Therapies for diseases of copper metabolism

Abstract

The disclosure relates to therapeutic methods comprising administering compounds that can inhibit prolyl hydroxylase and pharmaceutical compositions which comprise the prolyl hydroxylase inhibitors. In particular, the methods and pharmaceutical compositions described herein can be useful for treating a disease or condition of copper metabolism in subjects in need thereof, such as a subject having Wilson disease or Menkes disease.

Description

TITLE: THERAPIES FOR DISEASES OF COPPER METABOLISMCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to provisional application U.S. Serial No. 63 / 516,169, filed July 28, 2023, which is incorporated herein by reference in its entirety.GOVERNMENT SUPPORT

[0002] This invention was made with government support under R01 DK131190 awarded by the National Institutes of Health. The government has certain rights in the invention.SEQUENCE LISTING XML

[0003] The instant application contains a sequence listing, which has been submitted in XML file format by electronic submission and is hereby incorporated by reference in its entirety. The XML file, created on July 24, 2024, is named P14516WOOO.xml and is 8,751 bytes in size.TECHNICAL FIELD

[0004] The present disclosure relates to methods and compositions for treating a copper metabolism-associated disease.BACKGROUND

[0005] Copper (Cu) is an essential cofactor in enzymes that function in a wide range of biochemical processes. The importance of Cu lies in its ability to cycle between Cu1+and Cu2+oxidation states, which enables cuproenzymes to catalyze redox reactions. However, this same redox property makes Cu potentially toxic at elevated concentrations. Consequently, cells have evolved a sophisticated cadre of transporters and Cu-binding proteins to control the uptake, intracellular movement, storage and efflux of Cu, while preventing formation of the free ion within the cytoplasm. Whole body Cu homeostasis is maintained at two points of regulatory control: 1) Cu absorption across the small intestine, which is facilitated via the Cu-exporting fltype ATPase, ATP7A; and 2) Cu excretion from the liver into the bile, which is mediated by the related Cu- ATPase, ATP7B.

[0006] Mutations in ATP7A give rise to a lethal pediatric disorder of systemic Cu deficiency called Menkes disease, whereas mutations in ATP7B give rise to a disorder of hepatic Cu overload called Wilson disease. Menkes disease affects males due to its X-linked inheritance, and affected patients exhibit a range of defects that reflect systemic deficiencies in cuproenzyme activity, including neurological degeneration, seizure, hypotonia, hypothermia, hypopigmentation and connective tissue defects. Injections of Cu salts are the standard of care for Menkes patients, however, the prognosis rarely improves with such treatments and most patients die in early childhood. In Wilson disease patients, Cu accumulates primarily in the liver,with secondary elevations in plasma, urine, brain, and cornea. The Cu accumulation in Wilson patients is a progressive process that typically begins by age 6, with clinical symptoms of liver disease appearing in late adolescence. Treatments for Wilson disease include the Cu-chelator, penicillamine or trientine, which is taken with food to block the absorption of Cu in the intestine. However, such treatments cause significant neurological deterioration in -30% of patients that can lead to permanent impairment. Furthermore, because Cu chelators are ineffective in patients with advanced liver disease or fulminant liver failure, liver transplantation may be the only treatment option. Thus, there exists a need in the art for improved treatments for both Menkes disease and Wilson disease.SUMMARY

[0007] Certain aspects of the present disclosure are based on the surprising discovery that prolyl hydroxylase inhibitors activate ATP7A and ATP7B expression and that these drugs represent novel therapies for Wilson disease and Menkes disease.

[0008] Pharmaceutical compositions for the treatment of Menkes disease comprising a prolyl hydroxylase inhibitor and a copper salt are provided.

[0009] Methods of treating Wilson disease or Menkes disease in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising a prolyl hydroxylase inhibitor are also provided.

[0010] While multiple embodiments are disclosed, still other embodiments of the present disclosure will become apparent based on the detailed description, which shows and describes illustrative embodiments of the disclosure. Accordingly, the figures and detailed description are to be regarded as illustrative in nature and not restrictive.BRIEF DESCRIPTION OF THE FIGURES

[0011] The following drawings form part of the specification and are included to further demonstrate certain embodiments. In some instances, embodiments can be best understood by referring to the accompanying figures in combination with the detailed description presented herein. The description and accompanying figures may highlight a certain specific example, or a certain embodiment. However, one skilled in the art will understand that portions of the example or embodiment may be used in combination with other examples or embodiments.

[0012] FIG. 1A-C shows a genome-wide knockout screen identified the von Hippel Lindau (VHL) gene as a novel regulator of Cu tolerance in Cu-sensitive Cuscells. FIG. 1A shows the workflow of the CRISPR-based screen to identify gRNAs that confer Cu tolerance in Cuscells. The most enriched gRNAs pulled out of this screen were against CTR1 (a Cu importer) and VHLI (SEQ ID NOs: 1-5). FIG. IB shows sequence verification of mutations in CTR1 and VHLgenes (SEQ ID NOs: 6-9). To verify the results of the screen, Cuscells were subjected to CRISPR-Cas9 mediated disruption of CTR1 and VHL genes using independent gRNAs and then selected in IpM Cu. Single colonies were picked and expanded to derive clonally pure Cus / CTR1- and Cus / VHL- cell lines. Fragments of CTR1 and VHL genes corresponding to each gRNA were amplified by PCR and sequenced. FIG. 1C shows Cu resistance in Cus / CTR1- and Cus / VHL- cell lines was determined using a Crystal Violet growth assay and compared against Cusparental cells.

[0013] FIG. 2A-G shows CRISPR-targeted disruption of the VHL gene induced ATP7B expression at the mRNA and protein level. FIG. 2A shows detection ATP7B mRNA levels in both Cusand Cus / VHL- cell lines by quantitative real time PCR. Values are expressed as the mean ± SEM normalized against 18S rRNA (**** < 0.0001; n = 3). FIG. 2B shows immunoblot analysis of ATP7B and HIF-la expression in both Cusand Cus / VHL- cells. FIG. 2C shows immunofluorescence microscopy detection of ATP7B in both Cusand Cus / VHL- cell lines. Nuclei were stained with DAPI. FIG. 2D shows forced expression of VHL in Cus / VHL- cells reduces ATP7B expression. The Cus / VHL- cells were transiently transfected with low levels of a plasmid expressing HA-tagged VHL (VHL-HA). After 2 days, immunofluorescence microscopy was performed using anti-HA antibodies to identify VHL-HA expressing cells and ATP7B protein was detected using anti-ATP7B antibodies. Nuclei were stained with DAPI. Note that ATP7B expression is lost in transfected cells but not in cells that failed to be transfected with VHL-HA (arrowheads). FIG. 2E shows forced expression of ATP7B in Cuscells confers viability in basal medium. Cuscells are inviable in basal medium but rescued by transfection with an ATP7B-expression plasmid (pATP7B). As a control, Cuscells were shown to be viable in basal medium containing the Cu chelator, BCS (IpM). Cells were grown for 5 days and stained with Crystal violet. FIG. 2F-G shows quantitative real time PCR was performed to detect ATP7B and ATP7A mRNA levels in WT and VHL-null renal carcinoma cells (RENCA WT and RENCA VHL- cells). Values are expressed as the mean ± SEM normalized to 18S rRNA (****p < 0.0001; n = 3).

[0014] FIG. 3A-E shows hypoxia confers viability to Cuscells and induces ATP7B expression in vitro and in vivo. FIG. 3A shows Cuscells (2 x 104cells / well) in basal media were exposed to either 21%O2 or 1%O2 for 4 days and then fixed and stained with Crystal violet. FIG. 3B shows quantitative real time PCR analysis oiATP7B mRNA in Cuscells exposed for 24 h to either 21% O2 or 1% O2. Values are expressed as the mean ± SEM normalized to 18S rRNA (****p < 0.0001; n = 3). FIG. 3C shows immunoblot analysis of ATP7B and ATP7A proteins in the livers of C57BL / 6 mice exposed to 21% O2 or 10% O2 for 5 days. Ponceau staining was used asa control for protein loading. Each lane represents the liver of a different mouse. FIG. 3D shows ceruloplasmin (CP) activity in the serum of mice exposed for 5 days to either 21% O2 or 1% O2 (mean ± SEM; **p < 0.01. FIG. 3E shows Cu concentrations in the liver of mice exposed for 5 days to either 21% O2 or 1% O2 were determined by ICP-MS (mean ± SEM; *p < 0.05).

[0015] FIG. 4A-E shows Roxadustat confers viability to Cuscells and induces ATP7B expression in vitro and in vivo. FIG. 4A shows Roxadustat rescues the growth of Cuscells in basal medium. Cuscells were seeded in basal media containing either IpM BCS (positive control), 1% (v / v) DMSO (vehicle control), or the indicated concentrations of Roxadustat in DMSO. After 5 days, cells were fixed and stained with Crystal Violet to reveal surviving cells. FIG. 4B shows quantitative real time PCR was performed to detect ATP7B mRNA levels in Cuscells treated for the indicated times with 1% DMSO or Roxadustat (40pM). Values were normalized to 18S rRNA and expressed as the mean ± SEM (****p < 0.0001; n = 3). FIG. 4C shows immunoblot analysis of ATP7B in Cuscells treated for 120 h (5 days) in basal medium containing 40pM Roxadustat or 1% DMSO (for 24 h; prior to cell death). Tubulin was detected as a loading control. FIG. 4D-E shows Roxadustat induces hepatic expression of ATP7B and ATP7A in mice. Wild type C57BL / 6 mice (age 8 weeks) were injected intraperitoneally with Roxadustat (10 mg / kg) or 1% DMSO (vehicle) every 24 h over 5 days. Livers were subjected to immunoblot analysis of ATP7B. Ponceau staining was used as a control for protein loading. Each lane represents a different mouse liver.

[0016] FIG. 5A-D shows the Cu-sensitive phenotype of Cuscells is rescued by multiple prolyl hydroxylase inhibitors. Cuscells were treated with Roxadustat, Daprodustat, Vadadustat, or Enarodustat at the indicated concentrations for 5 days, then stained with Crystal Violet dye. The Cu chelator, BCS (IpM), was used as a positive control and DMSO was a vehicle control. FIG. 5A-B shows quantification of Cuscell rescue by Roxadustat and Daprodustat. FIG. 5C-D shows visualization of Crystal Violet stained Cuscells showing the rescue by Vadadustat and Enradustat.

[0017] FIG. 6A-D shows Prolyl hydroxylase inhibitors restore Cu-delivery to tyrosinase in cells lacking ATP7A. FIG. 6A shows ATP7A- B16 cells were incubated with DMSO or Roxadustat for the indicated time points followed by in-situ tyrosinase assay (L-DOPA oxidase activity). Wild type B16 cells were used as a positive control for tyrosinase activity. FIG. 6B shows quantitative analysis of tyrosinase activity in WT and ATP7A- B16 cells treated for 5 days with either DMSO (-) or 40 pM Roxadustat (+). Values are expressed as the mean ± SEM (****p < 0.0001). FIG. 6C shows immunoblot analysis of ATP7B protein expression in ATP7A- B16 cells treated for 5 days with 40pM Roxadustat. FIG. 6D shows in-situ tyrosinase assay inATP7A- B16 cells treated for 5 days with DMSO vehicle or the indicated concentrations of Daprodustat.

[0018] FIG. 7A-B shows Roxadustat activates Cu-dependent melanogenesis and enhances survival of the mobr mouse model of Menkes disease. FIG. 7A is a table showing five different treatments administered to mobr mice via subcutaneous injections on PND7 and PND10 and their survival at PND21 and PND90. FIG.7B shows images of mice from each treatment group at PND14 along with a wild type littermate control to illustrate the normal coat color of the C57BL6 strain. Note the hypopigmentation of control mobr mice and the ability of Roxadustat to potentiate the effect of Cu-HIS on melanogenesis.

[0019] FIG. 8A-I shows Roxadustat reduces liver pathology in a mouse model of Wilson disease. Three-month-old WT and ATP7B-KO mice were treated each day for 10 days via intraperitoneal injection with either DMSO (vehicle control) or Roxadustat (lOmg / kg) and then euthanized for analysis. FIG. 8A shows hematoxylin and eosin staining of liver tissue. Untreated DMSO control ATP7B-KO livers exhibit enlarged hepatocytes with pleomorphic nuclei, inflammatory infiltrates and steatosis (arrows). These pathological features were diminished in the livers of Roxadustat treated ATP7B-KO mice. FIG. 8B-C shows quantitative analysis of density and diameter of nuclei revealed significant normalization in the livers of ATP7B-KO mice treated with Roxadustat compared to DMSO. FIG. 8D-F shows concentrations of liver enzymes AST, ALP and ALT in the plasma of WT and ATP7B-KO mice treated with DMSO or Roxadustat. Note that Roxadustat significantly reduced these markers of liver damage. FIG. 8G shows immunoblot analysis of ATP7A in the livers of WT and ATP7B- KO mice treated with DMSO or Roxadustat, n= 3 / group. Ponceau S stain was used as a loading control. Each lane represents the liver of a different mouse. Note the expression of ATP7A in the liver of DMSO-treated ATP7B-KO mice compared to WT mice and that Roxadustat further enhanced ATP7A expression in both groups. FIG. 8H-I shows quantification of metallothionein MT-I and MT-II mRNA levels by real time PCR in the livers from WT and ATP7B-KO mice. As expected, MT-I and MT-II were elevated in the livers of control ATP7B-KO mice compared to WT, however, Roxadustat significantly elevated their expression further in the KO group.Values are expressed as the mean ± SEM (****p < 0.0001; n = 3).DETAILED DESCRIPTION

[0020] So that the present disclosure may be more readily understood, certain terms are first defined. 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 embodiments of the disclosure pertain. Many methods and materials similar, modified, or equivalent to thosedescribed herein can be used in the practice of the embodiments of the present disclosure without undue experimentation, the preferred materials and methods are described herein. In describing and claiming the embodiments of the present disclosure, the following terminology will be used in accordance with the definitions set out below.

[0021] It is to be understood that all terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting in any manner or scope. For example, as used in this specification and the appended claims, the singular forms "a," "an" and "the" can include plural referents unless the content clearly indicates otherwise. Similarly, the word “or” is intended to include “and” unless the context clearly indicate otherwise. The word “or” means any one member of a particular list and also includes any combination of members of that list. Further, all units, prefixes, and symbols may be denoted in its SI accepted form.

[0022] Numeric ranges recited within the specification are inclusive of the numbers defining the range and include each integer within the defined range. Throughout this disclosure, various embodiments of this disclosure are presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible sub-ranges, fractions, and individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6, and decimals and fractions, for example, 1.2, 3.8, P , and 43 / 4. This applies regardless of the breadth of the range.

[0023] As used herein, the term “administration” typically refers to the administration of a composition to a subject or system. Those of ordinary skill in the art will be aware of a variety of routes that may, in appropriate circumstances, be utilized for administration to a subject, for example a human subject. For example, in embodiments, administration may be ocular, oral, parenteral, topical, etc. In some particular embodiments, administration may be bronchial (e.g., by bronchial instillation), buccal, dermal (which may be or comprise, for example, one or more of topical to the dermis, intradermal, interdermal, transdermal, etc.), enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, within a specific organ (e. g. intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by intratracheal instillation), vaginal, vitreal, etc. In certain embodiments, administration may involve dosing that is intermittent (e.g., a plurality of doses separated in time) and / or periodic (e.g., individual doses separated by acommon period of time) dosing. In certain embodiments, administration may involve continuous dosing (e.g., perfusion) for at least a selected period of time.

[0024] As used herein, the term “dose(s)” means a quantity of the compound or a pharmaceutically acceptable salt, solvate, or hydrate thereof to be administered at one time. A dose may comprise a single unit dosage form, or alternatively may comprise more than a single unit dosage form (e.g., a single dose may comprise two tablets), or even less than a single unit dosage form (e.g., a single dose may comprise half of a tablet).

[0025] As used herein, the term “daily dose” means a quantity of the compound, or a pharmaceutically acceptable salt, solvate, or hydrate thereof that is administered in a 24-hour period. Accordingly, a daily dose may be administered all at once (i.e., once daily dosing) or alternatively the daily dosing may be divided such that administration of the compound is twice daily, three times daily, or even four times daily.

[0026] The term “excipient” refers to an inert or inactive substance added to a pharmaceutical composition to further facilitate administration of a compound.

[0027] As used herein, the terms “improve,” “increase” or “reduce,” or grammatical equivalents, indicate values that are relative to a baseline measurement, such as a measurement in the same individual prior to initiation of the treatment described herein, or a measurement in a control sample or subject (or multiple control samples or subjects) in the absence of the treatment described herein. A “control subject” is a subject afflicted with the same form of disease as the subject being treated, who is about the same age as the subject being treated.

[0028] The term “pharmaceutically acceptable”, as used herein, refers to substances that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0029] As used herein, the term “subject” refers to a human or any non-human animal (e.g., mouse, rat, rabbit, dog, cat, cattle, swine, sheep, horse or primate). A human includes pre- and post-natal forms. In certain embodiments, a subject is a human being. As used herein, the term “patient” refers to a human. In certain embodiments, a patient may be suffering from, and / or susceptible to a disease, disorder, and / or condition. A patient can be afflicted with or is susceptible to a disease or disorder but may or may not display symptoms of the disease or disorder. A subject can be a patient.

[0030] As used herein, the term “therapeutically effective amount” of a therapeutic agent means an amount that is sufficient, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, to treat, diagnose, prevent, and / or delay the onset of thesymptom(s) of the disease, disorder, and / or condition. It will be appreciated by those of ordinary skill in the art that a therapeutically effective amount is typically administered via a dosing regimen comprising at least one-unit dose.

[0031] As used herein, the term “treat,” “treatment,” or “treating” refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of and / or reduce incidence of one or more symptoms or features of a particular disease, disorder, and / or condition. Treatment may be administered to a subject who does not exhibit signs of a disease and / or exhibits only early signs of the disease for the purpose of decreasing the risk of developing pathology associated with the disease.

[0032] As used herein “hypoxia” refers to a deficiency of oxygen. A low oxygen condition is also referred to as a “hypoxic condition.”

[0033] A “hypoxia response” is a response by a cell and / or an organism to hypoxia. Hypoxia is one non-limiting way to induce a hypoxia response. A hypoxia response includes, but is not limited to, a physiological response (e.g., a systemic or pulmonary hemodynamic response, a change in the regulation of cellular metabolism, and up-regulation of genes (e.g., HIF responsive genes)) and a pathological response (e.g., pulmonary hypertension, cerebral ischemia, myocardial ischemia, and tumor angiogenesis). Non-limiting examples of systemic responses include pulmonary vasoconstriction, systemic vasodilation, increased cytosolic calcium concentration, and neurotransmitter release, for example, catecholamines, acetylcholine, and serotonin. Non-limiting examples of a response affecting the regulation of cellular metabolism include uncontrolled cell swelling, cell necrosis, impaired mitochondrial respiratory chain function, increased cellular glycolysis, decreased cellular energy consumption, and decreased cellular oxygen consumption. Other examples of a hypoxia response include increased ventilation, increased cardiac output, a switch from aerobic to anaerobic metabolism, promotion of improved vascularization, an increase of erythropoietin with augmented erythropoiesis, enhancement of the oxygen carrying capacity of the blood, reduced oxygen toxicity, increased or reduced reactive oxygen species, and increased or reduced oxidative stress. A hypoxia response may involve oxygen-responsive pathways to sense and to respond to changes in oxygen availability. For example, HIFs may respond to a low oxygen environment and activate one or more HIF -responsive genes.

[0034] “Normoxia” or a “normoxic condition” refers to a normal level of oxygen condition.

[0035] The “prolyl hydroxylase” enzymes hydroxylate alpha subunits of HIF at conserved proline residues. Hydroxylation and degradation occurs under normoxic conditions. Prolyl hydroxylase enzyme activity is inhibited under hypoxic conditions.

[0036] As used herein, the term “alkyl” means acyclic linear and branched hydrocarbon groups, e.g. “C1-C20 alkyl” refers to alkyl groups having 1-20 carbons. An alkyl group may be linear or branched. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl tert-pentyl, hexyl, isohexyl, and the like. Other alkyl groups will be readily apparent to those of skill in the art given the benefit of the present disclosure. An alkyl group may be unsubstituted or substituted with one or more substituent groups as described herein. For example, an alkyl group may be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen, haloalkyl (e.g., — CH2F and — CH2CI, as well as perhaloalkyl groups such as CF3 and CCI3), — CO2R', — CN, — OH, — OR', — NH2, — NHR', — N(R')2, — SR' or — SO2R', wherein each instance of R' independently is C1-C3 alkyl. In certain embodiments, the alkyl is unsubstituted. In certain embodiments, the alkyl is substituted (e.g., with 1, 2, 3, 4, 5, or 6 substituent groups as described herein).

[0037] The term “alkoxy,” as used herein, a monovalent substituent which consists of a linear or branched, substituted or unsubstituted, acyclic or cyclic alkyl group linked through an ether oxygen and having its free valence bond from the ether oxygen such as, for example methoxy, ethoxy, 1- and 2-propoxy, 1 -butoxy, 1,2-dimethylethoxy, 1- and 2-pentoxy-, 3 -hexoxy- and the like.

[0038] The term “alkylene,” as used herein, represents a saturated divalent straight or branched chain hydrocarbon group, e.g., a “C1-C10 alkylene” group having 1-10 carbons. Alkylene groups are exemplified by methylene, ethylene, isopropylene and the like. An alkylene may be unsubstituted or substituted with substituent groups as described herein. In certain embodiments, an alkylene group is unsubstituted

[0039] The terms “aryl” and “ar-”, used alone or as part of a larger moiety, e.g., “aralkyl”, “aralkoxy”, or “aryloxyalkyl”, refer to an optionally substituted Ce-i4 aromatic hydrocarbon moiety comprising one to three aromatic rings. For example, the aryl group is a Ce-io aryl group (i.e., phenyl and naphthyl). Aryl groups include, without limitation, optionally substituted phenyl, naphthyl, or anthracenyl. The terms “aryl” and “ar-”, as used herein, also include groups in which an aryl ring is fused to one or more cycloaliphatic rings to form an optionally substituted cyclic structure such as a tetrahydronaphthyl, indenyl, or indanyl ring. The term “aryl” may be used interchangeably with the terms “aryl group”, “aryl ring”, and “aromatic ring”.

[0040] As used herein, the term “benzyloxy” refers to a monovalent substituent which consists of a substituted or unsubstituted benzyl group linked through an ether oxygen and having its freevalence bond from the ether oxygen.

[0041] As used herein, the term “cycloalkyl” means a nonaromatic, saturated, cyclic group, e.g. “C3-C10 cycloalkyl.” In certain embodiments, a heterocyclyl is monocyclic. In certain embodiments, a heterocyclyl is polycyclic (e.g., bicyclic or tricyclic). In polycyclic cycloalkyl groups, individual rings can be fused, bridged, or spirocyclic. Examples of a cycloalkyl group include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbomanyl, bicyclo[3.2.1]octanyl, octahydro-pentalenyl, and spiro[4.5]decanyl, and the like. The term “cycloalkyl” may be used interchangeably with the term “carbocycle”. A cycloalkyl group may be unsubstituted or substituted with one or more substituent groups as described herein. For example, a cycloalkyl group may be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen ,— CO2R', — CN, —OH, —OR', — NH2, — NHR', — N(R')2, —SR' or— SO2R', wherein each instance of R' independently is C1-C3 alkyl. In certain embodiments, the cycloalkyl is unsubstituted. In certain embodiments, the cycloalkyl is substituted (e.g., with 1, 2, 3, 4, 5, or 6 substituent groups as described herein).

[0042] As used herein, the term “halo” or “halogen” means fluorine, chlorine, bromine, or iodine.

[0043] As used herein, the term “heterocyclyl” means a nonaromatic, cyclic structure having at least one of any type of heteroatom as ring atoms, having any degree of unsaturation, and excludes aromatic heterocyclic rings that are defined as “heteroaryl” herein. For example, a “3- to 10-membered heterocyclyl” refers to heterocyclics having 3-10 ring atoms that are carbon or heteroatoms as described herein. The one or more heteroatoms may be selected from nitrogen, sulfur, and oxygen. The term “heterocycle”, “heterocyclyl”, “heterocyclic”, “heterocycloalkyl,” and “heterocyclic ring” can be used interchangeably. A heterocyclyl group can be attached as a substituent via a carbon atom or a heteroatom (e.g. a nitrogen atom). In certain embodiments, a heterocyclyl is monocyclic. In certain embodiments, a heterocyclyl is polycyclic (e.g., bicyclic or tricyclic). Examples include 2-tetrahydrofuranyl, 3 -tetrahydrofuranyl, dihydrofuranyl, tetrahydropyranyl, pyranyl, thiopyranyl, 2-tetrahydrothiophenyl, 3 -tetrahydrothiophenyl, 2- morpholinyl, 3-morpholinyl, 4-morpholinyl, 2-thiomorpholinyl, 3-thiomorpholinyl, 4- thiomorpholinyl, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, 1-piperazinyl, 2-piperazinyl, 1- piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, l,3-oxazoIidin-3-yI, isothiazolidinyl, 1,3- thiazolidin-3-yl, l,2-pyrazolidin-2-yl, 1,3-pyrazolidin-l-yl, 1,2- tetrahydrothiazin-2-yl, 1,3- tetrahydrothiazin-3-yl, tetrahydrothiadiazinyl, 1,2- tetrahydrodiazin-2-yl, 1,3-tetrahydrodiazin-l- yl, tetrahydroazepinyl, piperizin-2-onyl, piperizin-3-onyl, 2-pyrrolinyl, 3-pyrrolinyl, imidazolidinyl, 2-imidazolidinyl, 1,4- dioxanyl, and 4-thiazolidinyl, and the like. A heterocyclylgroup may be unsubstituted or substituted with one or more substituent groups as described herein. For example, a heterocyclyl group may be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen, — CO2R', — CN, — OH, — OR', — NH2, — NHR', — N(R')2, — SR' or — SO2R', wherein each instance of R' independently is C1-C3 alkyl. In certain embodiments, the heterocyclyl is unsubstituted. In certain embodiments, the heterocyclyl is substituted (e.g., with 1, 2, 3, 4, 5, or 6 substituent groups as described herein).

[0044] As used herein, the term “heteroaryl” means an aromatic moiety having at least one of any type of heteroatom as ring atoms. For example, a “5- to 14-membered heteroaryl” refers to heteroaryls having 5-14 ring atoms that are carbon or heteroatoms as described herein. The one or more heteroatoms may be selected from nitrogen, sulfur, and oxygen. A heteroaryl group can be attached as a substituent via a carbon atom or a heteroatom (e.g. a nitrogen atom). In certain embodiments, a heteroaryl is monocyclic. In certain embodiments, a heteroaryl is polycyclic (e.g., bicyclic or tricyclic). In certain embodiments, polycyclic heteroaryls comprise a cyclic group that is nonaromatic (e.g., a heteroaryl fused to a cycloalkyl or a heterocyclyl group as described herein). The term “heteroaryl” may be used interchangeably with the term “heteroaryl ring” or the term “heteroaromatic”. Examples include 5-membered monocyclic rings such as pyrrolyl, imidazolyl, pyrazolyl, triazolyl, furyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, thiazolyl, isothiazolyl, and the like; and 6-membered monocyclic rings such as pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl and the like. For further examples, see, e.g., Katritzky, Handbook of Heterocyclic Chemistry. Further specific examples of heteroaryl rings include 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-pyrimidyl, 3 -pyridazinyl, 2- thiazolyl, 4-thiazolyl, 5-thiazolyl, 5-tetrazolyl, 2-triazolyl, 5-triazolyl, 2-thienyl, 3-thienyl and carbazolyl. The term “heteroaryl” also refers to rings that are optionally substituted. A heteroaryl group may be optionally substituted with one or more functional groups discussed below. Still other examples include indolyl, azaindolyl, benzimidazolyl, indazolyl, imidazopyridinyl, imidazopyrimidinyl, pyrrolopyrimidinyl, pyrrolopyridinyl, pyrazolopyridinyl, pyrazolopyrimidinyl, quinolyl, isoquinolyl, benzoxazolyl, benzathiazolyl, benzothiophenyl, benzofuranyl, and isobenzofuranyl, and the like. A heteroaryl group may be unsubstituted or substituted with one or more substituent groups as described herein. For example, a heteroaryl group may be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen ,—CO2R', — CN, —OH, —OR', — NH2, —NHR', — N(R')2, —SR' or— SO2R', wherein each instance of R' independently is C1-C3 alkyl. In certain embodiments, the heteroaryl is unsubstituted. In certain embodiments, the heteroaryl is substituted (e.g., with 1, 2, 3, 4, 5, or 6 substituent groups as described herein).Prolyl hydroxylase inhibitors

[0045] Hypoxia-inducible factor (HIF) is a heterodimeric transcription factor consisting of an oxygen-sensitive HIF-a subunit and a constitutively expressed HIF-P subunit. Under normoxic conditions, the HIF-a subunit is produced and continuously degraded with a tU of approximately 5 minutes. In response to hypoxic conditions, levels of HIFa are elevated in most cells because of a decrease in HIFa prolyl hydroxylation.

[0046] Prolyl hydroxylation of HIFa is accomplished by a family of proteins variously termed the prolyl hydroxylase domain-containing proteins (PHD1, 2, and 3), also known as HIF prolyl hydroxylases (HIF-PH-3, 2, and 1) or EGLN-2, 1, and 3, which use physiological levels of oxygen to hydroxylate specific proline residues of HIF-a, signaling proteasomal degradation of HIF-a (Ke et al. (2006) Mol. Pharmacol. 70 (5): 1469-80). Under hypoxic conditions the activity of prolyl hydroxylase is decreased, degradation of HIF-a is inhibited and cellular levels of HIF-a are increased. Stabilization of HIF-a allows it to translocate to the nucleus and dimerize with the HIF-P subunit where it binds to cis-acting hypoxia-response elements and activates the transcription of target genes involved in erythropoiesis and iron homeostasis. The prolyl hydroxylases thus function as critical sensors of cellular oxygen levels, and regulate oxygen homeostasis in response to hypoxia through inhibition of hydroxylation thus leading to stabilization of HIF-a, which in turn signals gene transcription and translation of proteins necessary for the cellular and organ responses to poor oxygenation (Myllylharju et al. (2013) Acta Physiol 208 (2): 148-65).

[0047] Provided herein are compounds for use in compositions and methods for treating and / or preventing a disease or condition of copper metabolism, compositions and methods for reducing mortality and morbidity related to a disease or condition of copper metabolism, and compositions and methods for reducing incidence, severity, or risk of a disease or condition of copper metabolism. Compounds that can be used with the compositions and formulations provided herein are prolyl hydroxylase inhibitors.

[0048] Non-limiting examples of prolyl hydroxylase inhibitors include vadadustat (AKB-6548), roxadustat (FG-4592), daprodustat (GSK-12788363), molidustat (BAY 85-3934), enarodustat (JTZ-951), or desidustat (ZYAN1), and compounds described in U.S. Pat. Nos. 8,759,345, 8,937,078, 8,796,263, 9,273,034, 8,530,404, 7,696,223, 7,629,357, 8,927,591, 8,269,008, 8,952,160, 8,952,160, 8,927,591, 8,921,389, 8,916,585, 8,703,795, 8,921,389, 7,662,854, and 9,040,522; in International Publication No. W02020 / 072645; in U.S. Provisional Pat. Application Nos. 62 / 947,170, 62 / 947,179, 62 / 992,585, 62 / 992,606, and 62 / 992,616; and in “Recent Advances in Developing Inhibitors for Hypoxia-Inducible Factor Prolyl Hydroxylasesand Their Therapeutic Implications” (Kim et al. Molecules 2015, 20, 20551-20568; see, e.g., any of the compounds described therein, including any of the compounds described in any of Tables 2, 3, or 4), each of which is hereby incorporated by reference in its entirety. In certain embodiments, a suitable compound is described in any of International Publication No. W02020 / 072645; and in U.S. Provisional Pat. Application Nos. 62 / 947,170, 62 / 947,179, 62 / 992,585, 62 / 992,606, and 62 / 992,616. Any known prolyl hydroxylase inhibitor may be used in the compositions and methods of the present disclosure.

[0049] Certain exemplary compounds are further described herein.Formula (I): Vadadustat and Related Compounds

[0050] Exemplary compounds that can be used in any of the compositions and methods described herein include those described in U.S. Pat. Nos. 7,811,595, 8,343,952, 8,323,671, 8,598,210, 8,722,895, 8,940,773, and 9,598,370; and in U.S. Publication No. US 20190192494A1, each of which is incorporated by reference in its entirety. In certain embodiments, a compound, or pharmaceutically acceptable salt thereof, is described in any of claims 1-32 of U.S. Pat. No. 7,811,595.

[0051] In certain embodiments, suitable compounds include a compound having a structure according to Formula (I),Formula (I), or a pharmaceutically acceptable salt thereof, wherein, R and R1are each independently:(i) hydrogen(ii) substituted or unsubstituted phenyl;(iii) substituted or unsubstituted heteroaryl; or(iv) substituted or unsubstituted alkyl; said substitution selected from:(i) Ci-C4alkyl;(ii) C3-C4 cycloalkyl;(iii) C1-C4 alkoxy;(iv) C3-C4 cycloalkoxy;(v) C1-C4 haloalkyl;(vi) C3-C4 halocycloalkyl;(vii) halogen;(viii) cyano;(ix) NHC(O)R4;(x) C(O)NR5aR5b;(xi) phenyl; and(xii) heteroaryl; or(xiii) two substituents are taken together to form a fused ring having from 5 to 7 atoms;R4is a C1-C4 alkyl or C3-C4 cycloalkyl;R5aand R5bare each independently selected from:(i) hydrogen;(ii) C1-C4 alkyl;(iii) C3-C4 cycloalkyl; or(iv) R5aand R5bare taken together to form a ring having from 3 to 7 atoms;R2is selected from:(i) OR6(ii) NR7aR7b; andR6is selected from hydrogen and C1-C4 alkyl or C3-C4 cycloalkyl;R7aand R7bare each independently selected from:(i) hydrogen;(ii) C1-C4 alkyl or C3-C4 cycloalkyl; or(iii) R7aand R7bare taken together to form a ring having from 3 to 7 atoms;R3is selected from hydrogen, methyl, and ethyl;L is a linking unit having a structure -[C(R8aR8b)]n-R8aand R8bare each independently selected from hydrogen, methyl and ethyl; n is an integer from 1 to 3; andR9is selected from hydrogen and methyl.

[0052] In certain embodiments, R is hydrogen.

[0053] In certain embodiments, R is unsubstituted phenyl. In certain embodiments, R is substituted phenyl. In certain embodiments, R is phenyl substituted with C1-C4 alkyl. In certain embodiments, R is phenyl substituted with C3-C4 cycloalkyl. In certain embodiments, R is phenyl substituted with C1-C4 alkoxy. In certain embodiments, R is phenyl substituted with C3- C4 cycloalkoxy. In certain embodiments, R is phenyl substituted with C1-C4 haloalkyl. In certainembodiments, R is phenyl substituted with C3-C4 halocycloalkyl. In certain embodiments, R is phenyl substituted with halogen. In certain embodiments, R is phenyl substituted with cyano. In certain embodiments, R is phenyl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R is phenyl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R is phenyl substituted with phenyl. In certain embodiments, R is phenyl substituted with heteroaryl. In certain embodiments, R is phenyl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0054] In certain embodiments, R is unsubstituted heteroaryl. In certain embodiments, R is substituted heteroaryl. In certain embodiments, R is heteroaryl substituted with C1-C4 alkyl. In certain embodiments, R is heteroaryl substituted with C3-C4 cycloalkyl. In certain embodiments, R is heteroaryl substituted with C1-C4 alkoxy. In certain embodiments, R is heteroaryl substituted with C3-C4 cycloalkoxy. In certain embodiments, R is heteroaryl substituted with Ci- C4 haloalkyl. In certain embodiments, R is heteroaryl substituted with C3-C4 halocycloalkyl. In certain embodiments, R is heteroaryl substituted with halogen. In certain embodiments, R is heteroaryl substituted with cyano. In certain embodiments, R is heteroaryl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R is heteroaryl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R is heteroaryl substituted with phenyl. In certain embodiments, R is heteroaryl substituted with heteroaryl. In certain embodiments, R is heteroaryl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0055] In certain embodiments, R is unsubstituted alkyl. In certain embodiments, R is substituted alkyl. In certain embodiments, R is alkyl substituted with C1-C4 alkyl. In certain embodiments, R is alkyl substituted with C3-C4 cycloalkyl. In certain embodiments, R is alkyl substituted with C1-C4 alkoxy. In certain embodiments, R is alkyl substituted with C3-C4 cycloalkoxy. In certain embodiments, R is alkyl substituted with C1-C4 haloalkyl. In certain embodiments, R is alkyl substituted with C3-C4 halocycloalkyl. In certain embodiments, R is alkyl substituted with halogen. In certain embodiments, R is alkyl substituted with cyano. In certain embodiments, R is alkyl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3- C4 cycloalkyl. In certain embodiments, R is alkyl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R is alkyl substituted with phenyl. In certain embodiments, R is alkyl substituted with heteroaryl. In certain embodiments, R is alkyl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0056] In certain embodiments, R is substituted or unsubstituted phenyl. In certain embodiments, R is chosen from 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2-chlorophenyl, 3 -chlorophenyl, and 4-chlorophenyl. In certain embodiments, R is chosen from 2- methoxyphenyl, 3 -methoxyphenyl, 4-methoxyphenyl, 2-ethoxyphenyl, 3 -ethoxy phenyl, 4- ethoxyphenyl, 2-iso-propoxyphenyl, 3-iso-propoxyphenyl, 4-iso-propoxyphenyl, 2-cyanophenyl,3 -cyanophenyl, and 4-cy anophenyl.

[0057] In certain embodiments, R is a unit having the formula:exemplary groups for R10are as described herein.

[0058] In certain embodiments, R10has the formula — C(O)NR5aR5b; R5aand R5bare each hydrogen or R5aand R5bare taken together to from a ring having 5 or 6 atoms. In certain embodiments, R5aand R5bare taken together to form a pyrrolidin-l-yl ring.

[0059] In certain embodiments, R10has the formula — NHC(O)R4; R4is a unit chosen from methyl, ethyl, n-propyl, iso-propyl, and cyclopropyl.

[0060] In certain embodiments, R10is a heteroaryl unit chosen from l,2,3,4-tetrazol-5-yl, [l,2,4]triazol-5-yl, imidazol-2-yl, furan-2-yl, furan-3-yl, thiophene-2-yl, and thiophene-3 -yl.

[0061] In certain embodiments, R is substituted or unsubstituted heteroaryl. In certain embodiments, R is a unit chosen from pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, isoquinolin-l-yl, isoquinolin-3-yl, and isoquinolin-4-yl.

[0062] In certain embodiments, R is a unit chosen from thiazol-2-yl, thiazol-4-yl, 1, 2,3,4- tetrazol-5-yl, [l,2,4]triazol-5-yl, imidazol-2-yl, furan-2-yl, furan-3-yl, thiophene-2-yl, and thiophene-3 -yl.

[0063] In certain embodiments, R1is hydrogen.

[0064] In certain embodiments, R1is unsubstituted phenyl. In certain embodiments, R1is substituted phenyl. In certain embodiments, R1is phenyl substituted with C1-C4 alkyl. In certain embodiments, R1is phenyl substituted with C3-C4 cycloalkyl. In certain embodiments, R1is phenyl substituted with C1-C4 alkoxy. In certain embodiments, R1is phenyl substituted with C3- C4 cycloalkoxy. In certain embodiments, R1is phenyl substituted with C1-C4 haloalkyl. In certain embodiments, R1is phenyl substituted with C3-C4 halocycloalkyl. In certain embodiments, R1is phenyl substituted with halogen. In certain embodiments, R1is phenyl substituted with cyano. In certain embodiments, R1is phenyl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R1is phenyl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R1is phenyl substituted with phenyl. In certain embodiments, R1is phenyl substituted with heteroaryl. In certain embodiments, R1is phenyl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0065] In certain embodiments, R1is unsubstituted heteroaryl. In certain embodiments, R1is substituted heteroaryl. In certain embodiments, R1is heteroaryl substituted with C1-C4 alkyl. In certain embodiments, R1is heteroaryl substituted with C3-C4 cycloalkyl. In certain embodiments, R1is heteroaryl substituted with C1-C4 alkoxy. In certain embodiments, R1isheteroaryl substituted with C3-C4 cycloalkoxy. In certain embodiments, R1is heteroaryl substituted with C1-C4 haloalkyl. In certain embodiments, R1is heteroaryl substituted with C3-C4 halocycloalkyl. In certain embodiments, R1is heteroaryl substituted with halogen. In certain embodiments, R1is heteroaryl substituted with cyano. In certain embodiments, R1is heteroaryl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R1is heteroaryl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R1is heteroaryl substituted with phenyl. In certain embodiments, R1is heteroaryl substituted with heteroaryl. In certain embodiments, R1is heteroaryl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0066] In certain embodiments, R1is unsubstituted alkyl. In certain embodiments, R1is substituted alkyl. In certain embodiments, R1is alkyl substituted with C1-C4 alkyl. In certain embodiments, R1is alkyl substituted with C3-C4 cycloalkyl. In certain embodiments, R1is alkyl substituted with C1-C4 alkoxy. In certain embodiments, R1is alkyl substituted with C3-C4 cycloalkoxy. In certain embodiments, R1is alkyl substituted with C1-C4 haloalkyl. In certain embodiments, R1is alkyl substituted with C3-C4 halocycloalkyl. In certain embodiments, R1is alkyl substituted with halogen. In certain embodiments, R1is alkyl substituted with cyano. In certain embodiments, R1is alkyl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3- C4 cycloalkyl. In certain embodiments, R1is alkyl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R1is alkyl substituted with phenyl. In certain embodiments, R1is alkyl substituted with heteroaryl. In certain embodiments, R1is alkyl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selectedfrom hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0067] In certain embodiments, R1is chosen from 2-methoxyphenyl, 3 -methoxyphenyl, 4- methoxyphenyl, 2-ethoxyphenyl, 3 -ethoxy phenyl, 4-ethoxyphenyl, 2-iso-propoxyphenyl, 3-iso- propoxyphenyl, 4-iso-propoxyphenyl, 2-cyanophenyl, 3 -cyanophenyl, and 4-cy anophenyl.

[0068] In certain embodiments, R2is OR6, wherein R6is hydrogen, C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R2is NR7aR7b, wherein R7aand R7bare each independently selected from the group consisting of hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R7aand R7bare taken together to form a ring having from 3 to 7 atoms.

[0069] In certain embodiments, R6is hydrogen. In certain embodiments, R6is methyl.

[0070] In certain embodiments, R2is — NR^R76; R7aand R7bare each independently hydrogen, methyl, or ethyl. In certain embodiments, R2is chosen from — NH2, — NHCH3, and — NfCFFh.

[0071] In certain embodiments, R3is hydrogen. In certain embodiments, R3is methyl. In certain embodiments, R3is ethyl.

[0072] In certain embodiments, L is a linking unit having a structure -[C(R8aR8b)]n-, wherein R8aand R8bare each independently selected from the group consisting of hydrogen, methyl and ethyl.

[0073] In certain embodiments, L is chosen from — CH2 — , — CH2CH2 — , and — C(CH3)2 — . In certain embodiments, L is — CH2 — . In certain embodiments, L is — CH2CH2 — . In certain embodiments, L is — C(CH3)2 — .

[0074] In certain embodiments, n is an integer from 1 to 3. In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3.

[0075] In certain embodiments, R9is hydrogen. In certain embodiments, R9is methyl.

[0076] In certain embodiments, a compound according to Formula (I) has the followingFormula (la), or a pharmaceutically acceptable salt thereof, whereinR is as defined anywhere herein.

[0077] In certain embodiments, a compound according to Formula (I) has the following structure:Formula (lb), or a pharmaceutically acceptable salt thereof, wherein R is as defined anywhere herein.

[0078] In certain embodiments, a compound according to Formula (I) has the following structure:Formula (Ic), or a pharmaceutically acceptable salt thereof, whereinR is as defined anywhere herein.

[0079] In certain embodiments, R is a heteroaryl unit chosen from thiazol-2-yl, thiazol-4-yl, l,2,3,4-tetrazol-5-yl, [l,2,4]triazol-5-yl, imidazol-2-yl, furan-2-yl, furan-3-yl, thiophene-2-yl, thiophene-3 -yl, pyridin-2-yl, pyri din-3 -yl, pyridin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, isoquinolin- 1-yl, isoquinolin-3-yl, and isoquinolin-4-yl.

[0080] In certain embodiments, a compound according to Formula (I) has the following structure:Formula (Id), or a pharmaceutically acceptable salt thereof, whereinR is selected from 2-chlorophenyl, 3 -chlorophenyl, 4-chlorophenyl, 2-cyanophenyl, 3- cyanophenyl, and 4-cy anophenyl; andR6is selected from hydrogen, methyl, and ethyl.

[0081] In certain embodiments, a compound is selected from the group consisting of:{[5-(3-Chloro-phenyl)-3-hydroxy-pyridine-2-carbonyl]-amino}-acetic acid methyl ester;{[5-(4-Chloro-phenyl)-3-hydroxy-pyridine-2-carbonyl]-amino}-acetic acid methyl ester; {[5-(2-Chloro-phenyl)-3-hydroxy-pyridine-2-carbonyl]-amino}-acetic acid methyl ester; {[5-(4-Fluoro-phenyl)-3-hydroxy-pyridine-2-carbonyl]-amino}-acetic acid methyl ester; [(3-Hydroxy-5-(4-methylphenyl)-pyridine-2-carbonyl)-amino]-acetic acid methyl ester;{[3-Hydroxy-5-(4-isopropyl-phenyl)-pyridine-2-carbonyl]-amino}-acetic acid methyl ester;{[5-(4-Ethyl-phenyl)-3-hydroxy-pyridine-2-carbonyl]-amino}-acetic acid methyl ester;{[3-Hydroxy-5-(3-trifluoromethyl-phenyl)-pyridine-2-carbonyl]-amino}-acetic acid methyl ester;{[5-(4-Cyano-phenyl)-3-hydroxy-pyridine-2-carbonyl]-amino}-acetic acid methyl ester; {[5-(3-Cyano-phenyl)-3-hydroxy-pyridine-2-carbonyl]-amino}-acetic acid methyl ester; {[5-(3-Carbamoyl-phenyl)-3-hydroxy-pyridine-2-carbonyl]-amino}-acetic acid methyl ester;({3-Hydroxy-5-[3-(pyrrolidine-l-carbonyl)-phenyl]-pyridine-2-carbonyl}-amino)-acetic acid methyl ester;({5-[3-(Cyclopropanecarbonyl-amino)-phenyl]-3-hydroxy-pyridine-2-carbonyl}-amino)- acetic acid methyl ester;({3-Hydroxy-5-[3-(2H-tetrazol-5-yl)-phenyl]-pyridine-2-carbonyl}-amino)-acetic acid methyl ester;[(5-Hydroxy-[3,3']bipyridinyl-6-carbonyl)-amino]-acetic acid methyl ester [(5'-Hydroxy-[2,3']bipyridinyl-6'-carbonyl)-amino]-acetic acid methyl ester; [(3-Hydroxy-5-pyrimidin-5-yl-pyridine-2-carbonyl)-amino]-acetic acid methyl ester;[(3-Hydroxy-5-isoquinolin-4-yl-pyridine-2-carbonyl)-amino]-acetic acid methyl ester; [(3-Hydroxy-5-thiazol-2-yl-pyridine-2-carbonyl)-amino]-acetic acid methyl ester; {[5-(3-Chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino}-acetic acid;{[5-(4-Chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino}-acetic acid;{[5-(2-Chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino}-acetic acid;{[5-(4-Fluorophenyl)-3-hydroxypyridine-2-carbonyl]amino}-acetic acid;[(3-Hydroxy-5-(4-methylphenyl)pyridine-2-carbonyl)amino]-acetic acid;{[5-(4-Ethylphenyl)-3-hydroxypyridine-2-carbonyl]amino}-acetic acid;{[3-Hydroxy-5-(4-isopropylphenyl)pyridine-2-carbonyl]amino}-acetic acid;{[3-Hydroxy-5-(3-trifluoromethylphenyl)pyridine-2-carbonyl]amino}-acetic acid;{[5-(4-Cyanophenyl)-3-hydroxypyridine-2-carbonyl]amino}-acetic acid;{[5-(3-Cyanophenyl)-3-hydroxypyridine-2-carbonyl]amino}-acetic acid;{[5-(5-Chloro-2-methylphenyl)-3-hydroxypyridine-2-carbonyl]amino}-acetic acid;{[3-Hydroxy-5-(4-isopropoxyphenyl)pyridine-2-carbonyl]amino}-acetic acid;({5-[3-(Cyclopropanecarbonylamino)phenyl]-3-hydroxy-pyridine-2-carbonyl}-amino)- acetic acid;({3-Hydroxy-5-[3-(pyrrolidine-l-carbonyl)phenyl]-pyridine-2-carbonyl}amino)-acetic acid;({3-Hydroxy-5-[3-(2H-tetrazol-5-yl)phenyl]-pyridine-2-carbonyl}-amino)-acetic acid;[(5'-Hydroxy-[2,3']bipyridinyl-6'-carbonyl)-amino]-acetic acid;[(5-Hydroxy-[3,3']bipyridinyl-6-carbonyl)-amino]-acetic acid;[(3-Hydroxy-5-pyrimidin-5-yl-pyridine-2-carbonyl)-amino]-acetic acid;[(3-Hydroxy-5-isoquinolin-4-yl-pyridine-2-carbonyl)-amino]-acetic acid;[(3-Hydroxy-5-thiazol-2-yl-pyridine-2-carbonyl)-amino]-acetic acid;{[5-(2,3-Dihydro-benzo[l,4]dioxin-6-yl)-3-hydroxy-pyridine-2-carbonyl]-amino}-acetic acid;5-(Chlorophenyl)-N-(2-amino-2-oxo-l,l-dimethylethyl)-3-hydroxylpyridin-2-yl amide;5-(Chl orophenyl)-N-(2-amino-2-oxoethyl)-3-hydroxylpyri din-2 -yl amide;5-(Chlorophenyl)-N-(2-amino-2-oxo-l -methylethyl)-3-hy droxylpyri din-2 -yl amide;5-(4-Methylphenyl)-N-(2-methylamino-2-oxoethyl)-3-hydroxylpyridin-2-yl amide;5-(3-Chlorophenyl)-N-(2-methylamino-2-oxoethyl)-3-hydroxylpyridin-2-yl amide;2-{[5-(3-Chloro-phenyl)-3-hydroxy-pyridine-2-carbonyl]-amino}-2-methyl-propionic acid methyl ester;2-{[5-(3 -Chi orophenyl)-3-hydroxy-pyridine-2-carbonyl]-amino}-2-methyl -propionic acid3-[(3-Hydroxy-5-(4-methylphenyl)-pyridine-2-carbonyl)-amino]-propionic acid ethyl ester;3-[(3-Hydroxy-5-(3-chlorophenyl)-pyridine-2-carbonyl)-amino]-propionic acid ethyl ester;3-{[5-(3-Chlorophenyl)-3-hydroxy-pyridine-2-carbonyl]-amino}-propionic acid;3-[(3-Hydroxy-5-(4-methylphenyl)-pyridine-2-carbonyl)-amino]-propionic acid;5-(Chl orophenyl)-N-(3-amino-3-oxo- l,l-dimethylpropyl)-3-hy droxylpyri din-2-yl amide; and5-(3-Chlorophenyl)-N-(2-dimethylamino-2-oxoethyl)-3-hydroxylpyridin-2-yl amide; or a pharmaceutically acceptable salt thereof.

[0082] In certain embodiments, compounds are salts comprising anions chosen from chloride, bromide, iodide, sulfate, bisulfate, carbonate, bicarbonate, phosphate, formate, acetate, propionate, butyrate, pyruvate, lactate, oxalate, malonate, maleate, succinate, tartrate, fumarate, and citrate.

[0083] In certain embodiments, compounds are salts comprising cations chosen from sodium, lithium, potassium, calcium, magnesium, and bismuth.

[0084] In certain embodiments, a compound is {[5-(3-chlorophenyl)-3-hydroxypyridine-2- carbonyl]amino}acetic acid (Compound 1), or a pharmaceutically acceptable salt thereof.Compound 1, also referred to as vadadustat or AKB-6548, has the following structure:

[0085] In certain embodiments, a compound is {[5-(3-fluorophenyl)-3-hydroxypyridine-2- carbonyl]amino}acetic acid (Compound la), or a pharmaceutically acceptable salt thereof.Compound la, also referred to as AKB-6899, has the following structure:Compound la (AKB-6899)Formula (II): Enarodustat and Related Compounds

[0086] Exemplary compounds that can be used in any of the compositions and methods described herein include those described in U.S. Pat. No. 8,283,465, U.S. Publication No.US20160145254A1, and U.S. Publication No. US20200017492A1, each of which is incorporated by reference in its entirety. In certain embodiments, a compound, or pharmaceutically acceptable salt thereof, is described in any of claims 1-30 of U.S. Pat. No. 8,283,465.

[0087] In certain embodiments, suitable compounds include a compound having a structure according to Formula (II),Formula (II), or a pharmaceutically acceptable salt thereof, wherein, R and R1are each independently:(i) hydrogen(ii) substituted or unsubstituted phenyl;(iii) substituted or unsubstituted heteroaryl; or(iv) substituted or unsubstituted alkyl; said substitution selected from:(i) C1-C4 alkyl;(ii) C3-C4 cycloalkyl;(iii) C1-C4 alkoxy;(iV) C3-C4 cycloalkoxy;(v) C1-C4 haloalkyl;(vi) C3-C4 halocycloalkyl;(vii) halogen;(viii) cyano;(ix) NHC(O)R4;(x) C(O)NR5aR5b;(xi) phenyl; and(xii) heteroaryl; or(xiii) two substituents are taken together to form a fused ring having from 5 to 7 atoms;R4is a C1-C4 alkyl or C3-C4 cycloalkyl;R5aand R5bare each independently selected from:(i) hydrogen;(ii) C1-C4 alkyl;(iii) C3-C4 cycloalkyl; or(iv) R5aand R5bare taken together to form a ring having from 3 to 7 atoms;R2is selected from:(i) OR6(ii) NR7aR7b; andR6is selected from hydrogen and C1-C4 alkyl or C3-C4 cycloalkyl;R7aand R7bare each independently selected from:(i) hydrogen;(ii) C1-C4 alkyl or C3-C4 cycloalkyl; or(iii) R7aand R7bare taken together to form a ring having from 3 to 7 atoms;R3is selected from hydrogen, methyl, and ethyl;L is a linking unit having a structure -[C(R8aR8b)]n-R8aand R8bare each independently selected from hydrogen, methyl and ethyl; n is an integer from 1 to 3; andR9is selected from hydrogen and methyl.

[0088] In certain embodiments, R is hydrogen.

[0089] In certain embodiments, R is unsubstituted phenyl. In certain embodiments, R is substituted phenyl. In certain embodiments, R is phenyl substituted with C1-C4 alkyl. In certain embodiments, R is phenyl substituted with C3-C4 cycloalkyl. In certain embodiments, R is phenyl substituted with C1-C4 alkoxy. In certain embodiments, R is phenyl substituted with C3- C4 cycloalkoxy. In certain embodiments, R is phenyl substituted with C1-C4 haloalkyl. In certain embodiments, R is phenyl substituted with C3-C4 halocycloalkyl. In certain embodiments, R is phenyl substituted with halogen. In certain embodiments, R is phenyl substituted with cyano. In certain embodiments, R is phenyl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R is phenyl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R is phenyl substituted with phenyl. In certain embodiments, R is phenyl substituted with heteroaryl. In certain embodiments, R is phenyl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0090] In certain embodiments, R is unsubstituted heteroaryl. In certain embodiments, R is substituted heteroaryl. In certain embodiments, R is heteroaryl substituted with C1-C4 alkyl. Incertain embodiments, R is heteroaryl substituted with C3-C4 cycloalkyl. In certain embodiments, R is heteroaryl substituted with C1-C4 alkoxy. In certain embodiments, R is heteroaryl substituted with C3-C4 cycloalkoxy. In certain embodiments, R is heteroaryl substituted with Ci- C4 haloalkyl. In certain embodiments, R is heteroaryl substituted with C3-C4 halocycloalkyl. In certain embodiments, R is heteroaryl substituted with halogen. In certain embodiments, R is heteroaryl substituted with cyano. In certain embodiments, R is heteroaryl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R is heteroaryl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R is heteroaryl substituted with phenyl. In certain embodiments, R is heteroaryl substituted with heteroaryl. In certain embodiments, R is heteroaryl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0091] In certain embodiments, R is unsubstituted alkyl. In certain embodiments, R is substituted alkyl. In certain embodiments, R is alkyl substituted with C1-C4 alkyl. In certain embodiments, R is alkyl substituted with C3-C4 cycloalkyl. In certain embodiments, R is alkyl substituted with C1-C4 alkoxy. In certain embodiments, R is alkyl substituted with C3-C4 cycloalkoxy. In certain embodiments, R is alkyl substituted with C1-C4 haloalkyl. In certain embodiments, R is alkyl substituted with C3-C4 halocycloalkyl. In certain embodiments, R is alkyl substituted with halogen. In certain embodiments, R is alkyl substituted with cyano. In certain embodiments, R is alkyl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3- C4 cycloalkyl. In certain embodiments, R is alkyl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R is alkyl substituted with phenyl. In certain embodiments, R is alkyl substituted with heteroaryl. In certain embodiments, R is alkyl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected fromhydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0092] In certain embodiments, R is substituted or unsubstituted alkyl. In certain embodiments, R is a Ci-Ce alkyl. In certain embodiments, R is a C3-C8 cycloalkyl. In certain embodiments, R is a C6-C14 aryl. In certain embodiments, R is a Ce-Cu aryl. In certain embodiments, R is a Ce- C14 aryl-Ci-Ce alkyl group.

[0093] In certain embodiments, R1is hydrogen.

[0094] In certain embodiments, R1is unsubstituted phenyl. In certain embodiments, R1is substituted phenyl. In certain embodiments, R1is phenyl substituted with C1-C4 alkyl. In certain embodiments, R1is phenyl substituted with C3-C4 cycloalkyl. In certain embodiments, R1is phenyl substituted with C1-C4 alkoxy. In certain embodiments, R1is phenyl substituted with C3- C4 cycloalkoxy. In certain embodiments, R1is phenyl substituted with C1-C4 haloalkyl. In certain embodiments, R1is phenyl substituted with C3-C4 halocycloalkyl. In certain embodiments, R1is phenyl substituted with halogen. In certain embodiments, R1is phenyl substituted with cyano. In certain embodiments, R1is phenyl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R1is phenyl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R1is phenyl substituted with phenyl. In certain embodiments, R1is phenyl substituted with heteroaryl. In certain embodiments, R1is phenyl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0095] In certain embodiments, R1is unsubstituted heteroaryl. In certain embodiments, R1is substituted heteroaryl. In certain embodiments, R1is heteroaryl substituted with C1-C4 alkyl. In certain embodiments, R1is heteroaryl substituted with C3-C4 cycloalkyl. In certain embodiments, R1is heteroaryl substituted with C1-C4 alkoxy. In certain embodiments, R1is heteroaryl substituted with C3-C4 cycloalkoxy. In certain embodiments, R1is heteroaryl substituted with C1-C4 haloalkyl. In certain embodiments, R1is heteroaryl substituted with C3-C4 halocycloalkyl. In certain embodiments, R1is heteroaryl substituted with halogen. In certain embodiments, R1is heteroaryl substituted with cyano. In certain embodiments, R1is heteroarylsubstituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R1is heteroaryl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R1is heteroaryl substituted with phenyl. In certain embodiments, R1is heteroaryl substituted with heteroaryl. In certain embodiments, R1is heteroaryl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0096] In certain embodiments, R1is unsubstituted alkyl. In certain embodiments, R1is substituted alkyl. In certain embodiments, R1is alkyl substituted with C1-C4 alkyl. In certain embodiments, R1is alkyl substituted with C3-C4 cycloalkyl. In certain embodiments, R1is alkyl substituted with C1-C4 alkoxy. In certain embodiments, R1is alkyl substituted with C3-C4 cycloalkoxy. In certain embodiments, R1is alkyl substituted with C1-C4 haloalkyl. In certain embodiments, R1is alkyl substituted with C3-C4 halocycloalkyl. In certain embodiments, R1is alkyl substituted with halogen. In certain embodiments, R1is alkyl substituted with cyano. In certain embodiments, R1is alkyl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3- C4 cycloalkyl. In certain embodiments, R1is alkyl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R1is alkyl substituted with phenyl. In certain embodiments, R1is alkyl substituted with heteroaryl. In certain embodiments, R1is alkyl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0097] In certain embodiments, R1is C1-C10 alkyl. In certain embodiments, R1is C3-C8 unsubstituted cycloalkyl. In certain embodiments, R1is substituted C3-C8 cycloalkyl. In certain embodiments, R1is C3-C8 cycloalkyl substituted with 1 to 5 substituents independently selectedfrom the group consisting of halogen, Ci-Ce alkyl, C3-C8 cycloalkyl, cyano, and Ci-Ce haloalkyl. In certain embodiments, R1is unsubstituted C3-C8 cycloalkenyl. In certain embodiments, R1is substituted C3-C8 cycloalkenyl. In certain embodiments, R1is C3-C8 cycloalkenyl substituted with 1 to 5 substituents independently selected from the group consisting of halogen, Ci-Ce alkyl, C3-C8 cycloalkyl, cyano, and Ci-Ce haloalkyl.

[0098] In certain embodiments, R1is unsubstituted Ce-Cu aryl. In certain embodiments, R1is substituted Ce-Cu aryl. In certain embodiments, R1is Ce-Cu aryl substituted with 1 to 5 substituents independently selected from the group consisting of halogen, Ci-Ce alkyl, C3-C8 cycloalkyl, cyano, and Ci-Ce haloalkyl. In certain embodiments, R1is unsubstituted heteroaryl, wherein the heteroaryl has, besides carbon atom, 1 to 6 hetero atoms selected from nitrogen atom, oxygen atom and sulfur atom. In certain embodiments, R1is substituted Ce-Cu heteroaryl, wherein the heteroaryl has, besides carbon atom, 1 to 6 hetero atoms selected from nitrogen atom, oxygen atom and sulfur atom. In certain embodiments, R1is heteroaryl substituted with 1 to 5 substituents independently selected from the group consisting of halogen, Ci-Ce alkyl, C3- Cs cycloalkyl, cyano, and Ci-Ce haloalkyl, and wherein the heteroaryl has, besides carbon atom, 1 to 6 hetero atoms selected from nitrogen atom, oxygen atom and sulfur atom.

[0099] In certain embodiments, R1is a Ce-Cu aryl-Ci-Ce alkyl group, wherein Ce-Cu aryl is optionally substituted by the same or different 1 to 5 substituents selected from the group consisting of halogen, Ci-Ce alkyl, C3-C8 cycloalkyl, cyano, and Ci-Ce haloalkyl. In certain embodiments, R1is a C3 — Cs cycloalkyl-Ci-Ce alkyl group, wherein C3-C8 cycloalkyl is optionally substituted with 1 to 5 substituents independently selected from the group consisting of halogen, Ci-Ce alkyl, C3-C8 cycloalkyl, cyano, and Ci-Ce haloalkyl. In certain embodiments, R1is a phenyl-Ci-Ce alkyl group.

[0100] In certain embodiments, R2is OR6, wherein R6is hydrogen, C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R2is NR7aR7b, wherein R7aand R7bare each independently selected from the group consisting of hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R7aand R7bare taken together to form a ring having from 3 to 7 atoms.

[0101] In certain embodiments, R3is hydrogen. In certain embodiments, R3is methyl. In certain embodiments, R3is ethyl.

[0102] In certain embodiments, L is a linking unit having a structure -[C(R8aR8b)]n-, wherein R8aand R8bare each independently selected from the group consisting of hydrogen, methyl and ethyl.

[0103] In certain embodiments, n is an integer from 1 to 3. In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3.

[0104] In certain embodiments, R9is hydrogen. In certain embodiments, R9is methyl.

[0105] In certain embodiments, a compound is [(7-hydroxy-5 -phenethyl [1,2,4] triazolo [1,5-a] pyridine-8-carbonyl)aminoacetic acid (Compound 2), or a pharmaceutically acceptable salt thereof. Compound 2, also referred to as enarodustat, or JTZ-951, has the following structure:Compound 2 (enarodustat, or JTZ-951)Formula (III): Desidustat and Related Compounds

[0106] Exemplary compounds that can be used in any of the compositions and methods described herein include those described in U.S. Pat. No. 9,394,300, and U.S. Publication No. US 20190359574A1, each of which is incorporated by reference in its entirety. In certain embodiments, a compound, or pharmaceutically acceptable salt thereof, is described in any of claims 1-10 of U.S. Pat. No. 9,394,300.

[0107] In certain embodiments, suitable compounds include a compound having a structure according to Formula (III),Formula (III), or a pharmaceutically acceptable salt thereof, wherein, R and R1are each independently:(i) hydrogen(ii) substituted or unsubstituted phenyl;(iii) substituted or unsubstituted heteroaryl; or(iv) substituted or unsubstituted alkyl; said substitution selected from:(i) Ci-C4alkyl;(ii) C3-C4 cycloalkyl;(iii) C1-C4 alkoxy;(iv) C3-C4 cycloalkoxy;(v) C1-C4 haloalkyl;(vi) C3-C4 halocycloalkyl;(vii) halogen;(viii) cyano;(ix) NHC(O)R4;(x) C(O)NR5aR5b;(xi) phenyl; and(xii) heteroaryl; or(xiii) two substituents are taken together to form a fused ring having from 5 to 7 atoms;R4is a C1-C4 alkyl or C3-C4 cycloalkyl;R5aand R5bare each independently selected from:(i) hydrogen;(ii) C1-C4 alkyl;(iii) C3-C4 cycloalkyl; or(iv) R5aand R5bare taken together to form a ring having from 3 to 7 atoms;R2is selected from:(i) OR6(ii) NR7aR7b; andR6is selected from hydrogen and C1-C4 alkyl or C3-C4 cycloalkyl;R7aand R7bare each independently selected from:(i) hydrogen;(ii) C1-C4 alkyl or C3-C4 cycloalkyl; or(iii) R7aand R7bare taken together to form a ring having from 3 to 7 atoms;R3is selected from hydrogen, methyl, and ethyl;L is a linking unit having a structure -[C(R8aR8b)]n-R8aand R8bare each independently selected from hydrogen, methyl and ethyl; n is an integer from 1 to 3; andR9is selected from hydrogen and methyl.

[0108] In certain embodiments, R is hydrogen.

[0109] In certain embodiments, R is unsubstituted phenyl. In certain embodiments, R is substituted phenyl. In certain embodiments, R is phenyl substituted with C1-C4 alkyl. In certain embodiments, R is phenyl substituted with C3-C4 cycloalkyl. In certain embodiments, R is phenyl substituted with C1-C4 alkoxy. In certain embodiments, R is phenyl substituted with C3- C4 cycloalkoxy. In certain embodiments, R is phenyl substituted with C1-C4 haloalkyl. In certain embodiments, R is phenyl substituted with C3-C4 halocycloalkyl. In certain embodiments, R is phenyl substituted with halogen. In certain embodiments, R is phenyl substituted with cyano. In certain embodiments, R is phenyl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R is phenyl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R is phenyl substituted with phenyl. In certain embodiments, R is phenyl substituted with heteroaryl. In certain embodiments, R is phenyl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0110] In certain embodiments, R is unsubstituted heteroaryl. In certain embodiments, R is substituted heteroaryl. In certain embodiments, R is heteroaryl substituted with C1-C4 alkyl. In certain embodiments, R is heteroaryl substituted with C3-C4 cycloalkyl. In certain embodiments, R is heteroaryl substituted with C1-C4 alkoxy. In certain embodiments, R is heteroaryl substituted with C3-C4 cycloalkoxy. In certain embodiments, R is heteroaryl substituted with Ci- C4 haloalkyl. In certain embodiments, R is heteroaryl substituted with C3-C4 halocycloalkyl. In certain embodiments, R is heteroaryl substituted with halogen. In certain embodiments, R is heteroaryl substituted with cyano. In certain embodiments, R is heteroaryl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R is heteroaryl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R is heteroaryl substituted with phenyl. In certain embodiments, R is heteroaryl substituted with heteroaryl. In certain embodiments, R is heteroaryl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0111] In certain embodiments, R is unsubstituted alkyl. In certain embodiments, R is substituted alkyl. In certain embodiments, R is alkyl substituted with C1-C4 alkyl. In certain embodiments, R is alkyl substituted with C3-C4 cycloalkyl. In certain embodiments, R is alkyl substituted with C1-C4 alkoxy. In certain embodiments, R is alkyl substituted with C3-C4 cycloalkoxy. In certain embodiments, R is alkyl substituted with C1-C4 haloalkyl. In certain embodiments, R is alkyl substituted with C3-C4 halocycloalkyl. In certain embodiments, R is alkyl substituted with halogen. In certain embodiments, R is alkyl substituted with cyano. In certain embodiments, R is alkyl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3- C4 cycloalkyl. In certain embodiments, R is alkyl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R is alkyl substituted with phenyl. In certain embodiments, R is alkyl substituted with heteroaryl. In certain embodiments, R is alkyl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0112] In certain embodiments, R1is hydrogen.

[0113] In certain embodiments, R1is unsubstituted phenyl. In certain embodiments, R1is substituted phenyl. In certain embodiments, R1is phenyl substituted with C1-C4 alkyl. In certain embodiments, R1is phenyl substituted with C3-C4 cycloalkyl. In certain embodiments, R1is phenyl substituted with C1-C4 alkoxy. In certain embodiments, R1is phenyl substituted with C3- C4 cycloalkoxy. In certain embodiments, R1is phenyl substituted with C1-C4 haloalkyl. In certain embodiments, R1is phenyl substituted with C3-C4 halocycloalkyl. In certain embodiments, R1is phenyl substituted with halogen. In certain embodiments, R1is phenyl substituted with cyano. In certain embodiments, R1is phenyl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R1is phenyl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected fromhydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R1is phenyl substituted with phenyl. In certain embodiments, R1is phenyl substituted with heteroaryl. In certain embodiments, R1is phenyl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0114] In certain embodiments, R1is unsubstituted heteroaryl. In certain embodiments, R1is substituted heteroaryl. In certain embodiments, R1is heteroaryl substituted with C1-C4 alkyl. In certain embodiments, R1is heteroaryl substituted with C3-C4 cycloalkyl. In certain embodiments, R1is heteroaryl substituted with C1-C4 alkoxy. In certain embodiments, R1is heteroaryl substituted with C3-C4 cycloalkoxy. In certain embodiments, R1is heteroaryl substituted with C1-C4 haloalkyl. In certain embodiments, R1is heteroaryl substituted with C3-C4 halocycloalkyl. In certain embodiments, R1is heteroaryl substituted with halogen. In certain embodiments, R1is heteroaryl substituted with cyano. In certain embodiments, R1is heteroaryl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R1is heteroaryl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R1is heteroaryl substituted with phenyl. In certain embodiments, R1is heteroaryl substituted with heteroaryl. In certain embodiments, R1is heteroaryl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0115] In certain embodiments, R1is unsubstituted alkyl. In certain embodiments, R1is substituted alkyl. In certain embodiments, R1is alkyl substituted with C1-C4 alkyl. In certain embodiments, R1is alkyl substituted with C3-C4 cycloalkyl. In certain embodiments, R1is alkyl substituted with C1-C4 alkoxy. In certain embodiments, R1is alkyl substituted with C3-C4 cycloalkoxy. In certain embodiments, R1is alkyl substituted with C1-C4 haloalkyl. In certainembodiments, R1is alkyl substituted with C3-C4 halocycloalkyl. In certain embodiments, R1is alkyl substituted with halogen. In certain embodiments, R1is alkyl substituted with cyano. In certain embodiments, R1is alkyl substituted with NHC(O)R4, wherein R4is a C1-C4 alkyl or C3- C4 cycloalkyl. In certain embodiments, R1is alkyl substituted with C(O)NR5aR5b, wherein R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl; or R5aand R5bare taken together to form a ring having from 3 to 7 atoms. In certain embodiments, R1is alkyl substituted with phenyl. In certain embodiments, R1is alkyl substituted with heteroaryl. In certain embodiments, R1is alkyl substituted with two substituents independently selected from the group consisting of C1-C4 alkyl, C3-C4 cycloalkyl, C1-C4 alkoxy, C3-C4 cycloalkoxy, C1-C4 haloalkyl, C3-C4 halocycloalkyl, NHC(O)R4, C(O)NR5aR5b, phenyl and heteroaryl, wherein the two substituents are taken together to form a fused ring having from 5 to 7 atoms, and wherein R4is a C1-C4 alkyl or C3-C4 cycloalkyl, R5aand R5bare each independently selected from hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R5aand R5bare taken together to form a ring having from 3 to 7 atoms.

[0116] In certain embodiments, R and R1are taken together to form a fused ring having from 5 to 7 atoms. In certain embodiments, R and R1are taken together to form a fused ring that is an aryl. In certain embodiments, R and R1are taken together to form a fused ring that is a phenyl.

[0117] In certain embodiments, R2is OR6, wherein R6is hydrogen, C1-C4 alkyl or C3-C4 cycloalkyl. In certain embodiments, R2is NR7aR7b, wherein R7aand R7bare each independently selected from the group consisting of hydrogen, C1-C4 alkyl, C3-C4 cycloalkyl, or R7aand R7bare taken together to form a ring having from 3 to 7 atoms.

[0118] In certain embodiments, R3is hydrogen. In certain embodiments, R3is methyl. In certain embodiments, R3is ethyl.

[0119] In certain embodiments, L is a linking unit having a structure -[C(R8aR8b)]n-, wherein R8aand R8bare each independently selected from the group consisting of hydrogen, methyl and ethyl.

[0120] In certain embodiments, n is an integer from 1 to 3. In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3.

[0121] In certain embodiments, R9is hydrogen. In certain embodiments, R9is methyl.

[0122] In certain embodiments, a compound is 2-(l-(cyclopropylmethoxy)-4-hydroxy-2-oxo- l,2-dihydroquinoline-3-carboxamido) acetic acid (Compound 3), or a pharmaceutically acceptable salt thereof. Compound 3, also referred to as desidustat, or ZYAN1, has the following structure:(desidustat, or ZYANl)Formula (IV): Roxadustat and Related Compounds

[0123] Still other compounds that can be useful in the compositions and methods described herein include those described in U.S. Pat. No. 7,323,475, which is incorporated by reference in its entirety. In certain embodiments, a compound, or a pharmaceutically acceptable salt thereof, is described in any of claims 1-46 of U.S. Pat. No. 7,323,475.

[0124] In certain embodiments, suitable compounds include a compound having a structure according to Formula (IV),Formula (IV), or a pharmaceutically acceptable salt thereof, wherein, q is zero or one;Ralis selected from the group consisting of hydrogen, alkyl, Substituted alkyl, alkoxy, substituted alkoxy, amino, substituted amino, aminoacyl, aryl, substituted aryl, halo, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, and — XRa6where X is oxygen, — S(O)n— , or — NRa7— where n is zero, one or two, Ra6is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted hetero cyclic, and Ra7is hydrogen, alkyl or aryl or, when X is — NRa7— , then Ra7and Ra6, together with the nitrogen atom to which they are bound, can be joined to form a heterocyclic or substituted heterocyclic group;Ra2and Ra3are independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxy, cyano. — S(O)n— N(Ra6)— Ra6where n is 0, 1, or 2, — NRa6C(O)NRa6Ra6, — XRa6where X is oxygen, — S(O)n— , or — NRa7— where n is zero, one or two, each Ra6is independently selected from the group consisting of hydrogen, alkyl, Substituted alkyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic provided that when when X is — SO — or — SO2 — , then Ra6is not hydrogen, and Ra7is selected from the group consisting of hydrogen, alkyl, aryl, or Ra2, Ra3together with the carbon atom pendent thereto, form an aryl substituted aryl, heteroaryl, or substituted heteroaryl;Ra4and Ra5are independently selected from the group consisting of hydrogen, halo, alkyl, substituted alkyl, alkoxy, Substituted alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl and — XRa6where X is oxygen, — S(O)n— , or — NRa7— where n is zero, one or two, Ra6is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and Ra7is hydrogen, alkyl or aryl or, when X is — NRa7— , then Ra7and Ra6, together with the nitrogen atom to which they are bound, can be joined to form a heterocyclic or Substituted heterocyclic group;Rais selected from the group consisting of hydrogen, alkyl and substituted alkyl;Rais selected from the group consisting of hydroxy, alkoxy, substituted alkoxy, acyloxy, cycloalkoxy, substituted cycloalkoxy, aryloxy, substituted aryloxy, heteroaryloxy, substituted heteroaryl oxy, aryl, — S(O) — Ral° wherein Ral° is selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl and n is zero, one or two; or a pharmaceutically acceptable salt, ester, or prodrug thereof; with the proviso that when Ra" is hydrogen and q is zero, then at least one of the following occurs:1) Rai is fluoro, bromo, iodo, alkyl, substituted alkyl, alkoxy, aminoacyl, Substituted alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, and — XRa6 where X is oxygen, — S(O)n — , or — NRa7 — where n is zero, one or two, Ra6 is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and Ra7 is hydrogen, alkyl or aryl; or2) Ra2 is substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, fluoro, bromo, iodo, cyano, — XRa6 where X is oxygen, — S(O)n — , or — NRa7 — where n is zero, one or two, Ra6 is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and Ra7is hydrogen, alkyl or aryl provided that: a) when Ra2is substituted alkyl such a substituent does not include trifluoromethyl; b) — XRa6is not alkoxy; and c) when — XRa6is substituted alkoxy such a substituent does not include benzyl or benzyl substituted by a substituent selected from the group consisting of (C1-C5) alkyl and (C1-C5) alkoxy or does not include a fluoroalkoxy substituent of the formula:— O— [CH2]X— CfH(2f+i-g)Fg where x is zero or one; f is an integer of from 1 to 5; and g is an integer of from 1 to (2f+l); or3) Ra3is substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, bromo, iodo, — XRa6where X is oxygen — S(O)n— , or — NRa7— where n is zero, one or two, Ra6is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and Ra7is hydrogen, alkyl or aryl provided that: a) when Ra3is substituted alkyl such a substituent does not include trifluoromethyl; b) — XRa6is not alkoxy; and c) when XRa6is substituted alkoxy such a substituent does not include benzyl or benzyl substituted by a substituent selected from the group consisting of (C1-C5) alkyl and (C — C) alkoxy or does not include a fluoroalkoxy substituent of the formula:— O— [CH2]X— CfH(2f+i-g)Fg where x is zero or one; f is an integer of from 1 to 5; and g is an integer of from 1 to (2f+l); or4) Ra4is iodo, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, — XRa6where X is oxygen — S(O)n— , or — NRa7— where n is zero, one or two, Ra6is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and Ra7is hydrogen, alkyl or aryl provided that: a) when Ra4is substituted alkyl such a substituent does not include trifluoromethyl; b) — XRa6is not alkoxy; andc) when XRa6is substituted alkoxy such a substituent does not include benzyl or benzyl substituted by a substituent selected from the group consisting of (C1-C5) alkyl and (C — C) alkoxy or does not include a fluoroalkoxy substituent of the formula:— O — [CH2]xCfH(2f+ 1 -g)Fgwhere x is zero or one; f is an integer of from 1 to 5; and g is an integer of from 1 to (2f+l); or5) Ra5is iodo, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, — XRa6where X is oxygen, — S(O)n— , or — NRa7— where n is zero, one or two, Ra6is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and Ra7is hydrogen, alkyl or aryl provided that: a) when Ra5is substituted alkyl such a substituent does not include trifluoromethyl; b) — XRa6is not alkoxy; and c) when XRa6is substituted alkoxy such a substituent does not include benzyl or benzyl substituted by a substituent selected from the group consisting of (C1-C5) alkyl and (C — C) alkoxy or does not include a fluoroalkoxy substituent of the formula:— O— [CH2]X— CfH(2f+i-g)Fg where x is zero or one; f is an integer of from 1 to 5; and g is an integer of from 1 to (2f+l); and with the further following proviso: that when Ral, Ra3, Ra4, and Ra5are hydrogen, then Ra2is not bromo.

[0125] In certain embodiments, a compound according to Formula (IV) has the following structure:Formula (IVa), or a pharmaceutically acceptable salt thereof, wherein q is zero or one;Rais selected from hydrogen and alkyl:Ralis selected from the group consisting of hydrogen, alkyl, Substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, halo, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, and — XRa6where X is oxygen, — S(O)n— , or — NRa7— where n is zero, one or two, Ra6is selected from the group consisting of alkyl, Substituted alkyl, aryl, sub stituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and Ra7is hydrogen, alkyl or aryl;Ra2and Ra3are independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxy, cyano, — XRa6where X is oxygen, — S(O)n— , or — NRa7— where n is zero, one or two, Ra6is selected from the group consisting of alkyl, Substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and Ra7is hydrogen, alkyl or aryl;Ra4and Ra5are independently selected from the group consisting of hydrogen, halo, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, and — XRa6where X is oxygen, — S(O)n— , or — NRa7— where n is zero, one or two, Ra6is selected from the group consisting of alkyl, Substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and Ra7is hydrogen, alkyl or aryl; or a pharmaceutically acceptable salt or prodrug thereof.

[0126] In certain embodiments, a compound according to Formula (IV) has the following structure:Formula (IVb),

[0127] or a pharmaceutically acceptable salt thereof, wherein,Ralis selected from the group consisting of hydrogen, halo, alkyl, substituted alkyl, alkoxy, substituted alkoxy, amino, substituted amino, aminoacyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, and — XRa6where X is oxygen, — S(O)n— , or — NRa7— where n is zero, one or two, Ra6is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and Ra7is hydrogen,alkyl or aryl or, when X is — NRa7— , then Ra7and Ra6, together with the nitrogen atom to which they are bound, can be joined to form a heterocyclic or substituted heterocyclic group; andRa2is selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxy, cyano, — S(O)n— N(Ra6)— Ra6where n is 0, 1 or 2, — NRa6C(O)NRa6Ra6, XRa6where X is oxygen, — S(O)n— , or — NRa7— where n is zero, one or two, each Ra6is independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, provided that when X is — SO — or — SO2— , then Ra6is not hydrogen, and Ra7is selected from the group consisting of hydrogen, alkyl, aryl, or Ra2, Ra3together with the carbon atom pendent thereto, form an aryl, substituted aryl, heteroaryl, or substituted heteroaryl; or a pharmaceutically acceptable salt, ester, or prodrug thereof; with the proviso that at least one of the following occurs:1) Ralis fluoro, bromo, iodo, alkyl, substituted alkyl, alkoxy, aminoacyl, substituted alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, and — XRa6where X is oxygen, — S(O)n— , or — NRa7— where n is zero, one or two, Ra6is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and Ra7is hydrogen, alkyl or aryl; or2) Ra2is substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, fluoro, bromo, iodo, cyano, — XRa6where X is oxygen, — S(O)n— , or — NRa7— where n is zero, one or two, Ra6is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and Ra7is hydrogen, alkyl or aryl provided that: a) when Ra2is substituted alkyl such a substituent does not include trifluoromethyl; b) — XRa6is not alkoxy; and c) when — XRa6is substituted alkoxy such a substituent does not include benzyl or benzyl substituted by a substituent selected from the group consisting of (C1-C5) alkyl and (C1-C5) alkoxy or does not include a fluoroalkoxy substituent of the formula:— O — [CH2]X— CfH(2f+i-g)Fgwhere x is zero or one; f is an integer of from 1 to 5; and g is an integer of from 1 to (2f+l).

[0128] In certain embodiments, a compound is (l-methyl-4-hydroxy-7-phenoxy-isoquinoline-3- carbonyl)-amino-acetic acid (Compound 4), or a pharmaceutically acceptable salt thereof.Compound 4, also referred to as roxadustat, or FG-4592, has the following structure:Compound 4 (roxadustat, or FG-4592)Formula (V): Daprodustat and Related Compounds

[0129] Still other compounds that can be useful in the compositions and methods described herein include those described in U.S. Pat. No. 8,324,208, which is incorporated by reference in its entirety. In certain embodiments, a compound, or a pharmaceutically acceptable salt thereof, is described in any of claims 1-16 of U.S. Pat. No. 8,324,208.

[0130] In certain embodiments, suitable compounds include a compound having a structure according to Formula (V),Formula (V), or a pharmaceutically acceptable salt thereof, whereinRbland Rb4are each independently selected from the group consisting of hydrogen, — NRb5Rb6, Ci-Cio alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C8cycloalkyl, C3-C8cycloalkyl-Ci-Cio alkyl, Cs-C8cycloalkenyl, Cs-C8cycloalkenyl- C1-C10 alkyl, C3-C8heterocycloalkyl, C3-C8heterocycloalkyl-Ci-Cio alkyl, aryl, aryl- C1-C10 alkyl, heteroaryl and heteroaryl- C1-C10 alkyl;Rb2is — NRb7Rb8or — ORb9;Rb3is H or C1-C4 alkyl; where Rb5and Rb6are each independently selected from the group consisting of hydrogen, C1-C10 alkyl, C3-C8cycloalkyl, C3-C8cycloalkyl-Ci-Cio alkyl, C3-C8heterocycloalkyl, C3-C8heterocycloalkyl-Ci-Cio alkyl, aryl, aryl- C1-C10 alkyl,heteroaryl and heteroaryl- C1-C10 alkyl, — C(O)Ci-C4 alkyl, — C(O)Cs — Ce cycloalkyl, — C(O)Cs — Ce heterocycloalkyl, — C(O)aryl, — C(O)heteroaryl and — S(O)2 C1-C4 alkyl, or, when Rb5and Rb6are attached to the same nitrogen, Rb5and Rb6taken together with the nitrogen to which they are attached forma 5- or 6- or 7-membered saturated ring optionally containing one other heteroatom selected from oxygen, nitrogen and sulphur, Rb7and Rb8are each independently selected from the group consisting of hydrogen, Ci- C10 alkyl, C2-C10 alkenyl, C3-C8 cycloalkyl, C3-C8 heterocycloalkyl, aryl, and heteroaryl, andRb9is H or a cation, or C1-C10 alkyl which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of C3-C6 cycloalkyl, heterocycloalkyl, aryl and heteroaryl;Xbis O or S; andY is O or S; where any carbon or heteroatom of Rbl, Rb2, Rb3, Rb4, Rb5, Rb6, Rb7, Rb8, Rb9is unsubstituted or is substituted with one or more substituents independently selected from Ci-Ce alkyl, Ci-Ce haloalkyl, halogen, — ORbl°, — NRb5Rb6, oxo, cyano, nitro, — C(O)Rbl°, — C(O)ORbl° — SRbl°, — S(O)Rbl°, — S(O)2Rbl°, — CONRb5Rb6, — N(Rb5)C(O)Rbl°, — N(Rb5)C(O)ORbl°, — OC(O)NRb5Rb6', — N(Rb5)C(O)NRb5Rb6, — SO2NRb5Rb6, — N(Rb5)SO2Rbl°, C2-C10 alkenyl, C2-C10 alkynyl, C3-C6cycloalkyl, C3-C6heterocycloalkyl, aryl, Ci-Ce alkyl-aryl, heteroaryl and Ci-Ce alkyl-heteroaryl, wherein Rb5and Rb6are the same as defined above and Rbl° is selected from hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl. — C(O)CiC4 alkyl, -C(O)aryl, -C(O)heteroaryl, — C(O)C3— C6cycloalkyl, — C(O)C3— C6heterocycloalkyl, — S(O)2Ci— C4alkyl, C3-C8cycloalkyl, C3-C8 heterocycloalkyl, Ce-Cu aryl, aryl-Ci-Cio alkyl, heteroaryl and heteroaryl-Ci-Cio alkyl.

[0131] In certain embodiments, Xbis O. In certain embodiments, Y is O.

[0132] In certain embodiments, Rbland Rb4are each independently selected from the group consisting of hydrogen, — NRb5Rb6, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C8 cycloalkyl, C3-C8 cycloalkyl-Ci-Cio alkyl, C5-C8 cycloalkenyl, C5-C8 cycloalkenyl- C1-C10 alkyl, C3-C8 heterocycloalkyl, C3-C8 heterocycloalkyl-Ci-Cio alkyl, aryl, aryl- C1-C10 alkyl, heteroaryl and heteroaryl- C1-C10 alkyl. In certain embodiments, Rblis C3-C8 cycloalkyl. In certain embodiments, Rb4is C3-C8 cycloalkyl.

[0133] In certain embodiments, Rb2is — ORb9, wherein Rb9is H or a cation. In certain embodiments, Rb2is OH.

[0134] In certain embodiments, Rb3is H.

[0135] In certain embodiments, a compound is N-(l,3-Dicyclohexyl-6-hydroxy-2,4-dioxo- l,2,3,4-tetrahydro-5-pyrimidinyl)carbonylglycine (Compound 5), or a pharmaceutically acceptable salt thereof. Compound 5, also referred to as daprodustat, or GSK-12788363, has the following structure:Compound 5 (daprodustat, or GSK-12788363)Formula (VI): Molidustat and Related Compounds

[0136] Still other compounds that can be useful in the compostions and methods described herein include those described in U.S. Pat. No. 8,389,520, which is incorporated by reference in its entirety. In certain embodiments, a compound, or a pharmaceutically acceptable salt thereof, is described in any of claims 1-10 of U.S. Pat. No. 8,389,520.

[0137] In certain embodiments, suitable compounds include a compound having a structure according to Formula (VI),or a pharmaceutically acceptable salt thereof, wherein Rclrepresents a heteroaryl group of the formula* denotes the linkage point with the dihydropyrazolone ring andRc4denotes hydrogen, fluorine, chlorine, bromine, cyano, C1-C4 alkyl, trifluoromethyl, hydroxymethyl, C1-C4 alkoxy, trifluoromethoxy, hydroxycarbonyl or C1-C4 alkoxy carbonyl;Rc2represents a heteroaryl group of the formulawherein# denotes the linkage point with the dihydropyrazolone ring andRc6, Rc6aand Rc6bare identical or different and independently of one another denote hydrogen or a substituent chosen from the group consisting of fluorine, chlorine, bromine, cyano, Ci-Ce alkyl, trifluoromethyl, hydroxyl, Ci-Ce alkoxy, trifluoromethoxy, amino, mono-Ci-C4 alkylamino, di-Ci-C4 alkylamino, hydroxycarbonyl, C1-C4 alkoxycarbonyl, 4-to 6-membered heterocycloalkyl, phenyl and 5-or 6-membered heteroaryl, whereinCi-Ce alkyl in its turn can be substituted by hydroxyl, C1-C4 alkoxy or amino and 4-to 6-membered heterocycloalkyl, phenyl and 5-or 6-membered heteroaryl in their turn can in each case be substituted once or twice in an identical or different manner by fluorine, chlorine, bromine, cyano, C1-C4 alkyl, trifluoromethyl, hydroxyl, C1-C4 alkoxy, trifluoromethoxy, oxo, amino, mono-Ci-C4 alkylamino, di-Ci-C4 alkylamino, hydroxycarbonyl or C1-C4 alkoxycarbonyl, andRc3represents hydrogen, or a salt thereof.

[0138] In certain embodiments, Rclis a heteroaryl group of the formulawherein* denotes the linkage point with the dihydropyrazolone ring, and wherein Rc4is hydrogen, fluorine, chlorine, bromine, cyano, C1-C4 alkyl, trifluoromethyl, hydroxymethyl, C1-C4 alkoxy, trifluoromethoxy, hydroxycarbonyl or C1-C4 alkoxy carbonyl.

[0139] In certain embodiments, Rc2is a heteroaryl group of the formulawherein # denotes the linkage point with the dihydropyrazolone ring and wherein Rc6, Rc6aand Rc6bare identical or different and independently of one another denote hydrogen or a substituent chosen from the group consisting of fluorine, chlorine, bromine, cyano, Ci-Ce alkyl, trifluoromethyl, hydroxyl, Ci-Ce alkoxy, trifluoromethoxy, amino, monoCid alkylamino, di-Ci-C4 alkylamino, hydroxycarbonyl, C1-C4 alkoxycarbonyl, 4-to 6- membered heterocycloalkyl, phenyl and 5-or 6-membered heteroaryl. In certain embodiments, a Ci-Ce alkyl is unsubstituted Ci-Ce alkyl. In certain embodiments, a Ci- Ce alkyl is substituted Ci-Ce alkyl. In certain embodiments, a Ci-Ce alkyl is substituted with hydroxyl, C1-C4 alkoxy or amino. In certain embodiments, a 4-to 6-membered heterocycloalkyl is unsubstituted. In certain embodiments, a 4-to 6-membered heterocycloalkyl is substituted. In certain embodiments, a 4-to 6-membered heterocycloalkyl is substituted with once or twice in an identical or different manner by fluorine, chlorine, bromine, cyano, C1-C4 alkyl, trifluoromethyl, hydroxyl, C1-C4 alkoxy, trifluoromethoxy, oxo, amino, mono-Ci-C4 alkylamino, di-Ci-C4 alkylamino, hydroxycarbonyl or C1-C4 alkoxy carbonyl. In certain embodiments, a phenyl is unsubstituted. In certain embodiments, a phenyl is substituted. In certain embodiments, a phenyl is substituted with once or twice in an identical or different manner by fluorine, chlorine, bromine, cyano, C1-C4 alkyl, trifluoromethyl, hydroxyl, C1-C4 alkoxy, trifluoromethoxy, oxo, amino, mono-Ci-C4 alkylamino, di-Ci-C4 alkylamino, hydroxycarbonyl or C1-C4 alkoxy carbonyl. In certain embodiments, a 5-or 6-membered heteroaryl is unsubstituted. In certain embodiments, a 5-or 6-membered heteroaryl is substituted. In certain embodiments, a 5-or 6-membered heteroaryl is substituted with once or twice in an identical or different manner by fluorine, chlorine, bromine, cyano, C1-C4 alkyl, trifluoromethyl, hydroxyl, C1-C4 alkoxy, trifluoromethoxy, oxo, amino, mono-Ci-C4 alkylamino, di-Ci-C4 alkylamino, hydroxycarbonyl or C1-C4 alkoxy carbonyl.

[0140] In certain embodiments, Rc3is hydrogen.

[0141] In certain embodiments, a compound 2-(6-Morpholin-4-ylpyrimidin-4-yl)-4-(lH-l,2,3- triazol-1- yl)-l,2-dihydro-3H-pyrazol-3-one (Compound 6), or a pharmaceutically acceptable salt thereof. Compound 6, also referred to as molidustat, or BAY 85-3934, has the following structure:Pharmaceutical Compositions

[0142] Pharmaceutical compositions may be used in the preparation of individual, single unit dosage forms. Pharmaceutical compositions and dosage forms provided herein comprise a prolyl hydroxylase inhibitor, such as a compound described herein ( e.g., a compound having a structure according to any one of Formulas (I)-(VI), or a compound such as vadadustat (AKB- 6548), roxadustat (FG-4592), daprodustat (GSK-12788363), molidustat (BAY 85-3934), enarodustat (JTZ-951), or desidustat (ZYAN1), or a pharmaceutically acceptable salt thereof). Pharmaceutical compositions and dosage forms can further comprise one or more excipients. Like the amounts and types of excipients, the amounts and specific types of active ingredients in a dosage form may differ depending on factors including, but not limited to, the route by which it is to be administered to patients.

[0143] In certain embodiments, a prolyl hydroxylase inhibitor may be administered by oral or parenteral route. As used herein, the term “parenteral” includes but not limited to subcutaneous, intradermal, intravascular injections, such as intravenous, intramuscular and any another similar injection or infusion technique; transdermal route; inhalation, and rectal route. In certain embodiments, a prolyl hydroxylase inhibitor may be administered orally, such as in a tablet or capsule formulation.

[0144] Pharmaceutical compositions that are suitable for oral administration can be provided as discrete dosage forms, such as, but not limited to, tablets (e.g., chewable tablets), caplets, capsules, and liquids (e.g., flavored syrups). Such dosage forms contain predetermined amountsof active ingredients, and may be prepared by methods of pharmacy well known to those skilled in the art.

[0145] Oral dosage forms provided herein are prepared by combining the active ingredients in an intimate admixture with at least one excipient according to conventional pharmaceutical compounding techniques. Excipients can take a wide variety of forms depending on the form of preparation desired for administration. For example, excipients suitable for use in oral liquid or aerosol dosage forms include, but are not limited to, water, glycols, oils, alcohols, flavoring agents, preservatives, and coloring agents. Examples of excipients suitable for use in solid oral dosage forms (e.g., powders, tablets, capsules, and caplets) include, but are not limited to, starches, sugars, micro-crystalline cellulose, diluents, granulating agents, lubricants, binders, and disintegrating agents.

[0146] In certain embodiments, oral dosage forms are tablets or capsules, in which case solid excipients are employed. In another embodiment, tablets can be coated by standard aqueous or non-aqueous techniques. Such dosage forms can be prepared by any of the methods of pharmacy. In general, pharmaceutical compositions and dosage forms are prepared by uniformly and intimately admixing the active ingredients with liquid carriers, finely divided solid carriers, or both, and then shaping the product into the desired presentation if necessary.

[0147] For example, a tablet can be prepared by compression or molding. Compressed tablets can be prepared by compressing in a suitable machine the active ingredients in a free-flowing form such as powder or granules, optionally mixed with an excipient.

[0148] Examples of excipients that can be used in oral dosage forms provided herein include, but are not limited to, binders, fillers, disintegrants, and lubricants. Binders suitable for use in pharmaceutical compositions and dosage forms include, but are not limited to, com starch, potato starch, or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethyl cellulose), polyvinyl pyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropyl methyl cellulose, (e.g., Nos. 2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof.

[0149] Suitable forms of microcrystalline cellulose include, but are not limited to, the materials sold as AVICEL-PH-101, AVICEL-PH-103 AVICEL RC-581, AVICEL-PH-105 (available from FMC Corporation, American Viscose Division, Avicel Sales, Marcus Hook, PA), and mixtures thereof. A specific binder is a mixture of microcrystalline cellulose and sodium carboxymethyl cellulose sold as AVICEL RC-581. Suitable anhydrous or low moistureexcipients or additives include AVICEL-PH-103™ and Starch 1500 LM. Other suitable forms of microcrystalline cellulose include, but are not limited to, silicified microcrystalline cellulose, such as the materials sold as PROSOLV 50, PROSOLV 90, PROSOLV HD90, PROSOLV 90 LM, and mixtures thereof.

[0150] Examples of fillers suitable for use in the pharmaceutical compositions and dosage forms provided herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof. The binder or filler in pharmaceutical compositions is, in certain embodiments, present in from about 50 to about 99 weight percent of the pharmaceutical composition or dosage form.

[0151] In certain embodiments, fillers may include, but are not limited to block copolymers of ethylene oxide and propylene oxide. Such block copolymers may be sold as POLOXAMER or PLURONIC, and include, but are not limited to POLOXAMER 188 NF, POLOXAMER 237 NF, POLOXAMER 338 NF, POLOXAMER 437 NF, and mixtures thereof.

[0152] In certain embodiments, fillers may include, but are not limited to isomalt, lactose, lactitol, mannitol, sorbitol xylitol, erythritol, and mixtures thereof.

[0153] Disintegrants may be used in the compositions to provide tablets that disintegrate when exposed to an aqueous environment. Tablets that contain too much disintegrant may disintegrate in storage, while those that contain too little may not disintegrate at a desired rate or under the desired conditions. Thus, a sufficient amount of disintegrant that is neither too much nor too little to detrimentally alter the release of the active ingredients may be used to form solid oral dosage forms. The amount of disintegrant used varies based upon the type of formulation, and is readily discernible to those of ordinary skill in the art. In certain embodiments, pharmaceutical compositions comprise from about 0.5 weight percent to about 15 weight percent of disintegrant, or from about 1 to about 5 weight percent of disintegrant.

[0154] Disintegrants that can be used in pharmaceutical compositions and dosage forms include, but are not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, povidone, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pre-gelatinized starch, other starches, clays, other algins, other celluloses, gums, and mixtures thereof.

[0155] Glidants that can be used in pharmaceutical compositions and dosage forms include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium stearyl fumarate, sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil, sunfloweroil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, and mixtures thereof. Additional glidants include, for example, a syloid silica gel (AEROSIL200, manufactured by W.R. Grace Co. of Baltimore, MD), a coagulated aerosol of synthetic silica (marketed by Degussa Co. of Plano, TX), CAB-O-SIL (a pyrogenic colloidal silicon dioxide product sold by Cabot Co. of Boston, MA), and mixtures thereof. If used at all, glidants may be used in an amount of less than about 1 weight percent of the pharmaceutical compositions or dosage forms into which they are incorporated.

[0156] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents, and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols, and fatty acid esters of sorbitan, and mixtures thereof.

[0157] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming, and preservative agents.

[0158] Suspensions, in addition to the active inhibitor(s) may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0159] In certain embodiments, a disease or condition of copper metabolism as described herein, may be treated by administering to a patient in need thereof from about 100 mg to about 1800 mg, about 100 mg to about 1500 mg, about 150 mg to about 1200 mg, about 150 mg to about 1450 mg, about 200 mg to about 1400 mg, about 250 mg to about 1350 mg, about 300 mg to about 1300 mg, about 350 mg to about 1250 mg, about 400 mg to about 1200 mg, about 450 mg to about 1150 mg, about 500 mg to about 1100 mg, about 550 mg to about 1050 mg, about 600 mg to about 1000 mg, about 650 mg to about 950 mg, about 700 mg to about 900 mg, about 850 mg to about 950 mg, about 800 mg to about 1000 mg, about 750 mg to about 1050 mg, about 700 mg to about 1100 mg, about 650 mg to about 1150 mg, about 600 mg to about 1200 mg, about 150 mg to about 1800 mg, about 300 mg to about 1800 mg, about 450 mg to about 1800 mg, about 600 mg to about 1800 mg, about 750 mg to about 1800 mg, or about 900 mg to about 1800 mg of a prolyl hydroxylase inhibitor. In certain embodiments, the dose of a prolylhydroxylase inhibitor is between 150 mg to 750 mg. In certain embodiments, the dose of a prolyl hydroxylase inhibitor is between 900 mg to 1800 mg. In certain embodiments, the dose of a prolyl hydroxylase inhibitor is between 150 mg to 1800 mg. In certain embodiments, the dose of a prolyl hydroxylase inhibitor is between 150 mg to 1000 mg. In certain embodiments, the dose of a prolyl hydroxylase inhibitor is about 100 mg, about 120 mg, about 140 mg, about 160 mg, about 180 mg, about 200 mg, about 220 mg, about 240 mg, about 260 mg, about 280 mg, about 300 mg, about 320 mg, about 340 mg, about 360 mg, about 380 mg, about 400 mg, about 420 mg, about 440 mg, about 460 mg, about 480 mg, about 500 mg, about 520 mg, about 540 mg, about 560 mg, about 580 mg, about 600 mg, about 620 mg, about 640 mg, about 660 mg, about 680 mg, about 700 mg, about 720 mg, about 740 mg, about 760 mg, about 780 mg, about 800 mg, about 820 mg, about 840 mg, about 860 mg, about 880 mg, about 900 mg, about 920 mg, about 940 mg, about 960 mg, about 980 mg, about 1000 mg, about 1020 mg, about 1040 mg, about 1060 mg, about 1080 mg, about 1100 mg, about 1120 mg, about 1140 mg, about 1160 mg, about 1180 mg, about 1200 mg, about 1220 mg, about 1240 mg, about 1260 mg, about 1280 mg, about 1300 mg, about 1320 mg, about 1340 mg, about 1360 mg, about 1380 mg, about 1400 mg, about 1420 mg, about 1440 mg, about 1460 mg, about 1480 mg, about 1500 mg, about 1650 mg, or about 1800 mg. In certain embodiments, such doses may be administered three times a week, once daily, twice daily, or three times daily.

[0160] In certain embodiments, the pharmaceutical composition may comprise a copper salt, such as copper acetate, copper carbonate, copper chloride, copper iodide, copper nitrate, copper oxide, copper selenite, copper sulfate, or copper pyrophosphate. The copper salt may also be a copper amino acid complex, such as copper aspartate, copper glutamate, copper glycinate, copper histidinate, copper lysinate, copper methionate, copper tyrosinate, or copper complexed to a mixture of amino acids. The copper salt may also be a copper carboxylic acid complex, such as copper ascorbate, copper citrate, copper formate, copper fumarate, copper gluconate, copper glutarate, copper malate, copper orotate, copper picolinate, copper sebacate, copper succinate, or copper tartrate. In certain embodiments, the copper salt is copper histidinate.Methods of Treatment

[0161] Described herein are therapeutic methods comprising increasing the activity of a hypoxia response in a subject. In particular, methods described herein can be useful for treating a disease or condition of copper metabolism in subjects in need thereof, such as a subject having Wilson disease or Menkes disease. In certain embodiments, the methods comprise administering a compound that can inhibit prolyl hydroxylase or a pharmaceutical composition described herein comprising a compound that can inhibit prolyl hydroxylase. For example, methods describedherein can be useful for treating Wilson disease or Menkes disease in a subject in need thereof, where the method comprises administering a compound that can inhibit prolyl hydroxylase.

[0162] Suitable prolyl hydroxylase inhibitors include, but are not limited to, those described herein, including compounds according to any one of Formulas (I)-(VI), such as vadadustat (AKB-6548), roxadustat (FG-4592), daprodustat (GSK-12788363), molidustat (BAY 85-3934), enarodustat (JTZ-951), or desidustat (ZYAN1), or any pharmaceutically acceptable salt thereof.

[0163] In certain embodiments, the methods described herein can treat Wilson disease, one or more conditions induced by or associated with Wilson disease, and / or one of more conditions in a patient having Wilson disease.

[0164] In certain embodiments, the methods described herein can treat Menkes disease, one or more conditions induced by or associated with Menkes disease, and / or one of more conditions in a patient having Menkes disease.

[0165] A disease or condition of copper metabolism as described herein may also be treated by hypoxia therapy. In certain embodiments, the methods comprise administering to a subject by inhalation a therapeutically effective amount of a therapeutic gas comprising between 5 to 20% O2. A hypoxia induction system can be used to prepare the therapeutic gas that is administered to the subject.

[0166] In a first example, a hypoxia induction system contains a first container containing a first gas comprising nitrogen and a second container containing a second gas comprising oxygen, and the oxygen-depleted air delivered to the subject is prepared by mixing the first gas and the second gas.

[0167] In a second example, a hypoxia induction system intakes ambient air, reduces the oxygen content of the intake air, to produce the oxygen-depleted air that is delivered to the subject. Colorado Altitude Training (CAT) provides a portable enclosure system that produces hypoxic air by this mechanism. CAT uses a high-flow hypoxic air delivery unit and a semi-sealed enclosure. The air unit draws in ambient room air and separates the oxygen molecules from the nitrogen molecules, creating the hypoxic or oxygen reduced air. Simulated altitudes of up to 14,500 feet can be achieved, and can be controlled to within 100 feet accuracy using CAT's digital control system. Hypoxico Altitude Training Systems devices generate hypoxic air in a manner similar to the CAT system. Simulated altitudes of about 21,500 feet can be achieved with the Hypoxico Altitude Training Systems.

[0168] In a third example, a hypoxia induction system intakes ambient air, adds nitrogen to the intake air, to produce the oxygen-depleted air that is delivered to the subject. The nitrogen can provided from, e.g., liquid nitrogen, cylinders of highly purified nitrogen (e.g., 100% nitrogen),or cylinders containing nitrogen (e.g., 80-95%) and oxygen (e.g., 5-20%). Nitrogen used to produce oxygen-depleted air can optionally be produced by pressure swing adsorption (PSA). Under high pressure, gases tend to be adsorbed to solid surfaces. When the pressure is reduced, the gas is desorbed. If air is passed under pressure through a vessel containing a carbon molecular sieve, oxygen gas is adsorbed and gas enriched in nitrogen and depleted in oxygen is produced. Dual step PSA can be used to produce a highly pure nitrogen gas: first, compressed air is forced through a carbon molecular sieve to produce nitrogen at a purity of approximately 98%; second, the nitrogen produced in the first step is forced into a second carbon molecular sieve and the nitrogen gas reaches a purity up to 99.999%. This nitrogen gas can be combined with ambient air to produce oxygen-depleted air that is delivered to the subject. Alternatively, if air is passed under pressure through a vessel containing an adsorbent bed of zeolite that attracts nitrogen more strongly than it does oxygen, part or all of the nitrogen will stay in the bed. When the bed reaches the end of its capacity to adsorb nitrogen, the pressure can be reduced to release the adsorbed nitrogen. This released nitrogen can be combined with ambient air to produce oxygen-depleted air that is delivered to the subject.

[0169] The therapeutic gas delivered to the subject can contain between 5 to 20% O2. The proportion of oxygen in the therapeutic gas can vary based upon factors including the age of the subject, the condition treated, and the duration of the hypoxic treatment. For example, the therapeutic gas can contain between 5 to 20% O2, between 6 to 18% O2, between 7 to 17% O2, between 8 to 16% O2, between 9 to 15% O2, between 10 to 15% O2, between 10 to 14% O2, between 10 to 13% O2, between 10 to 12% O2, or between 10 to 11% O2. In certain embodiments, the therapeutic gas contains about 5% O2, about 6% O2, about 7% O2, about 8% O2, about 9% O2, about 10% O2, about 11% O2, about 12% O2, about 13% O2, about 14% O2, about 15% O2, about 16% O2, about 17% O2, or about 18% O2.

[0170] The therapeutic gas can be administered to the subject by devices including but not limited to nasal prongs, a face mask, an enclosed tent or chamber (completely or semi-sealed), an intra-tracheal catheter, an endotracheal tube, or a tracheostomy tube. The use of nasal prongs or a face mask allow for greater mobility of the subject while being treated with the therapeutic gas.

[0171] The therapeutic gas can optionally be inhaled by the subject continuously over the course of a variety of time periods (e.g., at least 1 minute, at least 2 minutes, at least 3 minutes, at least 4 minutes, at least 5 minutes, at least 6 minutes, at least 7 minutes, at least 8 minutes, at least 9 minutes, at least 10 minutes, at least 15 minutes, at least 1 hour, at least 2 hours, at least 4 hours, at least 8 hours, at least 12 hours, at least 24 hours, at least 48 hours, or longer). In someinstances, administration of the therapeutic gas is intermittent (e.g., only during inspiration to save gas (sensing inspiration by a variety of methods, temperature, pressure, etc.)) or only for a few hours each day, wherein the therapeutic gas is inhaled (e.g., for a period of time specified above), followed by breathing ambient air, which is then followed by one or more additional periods of breathing the therapeutic gas.

[0172] The therapeutic gas can optionally be administered to a subject during sleep. This can be achieved by, for example, positioning a tent over a bed or placing a bed within an enclosed chamber. For children or infants, a tent can be positioned over a crib or a crib can be placed within an enclosed chamber.

[0173] A subject treated with the therapeutic gas can be monitored to measure the effects mediated by the treatment. For example, arterial oxygen saturation (SpO2) can be measured in the subject one or more times (continuously or intermittently) after administration of the therapeutic gas to the subject. The measured SpO2value can be used to feedback and adjust (e.g. automatically adjust) the oxygen content of the administered therapeutic gas so as to maintain SpO2in the subject in the range of 50-90% (e.g., 60-90%, 70-90%, 75-90%, 80-90%, 85-90%, or about 85%). In addition or alternatively, arterial partial oxygen pressure (Pat ) can be measured in the subject one or more times (continuously or intermittently) after administration of the therapeutic gas to the subject. The measured Pat value can be used to feedback and adjust (e.g. automatically adjust) the oxygen content of the administered therapeutic gas so as to maintain PaO2in the subject in the range of 25 mm Hg to 90 mm Hg (e.g., 25 mm Hg to 70 mm Hg, 25 mm Hg to 60 mm Hg, 25 mm Hg to 55 mm Hg, 25 mm Hg to 45 mm Hg, 25 mm Hg to 40 mm Hg, 25 mm Hg to 35 mm Hg, 35 mm Hg to 70 mm Hg, 35 mm Hg to 60 mm Hg, 45 mm Hg to 60 mm Hg, 50 mm Hg to 60 mm Hg, 55 mm Hg to 60 mm Hg, or about 55 mm Hg). In certain embodiments, SpO2and PaO2are both measured in the subject one or more times (continuously or intermittently) after administration of the therapeutic gas to the subject. The measured SpO2and PaO2values can be used to feedback and adjust (e.g. automatically adjust) the oxygen content of the administered therapeutic gas so as to maintain SpO2and PaO2in the subject in the range of 50-90% SpO2and 25 mm Hg to 70 mm Hg PaO2(e.g., 80-90% SpO2and 50 mm Hg to 60 mm Hg PaO2, 85-90% SpO2and 55 mm Hg to 60 mm Hg PaO2, or about 85% SpO2and about 55 mm Hg PaO2).

[0174] Blood hematocrit and / or circulating hemoglobin levels can be measured in a subject one or more times after administration of the therapeutic gas to the subject, wherein the detection of an increase in blood hematocrit and / or an increase in circulating hemoglobin levels after initiation of the treatment indicates that the treatment is having its desired effect. A decision ofwhether or how to continue the treatment can be based upon the measured blood hematocrit and / or circulating hemoglobin levels. For example, if the measured blood hematocrit and / or circulating hemoglobin levels exceed a pre-established target level, then the treatment may be terminated or the oxygen content of the administered therapeutic gas may be increased. In another example, if the measured blood hematocrit and / or circulating hemoglobin levels are below a pre-established target level, then the duration of the treatment may be extended and / or the oxygen content of the therapeutic gas may be decreased.

[0175] A disease or condition of copper metabolism may also be treated by causing a subject to breathe a therapeutically effective amount of air in a hypobaric chamber. In some instances, the hypobaric chamber has an atmospheric pressure equivalent to that between 1,500 meters to 10,000 meters above sea level (e.g., equivalent to 1,500 meters to 10,000 meters above sea level, equivalent to 2,000 meters to 8,000 meters above sea level, equivalent to 2,000 meters to 7,000 meters above sea level, equivalent to 2,000 meters to 6,000 meters above sea level, equivalent to 2,000 meters to 5,000 meters above sea level, equivalent to 2,000 meters to 4,000 meters above sea level, equivalent to 2,000 meters to 3,000 meters above sea level, equivalent to 2,000 meters to 2,500 meters above sea level, equivalent to 3,000 meters to 7,000 meters above sea level, equivalent to 3,000 meters to 6,000 meters above sea level, equivalent to 3,000 meters to 5,000 meters above sea level, equivalent to 3,000 meters to 4,000 meters above sea level, equivalent to 4,000 meters to 7,000 meters above sea level, equivalent to 4,000 meters to 6,000 meters above sea level, equivalent to 4,000 meters to 5,000 meters above sea level, equivalent to 5,000 meters to 8,000 meters above sea level, equivalent to 5,000 meters to 7,000 meters above sea level, equivalent to 5,000 meters to 6,000 meters above sea level, equivalent to 6,000 meters to 8,000 meters above sea level, equivalent to 6,000 meters to 7,000 meters above sea level, or equivalent to 7,000 meters to 8,000 meters above sea level).

[0176] In some subjects, such as some children or newborns, hypoxic breathing according to a method described herein may have adverse effects, such as causing pulmonary vasoconstriction, pulmonary hypertension, or shunting via the ductus arteriosus or a patent foramen ovale. To assist in dilating the pulmonary circulation and avoiding, or reducing the occurrence of, or alleviating potential adverse effects of hypoxic breathing, gaseous nitric oxide can optionally be administered to a subject in combination with breathing a therapeutic gas or breathing air in a hypobaric chamber to produce systemic hypoxia according to a method of treatment or prevention described herein. See Frostell et al. (1993) Anesthesiology 78:427-35. In certain embodiments where a subject is administered a therapeutic gas with reduced oxygen content, nitric oxide can optionally be administered within the therapeutic gas having reduced oxygencontent. In certain embodiments, lower body SpO2(e.g., toe) is measured in a subject (e.g., a child or infant) and nitric oxide is added to the inhaled therapeutic gas in an amount that causes the lower body SpO2to increase so as to decrease or eliminate the difference between lower body SpO2and upper body SpO2(e.g., as measured in a finger or ear).

[0177] The methods of the present disclosure may optionally include measuring copper levels in the body of a subject. Various means of measuring copper levels in the body are known in the art. In certain embodiments, free copper concentration in serum or plasma ultrafiltrate is measured by inductively coupled mass spectrometry to assist with diagnosis and monitoring of Wilson disease or Menkes disease. In certain embodiments, the free concentration in urine is determined. In certain embodiments, biliary secretion of copper is determined by measurement of copper concentration in feces. In certain embodiments, the copper content of hair is determined. In certain embodiments, the amount of free serum copper is determined as the amount of unbound copper circulating in the blood, which is copper unbound by Ceruloplasmin. It will be understood that this is the copper that is free to accumulate in the liver and other organs. In certain embodiments, Non-Ceruloplasmin-Bound Copper or NCC is determined by inductively coupled mass spectrometry or other methods known in the art.Embodiments

[0178] The following numbered embodiments also form part of the present disclosure:

[0179] 1. A pharmaceutical composition comprising a prolyl hydroxylase inhibitor and a copper salt.

[0180] 2. The pharmaceutical composition of embodiment 1, wherein the prolyl hydroxylase inhibitor comprises roxadustat, daprodustat, vadadustat, enarodustat, molidustat, desidustat, or a pharmaceutically acceptable salt thereof.

[0181] 3 The pharmaceutical composition of embodiment 1 or embodiment 2, wherein the copper salt comprises copper acetate, copper carbonate, copper chloride, copper iodide, copper nitrate, copper oxide, copper selenite, copper sulfate, copper pyrophosphate, copper aspartate, copper glutamate, copper glycinate, copper histidinate, copper lysinate, copper methionate, copper tyrosinate, copper ascorbate, copper citrate, copper formate, copper fumarate, copper gluconate, copper glutarate, copper malate, copper orotate, copper picolinate, copper sebacate, copper succinate, copper tartrate, or a combination thereof.

[0182] 4. The pharmaceutical composition of any one of embodiments 1-3, wherein the copper salt comprises copper histidinate.

[0183] 5. The pharmaceutical composition of any one of embodiments 1-4, further comprising an excipient.

[0184] 6. The pharmaceutical composition of embodiment 5, wherein the excipient comprises a glycol, oil, starch, sugar, alcohol, diluent, granulating agent, disintegrating agent, flavoring agent, lubricant, binder, preservative, coloring agent, or a combination thereof.

[0185] 7. The pharmaceutical composition of any one of embodiments 1-6, wherein the pharmaceutical composition is a powder, tablet, capsule, caplet, or liquid.

[0186] 8. A method of treating Wilson disease in a subject in need thereof, the method comprising: administering to the subject a pharmaceutical composition comprising a prolyl hydroxylase inhibitor.

[0187] 9. The method of embodiment 8, wherein the prolyl hydroxylase inhibitor comprises roxadustat, daprodustat, vadadustat, enarodustat, molidustat, desidustat, or a pharmaceutically acceptable salt thereof.

[0188] 10. The method of embodiment 8 or embodiment 9, further comprising an excipient.

[0189] 11. The method of embodiment 10, wherein the excipient comprises a glycol, oil, starch, sugar, alcohol, diluent, granulating agent, disintegrating agent, flavoring agent, lubricant, binder, preservative, coloring agent, or a combination thereof.

[0190] 12. The method of any one of embodiments 8-11, wherein the pharmaceutical composition is a powder, tablet, capsule, caplet, or liquid.

[0191] 13. The method of any one of embodiments 8-12, wherein the pharmaceutical composition is administered orally or parenterally.

[0192] 14. The method of any one of embodiments 8-13, wherein the pharmaceutical composition is administered daily.

[0193] 15. A method of treating Menkes disease in a subject in need thereof, the method comprising: administering to the subject a pharmaceutical composition comprising a prolyl hydroxylase inhibitor.

[0194] 16. The method of embodiment 15, wherein the prolyl hydroxylase inhibitor comprises roxadustat, daprodustat, vadadustat, enarodustat, molidustat, desidustat, or a pharmaceutically acceptable salt thereof.

[0195] 17. The method of embodiment 15 or embodiment 16, wherein the pharmaceutical composition further comprises a copper salt.

[0196] 18. The method of embodiment 17, wherein the copper salt comprises copper acetate, copper carbonate, copper chloride, copper iodide, copper nitrate, copper oxide, copper selenite, copper sulfate, copper pyrophosphate, copper aspartate, copper glutamate, copper glycinate, copper histidinate, copper lysinate, copper methionate, copper tyrosinate, copper ascorbate, copper citrate, copper formate, copper fumarate, copper gluconate, copper glutarate, coppermalate, copper orotate, copper picolinate, copper sebacate, copper succinate, copper tartrate, or a combination thereof.

[0197] 19. The method of embodiment 17 or embodiment 18, wherein the copper salt comprises copper histidinate.

[0198] 20. The method of any one of embodiments 15-19, further comprising an excipient.

[0199] 21. The method of embodiment 20, wherein the excipient comprises a glycol, oil, starch, sugar, alcohol, diluent, granulating agent, disintegrating agent, flavoring agent, lubricant, binder, preservative, coloring agent, or a combination thereof.

[0200] 22. The method of any one of embodiments 15-21, wherein the pharmaceutical composition is a powder, tablet, capsule, caplet, or liquid.

[0201] 23. The method of any one of embodiments 15-22, wherein the pharmaceutical composition is administered orally or parenterally.

[0202] 24. The method of any one of embodiments 15-23, wherein the pharmaceutical composition is administered daily.

[0203] All publications and patent applications mentioned in the specification are indicative of the level of skill of those skilled in the art to which this disclosure pertains. All publications and patent applications are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

[0204] Although the foregoing disclosure has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the appended claims.

[0205] The following examples are offered by way of illustration and not by way of limitation.EXAMPLESExample 1: Generation of an innovative Cu-sensitive cell line for cloning novel regulators of Cu homeostasis

[0206] The initial goal was to employ a genome-wide CRISPR / Cas9 knockout screen to identify gene disruptions that confer tolerance to Cu. To maximize the robustness of the screen, a Cu- sensitive cell line (hereafter Cuscells) was created by eliminating three proteins known to contribute to Cu tolerance - namely, the Cu exporter ATP7A, and the Cu-binding proteins, metallothionein I and II (MTI and MTII) Endogenous Cu levels in the basal medium (10% FBS in DMEM; ~1.7 pM Cu) were sufficient to kill Cuscells within -4 days; however, the cells could be propagated in the presence of a cell-impermeable Cu chelator, bathocuproinedisulfonate (BCS). Having successfully developed a novel Cu sensitive cell line, the CRISPR- based deletion screen to identify novel suppressors of Cu sensitivity was ready to proceed.Example 2: Identification of the von Hippel Lindau (VHL) gene as a novel regulator of Cu tolerance

[0207] Cuscells were transduced with a commercial lentiviral CRISPR library (GeCKOv2; Addgene) which contains a total of 130,209 gRNA constructs targeting 20,611 genes (6 gRNAs per gene). The transduced Cuscells were cultured for 10 days in basal media containing puromycin plus IpM BCS (control pool), or puromycin plus IpM CuCh (Cu-tolerant pool) (FIG. 1A). The gRNAs from both pools were then amplified by PCR and analyzed using next generation sequencing. Three gRNAs against the CTR1 gene, were most enriched by Cu selection. Since CTR1 encodes a high-affinity Cu importer, this result served as a positive control for the screen. CTR1 was validated as a hit in a separate experiment by showing that two independent gRNA constructs against the CTR1 gene conferred Cu tolerance when stably transfected into Cuscells. Colonies were picked to generate clonally pure Cus / C77?7- cell lines and sequence analysis of one cell line confirmed the insertion of a single nucleotide in the open reading frame of the Ctrl gene (FIG. IB). The Cu resistance profile of this Cus / C77?7- cell line was tested using a Crytal Violet assay and is shown (FIG. 1C). The second most enriched group of gRNAs were against the von Hippel Lindau gene (VHL which has no previously defined role in Cu tolerance. As with the Ctrl gene, two independent gRNAs targeting the VHL gene conferred Cu tolerance in Cuscells and sequence analysis of one clonally derived C C / VHL- cell line confirmed a 2-bp deletion in the VHL gene (FIG. IB). The Cu resistance profile of this Cus / 17 / / .- cell line is shown (FIG. 1C).Example 3: Disruption of the VHL gene stimulates ATP7B expression

[0208] The next objective was to identify the underlying mechanisms of Cu tolerance in Cus / VHL- cells. Since activation of ATP7B expression could potentially rescue the Cu-sensitive growth of Cuscells, it was hypothesized that ATP7B expression might be elevated in Cus / VHL- cells. Consistent with this hypothesis, ATP7B mRNA and protein were dramatically elevated in Cus / VHL- cells compared to parental Cuscells (FIG. 2A and FIG. 2B). Consistent with the loss of VHL function, the HIFla protein was stabilized in Cus / VHL- cells (FIG. 2B). Using immunofluorescence microscopy, ATP7B was barely detectable in Cuscells, whereas in Cus / VHL- cells ATP7B was detected strongly detected in the perinuclear region which is consistent with its reported location in the Golgi region of other cells (FIG. 2C). The elevated expression of ATP7B in Cus / VHL- cells was due to the loss of VHL because transient transfection of Cus / VHL- cells with an HA-tagged VHL plasmid resulted in the loss of ATP7Bstaining in VHL-HA positive cells (FIG. 2D), but not in cells that failed to express the plasmid (FIG. 2D; arrowheads). Stable transfection of Cuscells with an ATP7B expression plasmid conferred viability in basal medium (FIG. 2E), suggesting that the activation of ATP7B expression is responsible for the acquisition of Cu tolerance in Cus / VHL- cells. To verify whether the loss of VHL stimulates ATP7B expression other cell types, a wild type renal cancer cell line (RENCA / WT) as well as a derivative clone (RENCA / VHL-) in which the VHL gene was disrupted using CRISPR were obtained. Consistent with the findings in Cuscells, ATP7B mRNA was significantly elevated in RENCA / VHL- cells compared to RENCA / WT cells (FIG. 2F). Interestingly, ATP7A mRNA was also significantly upregulated in the RENCA / VHL- cells, although not to the same extent as ATP7B (FIG. 2G). Taken together, these results demonstrate that disruption of the VHL gene confers Cu tolerance due to upregulation of ATP7B expression.Example 4: Hypoxia stimulates hepatic expression of ATP7A and ATP7B in vivo

[0209] The VHL gene encodes a master regulator of oxygen metabolism and is part of the machinery that senses and responds to low oxygen conditions (hypoxia). The VHL protein works together with prolyl hydroxylases (PHDs) to constitutively degrade the transcription factor HIF1 / 2 (Hypoxia Inducible Factor) under normoxic conditions. However, the VHL-PHD machinery requires oxygen to function, and thus under hypoxic conditions HIF1 / 2 is not degraded and enters the nucleus to regulate HIF -responsive genes. Because VHL deletion mimics the effects of hypoxia, whether hypoxia confers viability to Cuscells in basal medium was tested. During normal physiology, most tissues are exposed to approximately 3% to 7% O2. Thus, whether exposure of Cuscells an atmosphere of 1% O2 could suppress the loss of viability in basal medium was investigated. Equal numbers of Cuscells were seeded in basal medium and grown for 4 days in an atmosphere of either 21%O2 (normoxia) or 1% O2 in the presence of 5% CO2. As expected, the Cuscells failed to propagate in normoxia, however, there was robust growth in hypoxic conditions (FIG. 3A). Exposure of Cuscells to 1% O2 for 24 h resulted in a significant increase in ATP7B mRNA levels compared to normoxia (FIG. 3B). To investigate whether ATP7B expression is responsive to hypoxia in vivo, immunoblot analysis of ATP7B in livers isolated from 8-week-old C57BL / 6 mice that were exposed to normoxia or hypoxia (10% O2) for four days was performed. Compared to normoxic mice, levels of ATP7B protein were markedly elevated in livers of hypoxic mice (FIG. 3C). Notably, hypoxia also increased the hepatic expression of ATP7A (FIG. 3C). Consistent with previous studies, hypoxia significantly increased ceruloplasmin activity in the serum (FIG. 3D). Furthermore, hypoxia elicited a small but significant reduction in hepatic Cu concentrations relative to normoxic levels (FIG. 3E). Taken together, these results demonstrate that hepatic expression of both ATP7A and ATP7Bare induced by hypoxia.Example 5: The prolyl hydroxylase inhibitor, Roxadustat, rescues Cuscells and stimulates expression of the ATP7B protein in vitro and in vivo

[0210] The results support the concept that loss of VHL or hypoxia increases the expression of ATP7A and ATP7B due to the stabilization of HIF1 / 2. HIF1 / 2 undergoes prolyl hydroxylation to mediate VHL-dependent degradation, which requires the activity of prolyl hydroxylases (PHDs). Thus, inhibition of PHD activity also results in the stabilization of HIF1 / 2. It was reasoned that PHD inhibitors might recapitulate the effect of VHL loss or hypoxia on ATP7A and ATP7B expression and confer Cu tolerance to Cuscells. The clinical-stage drug Roxadustat is an orally active PHD inhibitor that has been approved in multiple countries for the treatment of anemia. The treatment of Cuscells with Roxadustat conferred survival in basal medium (FIG. 4A) and triggered a strong induction of ATP7B expression at the level of mRNA and protein (FIG. 4B and FIG. 4C). These results indicate that Roxadustat-induced expression of ATP7B can rescue the Cu-sensitive phenotype of Cuscells by functionally substituting for the loss of ATP7A-mediated Cu export.

[0211] To test whether Roxadustat can induce ATP7B expression in vivo, Roxadustat was administered to 8-week-old C57BL / 6 mice every 24 h over 5 days via intraperitoneal injection (10 mg / kg). Immunoblot analysis revealed that Roxadustat induced marked increases in ATP7B protein in the liver compared to mice treated with DMSO vehicle control (FIG. 4D). Additionally, Roxadustat also induced ATP7A protein levels in the liver (FIG. 4E). Together with the earlier results, these findings support the working model that ATP7A and ATP7B proteins are regulated via the VHL / PHD / HIF pathway and suggest that hypoxia itself or hypoxia mimetics such as Roxadustat may provide novel therapeutic approaches to treat disorders of Cu homeostasis such as Menkes or Wilson disease.Example 6: The Cu-sensitive phenotype of Cuscells is rescued by other prolyl hydroxylase inhibitors

[0212] Next, whether the Cu-sensitive phenotype of Cuscells can be rescued by other clinically used prolyl-hydroxylases including Daprodustat, Vadadustat and Enarodustat was investigated. Each of these prolyl-hydroxylase inhibitors was able to rescue the growth of Cuscells when tested at a range of concentrations (FIG. 5).Example 7: Prolyl hydroxylase inhibitors restore Cu-delivery to the secretory pathway in cells lacking ATP7A

[0213] In addition to its role in Cu export, the ATP7A protein is also required for Cu transport into the secretory pathway to supply Cu to metalloenzymes that are synthesized within secretorycompartments such as tyrosinase, dopamine 0 hydroxylase and peptidylglycine a amidating monooxygenase. Hypoactivity of these metalloenzymes is a major contributor to the pathology of Menkes disease. To address if Roxadustat-induced expression of ATP7B might rescue cuproenzymes deficient in Menkes disease, a B16 melanoma model in which tyrosinase activity was abolished by CRISPR-mediated deletion of the ATP7A gene (B16 / ATP7A-) was used and whether Roxadustat can restore the activity of tyrosinase in these cells was tested. Roxadustat activated tyrosinase activity in B16 / ATP7A- cells (FIG. 6A and FIG. 6B) and increased expression of the ATP7B protein (FIG. 6C). These findings demonstrate that Roxadustat can induce ATP7B expression in cells lacking ATP7A to increase Cu bioavailability to Cu- dependent enzymes of the secretory pathway.Example 8: Roxadustat activates Cu-dependent melanogenesis and enhances survival of a mouse model of Menkes disease

[0214] The mottled Brindled (mobr) mouse harbors an in-frame deletion within the ATP7A gene and is a well-characterized model of Menkes disease. The mobr mice die at approximately postnatal day (PND)14 from defects attributable to hypoactivity of Cu-dependent enzymes. The widely used standard treatment for Menkes disease involves subcutaneous injections of copper histidinate (Cu-HIS), a natural complex of Cu found in blood plasma. Nonetheless, most Menkes patients treated with Cu-HIS die during infancy or early childhood, underscoring the urgent need for more effective therapies. As with Menkes patients, Cu-HIS does not prevent the postnatal lethality of mobr mice.

[0215] Based on the in vitro results, whether Cu-HIS might increase the survival of mobr mice if it was co-administered Roxadustat was tested. An experiment regimen was devised involving five different treatments administered to mobr mice via subcutaneous injections on PND7 and PND10 as follows:Group 1 : DMSO alone (vehicle control)Group 2: Cu-HIS (25 pg) + DMSOGroup 3: Cu-HIS (25 pg) + Roxadustat (10 mg / kg) Group 4: Cu-HIS (25 pg) + Roxadustat (15 mg / kg) Group 5: Roxadustat (15 mg / kg) + DMSO.

[0216] None of the mobr mice in Group 1 or Group 5 survived until weaning at PND21 (FIG. 7A). Of the Group 2 mice, only 28% survived to PND21 and none survived to PND90. Of the Group 3 mice, 50% survived to PND21 and 23% survived to PND90. Of the Group 4 cohort, 54% survived to PND21 and remained viable at PND90. Images of mice from each treatment group are shown at PND14 along with a wild type littermate C57BL / 6 control treated withDMSO to illustrate the black coat color (FIG. 7B). The mobr mice on the C57BL / 6 background are hypopigmented due to a lack of ATP7A-dependent metallation of tyrosinase, which is the rate limiting enzyme in the melanogenesis pathway. Note that DMSO alone (Group 1), or in combination with Roxadustat (Group 5), had little darkening effect on coat color (FIG. 7B). The Cu-HIS + DMSO (Group 2) treatment increased pigmentation compared to DMSO alone, however, the combination of Cu-HIS + Roxadustat (Group 4) generated the greatest darkening of coat color of the treatment groups (FIG. 7B). In summary, the in vitro and in vivo data suggest that in the absence of a functional ATP7A protein, Roxadustat has the potential to provide a therapeutic benefit in Menkes disease by 1) enhancing the expression of an alternative Cu transporter, ATP7B; 2) increasing Cu transport to enzymes such as tyrosinase within the secretory pathway; and 3) increasing the therapeutic efficacy of Cu-HIS in a mouse model of Menkes disease.Example 9: Roxadustat ameliorates liver pathology in a mouse model of Wilson disease

[0217] In addition to activating the expression of ATP7B, the results indicate that Roxadustat is also able to activate the expression of ATP7A in the liver of mice. Given the functional overlap of these Cu transporters, whether Roxadustat-induced may ameliorate the hepatic pathology caused by loss of function mutations in ATP7B in a mouse model of Wilson disease was tested. The ATP7B protein is primarily expressed in hepatocytes where it functions in Cu excretion from the body into the bile. Consequently, ATP7B-KO mice and patients with Wilson disease exhibit liver disease stemming from hepatic Cu overload. To test whether Roxadustat might provide a therapeutic benefit in Wilson disease, the therapeutic potential of Roxadustat (or DMSO vehicle) was evaluated in ATP7B-KO mice at 3 months old, an age at which there is significant Cu buildup and liver pathology. Roxadustat or DMSO control was administered daily for 10 days (10 mg / kg via IP injection) at which time mice were euthanized to evaluate biomarkers of liver pathology. Roxadustat significantly decreased elevated markers of liver pathology in ATP7B-KO mice including the size and number of hepatic nuclei (FIG. 8A-C), as well as plasma levels of AST, ALP and ALT (FIG. 8D-F). Notably, Roxadustat increased hepatic expression of ATP7A protein (FIG. 8G) as well as the mRNA levels of metallothioneins MT-I and MT-II (FIG. 8H and FIG. 81). Taken together, these data suggest that prolyl hydroxylase inhibitors such as Roxadustat have therapeutic potential as a therapy in Wilson disease by activating the expression of ATP7A and / or metallothioneins.

Claims

What is claimed is:

1. A pharmaceutical composition for the treatment of Menkes disease comprising: a prolyl hydroxylase inhibitor, and a copper salt.

2. The pharmaceutical composition of claim 1, wherein the prolyl hydroxylase inhibitor comprises roxadustat, daprodustat, vadadustat, enarodustat, molidustat, desidustat, or a pharmaceutically acceptable salt thereof.

3. The pharmaceutical composition of claim 1, wherein the copper salt comprises copper acetate, copper carbonate, copper chloride, copper iodide, copper nitrate, copper oxide, copper selenite, copper sulfate, copper pyrophosphate, copper aspartate, copper glutamate, copper glycinate, copper histidinate, copper lysinate, copper methionate, copper tyrosinate, copper ascorbate, copper citrate, copper formate, copper fumarate, copper gluconate, copper glutarate, copper malate, copper orotate, copper picolinate, copper sebacate, copper succinate, copper tartrate, or a combination thereof.

4. The pharmaceutical composition of claim 1, wherein the copper salt comprises copper histidinate.

5. The pharmaceutical composition of claim 1, further comprising an excipient.

6. The pharmaceutical composition of claim 5, wherein the excipient comprises a glycol, an oil, a starch, a sugar, an alcohol, a diluent, a granulating agent, a disintegrating agent, a flavoring agent, a lubricant, a binder, a preservative, a coloring agent, or a combination thereof.

7. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is a powder, tablet, capsule, caplet, or liquid.

8. A method of treating Wilson disease in a subject in need thereof, the method comprising: administering to the subject a pharmaceutical composition comprising a prolyl hydroxylase inhibitor.

9. The method of claim 8, wherein the prolyl hydroxylase inhibitor comprises roxadustat, daprodustat, vadadustat, enarodustat, molidustat, desidustat, or a pharmaceutically acceptable salt thereof.

10. The method of claim 8, further comprising an excipient.

11. The method of claim 10, wherein the excipient comprises a glycol, an oil, a starch, a sugar, an alcohol, a diluent, a granulating agent, a disintegrating agent, a flavoring agent, a lubricant, a binder, a preservative, a coloring agent, or a combination thereof.

12. The method of claim 8, wherein the pharmaceutical composition is a powder, tablet, capsule, caplet, or liquid.

13. The method of claim 8, wherein the pharmaceutical composition is administered orally or parenterally.

14. The method of claim 8, wherein the pharmaceutical composition is administered daily.

15. A method of treating Menkes disease in a subject in need thereof, the method comprising: administering to the subject a pharmaceutical composition comprising a prolyl hydroxylase inhibitor.

16. The method of claim 15, wherein the prolyl hydroxylase inhibitor comprises roxadustat, daprodustat, vadadustat, enarodustat, molidustat, desidustat, or a pharmaceutically acceptable salt thereof.

17. The method of claim 15, wherein the pharmaceutical composition further comprises a copper salt.

18. The method of claim 17, wherein the copper salt comprises copper acetate, copper carbonate, copper chloride, copper iodide, copper nitrate, copper oxide, copper selenite, copper sulfate, copper pyrophosphate, copper aspartate, copper glutamate, copper glycinate, copper histidinate, copper lysinate, copper methionate, copper tyrosinate, copper ascorbate, copper citrate, copper formate, copper fumarate, copper gluconate, copper glutarate, copper malate, copper orotate, copper picolinate, copper sebacate, copper succinate, copper tartrate, or a combination thereof.

19. The method of claim 17, wherein the copper salt comprises copper histidinate.

20. The method of claim 15, further comprising an excipient.

21. The method of claim 21, wherein the excipient comprises a glycol, an oil, a starch, a sugar, an alcohol, a diluent, a granulating agent, a disintegrating agent, a flavoring agent, a lubricant, a binder, a preservative, a coloring agent, or a combination thereof.

22. The method of claim 15, wherein the pharmaceutical composition is a powder, tablet, capsule, caplet, or liquid.

23. The method of claim 15, wherein the pharmaceutical composition is administered orally or parenterally.

24. The method of claim 15, wherein the pharmaceutical composition is administered daily.

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