Ectonucleotide pyrophosphatase / phosphodiesterase 1 (ENPP1) inhibitors for the treatment of hypophosphatasia

Abstract

Provided herein are methods for treating hypophosphatasia (HPP), comprising administering a therapeutically effective amount of an inhibitor of ectonucleotide pyrophosphatase / phosphodiesterase 1 (ENPP1).

Description

ECTONUCLEOTIDE PYROPHOSPHATASE / PHOSPHODIESTERASE 1 (ENPP1) INHIBITORS FOR THE TREATMENT OF HYPOPHOSPHATASIA CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the priority benefit of U.S. Provisional Application Nos. 63 / 493,684, filed March 31, 2023, and 63 / 495,994, filed April 13, 2023, both of which are hereby incorporated by reference in their entirety. FIELD

[0002] Provided herein are methods for treating hypophosphatasia (HPP), comprising administering a therapeutically effective amount of an inhibitor of ectonucleotide pyrophosphatase / phosphodiesterase 1 (ENPP1). BACKGROUND

[0003] Hypophosphatasia (HPP) is a rare inherited metabolic disorder caused by loss-of-function mutations in the ALPL gene, which encodes the tissue-nonspecific isozyme of alkaline phosphatase (TNSALP). Low TNSALP activity leads to the extracellular accumulation of phosphoethanolamine (PEA), inorganic pyrophosphate (PPi), and pyridoxal 5 phosphate (PLP) in blood or urine. PPi, an inhibitor of bone mineralization, is one of the key substrates involved in the pathogenesis of HPP. In HPP, excess PPi contributes to defective mineralization of bones (rickets or osteomalacia), premature loss of deciduous teeth, chronic bone and muscle pain, recurrent fractures, respiratory and muscle dysfunction and periodontal defects. Symptoms range from lethal or severe in the perinatal or infantile-onset forms of HPP, to modest or mild in the childhood and adult-onset forms.

[0004] The incidence of severe HPP ranges from about 1:100,000 to 1:300,000 live births. In certain populations the incidence is higher, such as in the Canadian Mennonites in whom the incidence is as high as about 1 in 2,500 births.

[0005] Hypophosphatasia presents a significant burden to affected individuals and their families. Therefore, identification of compounds, compositions, and methods that can be used to treat or prevent hypophosphatasia would fill an important unmet need. SUMMARY

[0006] Disclosed herein are methods of using an ENPP1 inhibitor for treating or preventing a soft bone disease. In some embodiments, the soft bone disease is hypophosphatasia. Ectonucleotide pyrophosphatase / phosphodiesterase 1 (ENPP1) is an ectoenzyme which hydrolyzes ATP into PPi and AMP in the extracellular space. PPi functions as a potent inhibitor of hydroxyapatite (HAP) crystalgrowth, which is the inorganic mineral component of calcium phosphate and critical for bone mineralization. As ENPP1 is the primary source of PPi, inhibition of this enzyme represents a promising therapeutic approach for the treatment of HPP. Normalizing extracellular PPi levels by targeting ENPP1 could provide significant relief from the severe symptoms of this debilitating disease. Targeted therapeutics that inhibit ENPP1 activity or neutralize PPi could lead to effective interventions for HPP.

[0007] In some embodiments, the hypophosphatasia is late-onset hypophosphatasia. In some embodiments, the hypophosphatasia is pediatric-onset hypophosphatasia.

[0008] In some embodiments, the hypophosphatasia is prenatal hypophosphatasia. In some embodiments, the hypophosphatasia is perinatal hypophosphatasia.

[0009] In some embodiments, the disease is infantile hypophosphatasia.

[0010] In some embodiments, the disease is childhood hypophosphatasia.

[0011] In some embodiments, the disease is adult hypophosphatasia.

[0012] In some embodiments, the disease is any form of hypophosphatasia, wherein the hypophosphatasia has dental involvement and / or premature tooth loss.

[0013] In some embodiments, the disease is odontohypophosphatasia.

[0014] In some embodiments, a method of treating a subject with a dental disorder is provided with an agent which inhibits ENPP1.

[0015] In some embodiments, a method of treating a subject with periodontal disease is provided with an agent which inhibits ENPP1.

[0016] In some embodiments, a method of treating hypophosphatasia is provided with an agent which inhibits ENPP1.

[0017] In some embodiments, a method of treating hypophosphatasia in a subject is provided, comprising administering to a subject in need thereof a therapeutically effective amount of a compound which inhibits hydrolysis of ATP via ENPP1 inhibition.

[0018] In some embodiments, a method of treating hypophosphatasia is provided, comprising administering to a subject in need thereof a therapeutically effective amount of a compound that inhibits ENPP1.

[0019] In some embodiments, the compound is a compound described in US20220135598A1, which is incorporated by reference herein. In some embodiments, the compound is of Formula I:or a pharmaceutically acceptable salt thereof, wherein: Ring C is 5- to 6-membered heteroaryl; Ring D is C6-aryl or 5- to 6-membered heteroaryl, wherein Ring D is fused to Ring C; A is hydrogen, C1-C6alkyl, or C6-aryl, each of which is optionally substituted with halogen; G is a bond, -CH2-, or -CH2-CH2-; Raand Rbare independently hydrogen or C1-C6alkyl; or Raand Rbare taken together with the atoms to which they are attached to form a C3-C6cycloalkyl ring; or any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a C4-C6cycloalkyl ring; L is a bond, linear or branched C1-C6alkylene, or linear or branched C2-C6alkenylene; t is 0 or 1; provided that when t is 0, then L is linear or branched C2-C6alkenylene; Z is -NRcS(O)2NH2, -NRcS(O)2CH3, -SO2NH2, -NRcC(O)CH3, -C(O)OH, -CONH2, -NRcCONH2, -CONH(OH), -B(OH)2, -P(O)(OH)2, -SO2OH, -NRcS(O)2CF3, -NRcS(O)2NHCH3, or -NRcCH2C6-aryl-S(O)2NH2. each R1and R2is independently oxo, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, adamantyl, 3- to 6-membered heterocyclyl, C6-aryl, 5- to 6-membered heteroaryl, -CN, halogen, C1-C6alkoxy, C1-C6haloalkoxy, -OR10, -SR10, -S(O)2R10, -S(O)2NR11R12, -NR10S(O)2R11, -NR11R12, -C(O)R10, -NR10C(O)R11, -NR10C(O)NR11R12, -C(O)OR10, -C(O)ONR11R12, or -C(O)NR11R12, wherein each is independently optionally substituted by R9;or two R2are taken together with the atoms to which they are attached to form a C5-C6cycloalkyl, 5- to 6-membered heterocyclyl, C6-aryl, or 5- to 6-membered heteroaryl, wherein each is independently optionally substituted by R9; each R9is independently selected from the group consisting of oxo, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, 3- to 6-membered heterocyclyl, C6-aryl, 5- to 6-membered heteroaryl, -CN, halogen, C1-C6alkoxy, C1-C6haloalkoxy, C1-C6haloalkyl, -OR13, -SR13, -S(O)2R13, -S(O)2NR14R15, -NR13S(O)2R14, -NR14R15, -C(O)R13, -NR13C(O)R14, -NR13C(O)NR14R15, -C(O)OR13, -C(O)ONR14R15, -C(O)NR14R15, and C1-C6alkyl optionally substituted by oxo, -OH, or halogen; each Rcis independently hydrogen, C1-C6alkyl, C3-C6cycloalkyl, -(C1-C6alkylene)C3-C6cycloalkyl, 3- to 6-membered heterocyclyl, -(C1-C6alkylene)3- to 6-membered heterocyclyl, -(C1-C6alkylene)C6-aryl, 5- to 6-membered heteroaryl, -(C1-C6alkylene)5- to 6-membered heteroaryl, C1-C6haloalkyl, -(C1-C6alkylene)OR13, -(C1-C6alkylene)SR13, -(C1-C6alkylene)S(O)2R13, -(C1-C6alkylene)S(O)2NR14R15, -(C1-C6alkylene)NR13S(O)2R14, -(C1-C6alkylene)NR14R15, -(C1-C6alkylene)C(O)R13, -(C1-C6alkylene)NR13C(O)R14, -(C1-C6alkylene)NR13C(O)NR14R15, -(C1-C6alkylene)C(O)OR13, -(C1-C6alkylene) C(O)ONR14R15, or -(C1-C6alkylene)-C(O)NR14R15, wherein each is independently optionally substituted with Rd; each Rdis independently selected from the group consisting of halogen, -OH, oxo, -CN, C1-C6haloalkyl, C1-C6alkoxy, C1-C6haloalkoxy, -COOH, and C1-C6alkyl optionally substituted with -OH, halogen, CN, or oxo; each R10, R11, and R12is independently hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, -(C1-C6alkylene)C3-C6cycloalkyl, 3- to 6-membered heterocyclyl, or -(C1-C6alkylene)3- to 6-membered heterocyclyl, wherein each of R10, R11and R12is independently optionally substituted by oxo, C2-C6alkenyl, C2-C6alkynyl, -CN, halogen, C1-C6alkoxy, or C1-C6alkyl optionally substituted by oxo, -OH, or halogen; or R11and R12are taken together with the atom(s) to which they are attached to form a 3-6 membered heterocyclyl optionally substituted by oxo, -OH, halogen, or C1-C6alkyl optionally substituted by oxo, -OH, or halogen; each R13, R14and R15is independently hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6haloalkyl, C3-C6cycloalkyl, or 3- to 6-membered heterocyclyl, wherein each of R10, R11, and R12is independently optionally substituted by oxo, -OH, C2-C6alkenyl, C2-C6alkynyl, -CN, halogen, or C1-C6alkyl optionally substituted by oxo, -OH, or halogen;or R14and R15are taken together with the atoms to which they are attached to form a 3-6 membered heterocyclyl optionally substituted by oxo, -OH, halogen, or C1-C6alkyl optionally substituted by oxo, -OH, or halogen; m is 0, 1 or 2; and n is 0, 1, 2, 3 or 4; provided that when Z is -NRcS(O)2CH3, -CONH2or -C(O)OH, G is a bond, and t is 1, then L is not a bond; and the compound is not 1-[7-(4-bromo-2,6-dimethylphenyl)-2,5-dimethyl-7H-pyrrolo[2,3- d]pyrimidin-4-yl]-3-azetidinebutanoic acid, 1-[7-(4-bromo-2,6-dimethylphenyl)-2,5-dimethyl-7H- pyrrolo[2,3-d]pyrimidin-4-yl]-3-azetidinepentanoic acid, or N-((1-(6-fluorobenzo[d]oxazol-2-yl)azetidin- 3-yl)methyl)-N-methylacetamide. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] FIG.1 illustrates the effects of compounds of the present disclosure on mineralization in Saos-2 cells.

[0021] FIG.2 illustrates the time-dependent effects of Compound 76 on the PPi levels in Sprague- Dawley rats 1, 4, and 8 hours after a single dose.

[0022] FIG.3 illustrates the time-dependent effects of Compound 76 on the PPi levels in mice 1 and 8 hours after the last dose of a continuous dosing for 5 days.

[0023] It will be recognized that some or all of the figures are schematic representations for the purpose of illustration. DETAILED DESCRIPTION Definitions

[0024] “Alkyl” refers to and includes saturated linear and branched univalent hydrocarbon structures and combination thereof, having the number of carbon atoms designated (i.e., C1-C10means one to ten carbons). Particular alkyl groups are those having 1 to 20 carbon atoms (a “ C1-C20alkyl”). More particular alkyl groups are those having 1 to 8 carbon atoms (a “C1-C8alkyl”), 3 to 8 carbon atoms (a “C3-C8alkyl”), 1 to 6 carbon atoms (a “C1-C6alkyl”), 1 to 5 carbon atoms (a “C1-C5alkyl”), or 1 to 4 carbon atoms (a “C1-C4alkyl”). Examples of alkyl include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, homologs and isomers of, for example, n- pentyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0025] “Alkenyl” as used herein refers to an unsaturated linear or branched univalent hydrocarbon chain or combination thereof, having at least one site of olefinic unsaturation (i.e., having at least one moiety of the formula C=C) and having the number of carbon atoms designated (i.e., C2-C10means two to ten carbon atoms). The alkenyl group may be in “cis” or “trans” configurations, or alternatively in “E” or “Z” configurations. Particular alkenyl groups are those having 2 to 20 carbon atoms (a “C2-C20alkenyl”), having 2 to 8 carbon atoms (a “C2-C8alkenyl”), having 2 to 6 carbon atoms (a “C2-C6alkenyl”), or having 2 to 4 carbon atoms (a “C2-C4alkenyl”). Examples of alkenyl include, but are not limited to, groups such as ethenyl (or vinyl), prop-1-enyl, prop-2-enyl (or allyl), 2-methylprop-1-enyl, but-1-enyl, but-2-enyl, but-3-enyl, buta-1,3-dienyl, 2-methylbuta-1,3-dienyl, homologs and isomers thereof, and the like.

[0026] “Alkylene” as used herein refers to the same residues as alkyl, but having bivalency. Particular alkylene groups are those having 1 to 6 carbon atoms (a “C1-C6alkylene”), 1 to 5 carbon atoms (a “C1-C5alkylene”), 1 to 4 carbon atoms (a “C1-C4alkylene”) or 1 to 3 carbon atoms (a “C1-C3 alkylene”). Examples of alkylene include, but are not limited to, groups such as methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), butylene (-CH2CH2CH2CH2-), and the like. “Alkenylene” as used herein refers to the same residues as alkenyl, but having bivalency.

[0027] “Alkynyl” as used herein refers to an unsaturated linear or branched univalent hydrocarbon chain or combination thereof, having at least one site of acetylenic unsaturation (i.e., having at least one moiety of the formula C≡C) and having the number of carbon atoms designated (i.e., C2-C10means two to ten carbon atoms). Particular alkynyl groups are those having 2 to 20 carbon atoms (a “C2-C20alkynyl”), having 2 to 8 carbon atoms (a “C2-C8alkynyl”), having 2 to 6 carbon atoms (a “C2-C6alkynyl”), or having 2 to 4 carbon atoms (a “C2-C4alkynyl”). Examples of alkynyl include, but are not limited to, groups such as ethynyl (or acetylenyl), prop-1-ynyl, prop-2-ynyl (or propargyl), but-1-ynyl, but-2-ynyl, but-3-ynyl, homologs and isomers thereof, and the like.

[0028] “Aryl” refers to and includes polyunsaturated aromatic hydrocarbon groups. Aryl may contain additional fused rings (e.g., from 1 to 3 rings), including additionally fused aryl, heteroaryl, cycloalkyl, and / or heterocyclyl rings. In one variation, the aryl group contains from 6 to 14 annular carbon atoms. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, biphenyl, and the like.

[0029] “Carbonyl” refers to the group C=O.

[0030] “Cycloalkyl” refers to and includes cyclic univalent hydrocarbon structures, which may be fully saturated, mono- or polyunsaturated, but which are non-aromatic, having the number of carbon atoms designated (e.g., C1-C10means one to ten carbons). Cycloalkyl can consist of one ring, such as cyclohexyl, or multiple rings, such as adamantly, but excludes aryl groups. In certain embodiments,cycloalkyl comprising more than one ring may be fused, spiro or bridged, or combinations thereof. In certain embodiments, cycloalkyl is a cyclic hydrocarbon having from 3 to 13 annular carbon atoms. In certain embodiments, cycloalkyl is a cyclic hydrocarbon having from 3 to 8 annular carbon atoms (a "C3- C8cycloalkyl"). Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, norbornyl, and the like.

[0031] “Halo” or “halogen” refers to elements of the Group 17 series having atomic number 9 to 85. In certain embodiments, halo groups include fluoro, chloro, bromo, and iodo. Where a residue is substituted with more than one halogen, it may be referred to by using a prefix corresponding to the number of halogen moieties attached, e.g., dihaloaryl, dihaloalkyl, trihaloaryl, etc. refer to aryl and alkyl substituted with two (“di”) or three (“tri”) halo groups, which may be but are not necessarily the same halo; thus 4- chloro-3-fluorophenyl is within the scope of dihaloaryl. An alkyl group in which each hydrogen is replaced with a halo group is referred to as a “perhaloalkyl.” In certain embodiments, a perhaloalkyl group is trifluoroalkyl (-CF3). Similarly, “perhaloalkoxy” refers to an alkoxy group in which a halogen takes the place of each H in the hydrocarbon making up the alkyl moiety of the alkoxy group. An example of a perhaloalkoxy group is trifluoromethoxy (-OCF3).

[0032] “Heteroaryl” refers to and includes unsaturated aromatic cyclic groups having from 1 to 10 annular carbon atoms and at least one annular heteroatom, including but not limited to heteroatoms such as nitrogen, oxygen and sulfur, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. A heteroaryl group can be attached to the remainder of the molecule at an annular carbon or at an annular heteroatom. Heteroaryl may contain additional fused rings (e.g., from 1 to 3 rings), including additionally fused aryl, heteroaryl, cycloalkyl, and / or heterocyclyl rings. Examples of heteroaryl groups include, but are not limited to imidazolyl, pyrrolyl, pyrazolyl, 1,2,4- triazolyl, thiophenyl, furanyl, thiazolyl, isothiazolyl, 1,3,4-thiadiazolyl oxazolyl, isoxazolyl, 1,3,4- oxadiazolyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl, indolyl, indazolyl, benzoimidazolyl, pyrrolopyridinyl, pyrrolopyridazinyl, pyrrolopyrimidinyl, pyrazolopyridinyl, pyrazolopyrimidinyl, imidazopyridinyl, purinyl, benzofuranyl, furopyridinyl, benzooxazolyl, benzothiophenyl, benzothiazolyl, oxazolopyridinyl, thiazolopyridinyl, thienopyridinyl, quinolinyl, quinolonyl, naphthyridinyl, quinazolinyl, pyridopyrimidinyl, cinnolinyl, or pyridopyridazinyl, and the like.

[0033] “Heterocycle” or “heterocyclyl” refers to a saturated or an unsaturated non-aromatic group having from 1 to 10 annular carbon atoms and from 1 to 4 annular heteroatoms, such as nitrogen, sulfur or oxygen, and the like, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. A heterocyclyl group may have a single ring or multiple condensed rings, but excludes heteroaryl groups. A heterocycle comprising more than one ring may be fused, spiroor bridged, or any combination thereof. In fused ring systems, one or more of the fused rings can be aryl or heteroaryl. Examples of heterocyclyl groups include, but are not limited to, aziridinyl, azetidinyl, oxetanyl, morpholinyl, thiomorpholinyl, azepanyl tetrahydropyranyl, dihydropyranyl, piperidinyl, piperazinyl, pyrrolidinyl, thiazolinyl, thiazolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, and the like.

[0034] “Oxo” refers to the moiety =O.

[0035] “Optionally substituted” unless otherwise specified means that a group may be unsubstituted or substituted by one or more (e.g., 1, 2, 3, 4, or 5) of the substituents listed for that group in which the substituents may be the same of different. In one embodiment, an optionally substituted group has one substituent. In another embodiment, an optionally substituted group has two substituents. In another embodiment, an optionally substituted group has three substituents. In another embodiment, an optionally substituted group has four substituents. In some embodiments, an optionally substituted group has 1 to 2, 2 to 5, 3 to 5, 2 to 3, 2 to 4, 3 to 4, 1 to 3, 1 to 4, or 1 to 5 substituents.

[0036] As used herein, the term “pharmaceutically acceptable” refers to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively non-toxic, i.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0037] “Pharmaceutically acceptable salt” is meant to include salts of the active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. When compounds as disclosed herein contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. When compounds as disclosed herein contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, phosphoric, partially neutralized phosphoric acids, sulfuric, partially neutralized sulfuric, hydroiodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like. Certain specific compounds of thepresent disclosure may contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17th Ed., Mack Publishing Company, Easton, Pa., (1985) and Journal of Pharmaceutical Science, 66:2 (1977), each of which is incorporated herein by reference in its entirety.

[0038] “Pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” refers to an excipient, carrier or adjuvant that can be administered to a subject, together with at least one compound, and which does not destroy the pharmacological activity thereof and is generally safe, nontoxic and neither biologically nor otherwise undesirable when administered in doses sufficient to deliver a therapeutic amount of the agent.

[0039] “Treatment” or “treating” is an approach for obtaining beneficial or desired results including clinical results. Beneficial or desired clinical results may include one or more of the following: a) inhibiting the disease or condition (e.g., decreasing one or more symptoms resulting from the disease or condition, and / or diminishing the extent of the disease or condition); b) slowing or arresting the development of one or more clinical symptoms associated with the disease or condition (e.g., stabilizing the disease or condition, preventing or delaying the worsening or progression of the disease or condition, and / or preventing or delaying the spread (e.g., metastasis) of the disease or condition); and / or c) relieving the disease, that is, causing the regression of clinical symptoms (e.g., ameliorating the disease state, providing partial or total remission of the disease or condition, enhancing effect of another medication, delaying the progression of the disease, increasing the quality of life, and / or prolonging survival.

[0040] “Prevention” or “preventing” means any treatment of a disease or condition that causes the clinical symptoms of the disease or condition not to develop. Compounds may, in some embodiments, be administered to a subject (including a human) who is at risk or has a family history of the disease or condition.

[0041] A “disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal’s health continues to deteriorate.

[0042] “Subject” refers to an animal, such as a mammal (including a human), that has been or will be the object of treatment, observation or experiment. The methods described herein may be useful in human therapy and / or veterinary applications. In some embodiments, the subject is a mammal. In one embodiment, the subject is a human.

[0043] The term “therapeutically effective amount” or “effective amount” of a compound described herein or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analog thereof means an amount sufficient to effect treatment when administered to asubject, to provide a therapeutic benefit such as amelioration of symptoms or slowing of disease progression. For example, a therapeutically effective amount may be an amount sufficient to decrease a symptom of a disease or condition of hypophosphatasia. The therapeutically effective amount may vary depending on the subject, and disease or condition being treated, the weight and age of the subject, the severity of the disease or condition, and the manner of administering, which can readily be determined by one or ordinary skill in the art.

[0044] As used herein, the term “NPP” or “ENPP” refers to ectonucleotide pyrophosphatase / phosphodiesterase.

[0045] As used herein the terms “alteration,” “defect,” “variation,” or “mutation” refer to a mutation in a gene in a cell that affects the function, activity, expression (transcription or translation) or conformation of the polypeptide it encodes. Mutations encompassed by the present disclosure can be any mutation of a gene in a cell that results in the enhancement or disruption of the function, activity, expression or conformation of the encoded polypeptide, including the complete absence of expression of the encoded protein and can include, for example, mis sense and nonsense mutations, insertions, deletions, frameshifts and premature terminations. Without being so limited, mutations encompassed by the present disclosure may alter splicing the mRNA (splice site mutation) or cause a shift in the reading frame (frameshift).

[0046] The methods described herein may be applied to cell populations in vivo or ex vivo. “In vivo” means within a living individual, as within an animal or human. In this context, the methods described herein may be used therapeutically in an individual. “Ex vivo” means outside of a living individual. Examples of ex vivo cell populations include in vitro cell cultures and biological samples including fluid or tissue samples obtained from individuals. Such samples may be obtained by methods well known in the art. Exemplary biological fluid samples include blood, cerebrospinal fluid, urine, and saliva. In this context, the compounds and compositions described herein may be used for a variety of purposes, including therapeutic and experimental purposes. For example, the compounds and compositions described herein may be used ex vivo to determine the optimal schedule and / or dosing of administration of a compound of the present disclosure for a given indication, cell type, individual, and other parameters. Information gleaned from such use may be used for experimental purposes or in the clinic to set protocols for in vivo treatment. Other ex vivo uses for which the compounds and compositions described herein may be suited are described below or will become apparent to those skilled in the art. The selected compounds may be further characterized to examine the safety or tolerance dosage in human or non-human subjects. Such properties may be examined using commonly known methods to those skilled in the art.

[0047] As used herein, “HPP” refers to hypophosphatasia.Methods of Treatment

[0048] Provided herein are methods of treating or preventing a soft bone disease, comprising administering a therapeutically effective amount of an ectonucleotide pyrophosphatase / phosphodiesterase 1 (ENPP1) inhibitor to a subject in need thereof. In certain embodiments, provided are methods of treating or preventing hypophosphatasia, comprising administering a therapeutically effective amount of an ectonucleotide pyrophosphatase / phosphodiesterase 1 (ENPP1) inhibitor to a subject in need thereof.

[0049] Hypophosphatasia (HPP) is an inherited metabolic disease, characterized by low activity of tissue non-specific alkaline phosphatase (TNAP). Low activity of TNAP leads to systematic accumulation of its substrates, namely inorganic pyrophosphate (PPi), a strong inhibitor of mineralization, and pyridoxal-5′-phosphate (PLP), a cofactor for several enzymes, which largely explain the musculoskeletal and systemic features of the disease. HPP is characterized by a wide spectrum of manifestations and severity, ranging from death in utero to dental complications only in children and adults, or asymptomatic carriers of ALPL mutations. Due to the relevant rarity of the disease, the lack of awareness of HPP among physicians and the absence of pathognomonic symptoms, especially in the mild adult form, there is still considerable delay in its diagnosis and management.

[0050] Also provided herein are methods of treating or preventing hypophosphatasia, comprising administering a therapeutically effective amount of a composition comprising an ENPP1 inhibitor and a pharmaceutically acceptable carrier to a subject in need thereof.

[0051] In some embodiments, the therapeutically active amount of ENPP1 inhibitor treats hypophosphatasia in a subject in need thereof by restoring normal circulating PPi levels.

[0052] In some embodiments, the therapeutically active amount of ENPP1 inhibitor treats hypophosphatasia in a subject in need thereof by modulating the phosphate / pyrophosphate (Pi / PPi) ratio.

[0053] In some embodiments, the therapeutically active amount of ENPP1 inhibitor treats hypophosphatasia in a subject in need thereof by promoting mineralization.

[0054] In some embodiments, the subject is suffering from prenatal hypophosphatasia.

[0055] In some embodiments, the subject is suffering from perinatal hypophosphatasia.

[0056] In some embodiments, the subject is suffering from infantile hypophosphatasia.

[0057] In some embodiments, the subject is suffering from childhood hypophosphatasia.

[0058] In some embodiments, the subject is suffering from adult hypophosphatasia.

[0059] In some embodiments, the subject is suffering from odontohypophosphatasia.

[0060] In some embodiments, the subject is suffering from periodontal disease.

[0061] In some embodiments, the subject has an alteration in the ALPL gene. In some embodiments, the alteration in the ALPL gene is a mutation. In some embodiments, the subject has an alteration in a gene that can regulate TNSALP activity. In some embodiments, the alteration in a gene that can regulate TNSALP activity is an alteration in RUNX2.

[0062] In some embodiments, the therapeutically effective amount of ENPP1 inhibitor is administered daily. In some embodiments, the therapeutically effective amount of ENPP1 inhibitor is administered two to three times a day.

[0063] In some embodiments, the therapeutically effective amount of ENPP1 inhibitor is formulated for oral administration.

[0064] In some embodiments, the therapeutically effective amount of ENPP1 inhibitor is formulated for subcutaneous administration.

[0065] In some embodiments, the therapeutically effective amount of ENPP1 inhibitor is formulated for intraperitoneal administration. Compounds

[0066] Provided herein are compounds that inhibit ENPP1, where such compounds may be used in the methods and compositions disclosed herein, such as treating or preventing a soft bone disease. Such compounds may be useful in the treatment of hypophosphatasia.

[0067] In certain embodiments, the ENPP1 inhibitor is a compound described in US20220135598A1. In certain embodiments, the compound is a compound of Formula I:or a pharmaceutically acceptable salt thereof, wherein:Ring C is 5- to 6-membered heteroaryl; Ring D is C6-aryl or 5- to 6-membered heteroaryl, wherein Ring D is fused to Ring C; A is hydrogen, C1-C6alkyl, or C6-aryl, each of which is optionally substituted with halogen; G is a bond, -CH2-, or -CH2-CH2-; Raand Rbare independently hydrogen or C1-C6alkyl; or Raand Rbare taken together with the atoms to which they are attached to form a C3-C6cycloalkyl ring; or any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a C4-C6cycloalkyl ring; L is a bond, linear or branched C1-C6alkylene, or linear or branched C2-C6alkenylene; t is 0 or 1; provided that when t is 0, then L is linear or branched C2-C6alkenylene; Z is -NRcS(O)2NH2, -NRcS(O)2CH3, -SO2NH2, -NRcC(O)CH3, -C(O)OH, -CONH2, -NRcCONH2, -CONH(OH), -B(OH)2, -P(O)(OH)2, -SO2OH, -NRcS(O)2CF3, -NRcS(O)2NHCH3, or -NRcCH2C6-aryl-S(O)2NH2. each R1and R2is independently oxo, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, adamantyl, 3- to 6-membered heterocyclyl, C6-aryl, 5- to 6-membered heteroaryl, -CN, halogen, C1-C6alkoxy, C1-C6haloalkoxy, -OR10, -SR10, -S(O)2R10, -S(O)2NR11R12, -NR10S(O)2R11, -NR11R12, -C(O)R10, -NR10C(O)R11, -NR10C(O)NR11R12, -C(O)OR10, -C(O)ONR11R12, or -C(O)NR11R12, wherein each is independently optionally substituted by R9; or two R2are taken together with the atoms to which they are attached to form a C5-C6cycloalkyl, 5- to 6-membered heterocyclyl, C6-aryl, or 5- to 6-membered heteroaryl, wherein each is independently optionally substituted by R9; each R9is independently selected from the group consisting of oxo, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, 3- to 6-membered heterocyclyl, C6-aryl, 5- to 6-membered heteroaryl, -CN, halogen, C1-C6alkoxy, C1-C6haloalkoxy, C1-C6haloalkyl, -OR13, -SR13, -S(O)2R13, -S(O)2NR14R15, -NR13S(O)2R14, -NR14R15, -C(O)R13, -NR13C(O)R14, -NR13C(O)NR14R15, -C(O)OR13, -C(O)ONR14R15, -C(O)NR14R15, and C1-C6alkyl optionally substituted by oxo, -OH, or halogen; each Rcis independently hydrogen, C1-C6alkyl, C3-C6cycloalkyl, -(C1-C6alkylene)C3-C6cycloalkyl, 3- to 6-membered heterocyclyl, -(C1-C6alkylene)3- to 6-membered heterocyclyl, -(C1-C6alkylene)C6-aryl, 5- to 6-membered heteroaryl, -(C1-C6alkylene)5- to 6-membered heteroaryl,C1-C6haloalkyl, -(C1-C6alkylene)OR13, -(C1-C6alkylene)SR13, -(C1-C6alkylene)S(O)2R13, -(C1-C6alkylene)S(O)2NR14R15, -(C1-C6alkylene)NR13S(O)2R14, -(C1-C6alkylene)NR14R15, -(C1-C6alkylene)C(O)R13, -(C1-C6alkylene)NR13C(O)R14, -(C1-C6alkylene)NR13C(O)NR14R15, -(C1-C6alkylene)C(O)OR13, -(C1-C6alkylene) C(O)ONR14R15, or -(C1-C6alkylene)-C(O)NR14R15, wherein each is independently optionally substituted with Rd; each Rdis independently selected from the group consisting of halogen, -OH, oxo, -CN, C1-C6haloalkyl, C1-C6alkoxy, C1-C6haloalkoxy, -COOH, and C1-C6alkyl optionally substituted with -OH, halogen, CN, or oxo; each R10, R11, and R12is independently hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, -(C1-C6alkylene)C3-C6cycloalkyl, 3- to 6-membered heterocyclyl, or -(C1-C6alkylene)3- to 6-membered heterocyclyl, wherein each of R10, R11and R12is independently optionally substituted by oxo, C2-C6alkenyl, C2-C6alkynyl, -CN, halogen, C1-C6alkoxy, or C1-C6alkyl optionally substituted by oxo, -OH, or halogen; or R11and R12are taken together with the atom(s) to which they are attached to form a 3-6 membered heterocyclyl optionally substituted by oxo, -OH, halogen, or C1-C6alkyl optionally substituted by oxo, -OH, or halogen; each R13, R14and R15is independently hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6haloalkyl, C3-C6cycloalkyl, or 3- to 6-membered heterocyclyl, wherein each of R10, R11, and R12is independently optionally substituted by oxo, -OH, C2-C6alkenyl, C2-C6alkynyl, -CN, halogen, or C1-C6alkyl optionally substituted by oxo, -OH, or halogen; or R14and R15are taken together with the atoms to which they are attached to form a 3-6 membered heterocyclyl optionally substituted by oxo, -OH, halogen, or C1-C6alkyl optionally substituted by oxo, -OH, or halogen; m is 0, 1 or 2; and n is 0, 1, 2, 3 or 4; provided that when Z is -NRcS(O)2CH3, -CONH2or -C(O)OH, G is a bond, and t is 1, then L is not a bond.

[0068] In some embodiments, the compound is not 1-[7-(4-bromo-2,6-dimethylphenyl)-2,5-dimethyl- 7H-pyrrolo[2,3-d]pyrimidin-4-yl]-3-azetidinebutanoic acid, 1-[7-(4-bromo-2,6-dimethylphenyl)-2,5- dimethyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-3-azetidinepentanoic acid, or acetamide, N-[[1-(6-fluoro-2- benzoxazolyl)-3-azetidinyl]methyl]-N-methyl.

[0069] In some embodiments, A is hydrogen. In some embodiments, A is C1-C6alkyl. In some embodiments, A is methyl. In some embodiments, A is C1-C6alkyl optionally substituted by halogen. In some embodiments, A is -CF3. In some embodiments, A is C6-aryl. In some embodiments, A is phenyl.

[0070] In some embodiments, G is a bond. In some embodiments, G is -CH2-. In some embodiments, G is -CH2-CH2-.

[0071] In some embodiments, Raand Rbare independently hydrogen or C1-C6alkyl. In some embodiments, Raand Rbboth are hydrogen. In some embodiments, any one of Raand Rbis hydrogen and other is C1-C6alkyl. In some embodiments, Raand Rbboth are C1-C6alkyl. In some embodiments, any one of Raand Rbis hydrogen and other is methyl. In some embodiments, any one of Raand Rbis hydrogen and other is ethyl. In some embodiments, Raand Rbboth are methyl.

[0072] In some embodiments, Raand Rbare taken together with the atoms to which they attached to form a C3-C6cycloalkyl ring. In some embodiments, Raand Rbare taken together with the atoms to which they attached to form a cyclopropyl ring.

[0073] In some embodiments, any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a C4-C6cycloalkyl. In some embodiments, any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a cyclobutyl ring. In some embodiments, any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a cyclopentyl ring. In some embodiments, any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a cyclohexyl ring.

[0074] In some embodiments, any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a C4-C6cycloalkyl ring, in such case the other one of Raor Rbis hydrogen. In some embodiments, any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a cyclobutyl ring, in such case the other one of Raor Rbis hydrogen. In some embodiments, any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a cyclopentyl ring, in such case the other one of Raor Rbis hydrogen. In some embodiments, any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a cyclohexyl ring, in such case the other one of Raor Rbis hydrogen.

[0075] In some embodiments, G is a bond, -CH2- or -CH2-CH2-; and any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a C4-C6cycloalkyl ring. In some embodiments, G is -CH2-; and any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a cyclobutyl ring. In some embodiments, G is -CH2-; and any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a cyclopentylring. In some embodiments, G is -CH2-CH2-; and any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a cyclohexyl ring.

[0076] In some embodiments, L is a bond, or C1-C6alkylene or C2-C6alkenylene. The C1-C6alkylene or C2-C6alkenylene can be linear or branched. In some embodiments, L is a bond. In some embodiments, L is a C1-C6alkylene. In some embodiments, L is -CH2-. In some embodiments, L is -C2H4-. In some embodiments, L is -C3H6-. In some embodiments, L is -CH(CH3)-. In some embodiments, L is -C(CH3)2-. In some embodiments, L is -CH(CH3)-CH2-. In some embodiments, L is -CH2-CH(CH3)-. In some embodiments, L is -C(CH3)2-CH2-. In some embodiments, L is -CH2-C(CH3)2-. In some embodiments, L is -CH(C2H5)-CH2-.

[0077] In some embodiments, L is a C2-C6alkenylene. The C2-C6alkenylene can be linear or branched. In some embodiments, L is a -CH=CH-. In some embodiments, L is -CH2-CH=CH-.

[0078] In some embodiments, t is 0 or 1. In some embodiments, t is 0. In some embodiments, t is 1.

[0079] In some embodiments, when t is 0 then Raand Rbare absent and L is C2-C6alkenylene. In some embodiments, when t is 0 then Raand Rbare absent and L is -CH=CH-.

[0080] In some embodiments, when t is 1 then Raand Rbare present and L is a bond or C1-C6alkylene. In some embodiments, when t is 1 then Raand Rbare present and L is a bond. In some embodiments, when t is 1 then Raand Rbare present and L is C1-C6alkylene. In some embodiments, when t is 1 then Raand Rbare present and L is -CH2-. In some embodiments, when t is 1 then Raand Rbare present and L is -CH2-CH2-. Wherein Raand Rbare independently selected from hydrogen or C1-C6alkyl.

[0081] In some embodiments, when t is 1 then Raand Rbare present and any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a C4-C6cycloalkyl ring, in such case other one of Raor Rbis hydrogen. In some embodiments, when t is 1 then Raand Rbare present and any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a cyclobutyl ring, in such case other one of Raor Rbis hydrogen.

[0082] In some embodiments, Z is selected from -NRcSO2NH2, -NRcS(O)2CH3, -SO2NH2, -NRcC(O)CH3, -C(O)OH, -CONH2, NRcCONH2, -CONH(OH), -B(OH)2, -P(O)(OH)2, -SO2OH, -NRcS(O)2CF3, -NRcS(O)2NHCH3, or -NRcCH2C6-aryl-S(O)2NH2. In some embodiments, Z is -NRcS(O)2NH2.In some embodiments, Z is -NRcS(O)2CH3.In some embodiments, Z is -SO2NH2.In some embodiments, Z is -NRcC(O)CH3.In some embodiments, Z is -C(O)OH. In some embodiments, Z is -CONH2.In some embodiments, Z is -NRcCONH2. In some embodiments, Z is -CONH(OH). In some embodiments, Z is -B(OH)2.In some embodiments, Z is -P(O)(OH)2. In some embodiments, Z is -SO2OH. In some embodiments, Z is -NRcS(O)2CF3. In some embodiments, Z is -NRcS(O)2NHCH3. In someembodiments, Z is -NRcCH2C6-aryl-S(O)2NH...In some embodiments, Rcis selected from hydrogen, C1-C6alkyl, C3-C6cycloalkyl or C1-C6haloalkyl. In some embodiments, Rcis methyl. In some embodiments, Rcis ethyl. In some embodiments, Rcis isopropyl. In some embodiments, Rcis n-propyl. In some embodiments, Rcis cyclopropyl. In some embodiments, Rcis cyclobutyl. In some embodiments, Rcis entyl. In some embodiments, Rcis . In some embodimeccyclopnts, R is . In some embodiments,

[0083] In some embodiments, Rcis -(C1-C6alkylene)C3-C6cycloalkyl optionally substituted with Rd. In some embodiments, Rcis 3- to 6-membered heterocyclyl optionally substituted with Rd. In some embodiments, Rcis -(C1-C6alkylene)3- to 6-membered heterocyclyl optionally substituted with Rd. In some embodiments, Rcis -(C1-C6alkylene)C6-aryl optionally substituted with Rd. In some embodiments, Rcis 5- to 6-membered heteroaryl optionally substituted with Rd. In some embodiments, Rcis -(C1-C6alkylene)5- to 6-membered heteroaryl optionally substituted with Rd. In some embodiments, Rcis -(C1-C6alkylene)OR13. In some embodiments, Rcis -(C1-C6alkylene)SR13. In some embodiments, Rcis -(C1-C6alkylene)S(O)2R13. In some embodiments, Rcis -(C1-C6alkylene)S(O)2NR14R15. In some embodiments, Rcis -(C1-C6alkylene)NR13S(O)2R14. In some embodiments, Rcis -(C1-C6alkylene)NR14R15. In some embodiments, Rcis -(C1-C6alkylene)C(O)R13. In some embodiments, Rcis -(C1-C6alkylene)NR13C(O)R14. In some embodiments, Rcis -(C1-C6alkylene)NR13C(O)NR14R15. In some embodiments, Rcis -(C1-C6alkylene)C(O)OR13. In some embodiments, Rcis -(C1-C6alkylene) C(O)ONR14R15. In some embodiments, Rcis -(C1-C6alkylene)-C(O)NR14R15.

[0084] In some embodiments, -C1-C6alkylene of any Rcgroup may be optionally substituted with Rd.

[0085] In some embodiments, -C1-C6alkylene of any Rcgroup is linear or branched.

[0086] In some embodiments, Rcis selected from the group consisting of methyl, ethyl, isopropyl, n-propyl, n-butyl, cyclopropyl, cyclobutyl, cyclopentyl,, , , , ,, , , wherein wavy lines denote the attachment points.

[0087] In some embodiments, Z is selected from the group consisting of, ,e the attachment points.

[0088] In some embodiments, Z is -NRcSO2NH2.

[0089] In some embodiments, Z is -NHSO2NH2. In some embodiments, Z is. In some embodiments, Z is. In some embodiments, Z is. In some embodiments, ZIn some embodiments, Z isIn some embodiments, Z is. In some embodiments, Z is

[0090] In some embodiments, Z is -NHS(O)2CH3.

[0091] In some embodiments, Z is -SO2NH2.

[0092] In some embodiments, Z is -NHC(O)CH3.

[0093] In some embodiments, Z is -C(O)OH.

[0094] In some embodiments, Z is -CONH2.

[0095] In some embodiments, Z is -NHCONH2.

[0096] In some embodiments, Z is -CONH(OH).

[0097] In some embodiments, Z is -B(OH)2.

[0098] In some embodiments, Z is -P(O)(OH)2.

[0099] In some embodiments, Z is -S(O)2OH.

[0100] In some embodiments, Z is -NRcS(O)2CF3.

[0101] In some embodiments, Z is -NRcS(O)2NHCH3.

[0102] In some embodiments, Z is -NRcCH2C6-aryl-S(O)2NH2.

[0103] In some embodiments, Z is -NHCH2C6-aryl-S(O)2NH2.

[0104] In some embodiments, Z is -N(CH3)CH2C6-aryl-S(O)2NH2.

[0105] In some embodiments, G, Ra, Rb, L, t and Z together is selected from the group consisting of, wherein wavy lines denote the attachment points.

[0106] In some embodiments, when any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a C4-C6cycloalkyl ring then L and Z together is selected from, wherein wavy lines denote the attachment points.

[0107] In some embodiments, when any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a C4-C6cycloalkyl ring then L and Z together is selected from, wherein wavy lines denote the attachment points.

[0108] In some embodiments, when any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a cyclobutyl ring (which forms with the azetidine an azaspiroheptane ring) then L and Z together is selected from the group consisting of

[0109] In some embodiments, Ring C is 5- to 6-membered heteroaryl optionally substituted with R1. In embodiments, Ring C is 5-membered heteroaryl optionally substituted with R1. In some embodiments, Ring C is 6-membered heteroaryl optionally substituted with R1. In some embodiments, Ring C is selected from the group consisting of imidazole, pyrazole, pyrrole, pyridine, pyrimidine, pyridone,pyrimidone, pyridazine, pyridazinone, and triazine, wherein each of which is optionally substituted with R1. In some embodiments, the R1is selected from hydrogen, oxo, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, 3- to 6-membered heterocyclyl, C6-aryl, 5- to 6-membered heteroaryl, -CN, -CONR11R12, or -NR11R12, wherein each of which is optionally substituted by R9. In some embodiments, the R1is selected from hydrogen, oxo, methyl, ethyl, ethylene, propynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantly, oxetanyl, tetrahydropyranyl, pyrazolyl, pyridyl, pyrimidyl, phenyl, -CONH2, -NH2, -CHF2, -CF3, -Cl, and -CN; wherein each of which is optionally further substituted with pyridyl, -F, -CF3, or -CHF2.

[0110] In some embodiments, Ring D is C6-aryl or 5- to 6-membered heteroaryl, each of which is optionally substituted with R2; wherein Ring D is fused to Ring C. In some embodiments, Ring D is C6- aryl optionally substituted with R2; wherein Ring D is fused to Ring C. In some embodiments, Ring D is 5- to 6-membered heteroaryl optionally substituted with R2; wherein Ring D is fused to Ring C. In some embodiments, Ring D is 5-membered heteroaryl optionally substituted with R2; wherein Ring D is fused to Ring C. In some embodiments, Ring D is 6-membered heteroaryl optionally substituted with R2; wherein Ring D is fused to Ring C. In some embodiments, Ring D is selected from the group consisting of phenyl, pyrrole, pyrazole, imidazole, pyridine, thiophene, and pyrimidine optionally substituted with R2; wherein each of which is fused to Ring C. In some embodiments, R2is selected from hydrogen, halogen, oxo, C1-C6alkyl, C1-C6alkoxy, C3-C6cycloalkyl, -OR10, -CN, and C6-aryl; wherein each of which is optionally substituted by R9. In some embodiments, R2is selected from hydrogen, fluorine, bromine, chloro, oxo, methyl, ethyl, isopropyl, methoxy, ethoxy, propoxy, phenyl, 4-methoxypheyl, -CN, -OCH2F, -OCH2OCH3, -(OCH2CH2)morpholine, 4-hydroxycyclohexyl, -CF3, cyclopropyl, and phenyl.

[0111] In some embodiments, any two of R2are taken together with the atoms to which they attached to form a C5-C6cycloalkyl, 5- to 6-membered heterocyclyl, C6-aryl, or 5- to 6-membered heteroaryl, wherein each of which is optionally substituted by R9. In some embodiments, any two of R2are taken together with the atoms to which they attached to form imidazole, dioxole and dihydro dioxine, wherein each of which is optionally substituted with methyl.

[0112] In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2.

[0113] In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

[0114] In some embodiments, Ring C, Ring D, R1, and R2together is selected from the group consisting of:lines denote the attachment points.

[0115] In some embodiments, Ring C, Ring D, R1, and R2together is selected from the group consisting of:; wherein wavy lines denote the attachment points.

[0116] It is understood that each description of A, G, Ra, Rb, L, Z, Ring C, Ring D, R1, R2, Rc, m, n, and t may be independently combined with each description of A, G, Ra, Rb, L, Z, Ring C, Ring D, R1, R2, Rc, m, n, and t the same as if each and every combination were specifically and individually listed.

[0117] In some embodiments, provided is a compound of Formula II:or a salt thereof, wherein A, Ra, Rb, L, Z, Ring C, Ring D, R1, R2, m, and n are each independently as detailed herein.

[0118] In some embodiments, provided is a compound of Formula III:or a salt thereof, wherein L, Z, Ring C, Ring D, R1, R2, m, and n are each independently as detailed herein.

[0119] In some embodiments, provided is a compound of Formula IV:or a salt thereof, wherein: X is N or CR1; Y is N or CR1; provided that at same time X and Y both are not N; and A, G, Ra, Rb, L, Z, Ring D, R1, R2, n, and t are each independently as detailed herein.

[0120] In some embodiments, provided is a compound of any one of Formula IV-1 to IV-11:IV-7 IV-8 IV-9IV-10 IV-11 or a salt thereof, wherein X1, X2, and X3are independently N, NR2, or CR2; provided that any one of X1, X2, and X3is NR2and others are N or CR2; and A, G, Ra, Rb, L, Z, R1, R2, and t are each independently as detailed herein.

[0121] In some embodiments, provided is a compound of Formula V:or a salt thereof, wherein X is N or CR1; Y is N or CR1; provided that at same time X and Y both are not N; and L, Z, Ring D, R1, R2, and n are each independently as detailed herein.

[0122] In some embodiments, provided is a compound of any one of Formula V-1 to V-11:or a salt thereof, wherein X1, X2, and X3are independently N, NR2, or CR2; provided that any one of X1, X2and X3is NR2and others are N or CR2; and L, Z, R1, and R2are each independently as detailed herein.

[0123] In some embodiments, provided is a compound of Formula VI: Ror a salt thereof, wherein Y1is N or NR1;Y2is N, NR1, or CR1; provided that any one of Y1and Y2is NR1and other one is other than NR1; and A, G, Ra, Rb, L, Z, R1, R2, and t are each independently as detailed herein.

[0124] In some embodiments, provided is a compound of Formula VII:or a salt thereof, wherein Z1is N or CR1; and A, G, Ra, Rb, L, Z, R1, R2, and t are each independently as detailed herein.

[0125] In some embodiments, provided is a compound of Formula VIII:or a salt thereof, wherein, A, G, Ra, Rb, L, Z, R1, R2, and t are each independently as detailed herein.

[0126] In some embodiments, provided is a compound of Formula IX:or a salt thereof, wherein Y1is N or NR1; Y2is N, NR1, or CR1; provided that any one of Y1and Y2is NR1and other one is other than NR1; and L, Z, R1, and R2are each independently as detailed herein.

[0127] In some embodiments, provided is a compound of Formula X:or a salt thereof, wherein Z1is N or CR1; and L, Z, R1, and R2are each independently as detailed herein.

[0128] In some embodiments, provided is a compound of Formula XI:or a salt thereof, wherein L, Z, R1and R2are each independently as detailed herein.

[0129] In some embodiments, provided is a compound of Formula XII:or a salt thereof, wherein, X is N or CR1; X4and X5is independently N or CR2; provided that at same time both are not N; and A, G, Ra, Rb, L, Z, R1, R2, and t are each independently as detailed herein.

[0130] In some embodiments, provided is a compound of any one of Formula XII-1 to XII-6:or a salt thereof, wherein A, G, Ra, Rb, L, Z, R1, R2, Rc, and t are each independently as detailed herein.

[0131] In some embodiments, provided is a compound of Formula XIII:or a salt thereof, wherein X is N or CR1; X4and X5is independently N or CR2; provided that at same time both are not N; and L, Z, R1, and R2are each independently as detailed herein.

[0132] In some embodiments, provided is a compound of any one of Formula XIII-1 to XIII-9:or a salt thereof, wherein L, Z, R1, R2, and Rcare each independently as detailed herein.

[0133] Also provided are salts of compounds referred to herein, such as pharmaceutically acceptable salts. The disclosure also includes any or all of the stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomers or other forms of the compounds described.

[0134] In some embodiments, the compound is selected from Table 1. It is understood that individual enantiomers and diastereomers are included in the generic compound structures shown in Table 1. Specific synthetic methods for preparing the compounds of Table 1 are provided in US20220135598A1. Table 1Kits

[0135] Provided herein are also kits that include a compound of the disclosure, or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analog thereof, and suitable packaging. In one embodiment, a kit further includes instructions for use. In one aspect, a kit includes a compound of the disclosure, or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analog thereof, and a label and / or instructions for use of the compounds in the treatment of the indications, including the diseases or conditions, described herein.

[0136] Provided herein are also articles of manufacture that include a compound described herein or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analog thereof in a suitable container. The container may be a vial, jar, ampoule, preloaded syringe, and intravenous bag. Pharmaceutical Compositions and Modes of Administration

[0137] Compounds provided herein are usually administered in the form of pharmaceutical compositions. Thus, provided herein are also pharmaceutical compositions that contain one or more of the compounds described herein or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analog thereof and one or more pharmaceutically acceptable vehicles selected from carriers, adjuvants and excipients. Suitable pharmaceutically acceptable vehicles may include, for example, inert solid diluents and fillers, diluents, including sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers and adjuvants. Such compositions are prepared in a manner well known in the pharmaceutical art. See, e.g., Remington’s Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa.17th Ed. (1985); and Modern Pharmaceutics, Marcel Dekker, Inc.3rd Ed. (G.S. Banker & C.T. Rhodes, Eds.).

[0138] The pharmaceutical compositions may be administered in either single or multiple doses. The pharmaceutical composition may be administered by various methods including, for example, rectal, buccal, intranasal and transdermal routes. In certain embodiments, the pharmaceutical composition may be administered by intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, or as an inhalant.

[0139] One mode for administration is parenteral, for example, by injection. The forms in which the pharmaceutical compositions described herein may be incorporated for administration by injection include, for example, aqueous or oil suspensions, or emulsions, with sesame oil, corn oil, cottonseed oil, or peanut oil, as well as elixirs, mannitol, dextrose, or a sterile aqueous solution, and similar pharmaceutical vehicles.

[0140] Oral administration may be another route for administration of the compounds described herein. Administration may be via, for example, capsule or enteric coated tablets. In making the pharmaceutical compositions that include at least one compound described herein or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analog thereof, the active ingredient is usually diluted by an excipient and / or enclosed within such a carrier that can be in the form of a capsule, sachet, paper or other container. When the excipient serves as a diluent, it can be in the form of a solid, semi-solid, or liquid material, which acts as a vehicle, carrier or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, sterile injectable solutions, and sterile packaged powders.

[0141] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methyl cellulose. The formulations can additionally include lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as methyl and propylhydroxybenzoates; sweetening agents; and flavoring agents.

[0142] The compositions that include at least one compound described herein or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analog thereof can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the subject by employing procedures known in the art. Controlled release drug delivery systems for oral administration include osmotic pump systems and dissolutional systems containing polymer-coated reservoirs or drug-polymer matrix formulations. Examples of controlled release systemsare given in U.S. Patent Nos.3,845,770; 4,326,525; 4,902,514; and 5,616,345. Another formulation for use in the methods disclosed herein employ transdermal delivery devices (“patches”). Such transdermal patches may be used to provide continuous or discontinuous infusion of the compounds described herein in controlled amounts. The construction and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. See, e.g., U.S. Patent Nos.5,023,252, 4,992,445 and 5,001,139. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents.

[0143] For preparing solid compositions such as tablets, the principal active ingredient may be mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound described herein or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug, or deuterated analog thereof. When referring to these preformulation compositions as homogeneous, the active ingredient may be dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules.

[0144] The tablets or pills of the compounds described herein may be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action, or to protect from the acid conditions of the stomach. For example, the tablet or pill can include an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer that serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.

[0145] Compositions for inhalation or insufflation may include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described herein. In some embodiments, the compositions are administered by the oral or nasal respiratory route for local or systemic effect. In other embodiments, compositions in pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizing device or the nebulizing device may be attached to a facemask tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered, preferably orally or nasally, from devices that deliver the formulation in an appropriate manner. Dosing

[0146] The specific dose level of a compound of the present application for any particular subject will depend upon a variety of factors including the activity of the specific compound employed, the age, bodyweight, general health, sex, diet, time of administration, route of administration, and rate of excretion, drug combination and the severity of the particular disease in the subject undergoing therapy. For example, a dosage may be expressed as a number of milligrams of a compound described herein per kilogram of the subject’s body weight (mg / kg). Dosages of between about 0.1 and 150 mg / kg may be appropriate. In some embodiments, about 0.1 and 100 mg / kg may be appropriate. In other embodiments a dosage of between 0.5 and 60 mg / kg may be appropriate. Normalizing according to the subject’s body weight is particularly useful when adjusting dosages between subjects of widely disparate size, such as occurs when using the drug in both children and adult humans or when converting an effective dosage in a non-human subject such as dog to a dosage suitable for a human subject.

[0147] The daily dosage may also be described as a total amount of a compound described herein administered per dose or per day. Daily dosage of a compound of Formula I may be between about 1 mg and 4,000 mg, between about 2,000 to 4,000 mg / day, between about 1 to 2,000 mg / day, between about 1 to 1,000 mg / day, between about 10 to 500 mg / day, between about 20 to 500 mg / day, between about 50 to 300 mg / day, between about 75 to 200 mg / day, or between about 15 to 150 mg / day.

[0148] When administered orally, the total daily dosage for a human subject may be between 1 mg and 1,000 mg, between about 1,000-2,000 mg / day, between about 10-500 mg / day, between about 50-300 mg / day, between about 75-200 mg / day, or between about 100-150 mg / day.

[0149] The compounds of the present application or the compositions thereof may be administered once, twice, three, or four times daily, using any suitable mode described above. Also, administration or treatment with the compounds may be continued for a number of days; for example, commonly treatment would continue for at least 7 days, 14 days, or 28 days, for one cycle of treatment. Treatment cycles may be alternated with resting periods of about 1 to 28 days, commonly about 7 days or about 14 days, between cycles. The treatment cycles, in other embodiments, may also be continuous.

[0150] In a particular embodiment, the method comprises administering to the subject an initial daily dose of about 1 to 800 mg of a compound described herein and increasing the dose by increments until clinical efficacy is achieved. Increments of about 5, 10, 25, 50, or 100 mg can be used to increase the dose. The dosage can be increased daily, every other day, twice per week, or once per week. Synthesis of the Compounds

[0151] The compounds may be prepared using the methods disclosed in US20220135598A1 which incorporated by reference in its entirety and routine modifications thereof, which will be apparent given the disclosure therein and methods well known in the art. Conventional and well-known syntheticmethods may be used in addition to the teachings therein. If available, reagents may be purchased commercially, e.g., from Sigma Aldrich or other chemical suppliers. EXAMPLES

[0152] The following examples are included to demonstrate specific embodiments of the disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques to function well in the practice of the disclosure, and thus can be considered to constitute specific modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the disclosure. Biological Assays The following examples describe biological assays for testing ENPP1 inhibitors of the present disclosure. Example 1: ENPP1 Inhibition assay using an ATP substrate

[0153] The ability of compounds of the present disclosure to inhibit ENPP1 was tested in a biochemical ENPP1 inhibition assay with ATP as the substrate. Test compound stock solutions were prepared in DMSO and then serially diluted into 10 concentrations by 3-fold dilution in a 384-well plate (ProxiPlate-384 Plus; PE# 6008280), using a TECAN EVO200 liquid handler.30 nL of stock was transferred into a 384-well plate using an Echo550 dispenser. DMSO was used as vehicle control. A 5 µL solution containing 120 pM ENPP1 enzyme (R&D# 6136-EN-10), 2 mM MgCl2, 1 mM TCEP pH 7.0, 50 mM Tris HCl and 0.005% Tween-80 was added to assay plate and incubated for 30 min at RT. A 5 µL mix containing 300 nM ATP (Promega#V915B) and 2 mM MgCl2, 1 mM TCEP pH7.0, 50 mM Tris HCl and 0.005% Tween80 were added to all wells. The final concentrations of ENPP1 enzyme and ATP were 60 pM and 300 nM, respectively, in a total volume of 10 µL. The plate was incubated for 90 min at RT. The reaction was stopped by adding 5 µL of stop solution containing 20 mM EDTA and 50 mM Tris- HCl, 0.1% Prionex, 6 nM AMP / GMP HiLyte647 Tracer (BellBrook Labs# 3020-10K) and 6 nM AMP / GMP antibody-Tb to all wells, and the plate was incubated for another 120 minutes. The generated signal was measured using an Envision instrument. Remaining activity (%) was calculated using the read conversion ratio (CR) following the equation: Remaining activity (%) =100 x (CRsample-CRLC) / (CRHC-CRLC)

[0154] IC50values were calculated using XLFit (equation 201) by floating both bottom and top.

[0155] Certain compounds disclosed in Table 1 were tested in the above assay and exhibited an IC50of less than 3 µM. IC50for certain compounds are shown in Table A, where the data is presented in ranges, in which “+++” < 100 nM, 100 nM ≤ “++” < 3000 nM, and “+” ≥ 3000 nM. Table AExample 2: Mineralization assay

[0156] The ability of compounds of the present disclosure to modulate mineralization of human bone- related cells was examined. Saos-2 cells (ATCC CAT# HTB-85) were plated in 24 well plate at a seeding density of 3.5 × 105cells / well and allowed to grow to 100% confluency over the following 1-2 days. Once cells were completely confluent, the media was replaced with fresh media supplemented with 1x Ascorbic Acid (Sigma # A4544, “AA”), 1x β-Glycerophosphate (Sigma # G9422, “BGP”) and varying concentrations of ENPP1 inhibitor. The day of addition of the mineralization components was considered to be Day 1. On Day 3, the media was gently aspirated with no wash followed by the addition of fresh media, mineralization components, and ENPP1 inhibitor at concentrations of 1 and 10 µM. On day 5, the media was gently aspirated and the cells were washed twice with PBS. The cells were then fixed with ice cold 70% ethanol for 1 hour at 4 °C. Cells were washed once with PBS and stained with Alizarin Red for 30 minutes at room temperature with gentle rotation. Stained cells were washed three times with water for 10 minutes at room temperature with gentle rotation. After imaging the cells, dye extraction reagent was added to each well and the cells incubated for 30 minutes at room temperature. Alizarin Red was quantified by measuring the absorbance at a wavelength of 405 nm using an Envision instrument.

[0157] As shown in FIG.1, compounds of the present disclosure have the ability to modulate mineralization. “AA+BGP” represents 1x Ascorbic Acid+1x β-Glycerophosphate. Compounds 131 and 134 showed high mineralization level in 1 µM than in 10 µM, respectively. Compounds 135 and 76 showed similar mineralization level in both 1 µM and 10 µM. Example 3: Impact of compounds on PPi levels in WT animals

[0158] The effect of an exemplary compound of the present disclosure on PPi levels in wild-type animals was examined. Compound 76 was orally dosed to 6-8-week old female SD rats (n=3) at 100mg / kg, in a single dose, and to 6-8-week old C57BL / 6J mice (n=6) at 100mg / kg, BID for 5 days. Plasma samples were taken at pre-dose and 1, 4 and 8 hours post-dose from rats, and at 1 and 8 hours post-last dose from mice in heparin SST tubes and spun down (7000 rpm x 10 min) immediately. For PPi analysis, the platelet removal step was performed by spinning down the plasma with a Spin-X column (0.22 μM, Costar 8160) immediately (7000 rpm x 2 min) and the eluted plasma was aliquoted / frozen down on dry ice immediately. Prior to PPi analysis, plasma samples were de-proteinated with a 10 kD cutoff column (Amicon #UFC501024) (4 ℃). PPi from filtered plasma was first converted to ATP by ATP sulfurylase (R&D# 7175-AS-020) in the presence of adenosine phosphosulfate (APS; Sigma# A5508). Reactions were incubated at 37°C for 30 minutes followed by 10 minutes at 90°C to deactivate ATP sulfurylase. The resulting ATP was then quantified with Bactiter Glo detection reagent (Promega G8230; Madison, WI, USA). To account for endogenous ATP present in the plasma, a blank reaction was run for each sample with heat-inactivated ATP sulfurylase. The luminescent signals from this reaction were subtracted from the total luminescent signals to calculate the plasma PPi levels.

[0159] Compound 76 is able to reduce the PPi levels in rats in a time-dependent manner after a single dose (1, 4 and 8 hours afterwards), as shown in FIG.2, as well as in mice 1 and 8 hours after the last dose of a continuous dosing for 5 days, as shown in FIG.3. Example 4: Mineralization assay in TNAP KO MC3T3-E1-C4 cells

[0160] Compounds of the present disclosure are tested in a mineralization assay in TNAP knockout MC3T3-E1-C4 cells. The osteoblastic cell line (MC3T3-E1), established from C57BL / 6 mouse calvaria (skull), has the ability to synthesize extracellular matrix (collagen) and differentiate into osteoblasts and osteocytes in vitro. To induce mineralization, cells are grown for five days in αMEM containing 50 μg / ml ascorbate followed by supplementation with 2.5 mM NaPO4or 5 mM β-glycerophosphate as described previously (Liu et al.2014, Bone.67: 81–94). After 5 days, the media is gently aspirated with no wash followed by the addition of fresh media, mineralization components, and ENPP1 inhibitors in different concentrations. After 7 days, the media is gently aspirated, and the cells are washed twice with PBS. The cells are then fixed with ice cold 70% ethanol for 1 hour at 4 °C. Cells are washed once with PBS andstained with Alizarin Red for 30 minutes at room temperature with gentle rotation. Stained cells are washed three times with water for 10 minutes at room temperature with gentle rotation. After imaging the cells, dye extraction reagent is added to each well and the cells incubated for 30 minutes at room temperature. Alizarin Red is quantified by measuring the absorbance at a wavelength of 405 nm using an Envision instrument. Example 5: ENPP1 assay in TNAP KO MC3T3-E1-C4

[0161] Cells are treated with varying concentrations of an ENPP1 inhibitor of the present disclosure and incubated for 5 days. After 5 days the monolayer of cells is rinsed with cold PBS twice and 150ul of lysis buffer is added to each well. The cells are lysed using freeze-thaw method and centrifuged at 13.3 rpm for 10 min at 4°C and supernatant is collected. Samples are incubated with assay buffer (100mM Tris (pH9.0), 500mM NaCl, 5mM MgCl2, 0.05% Triton X-100). To each well, 2mM pNP-TMP (Sigma# T4510) is added as substrate. The production of p-nitrophenol is analyzed kinetically by measuring the absorbance at 405 nm using a Molecular Devices (San Jose, CA, USA) SpectraMax M2 plate reader. The activity is calculated as the change in absorbance over time (mOD / min). Example 6: Effect of compounds on survival of Akp- / - mice

[0162] Akp- / - mice (n = 17-20) are treated with an ENPP1 inhibitor of the present disclosure orally with QD dosing or vehicle starting no later than 2 days after birth and continuing up to at least 25 days of age. Body weights are measured daily and survival is analyzed at the end of the study. PPi concentrations in the plasma are measured at different time points (e.g., 0, 12, 24, 48 and 96 hours) after the last dose. Bone abnormalities are assessed by X-ray 24 hours after the last dose on study day. Example 7: Effect of compounds on Akp2+ / −mice

[0163] Akp2+ / −mice (n = 15) are treated with an ENPP1 inhibitor of the present disclosure orally with QD daily for at least 25 days. Body weights are analyzed daily. PPi concentrations in the plasma are measured at different time points (0, 12, 24, 48 and 96 hours) after the last dose. Bone abnormalities are assessed by X-ray 24 hours after the last dose on study day.

[0164] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0165] The disclosure illustratively described herein may suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. Thus, for example, the terms “comprising,” “including,” “containing,” etc. shall be read expansively and without limitation. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding anyequivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the disclosure.

[0166] Thus, it should be understood that although the present disclosure has been specifically disclosed by certain embodiments and optional features, modification, improvement and variation may be resorted to by those skilled in the art, and that such modifications, improvements and variations are considered to be within the scope of this disclosure. The materials, methods, and examples provided here are exemplary, and are not intended as limitations on the scope of the disclosure.

[0167] Specific embodiments have been described broadly and generically herein. Each of the narrower species and subgeneric groupings also form part of the disclosure. This includes the generic description with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein.

[0168] All publications, patent applications, patents, and other references mentioned herein are expressly incorporated by reference in their entirety, to the same extent as if each were incorporated by reference individually. In case of conflict, the present specification, including definitions, will control.

[0169] It is to be understood that while the disclosure has been described in conjunction with the above embodiments, that the foregoing description and examples are intended to illustrate and not limit the scope of the disclosure. Other aspects, advantages and modifications within the scope of the disclosure will be apparent to those skilled in the art to which the disclosure pertains.

Claims

CLAIMS What is claimed is:

1. A method of treating or preventing hypophosphatasia in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I:or a pharmaceutically acceptable salt thereof, wherein: Ring C is 5- to 6-membered heteroaryl; Ring D is C6-aryl or 5- to 6-membered heteroaryl, wherein Ring D is fused to Ring C; A is hydrogen, C1-C6alkyl, or C6-aryl, each of which is optionally substituted with halogen; G is a bond, -CH2-, or -CH2-CH2-; Raand Rbare independently hydrogen or C1-C6alkyl; or Raand Rbare taken together with the atoms to which they are attached to form a C3-C6cycloalkyl ring; or any one of Raand Rb, and A are taken together along with the atoms to which they are attached to form a C4-C6cycloalkyl ring; L is a bond, linear or branched C1-C6alkylene, or linear or branched C2-C6alkenylene; t is 0 or 1; provided that when t is 0, then L is linear or branched C2-C6alkenylene;Z is -NRcS(O)2NH2, -NRcS(O)2CH3, -SO2NH2, -NRcC(O)CH3, -C(O)OH, -CONH2, -NRcCONH2, -CONH(OH), -B(OH)2, -P(O)(OH)2, -SO2OH, -NRcS(O)2CF3, -NRcS(O)2NHCH3, or -NRcCH2C6-aryl-S(O)2NH2. each R1and R2is independently oxo, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, adamantyl, 3- to 6-membered heterocyclyl, C6-aryl, 5- to 6-membered heteroaryl, -CN, halogen, C1-C6alkoxy, C1-C6haloalkoxy, -OR10, -SR10, -S(O)2R10, -S(O)2NR11R12, -NR10S(O)2R11, -NR11R12, -C(O)R10, -NR10C(O)R11, -NR10C(O)NR11R12, -C(O)OR10, -C(O)ONR11R12, or -C(O)NR11R12, wherein each is independently optionally substituted by R9; or two R2are taken together with the atoms to which they are attached to form a C5-C6cycloalkyl, 5- to 6-membered heterocyclyl, C6-aryl, or 5- to 6-membered heteroaryl, wherein each is independently optionally substituted by R9; each R9is independently selected from the group consisting of oxo, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, 3- to 6-membered heterocyclyl, C6-aryl, 5- to 6-membered heteroaryl, -CN, halogen, C1-C6alkoxy, C1-C6haloalkoxy, C1-C6haloalkyl, -OR13, -SR13, -S(O)2R13, -S(O)2NR14R15, -NR13S(O)2R14, -NR14R15, -C(O)R13, -NR13C(O)R14, -NR13C(O)NR14R15, -C(O)OR13, -C(O)ONR14R15, -C(O)NR14R15, and C1-C6alkyl optionally substituted by oxo, -OH, or halogen; each Rcis independently hydrogen, C1-C6alkyl, C3-C6cycloalkyl, -(C1-C6alkylene)C3-C6cycloalkyl, 3- to 6-membered heterocyclyl, -(C1-C6alkylene)3- to 6-membered heterocyclyl, -(C1-C6alkylene)C6-aryl, 5- to 6-membered heteroaryl, -(C1-C6alkylene)5- to 6-membered heteroaryl, C1-C6haloalkyl, -(C1-C6alkylene)OR13, -(C1-C6alkylene)SR13, -(C1-C6alkylene)S(O)2R13, -(C1-C6alkylene)S(O)2NR14R15, -(C1-C6alkylene)NR13S(O)2R14, -(C1-C6alkylene)NR14R15, -(C1-C6alkylene)C(O)R13, -(C1-C6alkylene)NR13C(O)R14, -(C1-C6alkylene)NR13C(O)NR14R15, -(C1-C6alkylene)C(O)OR13, -(C1-C6alkylene) C(O)ONR14R15, or -(C1-C6alkylene)-C(O)NR14R15, wherein each is independently optionally substituted with Rd; each Rdis independently selected from the group consisting of halogen, -OH, oxo, -CN, C1-C6haloalkyl, C1-C6alkoxy, C1-C6haloalkoxy, -COOH, and C1-C6alkyl optionally substituted with -OH, halogen, CN, or oxo; each R10, R11, and R12is independently hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C6cycloalkyl, -(C1-C6alkylene)C3-C6cycloalkyl, 3- to 6-membered heterocyclyl, or -(C1-C6alkylene)3- to 6-membered heterocyclyl, wherein each of R10, R11and R12is independently optionally substituted by oxo, C2-C6alkenyl, C2-C6alkynyl, -CN, halogen, C1-C6alkoxy, or C1-C6alkyl optionally substituted by oxo, -OH, or halogen;or R11and R12are taken together with the atom(s) to which they are attached to form a 3-6 membered heterocyclyl optionally substituted by oxo, -OH, halogen, or C1-C6alkyl optionally substituted by oxo, -OH, or halogen; each R13, R14and R15is independently hydrogen, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6haloalkyl, C3-C6cycloalkyl, or 3- to 6-membered heterocyclyl, wherein each of R10, R11, and R12is independently optionally substituted by oxo, -OH, C2-C6alkenyl, C2-C6alkynyl, -CN, halogen, or C1-C6alkyl optionally substituted by oxo, -OH, or halogen; or R14and R15are taken together with the atoms to which they are attached to form a 3-6 membered heterocyclyl optionally substituted by oxo, -OH, halogen, or C1-C6alkyl optionally substituted by oxo, -OH, or halogen; m is 0, 1 or 2; and n is 0, 1, 2, 3 or 4; provided that when Z is -NRcS(O)2CH3, -CONH2, or -C(O)OH, G is a bond, and t is 1, then L is not a bond; and the compound is not 1-[7-(4-bromo-2,6-dimethylphenyl)-2,5-dimethyl-7H-pyrrolo[2,3- d]pyrimidin-4-yl]-3-azetidinebutanoic acid, 1-[7-(4-bromo-2,6-dimethylphenyl)-2,5-dimethyl-7H- pyrrolo[2,3-d]pyrimidin-4-yl]-3-azetidinepentanoic acid, or N-((1-(6-fluorobenzo[d]oxazol-2-yl)azetidin- 3-yl)methyl)-N-methylacetamide.

2. The method of claim 1, wherein the compound is selected from Table 1, or a pharmaceutically acceptable salt thereof.

3. The method of claim 1 or 2, wherein the compound, or a pharmaceutically acceptable salt thereof, is formulated as a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier.

4. The method of any one of claims 1 to 3, wherein the compound, or a pharmaceutically acceptable salt thereof, is formulated for oral delivery.

5. The method of any one of claims 1 to 3, wherein the compound, or a pharmaceutically acceptable salt thereof, is formulated for subcutaneous delivery.

6. The method of any one of claims 1 to 3, wherein the compound, or a pharmaceutically acceptable salt thereof, is formulated for intraperitoneal delivery.

7. The method of any one of claims 1 to 6, wherein the compound, or a pharmaceutically acceptable salt thereof, is administered acutely to the subject.

8. The method of any one of claims 1 to 6, wherein the compound, or a pharmaceutically acceptable salt thereof, is administered chronically to the subject.

9. The method of any one of claims 1 to 8, wherein the subject is a mammal.

10. The method of claim 9, wherein the mammal is human.

11. The method of any one of claims 1 to 10, wherein the hypophosphatasia is selected from the group consisting of late-onset hypophosphatasia, prenatal hypophosphatasia, perinatal hypophosphatasia, infantile hypophosphatasia, childhood hypophosphatasia, adult hypophosphatasia, odontohypophosphatasia, and hypophosphatasia that has dental involvement and / or premature tooth loss.

12. The method of claim 11, wherein the hypophosphatasia is adult hypophosphatasia.

13. The method of claim 11, wherein the hypophosphatasia is late-onset hypophosphatasia.

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