedAttorney Docket No.: STAN-2201WO alkylcarbamoyl (-(CO)-N(C1-C24alkyl)2), mono-substituted arylcarbamoyl (-(CO)-NH-aryl), thiocarbamoyl (-(CS)-NH2), carbamido (-NH-(CO)-NH2), cyano (-C≡N), isocyano (-N+≡C-), cyanato (-O-C≡N), isocyanato (-O-N+≡C-), isothiocyanato (-S-C≡N), azido (-N=N+=N-), formyl (-(CO)-H), thioformyl (-(CS)-H), amino (-NH2), mono- and di-(C1-C24 alkyl)- substituted amino, mono- and di-(C5-C20aryl)-substituted amino, C2-C24alkylamido (-NH- (CO)-alkyl), C5-C20 arylamido (-NH-(CO)-aryl), imino (-CR=NH where R = hydrogen, C1- C24alkyl, C5-C20aryl, C6-C20alkaryl, C6-C20aralkyl, etc.), alkylimino (-CR=N(alkyl), where R = hydrogen, alkyl, aryl, alkaryl, etc.), arylimino (-CR=N(aryl), where R = hydrogen, alkyl, aryl, alkaryl, etc.), nitro (-NO2), nitroso (-NO), sulfo (-SO2-OH), sulfonato (-SO2-O-), C1-C24alkylsulfanyl (-S-alkyl; also termed "alkylthio"), arylsulfanyl (-S-aryl; also termed "arylthio"), C1-C24alkylsulfinyl (-(SO)-alkyl), C5-C20arylsulfinyl (-(SO)-aryl), C1-C24alkylsulfonyl (-SO2-alkyl), C5-C20 arylsulfonyl (-SO2-aryl), phosphono (-P(O)(OH)2), phosphonato (-P(O)(O-)2), phosphinato (-P(O)(O-)), phospho (-PO2), and phosphino (-PH2), mono- and di-(C1-C24 alkyl)-substituted phosphino, mono- and di-(C5-C20 aryl)-substituted phosphine. In addition, the aforementioned functional groups may, if a particular group permits, be further substituted with one or more additional functional groups or with one or more hydrocarbyl moieties such as those specifically enumerated above.

[0050] By "linking" or "linker" as in "linking group," "linker moiety," etc., is meant a linking moiety that connects two groups via covalent bonds. The linker may be linear, branched, cyclic or a single atom. Examples of such linking groups include alkyl, alkenylene, alkynylene, arylene, alkarylene, aralkylene, and linking moieties containing functional groups including, without limitation: amido (-NH-CO-), ureylene (-NH-CO-NH-), imide (-CO-NH- CO-) , epoxy (-O-), epithio (-S-), epidioxy (-O-O-), carbonyldioxy (-O-CO-O-), alkyldioxy (- O-(CH2)n-O-), epoxyimino (-O-NH-), epimino (-NH-), carbonyl (-CO-), etc. In certain cases, one, two, three, four or five or more carbon atoms of a linker backbone may be optionally substituted with a sulfur, nitrogen or oxygen heteroatom. The bonds between backbone atoms may be saturated or unsaturated, usually not more than one, two, or three unsaturated bonds will be present in a linker backbone. The linker may include one or more substituent groups, for example with an alkyl, aryl or alkenyl group. A linker may include, without limitations, poly(ethylene glycol) unit(s) (e.g., -(CH2-CH2-O)-); ethers, thioethers, amines, alkyls (e.g., (C1-C12)alkyl) , which may be straight or branched, e.g., methyl, ethyl, n-propyl, 1- methylethyl (iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), and the like. The linker backbone may include a cyclic group, for example, an aryl, a heterocycle or aAttorney Docket No.: STAN-2201WO cycloalkyl group, where 2 or more atoms, e.g., 2, 3 or 4 atoms, of the cyclic group are included in the backbone. A linker may be cleavable or non-cleavable. Any convenient orientation and / or connections of the linkers to the linked groups may be used.

[0051] When the term "substituted" appears prior to a list of possible substituted groups, it is intended that the term apply to every member of that group. For example, the phrase "substituted alkyl and aryl" is to be interpreted as "substituted alkyl and substituted aryl."

[0052] In addition to the disclosure herein, the term “substituted,” when used to modify a specified group or radical, can also mean that one or more hydrogen atoms of the specified group or radical are each, independently of one another, replaced with the same or different substituent groups as defined below.

[0053] In addition to the groups disclosed with respect to the individual terms herein, substituent groups for substituting for one or more hydrogens (any two hydrogens on a single carbon can be replaced with =O, =NR70, =N-OR70, =N2or =S) on saturated carbon atoms in the specified group or radical are, unless otherwise specified, -R60, halo, =O, -OR70, -SR70, -NR80R80, trihalomethyl, -CN, -OCN, -SCN, -NO, -NO2, =N2, -N3, -SO2R70, -SO2O–M+, -SO2OR70, -OSO2R70, -OSO2O–M+, -OSO2OR70, -P(O)(O–(substituted alkyl, cycloalkyl, heteroalkyl, heterocycloalkylalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl, each R70is independently hydrogen or R60; each R80is independently R70or alternatively, two R80’s, taken together with the nitrogen atom to which they are bonded, form a 5-, 6- or 7-membered heterocycloalkyl which may optionally include from 1 to 4 of the same or different additional heteroatoms selected from the group consisting of O, N and S, of which N may have -H or C1-C3 alkyl substitution; and each M+is a counter ion with a net single positive charge. Each M+may independently be, for example, an alkali ion, such as K+, Na+, Li+; an ammonium ion, such as+N(R60)4; or an alkaline earth ion, such as [Ca2+]0.5, [Mg2+]0.5, or [Ba2+]0.5(“subscript 0.5 means that one of the counter ions for such divalent alkali earth ions can be an ionized form of a compound of the invention and the other a typical counter ion such as chloride, or two ionized compounds disclosed herein can serve as counter ions for such divalent alkali earth ions, or a doubly ionized compound ofAttorney Docket No.: STAN-2201WO the invention can serve as the counter ion for such divalent alkali earth ions). As specific examples, -NR80R80is meant to include -NH2, -NH-alkyl, N-pyrrolidinyl, N-piperazinyl, 4N- methyl-piperazin-1-yl and N-morpholinyl.

[0054] In addition to the disclosure herein, substituent groups for hydrogens on unsaturated carbon atoms in “substituted” alkene, alkyne, aryl and heteroaryl groups are, unless otherwise specified, -R60, halo, -O-M+, -OR70, -SR70, -S–M+, -NR80R80, trihalomethyl, -CF3, -CN, -OCN, -SCN, -NO, -NO2, -N3, -SO2R70, -SO – 3 M+, -SO3R70, -OSO2R70, -OSO3–M+, -OSO3R70, -PO3-2(M+)2, -P(O)(OR70)O–M+, -P(O)(OR70) ,70 70 70 70– 2 -C(O)R , -C(S)R , -C(NR )R , -CO2 M+, -CO2R70, -C(S)OR70, -C(O)NR80R80, -C(NR70)NR80R80, -OC(O)R70, -OC(S)R70, -OCO2– M+, -OCO R70, -OC70 70 70 70 70 70– 2 (S)OR , -NR C(O)R , -NR C(S)R , -NR CO2 M+, -NR70CO2R70, -NR70C(S)OR70, -NR70C(O)NR80R80, -NR70C(NR70)R70and -NR70C(NR70)NR80R80, where R60, R70, R80and M+are as previously defined, provided that in case of substituted alkene or alkyne, the substituents are not -O-M+, -OR70, -SR70, or -S–M+.

[0055] In addition to the groups disclosed with respect to the individual terms herein, substituent groups for hydrogens on nitrogen atoms in “substituted” heteroalkyl and cycloheteroalkyl groups are, unless otherwise specified, -R60, -O-M+, -OR70, -SR70, -S-M+, -NR80R80, trihalomethyl, -CF3, -CN, -NO, -NO2, -S(O)2R70, -S(O)2O-M+, -S(O)2OR70, -OS(O)2R70, -OS( O)2O-M+, -OS(O)2OR70, -P(O)(O-)2(M+)2, -P(O)(OR70)O-M+, -P(O)(OR70)(OR70), -C(O)R70, - C(S)R70, -C(NR70)R70, -C(O)OR70, -C(S)OR70, -C(O)NR80R80, -C(NR70)NR80R80, -OC(O)R70, -OC(S)R70, -OC(O)OR70, -OC(S)OR70, -NR70C(O)R70, -NR70C(S)R70, -NR70C(O)OR70, -NR70C(S)OR70, -NR70C(O)NR80R80, -NR70C(NR70)R70and -NR70C(NR70)NR80R80, where R60, R70, R80and M+are as previously defined.

[0056] In addition to the disclosure herein, in a certain embodiment, a group that is substituted has 1, 2, 3, or 4 substituents, 1, 2, or 3 substituents, 1 or 2 substituents, or 1 substituent.

[0057] Unless indicated otherwise, the nomenclature of substituents that are not explicitly defined herein are arrived at by naming the terminal portion of the functionality followed by the adjacent functionality toward the point of attachment. For example, the substituent “arylalkyloxycarbonyl” refers to the group (aryl)-(alkyl)-O-C(O)-.Attorney Docket No.: STAN-2201WO

[0058] As to any of the groups disclosed herein which contain one or more substituents, it is understood, of course, that such groups do not contain any substitution or substitution patterns which are sterically impractical and / or synthetically non-feasible. In addition, the subject compounds include all stereochemical isomers arising from the substitution of these compounds.

[0059] In certain embodiments, a substituent may contribute to optical isomerism and / or stereo isomerism of a compound. Salts, solvates, hydrates, and prodrug forms of a compound are also of interest. All such forms are embraced by the present disclosure. Thus the compounds described herein include salts, solvates, hydrates, prodrug and isomer forms thereof, including the pharmaceutically acceptable salts, solvates, hydrates, prodrugs and isomers thereof. In certain embodiments, a compound may be a metabolized into a pharmaceutically active derivative.

[0060] Unless otherwise specified, reference to an atom is meant to include isotopes of that atom. For example, reference to H is meant to include1H,2H (i.e., D) and3H (i.e., T), and reference to C is meant to include12C and all isotopes of carbon (such as13C).

[0061] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present invention. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.

[0062] While the apparatus and method has or will be described for the sake of grammatical fluidity with functional explanations, it is to be expressly understood that the claims, unless expressly formulated under 35 U.S.C. §112, are not to be construed as necessarily limited in any way by the construction of "means" or "steps" limitations, but are to be accorded the full scope of the meaning and equivalents of the definition provided by the claims under the judicial doctrine of equivalents, and in the case where the claims are expressly formulated under 35 U.S.C. §112 are to be accorded full statutory equivalents under 35 U.S.C. §112.

[0063] Definitions of other terms and concepts appear throughout the detailed description.Attorney Docket No.: STAN-2201WO DETAILED DESCRIPTION

[0064] As summarized above, aspects of the present disclosure include compounds, compositions and methods for the inhibition of ENPP1 and / or ENPP3. Aspects of the methods include contacting a sample with an ENPP1 and / or ENPP3 inhibitor compound to inhibit cGAMP hydrolysis activity of ENPP1 and / or ENPP3. These compounds, compositions and methods find use in a variety of applications in which inhibition of ENPP1 and / or ENPP3 is desired.

[0065] Also provided are pharmaceutical compositions and methods for treating cancer using the subject ENPP1 and / or ENPP3 inhibitor compounds. Aspects of the methods include administering to a subject a therapeutically effective amount of an ENPP1 and / or ENPP3 inhibitor compound to inhibit the hydrolysis of cGAMP and treat the subject for cancer. Compounds

[0066] In one aspect, the invention provides a compound, or a salt thereof, of the invention. In an exemplary embodiment, the invention provides a compound, or a salt thereof, according to a Formula described herein. In an exemplary embodiment, the invention provides a compound, or a salt thereof, described herein.

[0067] In one aspect, the invention provides a compound, or a salt thereof, having a structure of Formula (I): wherein Q is selected from the wherein each R1and R2are each independentlysubstituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, and that R1and R2, along with the nitrogen to which they are attached, can be optionally attached to form a substituted or unsubstituted 3-Attorney Docket No.: STAN-2201WO to 15- membered ring system; Rais selected from the group ,Ry,consisting of -NR3R4, -OR3, and R3and that Rband Rc, along with the atoms to which they are attached, can be optionally attached to form a substituted or unsubstituted 5- to 8- membered ring system, wherein each R3and R4are each independently selected from the group consisting of H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl and that R3and R4, along with the nitrogen to which they are attached, can be optionally attached to form a substituted or unsubstituted 3- to 15- membered ring system. In an exemplary embodiment, Rais , wherein Rxis as described herein. In an exemplaryembodiment, Rais selected from the group ,and ; wherein Rx, Ry, and Rzare each independently selected from the group of H and substituted or unsubstituted alkyl.

[0068] In an exemplary embodiment, the compound, or a salt thereof, has a structure of Formula (I), wherein Q is selected from the group consisting of -NR1R2, -OR1, and R1, wherein each R1and R2are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, and that R1and R2, along with the nitrogen to which they are attached, can be optionally attached to form a substituted orAttorney Docket No.: STAN-2201WO unsubstituted 3- to 15- membered ring system; Rais selected from the group consisting of ; H andthe group consisting of -NR3R4, -OR3, and R3, and that Rband Rc, along with the atoms to which they are attached, can be optionally attached to form a substituted or unsubstituted 5- to 8- membered ring system, each R3and R4are each independently selected from the group consisting of H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, and that R3and R4, along with the nitrogen to which they are attached, can be optionally attached to form a substituted or unsubstituted 3- to 15- membered ring system. In an exemplary embodiment, Rb, Rc, and Rdare each independently selected from the group consisting of H and substituted or unsubstituted C1-C6 alkoxy. In an exemplary embodiment, Rdis H. In an exemplary embodiment, Q is H.

[0069] In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Raand Q are as described herein, and each Rb, Rc, and Rdare independently selected from the group consisting of H, halogen, substituted or unsubstituted C1-C6 alkoxy, -NR3R4, wherein R3and R4are each independently selected from H, substituted or unsubstituted C1-C6 alkyl, and substituted or unsubstituted 3- to 7- membered heterocycloalkyl, or R3and R4, along with the nitrogen to which they are attached, are optionally attached to form a substituted or unsubstituted 3- to 15- membered ring system. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Raand Q are as described herein, and each Rb, Rc, and Rdare independently selected from the group consisting of H, halogen, substituted or unsubstituted C1-C6alkoxy, -NH2, -NH(substituted or unsubstituted C1-C6alkyl), -N(substituted or unsubstituted C1-C6 alkyl)2, azetidinyl, -NH(azetidinyl), substituted or unsubstituted piperazinyl, substituted or unsubstituted morpholinyl, -NH(1,3-dihydroxyisopropyl), substituted or unsubstituted 2,7-diazaspiro[4.4]nonane, -NH(tetrahydrothiophene 1,1- dioxide), substituted or unsubstituted pyridinyl, substituted or unsubstituted pyrimidinyl, and -C(O)OH. In an exemplary embodiment, the compound, or a salt thereof, has a structureAttorney Docket No.: STAN-2201WO according to a formula herein, wherein Raand Q are as described herein, and each Rb, Rc, and Rdare independently selected from the group consisting of H, halogen, substituted or unsubstituted C1-C6alkoxy, -NH2, -NH(substituted or unsubstituted C1-C6alkyl), - N(substituted or unsubstituted C1-C6 alkyl)2, substituted or unsubstituted 5- and 6-membered heteroaryl, and -C(O)OH. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Ra, Rc, Rd, and Q are as described herein, Rbis substituted or unsubstituted 5- and 6-membered heteroaryl. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Ra, Rc, Rd, and Q are as described herein, Rbis substituted or unsubstituted pyridinyl or pyrimidinyl. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Raand Q are as described herein, and each Rb, Rc, and Rdare independently selected from the group consisting of H, Br, -OCH3, -NH2, -NH(CH3), - N(CH3)2, azetidinyl, -NH(azetidinyl), piperazinyl, morpholinyl, -NH(1,3- dihydroxyisopropyl), 2,7-diazaspiro[4.4]nonane, -NH(tetrahydrothiophene 1,1-dioxide), pyridinyl, pyrimidinyl, and -C(O)OH. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Raand Q are as described herein, and each Rb, Rc, and Rdis H or Br. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Raand Q are as described herein, and each Rb, Rc, and Rdis H or -OCH3. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Raand Q are as described herein, and each Rb, Rc, and Rdis Br or -OCH3. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Raand Q are as described herein, and each Rb, Rc, and Rdis -NH2or -OCH3. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Raand Q are as described herein, and each Rb, Rc, and Rdis -N(CH3)2or -OCH3. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Raand Q are as described herein, and each Rb, Rc, and Rdis - NH(CH3) or -OCH3. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Raand Q are as described herein, and each . In an exemplary embodiment, the compound, or a saltto a formula herein, wherein Raand Q are as describedAttorney Docket No.: STAN-2201WO herein, and each Rb, Rc, and Rdis H or . In an exemplary embodiment, the compound, or a salt thereof, has a to a formula herein, wherein Raand Qare as described herein, and each Rb, Rc, and Rdis H . In an exemplary embodiment, the compound, or a salt thereof, has a to a formula herein,wherein Raand Q are as described herein, and each Rb, Rc, and Rdis H or -C(O)OH.

[0070] In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Ra, Rb, Rc, and Rdare as described herein, and Q is H, substituted or unsubstituted morpholino, substituted or unsubstituted piperazinyl, substituted or unsubstituted pyrazolyl, substituted or unsubstituted triazolyl, or substituted or unsubstituted imidazolyl, substituted or unsubstituted -NH(azetidinyl), substituted or unsubstituted -NH(pyridinyl), substituted or unsubstituted -NH(methylpyrazolyl-substituted C1-C3 alkyl), substituted or unsubstituted -NH(oxetanyl), substituted or unsubstituted -N(C1- C3alkyl)2, substituted or unsubstituted di-(C1-C3alkyl) amino (C1-C3alkyl)2N-, and substituted or unsubstituted -NH(acetamide). In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Ra, Rb, Rc, and Rdare ,of Formula , wherein Ra, Rb, Rc, and Q are as described herein. In anor a salt thereof, has a structure according to a formula herein, wherein Rband Rcare each independently H or OR3, and Ra, R3, and Q are as described herein. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Rband Rcare each independently H or OR3,Attorney Docket No.: STAN-2201WO R3is H or substituted C1-C6alkyl (e.g. substituted C1-C3alkyl), and Raand Q are as described herein. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Rband Rcare each independently H or OR3, R3is H or unsubstituted C1-C6 alkyl (e.g. unsubstituted C1-C3 alkyl), and Raand Q are as described herein. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Rbis methoxy, and Ra, Rc, and Q are as described herein. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Rcis methoxy, and Ra, Rb, and Q are as described herein. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Rbis methoxy, Rcis methoxy, and Raand Q are as described herein. In an exemplary embodiment, for any of the embodiments in this Q are as described herein. In an exemplaryembodiment, for any of the embodiments in this as described herein.

[0072] In an exemplary embodiment, the compound, or a salt thereof, has a structureaccording to , wherein Rb, Rc, Rd, Q, Rx, and Ry are as describedherein. In ancompound, or a salt thereof, has a structure according to a formula herein, wherein Rxis H, Ryis H, and Rb, Rc, Rd, and Q are as described herein.

[0073] In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Ra, Rb, Rc, and Rdare as described herein, and Q is - NR1R2, wherein each R1and R2are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted orAttorney Docket No.: STAN-2201WO unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, and that R1and R2, along with the nitrogen to which they are attached, can be optionally attached to form a substituted or unsubstituted 3- to 15- membered ring system. In an exemplary embodiment, the compound, or a salt thereof, has a structure according to a formula herein, wherein Ra, Rb, Rc, and Rdare as described herein, and Q is -NR1R2, wherein R1and R2, along with the nitrogen to which they are attached, are optionally attached to form a substituted or unsubstituted 3- to 15- membered ring system.

[0074] In an exemplary embodiment, the compound, or a salt thereof, has a structure according to , wherein Rb, Rc, Q, Rx, and Ryare as described herein. In an compound, or a salt thereof, has a structureaccording to a formula herein, wherein Rxis H, Ryis H, and Rb, Rc, Rd, and Q are as described herein. In an exemplary embodiment, the compound has a structure of Formula that R1and R2are not both H. In an exemplaryor unsubstituted heterocycloalkyl. In an exemplary embodiment, R1is H, and R2is substituted or unsubstituted azetidinyl. In an exemplary embodiment, R1and R2, along with the nitrogen to which they are attached, are attached to form a substituted or unsubstituted 5- to 6- membered ring system. In an exemplary embodiment, R1and R2, along with the nitrogen to which they are attached, form a substituted or unsubstituted morpholino, substituted or unsubstituted piperazinyl, substituted or unsubstituted pyrazolyl, substituted or unsubstituted triazolyl, or substituted or unsubstituted imidazolyl. In an exemplary embodiment, R1and R2, along with the nitrogenAttorney Docket No.: STAN-2201WO to which they are attached, form a substituted or unsubstituted triazolyl, or substituted or unsubstituted imidazolyl. In an exemplary embodiment, the compound has a structure which HO O HO O HO P P orherein, or a prodrug thereof (e.g., as described herein), or a pharmaceutically acceptable salt thereof.

[0076] Aspects of the present disclosure include ENPP1 and / or ENPP3 inhibitor compounds (e.g., as described herein), salts thereof (e.g., pharmaceutically acceptable salts), and / or solvate, hydrate and / or prodrug forms thereof. In addition, it is understood that, in any compound described herein having one or more chiral centers, if an absolute stereochemistry is not expressly indicated, then each center may independently be of R-configuration or S- configuration or a mixture thereof. It will be appreciated that all permutations of salts, solvates, hydrates, prodrugs and stereoisomers are meant to be encompassed by the present disclosure.

[0077] In some embodiments, the subject ENPP1 and / or ENPP3 inhibitor compounds, or a prodrug form thereof, are provided in the form of pharmaceutically acceptable salts. Compounds containing an amine or nitrogen containing heteroaryl group may be basic in nature and accordingly may react with any number of inorganic and organic acids to form pharmaceutically acceptable acid addition salts. Acids commonly employed to form such salts include inorganic acids such as hydrochloric, hydrobromic, hydriodic, sulfuric andAttorney Docket No.: STAN-2201WO phosphoric acid, as well as organic acids such as para-toluenesulfonic, methanesulfonic, oxalic, para- bromophenylsulfonic, carbonic, succinic, citric, benzoic and acetic acid, and related inorganic and organic acids. Such pharmaceutically acceptable salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-l,4- dioate, hexyne-l,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephathalate, sulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β- hydroxybutyrate, glycollate, maleate, tartrate, methanesulfonate, propanesulfonates, naphthalene- 1 -sulfonate, naphthalene-2-sulfonate, mandelate, hippurate, gluconate, lactobionate, and the like salts. In certain specific embodiments, pharmaceutically acceptable acid addition salts include those formed with mineral acids such as hydrochloric acid and hydrobromic acid, and those formed with organic acids such as fumaric acid and maleic acid.

[0078] In some embodiments, the subject compounds are provided in a prodrug form. “Prodrug” refers to a derivative of an active agent that requires a transformation within the body to release the active agent. In certain embodiments, the transformation is an enzymatic transformation. Prodrugs are frequently, although not necessarily, pharmacologically inactive until converted to the active agent. “Promoiety” refers to a form of protecting group that, when used to mask a functional group within an active agent, converts the active agent into a prodrug. In some cases, the promoiety will be attached to the drug via bond(s) that are cleaved by enzymatic or non-enzymatic means in vivo. Any convenient prodrug forms of the subject compounds can be prepared, e.g., according to the strategies and methods described by Rautio et al. (“Prodrugs: design and clinical applications”, Nature Reviews Drug Discovery 7, 255-270 (February 2008)). In some cases, the promoiety is attached to a hydrophilic head group of the subject compounds. In some cases, the promoiety is attached to a hydroxy or carboxylic acid group of the subject compounds. In certain cases, the promoiety is an acyl or substituted acyl group. In certain cases, the promoiety is an alkyl or substituted alkyl group, e.g., that forms an ester functional group when attached to a hydrophilic head group of the subject compounds, e.g., a phosphonate ester, a phosphate ester, etc.Attorney Docket No.: STAN-2201WO

[0079] In some embodiments, the subject compound is a phosphonate ester or phosphate ester prodrug that can be transformed to a compound including a phosphonic acid or phosphonate, or a phosphate head group.

[0080] In some embodiments, the subject compounds, prodrugs, stereoisomers or salts thereof are provided in the form of a solvate (e.g., a hydrate). The term "solvate" as used herein refers to a complex or aggregate formed by one or more molecules of a solute, e.g. a prodrug or a pharmaceutically-acceptable salt thereof, and one or more molecules of a solvent. Such solvates are typically crystalline solids having a substantially fixed molar ratio of solute and solvent. Representative solvents include by way of example, water, methanol, ethanol, isopropanol, acetic acid, and the like. When the solvent is water, the solvate formed is a hydrate. Methods of Inhibiting ENPP1 and / or ENPP3

[0081] In another aspect, the invention provides a method of inhibiting ENPP1 and / or ENPP3, comprising: contacting the ENPP1 and / or the ENPP3 with a compound described herein, thereby inhibiting ENPP1 and / or ENPP3. In an exemplary embodiment, the compound is cell impermeable. In an exemplary embodiment, the compound is cell permeable. In an exemplary embodiment, the ENPP1 and / or the ENPP3 is in a cellular sample. In an exemplary embodiment, the sample comprises cGAMP. In an exemplary embodiment, the cGAMP levels are elevated in the cellular sample (e.g., relative to a control sample not contacted with the ENPP1 inhibitor).

[0082] In another aspect, the invention provides a method of treating an ENPP1 and / or ENPP3 associated disease in a mammal in need of treatment thereof, comprising: administering to the mammal a therapeutically effective amount of a compound described herein, thereby treating the ENPP1 and / or ENPP3 associated disease. In an exemplary embodiment, the ENPP1 and / or ENPP3 associated disease is cancer. In an exemplary embodiment, the cancer is selected from the group consisting of adrenal, liver, kidney, bladder, breast, colon, gastric, ovarian, cervical, uterine, esophageal, colorectal, prostate, pancreatic, lung (both small cell and non-small cell), thyroid, carcinomas, sarcomas, glioblastomas, melanoma and head and neck tumors. In an exemplary embodiment, the cancer is breast cancer. In an exemplary embodiment, the cancer is lymphoma. In an exemplary embodiment, the invention provides a compound described herein for use inAttorney Docket No.: STAN-2201WO treating cancer. In an exemplary embodiment, the invention provides the use of a compound described herein in the manufacture of a medicament for treating cancer.

[0083] In another aspect, the invention provides a method of modulating an immune response in a subject, the method comprising: administrating to a subject a therapeutically effective amount of a compound described herein to treat the subject for an inflammatory condition.

[0084] As summarized above, aspects of the present disclosure include ENPP1 and / or ENPP3 inhibitors, and methods of inhibition using the same. ENPP1 and / or ENPP3 are members of the ecto-nucleotide pyrophosphatase / phosphodiesterase (ENPP) family. As such, aspects of the subject methods include inhibition of the hydrolase activity of ENPP1 and / or ENPP3 against cGAMP. The inventors discovered that cGAMP can have significant extracellular biological functions, which can be enhanced by blocking extracellular degradation of cGAMP, e.g., hydrolysis by its degradation enzyme ENPP1 and / or ENPP3. In certain instances, the ENPP1 target of inhibition is extracellular, and the subject ENPP1 inhibiting compounds are cell-impermeable, and thus are not capable of diffusion into cells. As such, the subject methods can provide for selective extracellular inhibition of ENPP1’s hydrolase activity and increased extracellular levels of cGAMP. As such, in some cases, the ENPP1 and / or ENPP3 inhibiting compounds are compounds that inhibit the activity of ENPP1 and / or ENPP3 extracellularly. Experiments conducted by the inventors indicate that inhibiting the activity of ENPP1 and / or ENPP3 increases extracellular cGAMP and may consequently boost the STING pathway.

[0085] By inhibiting an ENPP1 and / or ENPP3 it is meant that the activity of the enzyme is decreased by 10% or more, such as 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more (e.g., relative to a control in any convenient in vitro inhibition assay). In some cases, inhibiting an ENPP1 and / or ENPP3 means decreasing the activity of the enzyme by a factor of 2 or more, such as 3 or more, 5 or more, 10 or more, 100 or more, or 1000 or more, relative to its normal activity (e.g., relative to a control as measured by any convenient assay).

[0086] In some cases, the method is a method of inhibiting ENPP1 and / or ENPP3 in a sample. The term “sample” as used herein relates to a material or mixture of materials, typically, although not necessarily, in fluid form, containing one or more components of interest.Attorney Docket No.: STAN-2201WO

[0087] In some embodiments, there is provided a method of inhibiting ENPP1 and / or ENPP3, the method comprising contacting a sample with a cell impermeable ENPP1 and / or ENPP3 inhibitor to inhibit cGAMP hydrolysis activity of ENPP1 and / or ENPP3. In some cases, the sample is a cellular sample. In some cases, the sample comprises cGAMP. In certain cases, the cGAMP levels are elevated in the cellular sample (e.g., relative to a control sample not contacted with the inhibitor). The subject methods can provide for increased levels of cGAMP. By “increased level of cGAMP” is meant a level of cGAMP in a cellular sample contacted with a subject compound, where the cGAMP level in the sample is increased by 10% or more, such as 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more, or even more, relative to a control sample that is not contacted with the agent.

[0088] In certain embodiments the ENPP1 and / or ENPP3 inhibitor is an inhibitor as defined herein.

[0089] In some embodiments, the subject compounds have an ENPP1 and / or ENPP3 inhibition profile that reflects activity against additional enzymes. In some embodiments, the subject compounds specifically inhibit ENPP1 and / or ENPP3 without undesired inhibition of one or more other enzymes.

[0090] In some embodiments, the compounds of the disclosure interfere with the interaction of cGAMP and ENPP1 and / or ENPP3. For example, the subject compounds may act to increase the extracellular cGAMP by inhibiting the hydrolase activity of ENPP1 and / or ENPP3 against cGAMP. Without being bound to any particular theory, it is thought that increasing extracellular cGAMP activates the STING pathway.

[0091] In some embodiments, the subject compounds inhibit ENPP1 and / or ENPP3, as determined by an inhibition assay, e.g., by an assay that determines the level of activity of the enzyme either in a cell-free system or in a cell after treatment with a subject compound, relative to a control, by measuring the IC50or EC50value, respectively. In certain embodiments, the subject compounds have an IC50 value (or EC50 value) of 10 µM or less, such as 3 µM or less, 1 µM or less, 500 nM or less, 300 nM or less, 200nM or less, 100 nM or less, 50 nM or less, 30 nM or less, 10 nM or less, 5 nM or less, 3 nM or less, 1 nM or less, or even lower.

[0092] As summarized above, aspects of the disclosure include methods of inhibiting ENPP1 and / or ENPP3. A subject compound (e.g., as described herein) may inhibit at an activity ofAttorney Docket No.: STAN-2201WO ENPP1 and / or ENPP3 in the range of 10% to 100%, e.g., by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more. In certain assays, a subject compound may inhibit its target with an IC50of 1 x 10-6M or less (e.g., 1 x 10-6M or less, 1 x 10-7M or less, 1 x 10-8M or less, 1 x 10-9M or less, 1 x 10-10M or less, or 1 x 10-11M or less).

[0093] The protocols that may be employed in determining ENPP1 and / or ENPP3 activity are numerous, and include but are not limited to cell-free assays, e.g., binding assays; assays using purified enzymes, cellular assays in which a cellular phenotype is measured, e.g., gene expression assays; and in vivo assays that involve a particular animal (which, in certain embodiments may be an animal model for a condition related to the target pathogen).

[0094] In some embodiments, the subject method is an in vitro method that includes contacting a sample with a subject compound that specifically inhibits ENPP1 and / or ENPP3. In certain embodiments, the sample is suspected of containing ENPP1 and / or ENPP3 and the subject method further comprises evaluating whether the compound inhibits ENPP1 and / or ENPP3.

[0095] In certain embodiments, the subject compound is a modified compound that includes a label, e.g., a fluorescent label, and the subject method further includes detecting the label, if present, in the sample, e.g., using optical detection.

[0096] In certain embodiments, the compound is modified with a support or with affinity groups that bind to a support (e.g. biotin), such that any sample that does not bind to the compound may be removed (e.g., by washing). The specifically bound ENPP1, if present, may then be detected using any convenient means, such as, using the binding of a labeled target specific probe, or using a fluorescent protein reactive reagent.

[0097] In another embodiment of the subject method, the sample is known to contain ENPP1 and / or ENPP3.

[0098] In some embodiments, the method is a method of reducing cancer cell proliferation, where the method includes contacting the cell with an effective amount of a subject ENPP1 inhibitor compound (e.g., as described herein) to reduce cancer cell proliferation. In certain cases, the subject ENPP1 and / or ENPP3 inhibitor compounds can act intracellularly.Attorney Docket No.: STAN-2201WO Methods of Treatment

[0099] Aspects of the present disclosure include methods for inhibiting the hydrolase activity of ENPP1 and / or ENPP3 against cGAMP provides for increased levels of cGAMP and / or downstream modulation (e.g., activation) of the STING pathway. The inventors have discovered that cGAMP can be present in the extracellular space and that ENPP1 and / or ENPP3 can control extracellular levels of cGAMP. The inventors have also discovered that cGAMP can have significant extracellular biological functions in vivo. The results described and demonstrated herein indicate that ENPP1 and / or ENPP3 inhibition according to the subject methods can modulate STING activity in vivo, and thus find use in the treatment of a variety of diseases, e.g., as a target for cancer immunotherapy. As such, the subject methods can provide for selective extracellular inhibition of ENPP1 and / or ENPP3 activity (e.g., hydrolase activity of cGAMP) to increase extracellular levels of cGAMP and activate the stimulator of interferon genes (STING) pathway. In some instances, the subject method is a method for increasing a STING mediated response in a subject. In some instances, the subject method is a method for modulating an immune response in a subject.

[0100] A “STING mediated response” refers to any response that is mediated by STING, including, but not limited to, immune responses, e.g., to bacterial pathogens, viral pathogens, and eukaryotic pathogens. See, e.g., Ishikawa et al. Immunity 29: 538-550 (2008); Ishikawa et al. Nature 461: 788-792 (2009); and Sharma et al. Immunity 35: 194-207 (2011). STING also functions in certain autoimmune diseases initiated by inappropriate recognition of self DNA (see, e.g., Gall et al. Immunity 36: 120-131 (2012), as well as for the induction of adaptive immunity in response to DNA vaccines (see, e.g., Ishikawa et al. Nature 461: 788- 792 (2009). By increasing a STING mediated response in a subject is meant an increase in a STING mediated response in a subject as compared to a control subject (e.g., a subject who is not administered a subject compound). In some cases, the subject is human and the subject compounds and methods provide for activation of human STING. In some cases, the STING mediated response includes modulation of an immune response. In some instances, the subject method is a method of modulating an immune response in a subject.

[0101] In some cases, the STING mediated response includes increasing the production of an interferon (e.g., a type I interferon (IFN), type III interferon (IFN)) in a subject. Interferons (IFNs) are proteins having a variety of biological activities, e.g., antiviral, immunomodulating and antiproliferative. IFNs are relatively small, species-specific, single chain polypeptides, produced by mammalian cells in response to exposure to a variety ofAttorney Docket No.: STAN-2201WO inducers such as viruses, polypeptides, mitogens and the like. Interferons protect animal tissues and cells against viral attack and are an important host defense mechanism. Interferons may be classified as Type-I, Type-II and Type-III interferons. Mammalian Type-I interferons of interest include IFN-α (alpha), IFN-β (beta), IFN-κ (kappa), IFN-δ (delta), IFN- ε (epsilon), IFN-τ (tau), IFN-ω (omega), and IFN-ζ (zeta, also known as limitin).

[0102] Interferons find use in the treatment of a variety of cancers since these molecules have anti-cancer activity that acts at multiple levels. Interferon proteins can directly inhibit the proliferation of human tumor cells. In some cases, the anti-proliferative activity is also synergistic with a variety of approved chemotherapeutic agents such as cisplatin, 5FU and paclitaxel. The immunomodulatory activity of interferon proteins can also lead to the induction of an anti-tumor immune response. This response includes activation of NK cells, stimulation of macrophage activity and induction of MHC class I surface expression, leading to the induction of anti-tumor cytotoxic T lymphocyte activity. In addition, interferons play a role in cross-presentation of antigens in the immune system. Moreover, some studies further indicate that IFN-β protein may have anti-angiogenic activity. Angiogenesis, new blood vessel formation, is critical for the growth of solid tumors. IFN-β may inhibit angiogenesis by inhibiting the expression of pro-angiogenic factors such as bFGF and VEGF. Interferon proteins may also inhibit tumor invasiveness by modulating the expression of enzymes, such as collagenase and elastase, which are important in tissue remodeling.

[0103] Aspects of the methods include administering to a subject with cancer a therapeutically effective amount of an ENPP1 and / or ENPP3 inhibitor to treat the subject for cancer. In some instances, the subject is one who is diagnosed with or suspected of having cancer. Any convenient ENPP1 and / or ENPP3 inhibitors can be used in the subject methods of treating cancer. In certain cases, the ENPP1 and / or ENPP3 inhibitor compound is a compound as described herein. In certain cases, the ENPP1 and / or ENPP3 inhibitor is a cell impermeable compound. In certain cases, the ENPP1 and / or ENPP3 inhibitor is a cell permeable compound. In certain cases, the cancer is a solid tumor cancer. In certain embodiments, the cancer is selected from adrenal, liver, kidney, bladder, breast, colon, gastric, ovarian, cervical, uterine, esophageal, colorectal, prostate, pancreatic, lung (both small cell and non-small cell), thyroid, carcinomas, sarcomas, glioblastomas, melanoma and various head and neck tumors. In some cases, the cancer is breast cancer. In some embodiments, the cancer is lymphoma.Attorney Docket No.: STAN-2201WO

[0104] Aspects of the methods include administering to a subject a therapeutically effective amount of a cell impermeable ENPP1 and / or ENPP3 inhibitor to inhibit the hydrolysis of cGAMP and treat the subject for cancer. In certain cases the cancer is a solid tumor cancer. In certain embodiments, the cancer is selected from adrenal, liver, kidney, bladder, breast, colon, gastric, ovarian, cervical, uterine, esophageal, colorectal, prostate, pancreatic, lung (both small cell and non-small cell), thyroid, carcinomas, sarcomas, glioblastomas, melanoma and various head and neck tumors. In certain embodiments, the cancer is breast cancer. In some instances, the cancer is lymphoma.

[0105] In an exemplary embodiment, the ENPP1 and / or ENPP3 associated disease is a calcification disorder (e.g. pseudoxanthoma elasticum (PXE), generalized arterial calcification of infancy (GACI), calcific aortic valve disease (CAVD), calcium pyrophosphate dihydrate deposition (CPPD) disease), disease associated with a viral infection, atherosclerosis-related disorder (e.g. myocardial infarction (MI), renal arteriosclerosis, retinopathy), and / or fibrotic disease.

[0106] As such, aspects of the method include contacting a sample with a subject compound (e.g., as described above) under conditions by which the compound inhibits ENPP1. Any convenient protocol for contacting the compound with the sample may be employed. The particular protocol that is employed may vary, e.g., depending on whether the sample is in vitro or in vivo. For in vitro protocols, contact of the sample with the compound may be achieved using any convenient protocol. In some instances, the sample includes cells that are maintained in a suitable culture medium, and the complex is introduced into the culture medium. For in vivo protocols, any convenient administration protocol may be employed. Depending upon the potency of the compound, the cells of interest, the manner of administration, the number of cells present, various protocols may be employed.

[0107] In some embodiments, the subject method is a method of treating a subject for cancer. In some embodiments, the subject method includes administering to the subject an effective amount of a subject compound (e.g., as described herein) or a pharmaceutically acceptable salt thereof. The subject compound may be administered as part of a pharmaceutical composition (e.g., as described herein).

[0108] In some embodiments, an “effective amount” is an amount of a subject compound that, when administered to an individual in one or more doses, in monotherapy or in combination therapy, is effective to inhibit ENPP1 and / or ENPP3 by about 20% (20%Attorney Docket No.: STAN-2201WO inhibition), at least about 30% (30% inhibition), at least about 40% (40% inhibition), at least about 50% (50% inhibition), at least about 60% (60% inhibition), at least about 70% (70% inhibition), at least about 80% (80% inhibition), or at least about 90% (90% inhibition), compared to the ENPP1 and / or ENPP3 activity in the individual in the absence of treatment with the compound, or alternatively, compared to the ENPP1 and / or ENPP3 activity in the individual before or after treatment with the compound.

[0109] In some embodiments, a “therapeutically effective amount” is an amount of a subject compound that, when administered to an individual in one or more doses, in monotherapy or in combination therapy, is effective to decrease tumor burden in the subject by about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%, compared to tumor burden in the individual in the absence of treatment with the compound, or alternatively, compared to the tumor burden in the subject before or after treatment with the compound. As used herein the term “tumor burden” refers to the total mass of tumor tissue carried by a subject with cancer.

[0110] In some embodiments, a “therapeutically effective amount” is an amount of a subject compound that, when administered to an individual in one or more doses, in monotherapy or in combination therapy, is effective to reduce the dose of radiotherapy required to observe tumor shrinkage in the subject by about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%, compared to the dose of radiotherapy required to observe tumor shrinkage in the individual in the absence of treatment with the compound.

[0111] In some embodiments, a “therapeutically effective amount” of a compound is an amount that, when administered in one or more doses to an individual having cancer, is effective to achieve a 1.5-log, a 2-log, a 2.5-log, a 3-log, a 3.5-log, a 4-log, a 4.5-log, or a 5- log reduction in tumor size.

[0112] In some embodiments, a single dose of a compound is administered. In other embodiments, multiple doses are administered. Where multiple doses are administered over a period of time, the compound can be administered twice daily (qid), daily (qd), every other day (qod), every third day, three times per week (tiw), or twice per week (biw) over a period of time. For example, a compound is administered qid, qd, qod, tiw, or biw over a period of from one day to about 2 years or more. For example, a compound is administered at any ofAttorney Docket No.: STAN-2201WO the aforementioned frequencies for one week, two weeks, one month, two months, six months, one year, or two years, or more, depending on various factors.

[0113] Administration of a therapeutically effective amount of a subject compound to an individual with cancer can result in one or more of: 1) a reduction in tumor burden; 2) a reduction in the dose of radiotherapy required to effect tumor shrinkage; 3) a reduction in the spread of a cancer from one cell to another cell in an individual; 4) a reduction of morbidity or mortality in clinical outcomes; 5) shortening the total length of treatment when combined with other anti-cancer agents; and 6) an improvement in an indicator of disease response (e.g., a reduction in one or more symptoms of cancer). Any of a variety of methods can be used to determine whether a treatment method is effective. For example, a biological sample obtained from an individual who has been treated with a subject method can be assayed.

[0114] Any of the compounds described herein can be utilized in the subject methods of treatment. In some cases, the compound that is utilized in the subject methods is not cell permeable. In some cases, the compound that is utilized in the subject methods has poor cell permeability.

[0115] In some embodiments, the compound specifically inhibits ENPP1 and / or ENPP3. In some embodiments, the compound modulates the activity of cGAMP. In some embodiments, the compound interferes with the interaction of ENPP1 and / or ENPP3 and cGAMP. In some embodiments, the compound results in activation of the STING pathway.

[0116] In some embodiments, the subject is mammalian. In certain instances, the subject is human. Other subjects can include domestic pets (e.g., dogs and cats), livestock (e.g., cows, pigs, goats, horses, and the like), rodents (e.g., mice, guinea pigs, and rats, e.g., as in animal models of disease), as well as non-human primates (e.g., chimpanzees, and monkeys). The subject may be in need of treatment for cancer. In some instances, the subject methods include diagnosing cancer, including any one of the cancers described herein. In some embodiments, the compound is administered as a pharmaceutical preparation.

[0117] In certain embodiments, the ENPP1 and / or ENPP3 inhibitor compound is a modified compound that includes a label, and the method further includes detecting the label in the subject. The selection of the label depends on the means of detection. Any convenient labeling and detection systems may be used in the subject methods, see e.g., Baker, “The whole picture,” Nature, 463, 2010, p977-980. In certain embodiments, the compound includes a fluorescent label suitable for optical detection. In certain embodiments, theAttorney Docket No.: STAN-2201WO compound includes a radiolabel for detection using positron emission tomography (PET) or single photon emission computed tomography (SPECT). In some cases, the compound includes a paramagnetic label suitable for tomographic detection. The subject compound may be labeled, as described above, although in some methods, the compound is unlabeled and a secondary labeling agent is used for imaging. Utility

[0118] The compounds and methods of the invention, e.g., as described herein, find use in a variety of applications. Applications of interest include, but are not limited to: research applications and therapeutic applications. Methods of the invention find use in a variety of different applications including any convenient application where inhibition of ENPP1 is desired.

[0119] The subject compounds and methods find use in a variety of research applications. The subject compounds and methods may be used in the optimization of the bioavailability and metabolic stability of compounds.

[0120] The subject compounds and methods find use in a variety of therapeutic applications. Therapeutic applications of interest include those applications in cancer treatment. As such, the subject compounds find use in the treatment of a variety of different conditions in which the inhibition and / or treatment of cancer in the host is desired. For example, the subject compounds and methods may find use in treating a solid tumor cancer (e.g., as described herein). Pharmaceutical Compositions

[0121] In another aspect, the invention provides a pharmaceutical composition comprising: a) the compound, or a pharmaceutically acceptable salt thereof, described herein; and b) one or more pharmaceutically acceptable excipients. In an exemplary embodiment, the pharmaceutical composition is for use in treating cancer.

[0122] The herein-discussed compounds can be formulated using any convenient excipients, reagents and methods. Compositions are provided in formulation with a pharmaceutically acceptable excipient(s). A wide variety of pharmaceutically acceptable excipients are known in the art and need not be discussed in detail herein. Pharmaceutically acceptable excipients have been amply described in a variety of publications, including, for example, A. Gennaro (2000) “Remington: The Science and Practice of Pharmacy,” 20th edition, Lippincott, Williams, & Wilkins; Pharmaceutical Dosage Forms and Drug Delivery Systems (1999) H.C.Attorney Docket No.: STAN-2201WO Ansel et al., eds., 7thed., Lippincott, Williams, & Wilkins; and Handbook of Pharmaceutical Excipients (2000) A.H. Kibbe et al., eds., 3rded. Amer. Pharmaceutical Assoc.

[0123] The compounds of the invention can be administered orally, subcutaneously, intramuscularly, parenterally, or other route. The compounds of the invention may be administered by the same route of administration or by different routes of administration. The compounds of the invention can be administered by any suitable means including, but not limited to, for example, oral, rectal, nasal, topical (including transdermal, aerosol, buccal and sublingual), vaginal, parenteral (including subcutaneous, intramuscular, intravenous and intradermal), intravesical or injection into an affected organ.

[0124] The subject compounds may be administered in a unit dosage form and may be prepared by any methods well known in the art. Such methods include combining the subject compound with a pharmaceutically acceptable carrier or diluent which constitutes one or more accessory ingredients. A pharmaceutically acceptable carrier is selected on the basis of the chosen route of administration and standard pharmaceutical practice. Each carrier must be "pharmaceutically acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject. This carrier can be a solid or liquid and the type is generally chosen based on the type of administration being used.

[0125] Any drug delivery device or system that provides for the dosing regimen of the instant disclosure can be used. A wide variety of delivery devices and systems are known to those skilled in the art. EXAMPLES

[0126] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use embodiments of the present disclosure, and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.

[0127] While the present invention has been described with reference to the specific embodiments thereof, it should be understood by those skilled in the art that various changesAttorney Docket No.: STAN-2201WO may be made and equivalents may be substituted without departing from the true spirit and scope of the invention. In addition, many modifications may be made to adapt a particular situation, material, composition of matter, process, process step or steps, to the objective, spirit and scope of the present disclosure. All such modifications are intended to be within the scope of the claims appended hereto. Example 1: Synthesis of Compounds

[0128] Compounds may be synthesized using any convenient method. Methods which can be adapted for use in preparing compounds of this disclosure includes the exemplary synthetic methods described by Li et al. in PCT application Nos. PCT / US2020 / 015968, filed January 30, 2020, and PCT / US2018 / 050018, filed September 7, 2018, the disclosure of each of which is herein incorporated by reference in its entirety. Many general references providing commonly known chemical synthetic schemes and conditions useful for synthesizing the disclosed compounds are also available (see, e.g., Smith and March, March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Fifth Edition, Wiley-Interscience, 2001; or Vogel, A Textbook of Practical Organic Chemistry, Including Qualitative Organic Analysis, Fourth Edition, New York: Longman, 1978). Reactions may be monitored by thin layer chromatography (TLC), LC / MS and reaction products characterized by LC / MS and1H NMR. Intermediates and final products may be purified by silica gel chromatography or by HPLC.

[0129] A general synthesis procedure for compounds of the invention is provided below:Attorney Docket No.: STAN-2201WO Intermediates a)

[0130] (4-(((6,7-Dimethoxy-3- methyl)phenyl)methanol. To a suspension of 4-chloro-6,7-(1000 mg, 1 Eq, 3.72 mmol) and N- ethyl-N-isopropylpropan-2-amine (2.88 g, 6 Eq, 22.3 mmol)in isopropanol (100 mL), (4- (aminomethyl)phenyl)methanol (1.53 g, 3 Eq, 11.2 mmol) was added portion-wise and the mixture was stirred at room temperature for 4 days with occasional sonication. Progress of the reaction was monitored by HPLC. Once completed, the solvent was evaporated to obtain a bright-yellow residue. It was used in the next step without purification. ESI+ = 369.95.

[0131] (4-(((3-Amino-6,7- methyl)phenyl)methanol. To asuspension of crude (4-(((6,7- 4-yl)amino)methyl)phenyl)methanol (1.38 g, 1 Eq, 3.72 mmol) in MeOH (60 mL) was added palladium on carbon (396 mg, 10% Wt, .1 Eq, 370 μmol) and the mixture evacuated and filled with hydrogen gas through a balloon (3x) and the mixture was stirred at 25 °C for 6 hour. After the reaction was complete, hydrogen gas was removed by bubbling nitrogen gas through the reaction mixture and it was filtered under positive nitrogen pressure through a pad of celite to remove the palladium. Crude reaction mixture was evaporated to dryness and used in the next step without further purification. ESI+ = 340.15.Attorney Docket No.: STAN-2201WO

[0132] (4-((7,8-Dimethoxy-1H- 1-yl)methyl)phenyl)methanol. To a solution of (4-(((3-amino-6,7-amino)methyl)phenyl)methanol (1.26 g, 1 Eq, 3.71 mmol) in ethanol (10 mL) was added triethoxymethane (2.00 g, 2 mL, 4.0 Eq, 0.01 mol) and the mixture was stirred at 110 °C for 3 hour. After the reaction was complete, the solvent was evaporated and the crude reaction mixture was purified by flash column chromatography (SiO2, 100% hexanes to 90% EtOAc in hexanes) to obtain the product (1.10 g, 85% over 3 steps) as a white solid. ESI+ = 350.10.

[0133] 1-(4-(Bromomethyl)benzyl)- imidazo[4,5-c]quinoline. To asolution of (4-((7,8-dimethoxy-1H- 1-yl)methyl)phenyl)methanol (600 mg, 1.0 Eq, 1.72 mmol) in dry DCM (30 mL) was added tribromophosphane (465 mg, 1.72 mL, 1 molar, 1 Eq, 1.72 mmol) and the mixture was stirred at 0 °C for 2 hour. The reaction mixture was warmed to room temperature and stirred for an additional 2 hours. After the reaction was complete, the solvent was evaporated and the crude mixture was purified by flash column chromatography (SiO2, 100% DCM to 10% MeOH in DCM) to obtain the product as a brown semisolid (468 mg, 66%). ESI+ 412.00, 413.95.

[0134] Diethyl (4-((7,8-Dimethoxy-c]quinolin-1- yl)methyl)benzyl)phosphonate. A solution of 1-(4-(bromomethyl)benzyl)-7,8-dimethoxy-1H-Attorney Docket No.: STAN-2201WO imidazo[4,5-c]quinoline (1200 mg, 1 Eq, 2.911 mmol) in triethyl phosphite (4.836 g, 10.0 Eq, 29.11 mmol) was stirred at 100 °C for 2 hour. After the reaction was complete, the reaction mixture was cooled to 0 °C, and the precipitate was filtered to obtain the crude product. It was used in the next step without purification. ESI+ 470.10. EtO O EtO P N

[0135] 1-(4-((Diethoxyphosphoryl) dimethoxy-1H-imidazo[4,5- c]quinoline 5-Oxide. A solution of1H-imidazo[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonate (180 mg, 1 Eq, 383 μmol) in anhydrous DCM (20 mL) was cooled to 0 °C and 3-chlorobenzoperoxoic acid (220 mg, 77% Wt, 2.56 Eq, 982 μmol) was added slowly, and the mixture was slowly warmed to room temperature and stirred for 20 hour. After the reaction was complete, the volatiles were evaporated and the product was purified by reverse phase flash chromatography (C18, 95% water in ACN to 100% ACN) to obtain the product as a yellow solid (126 mg, 68%). ESI+ 486.00. b) General procedure for the synthesis of diethyl phosphonate substituted quinolines “General Procedure b”

[0136] General procedure for thequinolines. To a solution of 1-(4- ((diethoxyphosphoryl)methyl)benzyl)-7,8-dimethoxy-1H-imidazo[4,5-c]quinoline 5-oxide (10 mg, 1 Eq, 21 μmol), N-ethyl-N-isopropylpropan-2-amine (21 mg, 8 Eq, 0.16 mmol), and amine (3.1 mg, 1.2 Eq, 25 μmol) in DCM (1 mL) was added bromotri(1-pyrrolidinyl) phosphonium hexafluorophosphate (38 mg, 4 Eq, 82 μmol) and the mixture was stirred at 25 °C for 72 hour. After the reaction was complete, DCM was evaporated, and the crude reaction mixture was purified by HPLC to obtain the product.Attorney Docket No.: STAN-2201WO

[0137] Diethyl (4-((7,8- imidazo[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonate.the product was obtained (5 mg, 45%) as a white solid. ESI+ 554.96.

[0138] tert-Butyl 4-(1-(4-( benzyl)-7,8-dimethoxy-1H- imidazo[4,5-c]quinolin-4-yl)general procedure, the product was obtained (11 mg, 63%) as a white solid. ESI+ 654.08.

[0139] Diethyl (4-((7,8- 1-yl)-1H-imidazo[4,5-c]quinolin-1-yl)methyl)benzyl)phosphonate. Following general procedure, the product was obtained (7 mg, 60%) as a white solid. ESI+ 536.91.Attorney Docket No.: STAN-2201WO

[0140] Diethyl (4-((4-(1H- 1H-imidazo[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonate.the product was obtained (6 mg, 50%) as a white solid. ESI+ 536.01.

[0141] tert-Butyl 3-((1-(4-( benzyl)-7,8-dimethoxy-1H- imidazo[4,5-c]quinolin-4-yl)Following general procedure, the product was obtained (7 mg, 50%) as a white solid. ESI+ 640.07. Compounds HO O P

[0142] General procedure for theacid from diethyl phosphonate. To a solution of diethylphosphonate esater (1 Eq) in DCM (1 mL) at 0 °C, was added bromotrimethylsilane (3 Eq), and the mixture was slowly warmed to room temperature and stirred for 20 hour. After the completion, the volatiles were evaporated, and the crude mixture was purified by HPLC.Attorney Docket No.: STAN-2201WO Compound 1

[0143] (4-((7,8-Dimethoxy-4- [4,5-c]quinolin-1-yl)methyl)benzyl)phosphonic Acid. procedure, the product was obtained (3 mg, 40%) as a white solid. ESI+ 498.95 ESI- 497.15.1H NMR (500 MHz, DMSO) δ 8.30 (s, 1H), 7.19 – 7.14 (m, 2H), 7.11 (s, 1H), 7.08 (s, 1H), 6.97 (d, J = 7.9 Hz, 2H), 5.82 (s, 2H), 4.11 (t, J = 4.7 Hz, 4H), 3.79 (m, 7H), 3.58 (s, 3H), 2.77 (d, J = 21.1 Hz, 2H). HO O HO P Compound 2NH

[0144] (4-((7,8-Dimethoxy-4-[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonic Acid. Following general procedure, the product was obtained (2 mg, 40%) as a white solid. ESI+ 498.00, ESI- 496.10. Compound 3

[0145] (4-((7,8-Dimethoxy-4-(1H-1H-imidazo[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonic Acid. Following general procedure, the product was obtained (6 mg, 60%) as a white solid. ESI+ 480.95, ESI- 478.90.1H NMR (500 MHz, DMSO) δ 9.80 (s, 1H), 8.69 (s, 1H), 8.36 (s, 1H), 7.52 (s, 1H), 7.30 (s, 1H), 7.21 (dd, J = 8.3, 2.3 Hz, 2H), 7.09 (d, J = 7.9 Hz, 2H), 6.03 (s, 2H), 3.90 (s, 3H), 3.69 (s, 3H), 2.86 (d, J = 21.3 Hz, 2H).Attorney Docket No.: STAN-2201WO Compound 4

[0146] (4-((4-(1H-Imidazol-1-yl)- [4,5-c]quinolin-1-yl)methyl)benzyl)phosphonic Acid. the product was obtained (2 mg, 40%) as a white solid. ESI+ 479.95.1H NMR (500 MHz, DMSO) δ 9.20 (s, 1H), 8.66 (d, J= 5 Hz 1H), 8.46 (d, J= 5 Hz 1H), 7.45 (s, 1H), 7.26 (s, 1H), 7.20 (m, 3H), 7.06 (d, J = 8 Hz, 2H), 5.99 (s, 2H), 3.87 (s, 3H), 3.73 (s, 3H), 2.86 (d, J = 20 Hz, 2H). [4,5-c]quinolin-1-yl)methyl)benzyl)phosphonic Acid. Following general procedure, the product was obtained (2 mg, 40%) as a white solid. ESI+ 484.00. ESI- 482.20. Compound 6

[0148] 4-((7,8-Dimethoxy-1H-1-yl)methyl)benzyl dihydrogen Phosphate. To a stirred solution of (4-((7,8-dimethoxy-1H-imidazo[4,5-c]quinolin-1- yl)methyl)phenyl)methanol (1 Eq) in anhydrous THF at -40 °C can be added diphosphoryl chloride (2 Eq), and the resulting mixture can be stirred at -40 °C for 20 min. The reaction can be quenched with water and treated with saturated sodium bicarbonate solution until pH ~ 8 keeping the mixture under stirring for an hour at room temperature. The solution can beAttorney Docket No.: STAN-2201WO extracted with ethyl acetate; the aqueous phase can be made acidic using a 1 N HCI solution and extracted several times with ethyl acetate. The combined organic phases can be washed with brine, dried over sodium sulfate, and concentrated to give 4-((7,8-dimethoxy-1H- imidazo[4,5-c]quinolin-1-yl)methyl)benzyl dihydrogen phosphate. Compound 7

[0149] (4-((7,8-Dimethoxy- quinolin-1-yl)methyl)benzyl)phosphonothioic O,O-Acid. To a solution of compound diethylphosphonate (1 eq) in toluene (100 mL) can be added Lawesson’s reagent (1.0 eq) under nitrogen atmosphere. The mixture can be stirred at 110 °C for 16 h. The progress of the reaction mixture can be monitored by TLC. Then the mixture can be filtered and concentrated under reduced pressure. The residue can be purified by silica gel chromatography to afford O, O-diethyl phosphonothioate. To a solution of O,O-diethyl phosphonothioate (1.5 eq) in MeCN(0.1 L) can be added DBU (5.0 eq). After being stirred at 0 °C for 10 mm, 1-(4- (bromomethyl)benzyl)-7,8-dimethoxy-1H-imidazo[4,5-c]quinoline (1.0 eq) can be added slowly. The mixture can be allowed to warm to room temperature and stirred for 1 h. The progress of the reaction mixture can be monitored by TLC. Then the mixture can be concentrated under reduced pressure and the residue can be purified by silica gel chromatography to afford O,O-diethyl (4-((7,8-dimethoxy-1H-imidazo[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonothioate.

[0150] A solution of O,O-diethyl (4-((7,8-dimethoxy-1H-imidazo[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonothioate (1.0 eq) DCM (10 mL) at 0 °C, can be added bromotrimethylsilane (3 Eq), and the mixture can be slowly warmed to room temperature and^stirred for 20 hour. After the completion, the volatiles can be evaporated, and the crude mixture can be purified by HPLC.Attorney Docket No.: STAN-2201WO Compound 8

[0151] P-(4-((7,8- quinolin-1-yl)methyl)benzyl) (2 equivalents) and triphenyl phosphine (2 Eq) can be combined in anhydrous DCM at 0°C and allowed to stir to room temperature for 30 minutes. Separately, (4-((7,8-dimethoxy-1H-imidazo[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonic acid (1 Eq) and ammonia (2 Eq) can be dissolved in anhydrous DCM and then added dropwise to the betaine solution. The reaction can be allowed to stir for 30 minutes. The crude reaction mixture can be then purified by HPLC. Compound 9

[0152] 2-(4-((7,8-Dimethoxy-1H-1-yl)methyl)phenyl)acetic Acid. A solution of 1-(4-(bromomethyl)benzyl)-7,8-dimethoxy-1H-imidazo[4,5-c]quinoline (1 Eq) in THF can be cooled to 0 °C and nBuLi (1.5 Eq) can be added slowly and stirred for 1 h. Then CO2can be bubbled through the solution for 30 min and the solution can be warmed to room temperature. The reaction mixture can be acidified with 1N HCl and the solvent can be evaporated. The product can be purified by HPLC.

[0153] ESI- 376.2.1H NMR (500 MHz, DMSO) δ 9.05 (s, 1H), 8.53 (s, 1H), 7.50 (s, 1H), 7.25 – 7.18 (m, 3H), 7.08 (d, J = 8.2 Hz, 2H), 5.99 (s, 2H), 3.88 (s, 3H), 3.65 (s, 3H), 3.51 (s, 2H).Attorney Docket No.: STAN-2201WO- in THF can be cooled to 0 °C and nBuLi (1.5 Eq) can be added slowly and stirred for 1 h. Then trimethoxyborane (1 Eq) can be added to the solution and stirred for 1 h at room temperature. The reaction mixture can be acidified with 1N HCl and the solvent can be evaporated. The product can be purified by HPLC.

[0155] Step 1: A solution of 1-(4-(bromomethyl)benzyl)-7,8-dimethoxy-1H-imidazo[4,5- c]quinoline (140 mg, 1 Eq, 340 μmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2- dioxaborolane) (129 mg, 1.5 Eq, 509 μmol), and potassium carbonate (141 mg, 3 Eq, 1.02 mmol) in 1,4-Dioxane (5 mL) was bubbled with nitrogen for 10 min followed by addition of tetrakis(triphenylphosphine)palladium(0) (39.2 mg, .1 Eq, 34.0 μmol) and the mixture was stirred at 100 °C for 16 hour in a sealed vial. After the reaction was complete, the solvent was evaporated and crude mixture (in DCM) was purified in TEA deactivated silica by wet loading in using flash chromatography (100% DCM to 20% MeOH in DCM) to obtain the product as brown liquid (60 mg, 38%).

[0156] Step 2: A solution of 7,8-dimethoxy-1-(4-((4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)methyl)benzyl)-1H-imidazo[4,5-c]quinoline (60 mg, 1 Eq, 0.13 mmol), methylboronic acid (200 mg, 26 Eq, 3.34 mmol), and hydrogen chloride (7 mg, 2 mL, 0.1 molar, 2 Eq, 0.2 mmol) in Acetone (2 mL) was stirred at 25 °C for 2 hour. After the reaction was complete, the solvent was evaporated and crude mixture was purified by HPLC to obtain the product as brown liquid (6 mg, 10%). ESI+ 378.2.1H NMR (500 MHz, DMSO) δ 9.30 (s, 1H), 8.67 (d, J = 1.9 Hz, 1H), 7.52 (s, 1H), 7.26 (s, 1H), 7.04 – 7.00 (m, 2H), 6.96 – 6.90 (m, 2H), 5.93 (s, 2H), 3.85 (s, 3H), 3.63 (s, 3H), 1.96 (s, 2H).Attorney Docket No.: STAN-2201WO phenyl)-N-in methanol (10 mL) was added to a stirred solution of hydroxylamine hydrochloride (94 g, 1.4 mmol) in methanol (5 mL)at 0°C. The reaction mixture was stirred at 0°C for 30 min. The resultant precipitate was removed by filtration. The filtrate was collected to provide a free hydroxylamine solution.50 mL of this solution was added to methyl 2-(4-((7,8-dimethoxy- 1H-imidazo[4,5-c]quinolin-1-yl)methyl)phenyl)acetate (20 mg, 1 Eq, 51 μmol) solution in Methanol (20 mL) at 0 °C. The reaction mixture was slowly warmed to room temperature and stirred for 2 h. A precipitate was formed. Then the reaction was chilled to -20 °C overnight for complete precipitation. The precipitate was collected by filtration as the pure product (7 mg, 30%). ESI+ 393.2.1H NMR (500 MHz, DMSO) δ 10.58 (d, J = 1.6 Hz, 1H), 9.05 (s, 1H), 8.76 (s, 1H), 8.53 (s, 1H), 7.49 (s, 1H), 7.25 – 7.19 (m, 3H), 7.08 (d, J = 8.1 Hz, 2H), 5.98 (s, 2H), 3.88 (s, 3H), 3.67 (s, 3H), 3.23 (s, 2H). Compound 12

[0158] Diethyl (4-((7,8-4-yloxy)-1H-imidazo[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonate. Following the general procedure, 1-(4- ((diethoxyphosphoryl)methyl)benzyl)-7,8-dimethoxy-1H-imidazo[4,5-c]quinoline 5-oxide (1 Eq) can be treated with N-ethyl-N-isopropylpropan-2-amine (3 Eq), and piperidin-4-ol (1.2 Eq)^in DCM (1 mL) can be added bromotri(1-pyrrolidinyl) phosphonium hexafluorophosphate (2 Eq) and the reaction mixture can be stirred 20 h at room temperature. After the reaction is complete, solvent can be evaporated and the residue can be purified by HPLC.Attorney Docket No.: STAN-2201WO Compound 13

[0159] (4-((7,8-Dimethoxy- -1H-imidazo[4,5-c]quinolin-1-yl)methyl)benzyl) a of diethyl (4-((7,8-dimethoxy-4- (piperidin-4-yloxy)-1H-imidazo[4,5-c]quinolin-1-yl)methyl)benzyl)phosphonate (1 Eq) in DCM (1 mL) at 0 °C, can be added bromotrimethylsilane (3 Eq), and the mixture can be slowly warmed to room temperature and^stirred for 20 hours. After the completion, the volatiles can be evaporated, and the crude mixture can be purified by HPLC. Compound 14

[0160] Diethyl (4-((4-imidazo[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonate. A solution of 1-(4-((diethoxyphosphoryl)methyl)benzyl)-7,8- dimethoxy-1H-imidazo[4,5-c]quinoline 5-oxide (1 Eq) in anhydrous DCM was treated with phosphorus oxybromide (1.2 Eq) at 0 °C and it can be stirred for 20 h at room temperature. After the reaction is complete, solvent can be evaporated and the residue can be purified by flash chromatography.

[0161] Diethyl (4-((7,8-Dimethoxy-4-(piperidin-4-yl)-1H-imidazo[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonate. To a solution of diethyl (4-((4-bromo-7,8-dimethoxy-1H- imidazo[4,5-c]quinolin-1-yl)methyl)benzyl)phosphonate (1 Eq), piperidine-4-boronic acid pinacol ester hydrochloride (1 Eq), and potassium carbonate (2.01 Eq) in acetonitrile (5 mL) and Water (1 mL) can be bubbled with nitrogen for 10 minutes followed by the addition of 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride (0.1 Eq) and the mixture can be stirred at 90 °C for 15 hour. The reaction mixture can be extracted with EtOAc (3×5 mL), dried over magnesium sulfate, and purified through flash column chromatography to obtain the product.Attorney Docket No.: STAN-2201WO

[0162] (4-((7,8-Dimethoxy-4-(piperidin-4-yl)-1H-imidazo[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonic Acid. To a solution of diethyl (4-((7,8-dimethoxy-4- (piperidin-4-yl)-1H-imidazo[4,5-c]quinolin-1-yl)methyl)benzyl)phosphonate. (1 Eq) in DCM (1 mL) at 0 °C, can be added bromotrimethylsilane (3 Eq), and the mixture can be slowly warmed to room temperature and^stirred for 20 hour. After the completion, the volatiles can be evaporated, and the crude mixture can be purified by HPLC to produce compound 14.497.15.1H NMR (500 MHz, DMSO) δ 8.30 (s, 1H), 7.19 – 7.14 (m, 2H), 7.11 (s, 1H), 7.08 (s, 1H), 6.97 (d, J = 7.9 Hz, 2H), 5.82 (s, 2H), 4.11 (t, J = 4.7 Hz, 4H), 3.79 (m, 7H), 3.58 (s, 3H), 2.77 (d, J = 21.1 Hz, 2H).Attorney Docket No.: STAN-2201WO (s, ,(500 MHz, D2O) δ 8.22 (s, 1H), 7.14 (dd, J = 7.9, 2.3 Hz, 2H), 7.11 (s, 2H), 7.06 (s, 1H), 6.89 (d, J = 7.9 Hz, 2H), 5.68 (s, 2H), 4.13 (s, 2H), 3.74 (s, 3H), 3.59 (s, 3H), 2.55 (d, J = 19.3 Hz, 2H).Attorney Docket No.: STAN-2201WO ,yl)methyl)benzyl)phosphonate (20.0 mg, 1 Eq, 41.0 μmol), tert-butyl methylcarbamate (10.7 mg, 2 Eq, 81.9 μmol), cesium carbonate (66.7 mg, 5 Eq, 205 μmol), and 2- (dicyclohexylphosphanyl)-2',4',6'-tris(isopropyl)biphenyl (7.81 mg, .4 Eq, 16.4 μmol) in THF (20 mL) was added Pd2(dba)3 (7.50 mg, .2 Eq, 8.19 μmol) and the resulting mixture heated at 70 °C for 7 h. The mixture was filtered through Celite, and the filtrate was treated with water and extracted with EtOAc. The organic layer was washed with brine, dried over MgSO4, and concentrated under vacuum. The residue was purified by reverse-phase flashAttorney Docket No.: STAN-2201WO column chromatography (C18, Acetonitrile:water) to yield the product (11 mg, 50%) as a pale yellow solid.

[0171] STEP 2: To a solution of tert-butyl (1-(4-((diethoxyphosphoryl)methyl)benzyl)-1H- imidazo[4,5-c]quinolin-7-yl)(methyl)carbamate (20 mg, 1 Eq, 37 μmol) in Dichloromethane (1 mL) at 0 °C was added bromotrimethylsilane (17 mg, 3 Eq, 0.11 mmol) and the mixture was slowly warmed to room temperature and stirred for 20 hour. After the completion, the volatiles were evaporated and the crude mixture was purified by HPLC to obtain the product as a white solid (3 mg, 40%). –ESI+ 506.2.1H NMR (500 MHz, DMSO) δ 9.41 (s, 1H), 9.32 (d, J = 2.6 Hz, 1H), 8.74 – 8.68 (m, 1H), 8.49 (s, 1H), 8.16 (dd, J = 4.6, 1.5 Hz, 1H), 7.37 (dd, J = 8.4, 4.6 Hz, 1H), 7.26 (s, 1H), 7.22 (dd, J = 8.3, 2.3 Hz, 2H), 7.15 (s, 1H), 7.07 (d, J = 8.0 Hz, 2H), 5.93 (s, 2H), 3.85 (s, 3H), 3.63 (s, 3H) 2.90 (d, J = 21.5 Hz, 2H).Attorney Docket No.: STAN-2201WO – J(500 MHz, D2O) δ 9.18 – 9.14 (m, 1H), 8.66 (s, 1H), 7.72 (s, 1H), 7.45 (s, 1H), 7.40 (d, J =Attorney Docket No.: STAN-2201WO 7.3 Hz, 2H), 7.18 (d, J = 7.8 Hz, 2H), 6.02 (s, 2H), 3.95 (s, 3H), 2.92 (s, 6H), 2.82 (d, J = 19.8 Hz, 2H). = ,(500 MHz, D2O) δ 9.33 (d, J = 0.7 Hz, 1H), 8.68 (s, 1H), 8.62 (d, J = 0.8 Hz, 1H), 8.35 (d, J = 9.4 Hz, 1H), 7.70 (d, J = 2.5 Hz, 1H), 7.66 (dd, J = 9.3, 2.6 Hz, 1H), 7.48 – 7.46 (m, 2H), 7.24 (d, J = 7.9 Hz, 2H), 6.09 (s, 2H), 3.51 – 3.46 (m, 4H), 3.15 (t, J = 5.1 Hz, 4H), 2.93 (s, 2H).Attorney Docket No.: STAN-2201WO =(500 MHz, D2O) δ 8.69 (s, 1H), 8.03 (d, J = 4.5 Hz, 1H), 7.45 (dd, J = 18.2, 9.1 Hz, 1H), 7.11 (dd, J = 8.4, 2.2 Hz, 2H), 6.95 – 6.86 (m, 1H), 6.86 – 6.83 (m, 2H), 6.77 (ddd, J = 24.3, 9.2, 2.4 Hz, 1H), 5.46 (s, 2H), 3.91 – 3.67 (m, 1H), 3.68 – 3.62 (m, 2H), 3.61 – 3.49 (m, 1H), 3.32 – 3.05 (m, 1H), 2.64 (d, J = 19.8 Hz, 2H).general preparation c. ESI+ 487.2.1H NMR (500 MHz, D2O) δ 8.67 (s, 1H), 7.98 (s, 1H), 7.34 (d, J = 9.0 Hz, 1H), 7.16 – 7.10 (m, 2H), 6.88 (d, J = 7.9 Hz, 2H), 6.71 (d, J = 2.4 Hz, 1H), 6.63 (dd, J = 9.0, 2.5 Hz, 1H), 5.34 (s, 2H), 4.27 (t, J = 6.5 Hz, 1H), 3.64 (dd, J = 14.1, 7.2 Hz, 1H), 3.41 – 3.35 (m, 1H), 3.28 (dt, JAttorney Docket No.: STAN-2201WO = 13.8, 8.1 Hz, 1H), 3.03 (dd, J = 14.1, 5.9 Hz, 1H), 2.70 (d, J = 20.0 Hz, 2H), 2.62 (dt, J = 13.6, 6.8 Hz, 1H), 2.23 (dt, J = 14.0, 7.6 Hz, 1H), 0.83 (t, J = 7.0 Hz, 1H). Compound 34

[0184] This compound was preparation c. ESI+ 478.3.1H NMR(500 MHz, D2O) δ 8.47 (s, 1H), 7.71 (s, 1H), 7.20 – 7.14 (m, 2H), 6.92 (d, J = 7.9 Hz, 2H), 6.88 (d, J = 9.0 Hz, 1H), 6.25 (s, 1H), 5.94 (d, J = 8.9 Hz, 1H), 4.98 (s, 2H), 3.17 – 3.10 (m, 2H), 2.91 (d, J = 10.8 Hz, 3H), 2.84 (d, J = 12.0 Hz, 1H), 2.76 (d, J = 9.4 Hz, 1H), 2.73 – 2.65 (m, 3H), 1.78 – 1.68 (m, 4H). Compound 35

[0185] This compound wasgeneral preparation c. ESI+453.2.1H NMR (500 MHz, D2O) δ 8.61 (s, 1H), 7.95 (s, 1H), 7.20 – 7.12 (m, 3H), 6.89 (d, J = 7.9 Hz, 2H), 6.58 (d, J = 2.2 Hz, 1H), 6.46 (d, J = 9.1 Hz, 1H), 5.27 (s, 2H), 3.60 (s, 1H), 3.54 – 3.48 (m, 1H), 3.39 – 3.33 (m, 1H), 3.21 (s, 1H), 2.90 (d, J = 8.2 Hz, 1H), 2.68 (d, J = 19.9 Hz, 2H), 1.94 (s, 4H).Attorney Docket No.: STAN-2201WO((7-bromo-1H-imidazo[4,5-c]quinolin-1-yl)methyl)benzyl)phosphonate (40.0 mg, 1 Eq, 81.9 μmol), potassium carbonate (56.6 mg, 5 Eq, 410 μmol) in 1,2-dimethoxyethane (4 mL) and water (0.2 mL) was bubbled with nitrogen for 5 min. Then Pd(dppf)2Cl2 (12.0 mg, .2 Eq, 16.4 μmol) was added to the reaction and it was stirred at 90 °C for 14 hour. After the reaction was complete, it was purified by reverse phase flash chromatography (C18, acetonitrile and water) to obtain the product (28 mg, 70%) as a white powder.

[0187] Step 2. To a solution of diethyl (4-((7-(pyridin-4-yl)-1H-imidazo[4,5-c]quinolin-1- yl)methyl)benzyl)phosphonate (20 mg, 1 Eq, 41 μmol) was added concentrated bromotrimethylsilane (17 mg, 0.5 mL, 37% Wt, 1 Eq, 41 μmol) and the mixture was slowly warmed to room temperature and stirred for 20 hour. After the completion, the volatiles were evaporated and the crude mixture was purified by HPLC to obtain the product as a white solid (7 mg, 40%). ESI+ 431.2.1H NMR (500 MHz, D2O) δ 8.74 (s, 1H), 8.13 (d, J = 6.0 Hz, 3H), 7.53 – 7.48 (m, 2H), 7.33 (dd, J = 8.4, 2.2 Hz, 2H), 7.06 (t, J = 7.4 Hz, 4H), 6.98 (d, J = 8.6 Hz, 1H), 5.53 (s, 2H), 2.82 (d, J = 19.8 Hz, 2H).

[0188] Step 1. To a flame-dried 20 mL vial was added the diethyl (4-((7-bromo-1H- imidazo[4,5-c]quinolin-1-yl)methyl)benzyl)phosphonate (50 mg, 1 Eq, 0.10 mmol), trans- bis(acetato)bis[o-(di-o-tolylphosphino)benzyl]dipalladium(II) (9.6 mg, 0.1 Eq, 10 μmol) , N,N-dimethylpyridin-4-amine (25 mg, 2 Eq, 0.20 mmol), and molybdenum hexacarbonyl (14Attorney Docket No.: STAN-2201WO mg, 0.5 Eq, 51 μmol). The vial was evacuated under vacuum and purged with nitrogen (3 times). Dry 1,4-dioxane (2 mL), methanol (0.33 g, 100 Eq, 10 mmol), and N-ethyl-N- isopropylpropan-2-amine (26 mg, 2 Eq, 0.20 mmol) was added via syringe and the vial was sealed and the reaction mixture was heated to 120 °C for 12 h in a microwave reactor. Then the vial was allowed to cool down to room temperature and the residual solvent was removed under vacuum and the crude material was purified by reverse phase flash column chromatography (C18, ACN:water) to obtain the product as a colorless oil (22 mg, 46%).

[0189] Step 2. To a solution of methyl 1-(4-((diethoxyphosphoryl)methyl)benzyl)-1H- imidazo[4,5-c]quinoline-7-carboxylate (20 mg, 1 Eq, 43 μmol) was added concentrated bromotrimethylsilane (18 mg, 0.5 mL, 37% Wt, 1 Eq, 43 μmol) and the mixture was slowly warmed to room temperature and stirred for 20 hour. After the completion, the volatiles were evaporated and the crude mixture was purified by HPLC to obtain the product as a white solid (3 mg, 40%). ESI+ 398.1.1H NMR (500 MHz, D2O) δ 9.10 (s, 1H), 8.49 (d, J = 1.7 Hz, 1H), 8.36 (s, 1H), 8.04 (d, J = 8.6 Hz, 1H), 7.93 (dd, J = 8.7, 1.7 Hz, 1H), 7.24 (dd, J = 8.3, 2.3 Hz, 2H), 7.05 (d, J = 8.0 Hz, 2H), 5.83 (s, 2H), 2.77 (d, J = 19.8 Hz, 2H). Compound 38

[0190] This compound wasthe same procedure as compound 36. ESI+ 432.1.1H NMR (500 MHz, D2O) δ 8.64 (d, J = 8.1 Hz, 2H), 8.33 (s, 2H), 8.06 (s, 1H), 7.47 (m, 1H), 7.41 (s, 1H), 7.24 (d, J = 7.9 Hz, 2H), 6.98 (d, J = 7.9 Hz, 2H), 6.74 (s, 1H), 5.46 (s, 2H), 2.76 (d, J = 20.0 Hz, 2H). Example 2: Assessing Compound Activity

[0191] Compounds were assessed in an ENPP1 / ENPP3 enzyme activity assay using cGAMP as a substrate. Methods used to assess the subject compounds include those described by Mardjuki et al. J. Biol. Chem. (2020) 295(15):4881-92. An exemplary method is set forth below.Attorney Docket No.: STAN-2201WO Materials:

[0192] Mouse ENPP1: Expressed and purified according to Carozza et al. PNAS (2022) 119(21):e2119189119. Mouse ENPP3: Expressed and purified according to Mardjuki et al. bioRxiv 2024.01.12.575449 or, as with mouse ENPP1, according to Carozza et al. PNAS (2022) 119(21):e2119189119. Human ENPP1 (R&D Systems) or expressed and purified, as with mouse ENPP1, according to Carozza et al. PNAS (2022) 119(21):e2119189119. Human ENPP3 (Acro Biosystems) or expressed and purified, as with mouse ENPP1, according to Carozza et al. PNAS (2022) 119(21):e2119189119. cGAMP: Synthesized and purified according to Li et al. Nat. Chem. Biol. (2014) 10:1043-8. Polyphosphate:AMP phosphotransferase (PAP): The PAP gene (GenBank: AB092983.1) was synthesized (Integrated DNA Technologies) and cloned into the pTB146 vector with a His-SUMO C- terminal tag. BL21(DE3) cells transformed with the plasmid were grown and induced at OD600 = 1 with 0.75 mM IPTG at 16°C overnight. Cells were resuspended in a buffer containing 50 mM Tris pH 7.5, 400 mM NaCl, 10 mM imidazole, 2 mM DTT, protease inhibitor (Roche) and lysed with two freeze-thaw cycles and sonication. All subsequent steps were performed at 4°C. Lysate was cleared by centrifugation at 40,000 rcf for 1 hour and the supernatant was incubated with HisPur cobalt resin (Thermo Fisher Scientific) for 2 hours. Resin was washed twice with 30 mL of buffer containing 50 mM Tris pH 7.5, 150 mM NaCl and the protein was eluted with 50 mM Tris pH 7.5, 150 mM NaCl, 600 mM imidazole. Anion exchange chromatography (HiTrap Q HP) was performed. Rabbit Myokinase or Yeast Myokinase (MilliporeSigma). CellTiterGlo (Promega). Exemplary Procedure for human ENPP1 or human ENPP3 enzyme activity assay:

[0193] 0.125 nM human ENPP1 or human ENPP3 was incubated with 7.5 µM cGAMP and 5-fold serial dilutions of compounds in buffer containing 50 mM Tris pH 7.6, 250 nM NaCl, 500 µM CaCl2, and 1 µM ZnCl2 (total reaction volume = 20 µL) at room temperature overnight (~19-21 hours), after which the reactions were heat inactivated at 95°C for 10 minutes. The AMP degradation product was converted to ATP, which was detected using luciferase. To achieve this, an enzyme mixture of polyphosphate:AMP phosphotransferase (PAP) and myokinase was prepared according to Goueli et al. in EP2771480. Briefly, PAP was diluted to 2 mg / mL in a buffer containing 50 mM Tris pH 7.5, 0.1% NP-40. Myokinase was diluted to 2 KU / mL in a buffer containing 3.2 mM ammonium sulfate pH 6.0, 1 mM EDTA, and 4 mM polyphosphate. The heat-inactivated ENPP1 reaction was incubated with PAP (0.01 µg / µL) and myokinase (0.0075 U / µL) in buffer containing 40 mM Tris pH 7.5,Attorney Docket No.: STAN-2201WO 0.05 mg / mL Prionex, 5 mM MgCl2, 20 µM polyphosphate, and 0.15 g / L phenol red (for ease of pipetting) for 3 hours (total reaction volume = 20 µL). CellTiterGlo (10 µL) was added to the reaction according to the manufacturer's protocol and luminescence was measured. Data were normalized to 100% enzyme activity (no compound) and 0% enzyme activity (no enzyme) before performing a nonlinear regression with variable slope (four parameters) to obtain IC50 values (GraphPad Prism 10.2.2).

[0194] IC50 data is provided in the table below. Compound huENPP1 IC50 huENPP3 IC50 1 A C A0 nM; ND = Not DeterminedAttorney Docket No.: STAN-2201WO Example 3: Demonstration of inhibition of extracellular ENPP1 or ENPP3 in a cellular context.

[0195] Inhibitors of ENPP1 and / or ENPP3 could increase STING activation by inhibiting degradation of cGAMP when treated extracellularly in a cellular context.

[0196] hTERT RPE-1 cells can be transfected with a human ENPP1 or human ENPP3 expression plasmid. hTERT RPE-1 cells can be purchased from ATCC. hTERT RPE-1 cells can be plated in tissue culture treated plates in DMEM (Gibco) supplemented with 10% FBS (Atlanta Biologics) (v / v) and 100 U / mL penicillin-streptomycin (ThermoFisher).12-24 hours following plating, cells can be transfected with Fugene 6 (Promega) according to manufacturer’s instructions plus indicated concentrations of plasmid DNA.24 hours following transfection, cells can be treated with 20 µM cGAMP with or without inhibitor of ENPP1 and / or ENPP3.8-16 hours later, total RNA can be extracted, and real-time RT-PCR can be performed to measure transcript levels of interferon stimulated genes IFNB, IFIT2, IFIT3, CXCL10, OAS1, and housekeeping gene GAPDH. Data can be normalized to GAPDH and fold induction can be calculated using ΔΔCt.

[0197] Notwithstanding the appended claims, the disclosure is also defined by the following clauses:

[0198] 1. A compound, or a salt thereof, having a structure of (I), Q is selected from the group consisting of -NR1R2, -OR1, and R1,R2are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, and that R1and R2, along with the nitrogen to which they are attached, can be optionally attached to form a substituted or unsubstituted 3- to 15- membered ring system; Rais selected from the group ,Attorney Docket No.: STAN-2201WO ; wherein Rx, Ryand Rzare and substituted or unsubstitutedthe group consisting of - NR3R4, -OR3, and R3, and that Rband Rc, along with the atoms to which they are attached, can be optionally attached to form a substituted or unsubstituted 5- to 8- membered ring system, wherein each R3and R4are each independently selected from the group consisting of H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, and that R3and R4, along with the nitrogen to which they are attached, can be optionally attached to form a substituted or unsubstituted 3- to 15- membered ring system.

[0199] 2. The compound of clause 1, or a salt thereof, wherein Rdis H.

[0200] 3. The compound of clause 1, or a salt thereof, having a structure of Formula (II): .of clause 1, or a salt thereof, having a structure of Formula (IV): .Attorney Docket No.: STAN-2201WO

[0202] 5. The compound of clause 1, or a salt thereof, having a structure of Formula (V): .

[0203] 6. The compound of that R1and R2are not both H.

[0204] 7. The compound of clause 5, or a salt thereof, wherein R1is H, and R2is substituted or unsubstituted heterocycloalkyl.

[0205] 8. The compound of clause 5, or a salt thereof, wherein R1and R2, along with the nitrogen to which they are attached, are attached to form a substituted or unsubstituted 5- to 6- membered ring system.

[0206] 9. The compound of clause 5, or a salt thereof, wherein R1and R2, along with the nitrogen to which they are attached, form a substituted or unsubstituted morpholino, substituted or unsubstituted piperazinyl, substituted or unsubstituted triazolyl, or substituted or unsubstituted imidazolyl.

[0207] 10. The compound of clause 1, or a salt thereof, having a structure which is HO O HO P orAttorney Docket No.: STAN-2201WO

[0208] 11. A pharmaceutical composition comprising: a) the compound, or a pharmaceutically acceptable salt thereof, of any of the preceding clauses; and b) one or more pharmaceutically acceptable excipient(s).

[0209] 12. The composition of clause 11, for use in treating cancer.

[0210] 13. A method of inhibiting ENPP1 and / or ENPP3, comprising: contacting the ENPP1 and / or the ENPP3 with the compound of any of the preceding clauses, thereby inhibiting ENPP1 and / or ENPP3.

[0211] 14. A method of treating an ENPP1 and / or ENPP3 associated disease in a mammal in need of treatment thereof, comprising: administering to the mammal a therapeutically effective amount of the compound of any of the preceding clauses, thereby treating the ENPP1 and / or ENPP3 associated disease.

[0212] 15. The method of clause 14, wherein the ENPP1 and / or ENPP3 associated disease is cancer.

[0213] 16. The method of clause 14, wherein the cancer is selected from the group consisting of adrenal, liver, kidney, bladder, breast, colon, gastric, ovarian, cervical, uterine, esophageal, colorectal, prostate, pancreatic, lung (both small cell and non-small cell), thyroid, carcinomas, sarcomas, glioblastomas, melanoma and head and neck tumors.

[0214] 17. The method of clause 16, wherein the cancer is breast cancer.

[0215] 18. A compound according to any one of clauses 1-10 for use in treating cancer.

[0216] 19. Use of a compound according to any one of clauses 1-10 in the manufacture of a medicament for treating cancer.

[0217] 20. A method of modulating an immune response in a subject, the method comprising: administrating to a subject a therapeutically effective amount of the compound according to any one of clauses 1-10 to treat the subject for an inflammatory condition.

[0218] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it is readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.Attorney Docket No.: STAN-2201WO

[0219] Accordingly, the preceding merely illustrates the principles of the invention. It will be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are principally intended to aid the reader in understanding the principles of the invention and the concepts contributed by the inventors to furthering the art and are to be construed as being without limitation to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and embodiments of the invention as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Additionally, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless of structure. The scope of the present invention, therefore, is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of present invention is embodied by the following.

[0220] The scope of the present invention, therefore, is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of present invention is embodied by the appended claims. In the claims, 35 U.S.C. §112(f) or 35 U.S.C. §112(6) is expressly defined as being invoked for a limitation in the claim only when the exact phrase "means for" or the exact phrase "step for" is recited at the beginning of such limitation in the claim; if such exact phrase is not used in a limitation in the claim, then 35 U.S.C. §112(f) or 35 U.S.C. §112(6) is not invoked.

Claims

Attorney Docket No.: STAN-2201WO What is claimed is:

1. A compound, or a salt thereof, having a structure of Formula (I): whereinQ is selected from the group - - and R1wherein each R1and R2are each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl and that R1and R2, along with the nitrogen to which they are attached, can be optionally attached to form a substituted or unsubstituted 3- to 15- membered ring system; Rais selected from the group ,, and ;each independently selected from the group consisting of H and substituted or unsubstituted alkyl; and Rb, Rcand Rdare each independently selected from the group consisting of -NR3R4, -OR3, and R3and that Rband Rc, along with the atoms to which they are attached, can be optionally attached to form a substituted or unsubstituted 5- to 8- membered ring system wherein each R3and R4are each independently selected from the group consisting of H, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstitutedAttorney Docket No.: STAN-2201WO heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl and that R3and R4, along with the nitrogen to which they are attached, can be optionally attached to form a substituted or unsubstituted 3- to 15- membered ring system.

2. The compound of claim 1, or a salt thereof, wherein Rdis H.

3. The compound of claim 1, or a salt thereof, having a structure of Formula (II): .

4. The compound of claim 1,a structure of Formula (IV): .

5. The compound of claim 1,a structure of Formula (V): .

6. The compound of claimR1and R2are not both H.

7. The compound of claim 5, or a salt thereof, wherein R1is H, and R2is substituted or unsubstituted heterocycloalkyl.Attorney Docket No.: STAN-2201WO 8. The compound of claim 5, or a salt thereof, wherein R1and R2, along with the nitrogen to which they are attached, are attached to form a substituted or unsubstituted 5- to 6- membered ring system.

9. The compound of claim 5, or a salt thereof, wherein R1and R2, along with the nitrogen to which they are attached, form a substituted or unsubstituted morpholino, substituted or unsubstituted piperazinyl, substituted or unsubstituted triazolyl, or substituted or unsubstituted imidazolyl.

10. The compound of claim 1, or a salt thereof, having a structure which is ora) the compound, or a pharmaceutically acceptable salt thereof, of any of the preceding claims; and b) one or more pharmaceutically acceptable excipient(s).

12. The composition of claim 11, for use in treating cancer.

13. A method of inhibiting ENPP1 and / or ENPP3, comprising: contacting the ENPP1 and / or the ENPP3 with the compound of any of the preceding claims, thereby inhibiting ENPP1 and / or ENPP3.Attorney Docket No.: STAN-2201WO 14. A method of treating an ENPP1 and / or ENPP3 associated disease in a mammal in need of treatment thereof, comprising: administering to the mammal a therapeutically effective amount of the compound of any of the preceding claims, thereby treating the ENPP1 and / or ENPP3 associated disease.

15. The method of claim 14, wherein the ENPP1 and / or ENPP3 associated disease is cancer.

16. The method of claim 14, wherein the cancer is selected from the group consisting of adrenal, liver, kidney, bladder, breast, colon, gastric, ovarian, cervical, uterine, esophageal, colorectal, prostate, pancreatic, lung (both small cell and non-small cell), thyroid, carcinomas, sarcomas, glioblastomas, melanoma and head and neck tumors.

17. The method of claim 16, wherein the cancer is breast cancer.

18. A compound according to any one of claims 1-10 for use in treating cancer.

19. Use of a compound according to any one of claims 1-10 in the manufacture of a medicament for treating cancer.

20. A method of modulating an immune response in a subject, the method comprising: administering to a subject a therapeutically effective amount of the compound according to any one of claims 1-10 to treat the subject for an inflammatory condition.

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