WO2021127273A1

WO2021127273A1 - 1 -((6-oxo-1,6-dihydropyridazin-4-yl)methyl)piperazine and 1 -((6-oxo-1,6-dihydropyrimidin-4-yl)methyl)piperazine derivatives as prolyl hydroxylase, hif-1 alpha and pgk modulators for use in treating inflammatory diseases, cancer or infections

Info

Publication number: WO2021127273A1
Application number: PCT/US2020/065747
Authority: WO
Inventors: Craig Coburn; Martin W. Rowbottom
Original assignee: Gb004, Inc.
Priority date: 2019-12-20
Filing date: 2020-12-17
Publication date: 2021-06-24

Abstract

Described herein are compounds of formulae (I), (lla) and (Mb) as prolyl hydroxylase inhibitors, HIF-lalpha stabilizers and PGK up regulators for use in treating inflammatory diseases, cancer or infections: Preferred compounds are e.g. tert-butyl-4-((l-benzyl-6-oxo-l,6- dihydropyridazin-4-yl)methyl)piperazine-l-carboxylate and tert-butyl-4-((l-benzyl-6-oxo-l,6-dihydropyrimidin-4-y I) methyl) piperazine-l-carboxylate derivatives and related compounds. Exemplary compounds are e.g. compound 1A: and compound 2A: The present description discloses the synthesis and characterisation of exemplary compounds as well as pharmacological data thereof (e.g. pages 22 to 25 and 55 to 70; examples 1 to 5; compounds 1A to 2V; tables 1 and 2).

Description

1 -((6-OXO-1 ,6-DIHYDROPYRIDAZIN-4-YL)METHYL)PIPERAZINE AND -((6-OXO-1 ,6-DIHYDROPYRIMIDIN-4-YL)METHYL)PIPERAZINE DERIVATIVES AS PROLYL HYDROXYLASE, HIF-1 ALPHA

AND PGK MODULATORS FOR USE IN TREATING INFLAMMATORY DISEASES,

CANCER OR INFECTIONS

BACKGROUND OF THE DISCLOSURE

[0001] The present invention generally relates to prolyl hydroxylase inhibitors that can stabilize hypoxia inducible factor-1 alpha (HIF-la), as well as to pharmaceutical compositions containing the same and to methods of their use and preparation.

BACKGROUND

[0002] Transcription factor Hypoxia-Inducible Factor 1 (HIF-1) is one of the key regulators of oxygen homeostasis and the physiological responses to low oxygen levels (hypoxia). HIF-1 is a heterodimeric protein comprising two subunits, HIF-la and HIF- 1b, that bind to the hypoxia response element (HRE).

[0003] Under healthy conditions, HIF-la is degraded by one of several 4-proiyl hydroxylase enzymes, inter alia, HIFPH2 (or EGLN1 or PHD2) and HIFPH3 (or F.GLN3 of PHDS). When cells undergo hypoxia (oxygen concentrations below 6%), this enzymatic transformation slows or stops and HIF-la levels increase. At elevated levels, HIF-la combines with HIF-Ib to form the active transcription factor complex HIF-1, which can then activate any of several biological pathways to alleviate the hypoxia, including, inter alia, angiogenic, erythropoietic (EPO), glucose metabolism (PGK), matrix alteration, and enhanced capacity of phagocytes to respond to pathogens. Many HiF- 1-regulated genes have been identified, including genes coding for proteins involved in angiogenesis, energy metabolism, erythropoiesis, cell proliferation and viability, vascular remodeling and vasomotor responses. Therefore, modulation of HIF activation in ceils is critical to preventing, controlling, curing, or otherwise affecting a wide array of diseases, disease states and conditions.

[0004] in patients requiring increased tissue oxygen, e.g. due to peripheral vascular disease (PVD), inhibiting the H i FI enzymes should stimulate the body's own angiogenic response without the consequences of oxygen deficiency. In diseases of Ischemia, inter alia, CAD and anemia, stimulation of angiogenic, erythropoietic, and metabolism adaption can provide therapeutic benefits. Up regulation of HIF-la also provides a method for enhancing immunity, for example, by increasing the capacity of phagocytes.

[000S] There is a need for methods for controlling HiF-la activity and the 4-prolyl hydroxylase enzymes that degrade HIF-Ia. Inhibitors of 4-proiyl hydroxylase enzymes provide methods for altering HIF-io. levels and thus provide methods for treating a variety of diseases or disease states.

[0006] Described herein are compounds that stabilize HIF-la and methods for using them to treat diseases, conditions, syndromes, and the like, that are affected by the level of hypoxia- inducible transcription factors. Regulation of these factors both during normoxia and hypoxia may provide methods for re-balancing or regulating one or more biological pathways associated with abnormal conditions, such as but not limited to invasion of the body by pathogens, (e.g. bacteria, fungi, viruses, parasites), abnormal cellular regulation, he., cancer, ischemia, heart attack, stroke and chronic lung disease and the side effects of vaccination.

BRIEF SUMMARY

[0007] In one embodiment, compounds are provided having the structure of Formula

(I):

or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof, wherein:

X is -N=CR^X- or -CR^X=N-;

R^x is H or halogen;

W is -CH2-, -CH2-CH2-, -CH2-CH2-CH2- or -CH2-CH2-CH2-CH2-;

Z is -COOH, -COOMe, -COOEt or

R^z is halogen, CH₃, CF₃, COOH, COOMe, OMe, CN or two adjacent R^z groups with the C atoms to which they are attached form a 5 or 6-membered ring; n is 0, 1 or 2;

T is a bond, -CH2- or -CH2-CH2-;

Y is -COOH, -COOMe, -COOEt or

R^Y is halogen, CH₃, CF₃, COOH, COOMe, OMe, CN or two adjacent R^Y groups with the C atoms to which they are attached form a 5 or 6-membered ring; and n' is 0, 1 or 2.

[000S] In another embodiment, compounds are provided having the structure listed in Table 1, or a pharmaceutically acceptable isomer, racemate, hydrate, solvate, isotope, or salt thereof.

[0009] In another embodiment, a composition is provided comprising a compound having the structure of Formula (I), Formula (lla) or Formula (lib), or a pharmaceutically acceptable isomer, racemate, hydrate, solvate, isotope, or salt thereof, and a pharmaceutically acceptable carrier, diluent, or excipient.

[0010] In another embodiment, a use is provided, of a compound of Formula (I), Formula (lla) or Formula (Mb) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof in the manufacture of a medicament. [0011] In another embodiment, a method for inhibiting hypoxia-inducible factor 1-a (HIF-la) activity is provided, comprising contacting the HIF-la with an effective amount of a compound having the structure of Formula (I), Formula (lla) or Formula (Mb) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof

[0012] In another embodiment, a method for inhibiting hypoxia-inducible factor 1-a (HIF-la) activity in a subject is provided, comprising administering to the subject an effective amount of a compound having the structure of Formula (I), Formula (I la) or Formula (Mb) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof

[0013] In another embodiment, a method for up regulating phosphoglycerate kinase (PGK) activity is provided, comprising contacting the PGK with an effective amount of a compound of Formula (I), Formula (I la) or Formula (Mb) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof. [0014] In another embodiment, a method for increasing the immune response in a subject is provided, comprising administering to the subject an effective amount of a compound of Formula (I), Formula (lla) or Formula (Mb) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof. [001S] In another embodiment, a method for treating a subject with an inflammatory epithelial disease is provided, comprising administering to the subject an effective amount of a compound of Formula (I), Formula (lla) or Formula (Mb) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof.

[0016] In another embodiment, a method for treating an infection in a subject, comprising administering to the subject an effective amount of a compound of Formula (I), Formula (lla) or Formula (Mb) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof.

DETAILED DESCRIPTION

[0017] The present invention relates to pyridazinone and pyrimidinone compounds, pharmaceutical compositions containing them, and methods of using them for the treatment of disease states, disorders, and conditions mediated by prolyl hydroxylase activity.

[0018] As used herein, "alkyl" means a straight chain or branched saturated hydrocarbon group. "Lower alkyl" means a straight chain or branched alkyl group having from 1 to 8 carbon atoms, in some embodiments from 1 to 6 carbon atoms, in some embodiments from 1 to 4 carbon atoms, and in some embodiments from 1 to 2 carbon atoms. Examples of straight chain lower alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched lower alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and 2,2-dimethylpropyl groups.

[0019] "Alkenyl" groups include straight and branched chain and cyclic alkyl groups as defined above, except that at least one double bond exists between two carbon atoms. Thus, alkenyl groups have from 2 to about 20 carbon atoms, and typically from 2 to 12 carbons or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to -CH=CH2, -CH=CH(CH₃), -CH=C(CH₃)₂, -C(CH₃)=CH₂, -C(CH₃)=CH(CH₃), -C(CH₂CH₃)=CH₂, -CH=CHCH₂CH₃, -CH=CH(CH₂)₂CH₃, -CH=CH(CH₂)₃CH₃, -CH=CH(CH₂)4CH₃, vinyl, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among others.

[0020] "Alkynyl" groups include straight and branched chain alkyl groups, except that at least one triple bond exists between two carbon atoms. Thus, alkynyl groups have from 2 to about 20 carbon atoms, and typically from 2 to 12 carbons or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to -CºCH, -CºC(CH₃), -CºC(CH₂CH₃), -CH₂CºCH, -CH₂CºC(CH₃), and -CH₂CºC(CH₂CH₃), among others.

[0021] As used herein, "alkylene" means a divalent alkyl group. Examples of straight chain lower alkylene groups include, but are not limited to, methylene (i.e., -CH₂-), ethylene (i.e., -CH₂CH₂-), propylene (i.e., -CH₂CH₂CH₂-), and butylene (i.e., -CH₂CH₂CH₂CH₂-). AS used herein, "heteroalkylene" is an alkylene group of which one or more carbon atoms is replaced with a heteroatom such as, but not limited to, N, O, S, or P.

[0022] "Alkoxy" refers to an alkyl as defined above joined by way of an oxygen atom (i.e., -O-alkyl). Examples of lower alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, n-butoxy, isopropoxy, sec-butoxy, tert-butoxy, and the like. [0023] The terms "carbocyclic" and "carbocycle" denote a ring structure wherein the atoms of the ring are carbon. Carbocycles may be monocyclic or polycyclic. Carbocycle encompasses both saturated and unsaturated rings. Carbocycle encompasses both cycloalkyl and aryl groups. In some embodiments, the carbocycle has 3 to 8 ring members, whereas in other embodiments the number of ring carbon atoms is 4, 5, 6, or 7. Unless specifically indicated to the contrary, the carbocyclic ring can be substituted with as many as N substituents wherein N is the size of the carbocyclic ring with for example, alkyl, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0024] "Cycloalkyl" groups are alkyl groups forming a ring structure, which can be substituted or unsubstituted. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, the cycloalkyl group has 3 to 8 ring members, whereas in other embodiments the number of ring carbon atoms range from 3 to 5, 3 to 6, or 3 to 7. Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like. Cycloalkyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined above. Representative substituted cycloalkyl groups can be mono-substituted or substituted more than once, such as, but not limited to, 2,2-, 2,3-, 2,4- 2,5- or 2,6- disubstituted cyclohexyl groups or mono-, di- or tri-substituted norbornyl or cycloheptyl groups, which can be substituted with, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0025] "Aryl" groups are cyclic aromatic hydrocarbons that do not contain heteroatoms. Thus, aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments, aryl groups contain 6-14 carbons in the ring portions of the groups. The terms "aryl" and "aryl groups" include include fused rings wherein at least one ring, but not necessarily all rings, are aromatic, such as fused aromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, and the like).

[0026] "Carbocyclealkyl" refers to an alkyl as defined above with one or more hydrogen atoms replaced with carbocycle. Examples of carbocyclealkyl groups include but are not limited to benzyl and the like.

[0027] As used herein, "heterocycle" or "heterocyclyl" groups include aromatic and non-aromatic ring compounds (heterocyclic rings) containing 3 or more ring members, of which one or more is a heteroatom such as, but not limited to, N, O, S, or P. A heterocycle group as defined herein can be a heteroaryl group or a partially or completely saturated cyclic group including at least one ring heteroatom. In some embodiments, heterocycle groups include 3 to 20 ring members, whereas other such groups have 3 to 15 ring members. At least one ring contains a heteroatom, but every ring in a polycyclic system need not contain a heteroatom. For example, a dioxolanyl ring and a benzdioxolanyl ring system (methylenedioxyphenyl ring system) are both heterocycle groups within the meaning herein. A heterocycle group designated as a C2-heterocycle can be a 5-membered ring with two carbon atoms and three heteroatoms, a 6-membered ring with two carbon atoms and four heteroatoms and so forth. Likewise, a C4-heterocycle can be a 5-membered ring with one heteroatom, a 6- membered ring with two heteroatoms, and so forth. The number of carbon atoms plus the number of heteroatoms sums up to equal the total number of ring atoms. A saturated heterocyclic ring refers to a heterocyclic ring containing no unsaturated carbon atoms.

[0028] "Heteroaryl" groups are aromatic ring compounds containing 5 or more ring members, of which, one or more is a heteroatom such as, but not limited to, N, O, and S. A heteroaryl group designated as a C2-heteroaryl can be a 5-membered ring with two carbon atoms and three heteroatoms, a 6-membered ring with two carbon atoms and four heteroatoms and so forth. Likewise, a C4-heteroaryl can be a 5-membered ring with one heteroatom, a 6-membered ring with two heteroatoms, and so forth.

The number of carbon atoms plus the number of heteroatoms sums up to equal the total number of ring atoms. Heteroaryl groups include, but are not limited to, groups such as pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, quinoxalinyl, and quinazolinyl groups. The terms "heteroaryl" and "heteroaryl groups" include fused ring compounds such as wherein at least one ring, but not necessarily all rings, are aromatic, including tetrahydroquinolinyl, tetrahydroisoquinolinyl, indolyl and 2,3-dihydro indolyl.

[0029] "Heterocyclealkyl" refers to an alkyl as defined above with one or more hydrogen atoms replaced with heterocycle. Examples of heterocyclealkyl groups include, but are not limited to, morpholinoethyl and the like.

[0030] "Halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.

[0031] "Haloalkyl" refers to an alkyl as defined above with one or more hydrogen atoms replaced with halogen. Examples of lower haloalkyl groups include, but are not limited to, -CF3, -CH2CF3, and the like.

[0032] "Haloalkoxy" refers to an alkoxy as defined above with one or more hydrogen atoms replaced with halogen. Examples of lower haloalkoxy groups include, but are not limited to -OCF3, -OCH2CF3, and the like.

[0033] "Hydroxyalkyl" referes to an alkyl as defined above with one or more hydrogen atoms replaced with -OH. Examples of lower hydroxyalkyl groups include, but are not limited to -CH2OH, -CH2CH2OH, and the like.

[0034] As used herein, the term "optionally substituted" refers to a group (e.g., an alkyl, carbocycle, or heterocycle) having 0, 1, or more substituents, such as 0-25, 0-20, 0-10 or 0-5 substituents. Substituents include, but are not limited to -OR^a, -NR^aR^b, -S(0)₂R^a or -S(0)₂OR^a, halogen, cyano, alkyl, haloalkyl, alkoxy, carbocycle, heterocycle, carbocyclalkyl, or heterocyclealkyl, wherein each R^a and R^b is, independently, H, alkyl, haloalkyl, carbocycle, or heterocycle, or R^a and R^b, together with the atom to which they are attached, form a 3-8 membered carbocycle or heterocycle. [0035] Described herein are compounds having the structure of Formula (I):

Formula (I) or pharmaceutically acceptable salts, solvates, hydrates, isomers, tautomers, racemates or isotopes thereof, wherein:

X is -N=CR^X- or -CR^X=N-;

R^x is H or halogen;

W is -CH2-, -CH2-CH2-, -CH2-CH2-CH2- or -CH2-CH2-CH2-CH2-;

Z is -COOH, -COOMe, -COOEt or

R^z is halogen, CH3, CF3, COOH, COOMe, OMe, CN or two adjacent R^z groups with the C atoms to which they are attached form a 5 or 6-membered ring; n is 0, 1 or 2;

T is a bond, -CH2- or -CH2-CH2-;

Y is -COOH, -COOMe, -COOEt or

[0036] In some embodiments, R^x is H. In other embodiments, R^x is halogen. In other embodiments, R^x is fluorine or chlorine.

[0037] In some embodiments, W is -CH₂-. In other embodiments, W is -CH₂-CH₂-. In other embodiments, W is -CH₂-CH₂-CH₂-. In other embodiments, W is -CH₂-CH₂-CH₂- CH2-.

[0038] In some embodiments, Z is -COOH. In other embodiments, Z is -COOMe. In other embodiments, Z is -COOEt. In other embodiments, Z is

In some embodiments, R^z is halogen, CH3, CF3. In other embodiments, R^z is halogen. In other embodiments, R^z is fluorine or chlorine. In other embodiments, R^z is CH₃ or CF₃. In other embodiments, n is 1. In other embodiments, n is 2. In other embodiments, two adjacent R^z groups with the C atoms to which they are attached form a 5 or 6- membered ring. In other embodiments, two adjacent R^z groups with the C atoms to which they are attached form a 5-membered ring. In other embodiments, two adjacent R^z groups with the C atoms to which they are attached form a 6-membered ring.

[0039] In some embodiments, T is a bond. In other embodiments, T is -CH2-. In other embodiments, T is -CH2-CH2-.

[0040] In some embodiments, Y is -COOH, -COOMe or -COOEt. In other embodiments, Y is -COOH. In other embodiments, Y is -COOMe. In other embodiments, Y is -COOEt.

In other embodiments, Y is

Boc

In other embodiments, Y is

In some embodiments, R^Y is halogen, CH3, CF3. In other embodiments, R^Y is halogen. In other embodiments, R^Y is fluorine or chlorine. In other embodiments, R^Y is CH3 or CF3. In other embodiments, n' is 1. In other embodiments, n' is 2. In other embodiments, n' is 2 and the two R^Y groups are the same. In other embodiments, n' is 2 and the two R^Y groups are different. In other embodiments, two adjacent R^Y groups with the C atoms to which they are attached form a 5 or 6-membered ring. In other embodiments, two adjacent R^Y groups with the C atoms to which they are attached form a 5-membered ring. In other embodiments, two adjacent R^Y groups with the C atoms to which they are attached form a 6-membered ring.

[0041] Also described herein are compounds having the structure of Formula (lla):

or pharmaceutically acceptable salts, solvates, hydrates, isomers, tautomers, racemates or isotopes thereof, wherein:

R^x is H or halogen;

W is -CH2-, -CH2-CH2-, -CH2-CH2-CH2- or -CH2-CH2-CH2-CH2-;

Z is -COOH, -COOMe, -COOEt or

T is a bond, -CH₂- or -CH2-CH2-;

Y is -COOH, -COOMe, -COOEt or

[0042] In some embodiments, R^x is H. In other embodiments, R^x is halogen. In other embodiments, R^x is fluorine or choline. [0043] In other embodiments, W is -CH2- .In further embodiments, W is -CH2-CH2-. In further embodiments, W is -CH2-CH2-CH2-. In further embodiments, W is -CH2-CH2-CH2- CH2-.

[0044] In some embodiments, Z is -COOH. In other embodiments, Z is -COOMe. In other embodiments, Z is -COOEt. In other embodiments, Z is

In some embodiments, R^z is halogen, CH3, CF3. In other embodiments, R^z is halogen. In other embodiments, R^z is fluorine or chlorine. In other embodiments, R^z is CH3 or CF3. In other embodiments, n is 1. In other embodiments, n is 2. In other embodiments, two adjacent R^z groups with the C atoms to which they are attached form a 5 or 6- membered ring. In other embodiments, two adjacent R^z groups with the C atoms to which they are attached form a 5-membered ring. In other embodiments, two adjacent R^z groups with the C atoms to which they are attached form a 6-membered ring.

[0045] In some embodiments, T is a bond. In other embodiments, T is -CFh-.ln other embodiments, T is -CH2-CH2-.

[0046] In some embodiments, Y is -COOH, -COOMe or -COOEt. In other embodiments, Y is -COOH. In other embodiments, Y is -COOMe. In other embodiments, Y is -COOEt.

In other embodiments, Y is

In some embodiments, R^Y is halogen, CH₃, CF₃. In other embodiments, R^Y is halogen. In other embodiments, R^Y is fluorine or chlorine. In other embodiments, R^Y is CH₃ or CF₃. In other embodiments, n' is 1. In other embodiments, n' is 2. In other embodiments, n' is 2 and the two R^Y groups are the same. In other embodiments, n' is 2 and the two R^Y groups are different. In other embodiments, two adjacent R^Y groups with the C atoms to which they are attached form a 5 or 6-membered ring. In other embodiments, two adjacent R^Y groups with the C atoms to which they are attached form a 5-membered ring. In other embodiments, two adjacent R^Y groups with the C atoms to which they are attached form a 6-membered ring.

[0047] Also described herein are compounds having the structure of Formula (lib):

R^x is H or halogen;

W is -CH2-, -CH2-CH2-, -CH2-CH2-CH2- or -CH2-CH2-CH2-CH2-;

Z is -COOH, -COOMe, -COOEt or

T is a bond, -CH2- or -CH2-CH2-;

Y is -COOH, -COOMe, -COOEt or

[0048] In some embodiments, R^x is H. In other embodiments, R^x is halogen. In other embodiments, R^x is fluorine or choline.

[0049] In other embodiments, W is -CH₂- .In further embodiments, W is -CH₂-CH₂-. In further embodiments, W is -CH₂-CH₂-CH₂-. In further embodiments, W is -CH₂-CH₂-CH₂- CH₂-.

[0050] In some embodiments, Z is -COOH. In other embodiments, Z is -COOMe. In other embodiments, Z is -COOEt. In other embodiments, Z is

In some embodiments, R^z is halogen, CH₃, CF₃. In other embodiments, R^z is halogen. In other embodiments, R^z is fluorine or chlorine. In other embodiments, R^z is CH₃ or CF₃. In other embodiments, n is 1. In other embodiments, n is 2. In other embodiments, two adjacent R^z groups with the C atoms to which they are attached form a 5 or 6- membered ring. In other embodiments, two adjacent R^z groups with the C atoms to which they are attached form a 5-membered ring. In other embodiments, two adjacent R^z groups with the C atoms to which they are attached form a 6-membered ring.

[0051] In some embodiments, T is a bond. In other embodiments, T is -CH₂-.ln other embodiments, T is -CH₂-CH₂-.

[0052] In some embodiments, Y is -COOH, -COOMe or -COOEt. In other embodiments, Y is -COOH. In other embodiments, Y is -COOMe. In other embodiments, Y is -COOEt.

In other embodiments, Y is

[0053] Also described herein are compounds, or pharmaceutically acceptable salts, solvates, hydrates, isomers, tautomers, racemates, or isotopes thereof, selected from:

[0054] Also described herein are compounds having the structure of Formula (I). In other embodiments, compounds are provided which are pharmaceutically acceptable salts of Formula (I). In other embodiments, compounds are provided which are solvates of Formula (I). In other embodiments, compounds are provided which are hydrates of Formula (I). In other embodiments, compounds are provided which are isomers of Formula (I). In other embodiments, compounds are provided which are tautomers of Formula (I). In other embodiments, compounds are provided which are racemates of Formula (I). In other embodiments, compounds are provided which are isotopes of Formula (I).

[0G5S] Also described herein are compounds having the structure of Formula (lla). In other embodiments, compounds are provided which are pharmaceutically acceptable salts of Formula (lla). In other embodiments, compounds are provided which are solvates of Formula (lla). In other embodiments, compounds are provided which are hydrates of Formula (lla). In other embodiments, compounds are provided which are isomers of Formula (lla). In other embodiments, compounds are provided which are tautomers of Formula (lla). In other embodiments, compounds are provided which are racemates of Formula (lla). In other embodiments, compounds are provided which are isotopes of Formula (lla).

[0056] Also described herein are compounds having the structure of Formula (lib). In other embodiments, compounds are provided which are pharmaceutically acceptable salts of Formula (Mb). In other embodiments, compounds are provided which are solvates of Formula (Mb). In other embodiments, compounds are provided which are hydrates of Formula (Mb). In other embodiments, compounds are provided which are isomers of Formula (Mb). In other embodiments, compounds are provided which are tautomers of Formula (Mb). In other embodiments, compounds are provided which are racemates of Formula (Mb). In other embodiments, compounds are provided which are isotopes of Formula (Mb).

[0057] Also described herein are pharmaceutical compositions comprising compounds having the structure of Formula (I), Formula (lla) or Formula (Mb) or pharmaceutically acceptable salts, solvates, hydrates, isomers, tautomers, racemates, or isotopes thereof, and at least one pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical compositions comprise compounds having the structure of Formula (I), or pharmaceutically acceptable salts, solvates, hydrates, isomers, tautomers, racemates, or isotopes thereof, and at least one pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical compositions comprise compounds having the structure of Formula (lla) or pharmaceutically acceptable salts, solvates, hydrates, isomers, tautomers, racemates, or isotopes thereof, and at least one pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical compositions comprise compounds having the structure of Formula (lib) or pharmaceutically acceptable salts, solvates, hydrates, isomers, tautomers, racemates, or isotopes thereof, and at least one pharmaceutically acceptable excipient.

[0858] Also described herein are uses of a compound Formula (I), Formula (I la) or Formula (Mb) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof, in the manufacture of a medicament. In some embodiments the medicament is for the treatment of an inflammatory disease. In some embodiments the medicament is for the treatment of cancer. In some embodiments the medicament is for the treatment of or an infection. In some embodiments the medicament is for the treatment of Irritable Bowel Syndrome (IBS), Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, diversion colitis, Behget's syndrome or indeterminate colitis.

[0859] Also described herein are methods of inhibiting hypoxia-inducible factor 1-a (H!F-i ) activity, comprising contacting the HSF-la with an effective amount of a compound of Formula (I), Formula (I la) or Formula (Mb) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof. [0880] Also described herein are methods of inhibiting hypoxia-inducible factor 1-s (HIF-Ia) activity in a subject, comprising administering to the subject an effective amount of a compound of a Formula (I), Formula (lla) or Formula (Mb) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof.

[0861] Also described herein are methods of up regulating of phosphoglycerate kinase (PGK) activity, comprising contacting the PGK with an effective amount of a compound of Formula (I), Formula (lla) or Formula (Mb) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof.

[0882] Also described herein are methods of increasing the immune response in a subject, comprising administering to the subject an effective amount of a compound of Formula (I), Formula (lla) or Formula (Mb) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof. In some embodiments, the subject has a medical condition causing a decreased cellular immunity. In some embodiments, the medical condition is an inflammatory disease. In some embodiments, the medical condition is cancer. In some embodiments, the medical condition is an infection. In some embodiments, the medical condition is Irritable Bowel Syndrome (IBS), Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, diversion colitis, Behget's syndrome or indeterminate colitis.

[0083] Also described herein are methods of treating a subject with an inflammatory epithelial disease, comprising administering to the subject an effective amount of a compound of Formula (I), Formula (I la) or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof. In some embodiments, the inflammatory epithelial disease is a disease affecting the Gl tract. In some embodiments, the inflammatory epithelial disease is a disease affecting the respiratory tract. In some embodiments, the inflammatory epithelial disease is a disease affecting mucosa. In some embodiments, the inflammatory epithelial disease is a disease affecting the skin. In some embodiments, the inflammatory epithelial disease is a disease affecting the lining of major organs and endocrine glands or vascular tissue. In some embodiments, the inflammatory epithelial disease is a disease affecting intestinal epithelial tissue. In some embodiments, the inflammatory epithelial disease is Irritable Bowel Syndrome (IBS). In some embodiments, the inflammatory epithelial disease is Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, diversion colitis, Behget's syndrome or indeterminate colitis. In some embodiments, the inflammatory epithelial disease is Crohn's disease. In some embodiments, the inflammatory epithelial disease is ulcerative colitis. In some embodiments, the methods of treating a subject with an inflammatory epithelial disease, further comprise administering at least one additional therapeutic agent. In some embodiments, the additional therapeutic agent is an anti-inflammatory, an immune system suppressor, an antibiotic, an anti-diarrheal or a pain reliever. In some embodiments, the anti-inflammatory is a corticosteroid, an aminosalicylate, mesalamine (Asacol HD, Delzicol), balsalazide (Colazal) or olsalazine (Dipentum). In some embodiments, the immunosuppressant is azathioprine (Azasan, Imuran), mercaptopurine (Purinethol, Purixan), cyclosporine (Gengraf, Neoral, Sandimmune) or methotrexate (Trexall). In some embodiments, the immune system suppressor is infliximab (Remicade), adalimumab (Humira), golimumab (Simponi), natalizumab (Tysabri), vedolizumab (Entyvio) or ustekinumab (Stelara). In some embodiments, the antibiotic is ciprofloxacin (Cipro) or metronidazole (Flagyl). In some embodiments, the anti-diarrheal is a fiber supplement, psyllium powder (Metamucil), methylcellulose (Citrucel) or loperamide (Imodium A-D). In some embodiments, the pain reliever is acetaminophen (Tylenol).

[0064] Also described herein are methods of treating an infection in a subject, comprising administering to the subject an effective amount of a compound of Formula (I), Formula (I la) or Formula (lib), or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof. In some embodiments, the infection is caused by bacteria. In some embodiments, the bacterium is Staphylococcus aureus , rnethieil!in resistant Staphylococcus aureu , Streptococcus pyogenes, Pseudomonas aeruginosa, or Adnetobacter haumannii. In some embodiments, the infection is caused by a virus. In some embodiments, the infection is caused by a fungus. In some embodiments, the infection is caused by a yeast. In some embodiments, the infection is caused by a parasite. In some embodiments, the methods of treating an infection in a subject, further comprise administering an antimicrobial agent.

[0065] Representative compounds having the structure of any one of Formulas (I), (I la) or (Mb), as applicable, include the compounds listed in table 1, as well as pharmaceutically acceptable isomers, racemates, hydrates, solvates, isotopes, or salts thereof. Table 1: Representative compounds

[0066] "Isomer" as used herein to encompasses all chiral, diastereomeric or racemic forms of a structure, unless a particular stereochemistry or isomeric form is specifically indicated. Such compounds can be enriched or resolved optical isomers at any or all asymmetric atoms as are apparent from the depictions, at any degree of enrichment. Both racemic and diastereomeric mixtures, as well as the individual optical isomers can be synthesized so as to be substantially free of their enantiomeric or diastereomeric partners, and these are all within the scope of certain embodiments of the disclosure. The isomers resulting from the presence of a chiral center comprise a pair of non-superimposable isomers that are called "enantiomers." Single enantiomers of a pure compound are optically active (i.e., they are capable of rotating the plane of plane polarized light and designated R or S).

[0067] "Isolated optical isomer" means a compound which has been substantially purified from the corresponding optical isomer(s) of the same formula. For example, the isolated isomer may be at least about 80%, at least 80% or at least 85% pure. In other embodiments, the isolated isomer is at least 90% pure or at least 98% pure, or at least 99% pure by weight.

[0068] "Substantially enantiomerically or diasteromerically" pure means a level of enantiomeric or diastereomeric enrichment of one enantiomer with respect to the other enantiomer or diasteromer of at least about 80%, and more specifically in excess of 80%, 85%, 90%, 95%, 98%, 99%, 99.5% or 99.9%.

[0069] As used herein the terms "racemate" and "racemic mixture" refer to an equal mixture of two enantiomers. A racemate is labeled "(±)" because it is not optically active (i.e., will not rotate plane-polarized light in either direction since its constituent enantiomers cancel each other out).

[0070] Unless stated to the contrary, a formula with chemical bonds shown only as solid lines and not as wedges or dashed lines contemplates each possible isomer, e.g., each enantiomer, diastereamer, and meso compound, and a mixture of isomers, such as a racemic or scalemic mixture.

[0071] A "hydrate" is a compound that exists in combination with water molecules. The combination can include water in stoichiometric quantities, such as a monohydrate or a dihydrate, or can include water in random amounts. As the term is used herein a "hydrate" refers to a solid form; that is, a compound in a water solution, while it may be hydrated, is not a hydrate as the term is used herein.

[0077] A "solvate" is similar to a hydrate except that a solvent other that water is present. For example, methanol or ethanol can form an "alcoholate", which can again be stoichiometric or non-stoichiometric. As the term is used herein a "solvate" refers to a solid form; that is, a compound in a solvent solution, while it may be solvated, is not a solvate as the term is used herein.

[0073] "Isotope" refers to atoms with the same number of protons but a different number of neutrons, and an isotope of a compound having the structure of any one of Formulas (I), (I la) or (Mb) includes any such compound wherein one or more atoms are replaced by an isotope of that atom. For example, carbon 12, the most common form of carbon, has six protons and six neutrons, whereas carbon 13 has six protons and seven neutrons, and carbon 14 has six protons and eight neutrons. Hydrogen has two stable isotopes, deuterium (one proton and one neutron) and tritium (one proton and two neutrons). While fluorine has a number of isotopes, fluorine 19 is longest-lived. Thus, an isotope of a compound having the structure of any one of Formulas (I), (I la) or (Mb) includes, but is not limited to, compounds having the structure of any one of Formulas (I), (lla) or (Mb) wherein one or more carbon 12 atoms are replaced by carbon-13 and/or carbon-14 atoms, wherein one or more hydrogen atoms are replaced with deuterium and/or tritium, and/or wherein one or more fluorine atoms are replaced by fluorine-19.

[0074] "Salt" generally refers to an organic compound, such as a carboxylic acid or an amine, in ionic form, in combination with a counter ion. For example, salts formed between acids in their anionic form and cations are referred to as "acid addition salts". Conversely, salts formed between bases in the cationic form and anions are referred to as "base addition salts."

[0075] The term "pharmaceutically acceptable" refers an agent that has been approved for human consumption and is generally non-toxic. For example, the term "pharmaceutically acceptable salt" refers to nontoxic inorganic or organic acid and/or base addition salts (see, e.g., Lit et al., Salt Selection for Basic Drugs, Int. J. Pharm., 33, 201-217, 1986) (incorporated by reference herein).

Pharmaceutically acceptable base addition salts of compounds of the disclosure include, for example, metallic salts including alkali metal, alkaline earth metal, and transition metal salts such as, for example, calcium, magnesium, potassium, sodium, and zinc salts. Pharmaceutically acceptable base addition salts also include organic salts made from basic amines such as, for example, N,N'dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N- methylglucamine), and procaine.

Pharmaceutically acceptable acid addition salts may be prepared from an inorganic acid or from an organic acid. Examples of inorganic acids include hydrochloric, hydrobromic, hydriodic, nitric, carbonic, sulfuric, and phosphoric acids. Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, aromatic aliphatic, heterocyclic, carboxylic, and sulfonic classes of organic acids, examples of which include formic, acetic, trifluoroacetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic, mandelic, hippuric, malonic, oxalic, embonic (pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, panthothenic, trifluoromethanesulfonic, 2-hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic, cyclohexylaminosulfonic, stearic, alginic, hydroxybutyric, salicylic, -galactaric, and galacturonic acid.

[0078] Although pharmaceutically unacceptable salts are not generally useful as medicaments, such salts may be useful, for example as intermediates in the synthesis of compounds having the structure of any one of Formulas (I), (lla) or (Mb), for example in their purification by recrystallization.

[0077] In certain embodiments, the disclosure provides a pharmaceutical composition comprising a compound having the structure of any one of Formulas (I), (lla) or (Mb), or a pharmaceutically acceptable isomer, racemate, hydrate, solvate, isotope, or salt thereof, together with at least one pharmaceutically acceptable carrier, diluent, or excipient. For example, the active compound will usually be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier which can be in the form of an ampoule, capsule, sachet, paper, or other container. When the active compound is mixed with a carrier, or when the carrier serves as a diluent, it can be solid, semi-solid, or liquid material that acts as a vehicle, excipient, or medium for the active compound. The active compound can be adsorbed on a granular solid carrier, for example contained in a sachet. Some examples of suitable carriers are water, salt solutions, alcohols, polyethylene glycols, polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatin, lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugar, cyclodextrin, amylose, magnesium stearate, talc, gelatin, agar, pectin, acacia, stearic acid, or lower alkyl ethers of cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid monoglycerides and diglycerides, pentaerythritol fatty acid esters, polyoxyethylene, hydroxymethylcellulose, and polyvinylpyrrolidone. Similarly, the carrier or diluent can include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax.

[0078] As used herein, the term "pharmaceutical composition" refers to a composition containing one or more of the compounds described herein, or a pharmaceutically acceptable isomer, racemate, hydrate, solvate, homolog or salt thereof, formulated with a pharmaceutically acceptable carrier, which can also include other additives, and manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gelcap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or in any other formulation described herein. Conventional procedures and ingredients for the selection and preparation of suitable formulations are described, for example, in Remington: The Science and Practice of Pharmacy, 21st Ed., Gennaro, Ed., Lippencott Williams & Wilkins (2005) and in The United States Pharmacopeia: The National Formulary (USP 36 NF31), published in 2013.

[0079] In another embodiment, there are provided methods of making a composition of a compound described herein including formulating a compound of the disclosure with a pharmaceutically acceptable carrier or diluent. In some embodiments, the pharmaceutically acceptable carrier or diluent is suitable for oral administration. In some such embodiments, the methods can further include the step of formulating the composition into a tablet or capsule. In other embodiments, the pharmaceutically acceptable carrier or diluent is suitable for parenteral administration. In some such embodiments, the methods further include the step of lyophilizing the composition to form a lyophilized preparation.

[0080] As used herein, the term "pharmaceutically acceptable carrier" refers to any ingredient other than the disclosed compounds, or a pharmaceutically acceptable isomer, racemate, hydrate, solvate, homolog or salt thereof (e.g., a carrier capable of suspending or dissolving the active compound) and having the properties of being nontoxic and non-inflammatory in a patient. Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspensing or dispersing agents, sweeteners, or waters of hydration. Exemplary excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol. [0081] The formulations can be mixed with auxiliary agents which do not deleteriously react with the active compounds. Such additives can include wetting agents, emulsifying and suspending agents, salt for influencing osmotic pressure, buffers and/or coloring substances, preserving agents, sweetening agents, or flavoring agents. The compositions can also be sterilized if desired.

[0082] The route of administration can be any route which effectively transports the active compound of the disclosure to the appropriate or desired site of action, such as oral, nasal, pulmonary, buccal, subdermal, intradermal, transdermal, or parenteral, e.g., rectal, depot, subcutaneous, intravenous, inhalation of a dry powder form or a nebulized form, intraurethral, intramuscular, intranasal, ophthalmic solution, or an ointment, the oral route being preferred.

[0083] Dosage forms can be administered once a day, or more than once a day, such as twice or thrice daily. Alternatively, dosage forms can be administered less frequently than daily, such as every other day, or weekly, if found to be advisable by a prescribing physician. Dosing regimens include, for example, dose titration to the extent necessary or useful for the indication to be treated, thus allowing the patient's body to adapt to the treatment and/or to minimize or avoid unwanted side effects associated with the treatment. Other dosage forms include delayed or controlled- release forms. Suitable dosage regimens and/or forms include those set out, for example, in the latest edition of the Physicians' Desk Reference, incorporated herein by reference.

[0084] As used herein, the term "administering" or "administration" refers to providing a compound, a pharmaceutical composition comprising the same, to a subject by any acceptable means or route, including (for example) by oral, parenteral (e.g., intravenous), inhaled, or topical administration.

[0085] As used herein, the term "treatment" refers to an intervention that ameliorates a sign or symptom of a disease or pathological condition. As used herein, the terms "treatment", "treat" and "treating," with reference to a disease, pathological condition or symptom, also refers to any observable beneficial effect of the treatment. The beneficial effect can be evidenced, for example, by a delayed onset of clinical symptoms of the disease in a susceptible subject, a reduction in severity of some or all clinical symptoms of the disease, a slower progression of the disease, a reduction in the number of relapses of the disease, an improvement in the overall health or well-being of the subject, or by other parameters well known in the art that are specific to the particular disease. A prophylactic treatment is a treatment administered to a subject who does not exhibit signs of a disease or exhibits only early signs, for the purpose of decreasing the risk of developing pathology. A therapeutic treatment is a treatment administered to a subject after signs and symptoms of the disease have developed. [0086] As used herein, the term "subject" refers to an animal (e.g., a mammal, such as a human). A subject to be treated according to the methods described herein may be one who has been diagnosed with a neurodegenerative disease involving demyelination, insufficient myelination, or underdevelopment of a myelin sheath, e.g., a subject diagnosed with multiple sclerosis or cerebral palsy, or one at risk of developing the condition. Diagnosis may be performed by any method or technique known in the art. One skilled in the art will understand that a subject to be treated according to the present disclosure may have been subjected to standard tests or may have been identified, without examination, as one at risk due to the presence of one or more risk factors associated with the disease or condition.

[0087] As used herein, the term "effective amount" refers to a quantity of a specified agent sufficient to achieve a desired effect in a subject being treated with that agent. Ideally, an effective amount of an agent is an amount sufficient to inhibit or treat the disease without causing substantial toxicity in the subject. The effective amount of an agent will be dependent on the subject being treated, the severity of the affliction, and the manner of administration of the pharmaceutical composition. Methods of determining an effective amount of the disclosed compound sufficient to achieve a desired effect in a subject will be understood by those of skill in the art in light of this disclosure.

[0088] As used herein, the term "antimicrobial" or "antimicrobial agent" refers to the ability to treat or control (e.g., reduce, prevent, inhibit, break-down, or eliminate) microorganism growth or survival at any concentration. Similarly, the terms "antibacterial," "antiviral," and "antifungal" respectively mean the ability to treat or control (e.g., reduce, prevent, inhibit, break-down, or eliminate) bacterial, viral, and fungal growth or survival at any concentration.

[0089] As used herein, the term "chemotherapeutic agent" includes any other pharmaceutically active compound that can be used in conjunction with the disclosed HIF-Ia prolyl hydroxylase inhibitors. HIF1 Stabilization in Ceils

[0090] H!F-Ia is targeted for destruction via prolyl hydroxylation, an oxygen- dependent modification that signals for recognition by the E3 ubiquitin iigase complex containing the von Hippel-Lindau tumor suppressor (VHL). Three prolyl hydroxylases formerly referred to in the literature as EGLN1, EGLN2, and EGLN3 (also known as, have been identified in mammals, among which, EGLN1 {also known as HIFPH2 or PHD2), and EGLN3 {also known as HIFPH3 or PHD3), are hypoxia-inducible at their mRNA levels in a HIF-la dependent manner. HIF-la levels are controlled by these prolyl-4- hydroxylases by hydroxylating the HIF-la proiine residues Pro-402 and Pro- 564 in humans {Ivan, IVL et ak, (2001) "HIF-la targeted for VHL-mediated destruction by proiine hydroxylation: Implications for O? sensing." Science 292, 464-468; Jaakkola, P. et aL, {2001) "Targeting of HIF-la to the von Hippel-Lindau ubiquityiation complex by O^-reguiated prolyl hydroxylation." Science 292, 468-472; and Masson, N. et aL, (2001) "independent function of two destruction domains in hypoxia-inducible factor- a chains activated by prolyl hydroxylation." EMRO 1 20, 5197-5206). Under hypoxia conditions, EG INI and FGL.N3 activity is suppressed

[0091] Stimulated by a build up of the cellular concentration of HIF-la is the production of Phosphogiycerate Kinase (PGK) and Vascular Endothelial Growth Factor (VEGF). it has been shown that stimulation of VEGF induces the formation of functional neo-vessels in the mouse cornea and enhanced blood flow' in a dog model of coronary artery disease. The HIF-la prolyl hydroxylase inhibitors of the present disclosure provide enhancement in the expression of multiple hypoxia inducible genes including VEGF, GAPDH and erythropoietin (EPO). Additionally, the HIF-la prolyl hydroxylase inhibitors of the present disclosure provide enhanced the accumulation of HIFl-a in the cytoplasm and nucleus. Transgenic mice expressing a constitutive!y active HIF-la in the skin have increased dermal vascularity and had a 13-fold increase in VEGF levels Wounds

[0092] Chronic, non-healing wounds are a major cause of prolonged morbidity in the aged human population. This is especially the case in bedridden or diabetic patients who develop severe, non -healing skin ulcers. In many of these cases, the delay in healing is a result of inadequate blood supply either as a result of continuous pressure or of vascular blockage. Poor capillary circulation due to small artery atherosclerosis or venous stasis contributes to the failure to repair damaged tissue. Such tissues are often infected with microorganisms that proliferate unchallenged by the innate defense systems of the body which require well vascularized tissue to effectively eliminate pathogenic organisms. As a result, mos therapeutic intervention centers on restoring blood flow to ischemic tissues thereby allowing nutrients and immunological factors access to the site of the wound.

The present disclosure relates to methods for treating wounds and promoting wound healing in a subject comprising, administering to a subject in need of treatment an effective amount of one or more of the disclosed compounds.

The present disclosure relates to the use of one or more of the disclosed compounds for use in making a medicament for treating wounds and promoting wound healing. Antimicrobial

[0093] The hypoxia-responsive transcription factor HIF-lot is essential for regulation of inflammation in vivo. As such, it has been discovered (Peyssonnaux C. et a!., "HIF-la expression regulates the bactericidal capacity of phagocytes” J. Clinical Investigation 115(7), pp 1808-1815 (2005)) that bacterial infection induces HIF-la expression in myeloid cells even under normoxic conditions, and that HIF-la regulates the generation of critical molecular effectors of immune defense including granule proteases, antimicrobial peptides, nitric oxide, and TMF-a Bacterial infection induces a subset of HIF-la target genes specifically related to microbial killing, thereby demonstrating that HIF-la has an essential function in innate immunity distinct from hypoxic response. Therefore, HIF-la function is critical for myeloid cell bactericidal activity and the ability of the host to limit systemic spread of infection from an initial tissue focus increased activity of the HIF-la pathway through vHL deletion supports myeloid cell production of defense factors and improves bactericidal capacity. The disclosed compounds induce HIF-la activity and can also boost bacterial killing and HO production in a HIF-la-specific fashion. These discoveries provide methods for enhancing innate immune responses to microbial, for example, bacterial, infection. [0094] in some embodiments, the disclosed compounds can increase the stabilization of HiF-1 protein by acting directly or indirectly on one or more cellular processes which act to destabilize or to metabolize cellular components that stabilize the presence of HIF-1 protein, protect it from inhibition, or to increase the activity of the protein. Alternatively, the disclosed compounds can increase the activity of the HiF-1 protein by inhibiting or otherwise blocking the activity of compounds that inhibit the activity of the HIF-1 protein. As such, disclosed herein is a method for improving the treatment of microbial infections by administering a substance that increases the activity or level of at least one HIF-1 protein in a subject suffering from the microbial infection or at increased risk of microbial infection.

[0095] in one aspect, disclosed herein are methods for modulating the activity of at least one HIF-1 protein. As such, the disclosed methods comprise contacting at least one HIF-1 protein or HiF-1 interacting protein with one or more of the disclosed compounds that modulate the activity of the HIF-1 protein, or causing contact between the protein and substance. In the embodiment, the contacting is accomplished in vitro. In another embodiment, the contacting is accomplished in vivo. In a further embodiment, the contacting is accomplished ex vivo.

[0096] In another aspect, disclosed herein is a method of treating a subject infected or at risk of infection by a microbial agent comprising administering to a subject a therapeutically effective amount of one or more of the disclosed compounds. In one embodiment, the compound increases the amount or activity of HiF-1. In another embodiment, the microbial agent is a pathogen. Iterations of this embodiment related to pathogens includes, bacteria, fungi, protozoa, viruses, yeasts, and the like. A yet further iteration of this aspect relates to a method for treating a subject infected by or at risk of infection by a microbial agent comprising, increasing the microbial pathogen- killing activity of the subject's immune ceils.

[0097] One method for increasing the stabilization of HIF-1 is to inhibit the activity of 4-proiyl hydroxylase enzymes that begin the cellular break down of HIF-Ia thereby preventing HIF-la from combining with HSF-Ib to form H!F-1. As such, disclosed herein are methods for increasing the cellular response to disease states such as infection, i.e., presence of a pathogen such as a bacterium, a virus, a parasite, a yeast, a fungus, and the like by increasing phagocytosis. Also disclosed herein are methods for treating cancer by increasing the cellular immune response, for example, by stabilizing HIF-1, thereby increasing the ability of the body to reduce tumor size. Further disclosed herein are methods for treating diseases wherein an immune response can be stimulated by vaccination.

[0098] The present disclosure further relates to the use of one or more of the HIF-la prolyl hydroxylase inhibitors disclosed herein for making a medicament for treating anemia.

[0099] The present disclosure further relates to the use of one or more of the HIF-la prolyl hydroxylase inhibitors disclosed herein for making a medicament for treating increasing cellular immunity.

[0100] The present disclosure further relates to the use of one or more of the HIF-la prolyl hydroxylase inhibitors disclosed herein for making a medicament for treating cancer.

[0101] The present disclosure further relates to the use of one or more of the HIF-la prolyl hydroxylase inhibitors disclosed herein for making a medicament for increasing HiF-1 stabilization.

[0102] The present disclosure further relates to the use of one or more of the HIF-la prolyl hydroxylase inhibitors disclosed herein for making a medicament for treating peripheral vascular disease.

[0103] The present disclosure further relates to the use of one or more of the HIF-la. prolyl hydroxylase inhibitors disclosed herein for making a medicament for treating wounds.

[0104] The present disclosure further relates to the use of one or more of the HIF-la prolyl hydroxylase inhibitors disclosed herein for making a medicament that is an antimicrobial. [0105] The present disclosure further relates to the use of one or more of the HIF-la prolyl hydroxylase inhibitors disclosed herein for making a medicament for treating atherosclerotic lesions.

[0106] The present disclosure further relates to the use of one or more of the HIF-la prolyl hydroxylase inhibitors disclosed herein for making a medicament for treating diabetes.

[0107] The present disclosure further relates to the use of one or more of the HIF-la prolyl hydroxylase inhibitors disclosed herein for making a medicament for treating hypertension.

[OIOS] The present disclosure further relates to the use of one or more of the HIF-la prolyl hydroxylase inhibitors disclosed herein for making a medicament for treating a disease affected by the level of vascular endothelial growth factor (VEGF), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and erythropoietin (EPO). [0109] The present disclosure further relates to the use of one or more of the HIF-la prolyl hydroxylase inhibitors disclosed herein for making a medicament for treating Crohn's disease., ulcerative colitis, psoriasis, sarcoidosis, rheumatoid arthritis, hemangiomas, Os!er-Weber-Rendu disease, or hereditary hemorrhagic telangiectasia, solid or blood borne tumors and acquired immune deficiency syndrome.

[0110] The present disclosure further relates to the use of one or more of the HIF-la prolyl hydroxylase inhibitors disclosed herein for making a medicament for treating a disorder chosen from diabetic retinopathy, macular degeneration, cancer, sickle cell anemia, sarcoid, syphilis, pseudoxanthoma elasticum, Paget's disease, vein occlusion, artery occlusion, carotid obstructive disease, chronic uveitis/vitritis, mycobacterial infections, Lyme's disease, systemic lupus erythernatosis, retinopathy of prematurity, Eales' disease, Behcet's disease, infections causing a retinitis or choroiditis, presumed ocular histoplasmosis, Best's disease, myopia, optic pits, Stargardt's disease, pars planitis, chronic retinal detachment, hyperviscosity syndrome, toxoplasmosis, trauma and post-laser complications, diseases associated with rubeosis, and proliferative vitreoretinopathy. [0111] The disclosed compositions and the form of pharmaceutical preparations comprising the HiF-lct prolyl hydroxylase inhibitors alone, or in combination with another drug or other therapeutic agent, inter alia, chemotherapeutic agent or chemotherapeutic compound, can vary according to the intended route of administration.

[0112] Orally administered preparations can be in the form of solids, liquids, emulsions, suspensions, or gels, or in dosage unit form, for example as tablets or capsules. Tablets can be compounded in combination with other ingredients customarily used, such as tale, vegetable oils, polyols, gums, gelatin, starch, and other carriers. The HiF-Ia prolyl hydroxylase inhibitors can be dispersed in or combined with a suitable liquid carrier in solutions, suspensions, or emulsions.

[0113] Parenteral compositions intended for injection, either subcutaneously, intramuscularly, or Intravenously, can be prepared as liquids or solid forms for solution in liquid prior to injection, or as emulsions. Such preparations are sterile, and liquids to be injected intravenously should be isotonic. Suitable excipients are, for example, water, dextrose, saline, and glycerol.

[0114] Administration of pharmaceutically acceptable salts of the substances described herein is included within the scope of the present disclosure. Such salts can be prepared from pharmaceutically acceptable non-toxic bases including organic bases and inorganic bases. Salts derived from inorganic bases include sodium, potassium, lithium, ammonium, calcium, magnesium, and the like. Saits derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, basic amino acids, and the like. For a helpful discussion of pharmaceutical salts, see S. M. Berge et. ak, Journal of Pharmaceutical Sciences 66:1-19 (1977) the disclosure of which is hereby incorporated by reference. [0115] Substances for injection can be prepared in unit dosage form in ampules, or in muitidose containers. The HSF-la prolyl hydroxylase inhibitors or compositions comprising one or more HIF-la prolyl hydroxylase inhibitors to be delivered can be present in such forms as suspensions, solutions, or emulsions in oily or preferably aqueous vehicles. Alternatively, the salt of the HIF-la prolyl hydroxylase inhibitor can be in iyophi!ized form for reconstitution, at the time of delivery, with a suitable vehicle, such as sterile pyrogen-free water. Both liquids as we 11 as lyophi!ized forms that are to he reconstituted will comprise agents, preferably buffers, in amounts necessary to suitably adjust the pH of the injected solution. For any parenteral use, particularly if the formulation is to be administered intravenously, the total concentration of solutes should be controlled to make the preparation isotonic, hypotonic, or weakly hypertonic. Nonionic materials, such as sugars, are preferred for adjusting tonicity, and sucrose is particularly preferred. Any of these forms can further comprise suitable formulatory agents, such as starch or sugar, glycerol or saline. The compositions per unit dosage, whether liquid or solid, can contain from 0.1% to 99% of polynucleotide material.

Methods Relating to Stabilization of HIF-1

[0116] The eradication of invading microorganisms depends initially on innate immune mechanisms that preexist in all individuals and act within minutes of infection. Phagocytic cell types, including macrophages and neutrophils, play a key role in innate immunity because they can recognize, ingest, and destroy many pathogens without the aid of an adaptive immune response. The effectiveness of myeloid ceils in innate defense reflects their capacity to function in low oxygen environments.

Whereas in healthy tissues oxygen tension is generally 20-70 mm MG (i.e. 2.5-9% oxygen), much lower levels (<1% oxygen) have been described in wounds and necrotic tissue foci [Arnold et al., BrJ Exp Pathol 68, 569 (1987); Vogelberg & Konig, Clin Investig 71, 466 (1993); Negus et al., Am J Pathol 150, 1723 (1997)). it has also been shown {ZinkernageS A. 5. et al., "Pharmacologic Augmentation of Hypoxia-inducible Factor-la with Mirnosine Boosts the Bactericidal Capacity of Phagocytes” .1 Infectious Diseases {2QQ8):197: 214-217) that the HIF-Ia agonist mirnosine can boost the capacity of human phagocytes and whole blood to kill the leading pathogen Staphylococcus aureus in a dose-dependent fashion and reduce the lesion size in a murine model of 5. aureus skin infection.

[0117] Macrophages are one population of effector cells involved in immune responses. Their role in natural immunity includes mediation of phagocytosis, as well as release of cytokines and cytotoxic mediators. They also facilitate the development of acquired immunity through antigen presentation and release of immunomodulatory cytokines. Although macrophages are immune effectors, they are also susceptible to infection by agents such as bacteria, protozoa, parasites, and viruses (The Macrophage, C. E. Lewis & J.O'D. McGee eds., i RL Press at Oxford University Press, New York, N.Y., 1992). Viruses capab e of infecting macrophages include several RNA viruses such as measles virus (MV) (e.g., Joseph et aL, .L Virol 16, 1638-4649, 1975), respiratory syncytial virus (RSV) (Midulla et ak, Am Rev Respir. Dis 140, 771-777, 1989), and human immunodeficiency virus type 1 (HIV-1) (Meitzer and Gendelman, in Macrophage Biology and Activation, S. W. Russeii and S. Gordon, eds., Springer-Verlag, New York, N.Y., pp. 239-263(1992: Potts et aL, Virology 175, 465-476, 1990).

[0118] Disclosed herein is a method for increasing HIF-1 stabilization in a cell, comprising contacting a cell in vivo, in vitro, or ex vivo with an effective amount of one or more of the disclosed HIF-Ia prolyl hydroxylase inhibitors.

[0119] Also disclosed herein are methods for increasing the cellular immune response of a human or mammal in need of increased cellular immunity, comprising administering to a human or mammal in need with an effective amount of one or more of the disclosed HSF-Ia prolyl hydroxylase inhibitors.

[0120] Further disclosed herein are methods for increasing the cellular immune response of a human or mammal diagnosed with a medical condition causing a decreased cellular immunity, comprising administering to a human or mammal in need with an effective amount of one or more of the disclosed HiF-lct prolyl hydroxylase inhibitors.

[0121] Yet further disclosed herein are methods for increasing the cellular immune response of a human or mammal diagnosed with a medical condition causing a decreased cellular immunity, comprising administering to a human or mammal in need with an effective amount of one or more of the disclosed H IF- let prolyl hydroxylase inhibitors.

[0122] Still further disclosed herein are methods for increasing the cellular immune response of a human or mammal having a medical condition causing a decreased cellular immunity, comprising administering to a human or mammal in need with an effective amount of one or more of the disclosed HIF-la prolyl hydroxylase inhibitors. [0123] As such, the one or more HIF-la prolyl hydroxylase inhibitor and any co administered compounds can be administered or contacted with a cell topically, bucealiy, orally, intradermally, subcutaneously, mucosa!ly in the eye, vagina, rectum, and nose, intravenously, and intramuscularly.

Methods Relating to the Treatment of Cancer

[0124] As used herein cancer is defined herein as "an abnormal growth of cells which tend to proliferate in an uncontrolled way and, in some cases, to metastasize." As such, both metastatic and non-metastatic cancers can be treated by the disclosed methods.

[0125] Disclosed are methods for treating cancer in a human or mammal, comprising administering to a human or mammal with a cancer with an effective amount of one or more of the disclosed HIF-Ia prolyl hydroxylase inhibitors.

[0126] Also disclosed herein are methods for treating a human or mammal diagnosed with cancer, co-adminis†.ering to a human or mammal one or more chemotherapeutic agent or chemotherapeutic compound together with one or more of the disclosed HIF- la prolyl hydroxylase inhibitors.

[0127] The following are non-limiting examples of malignant and non-ma!ignant cancers. Acute Lymphoblastic; Acute Myeloid Leukemia; Adrenocortical Carcinoma; Adrenocortical Carcinoma, Childhood; Appendix Cancer; Basal Cell Carcinoma; Bile Duct Cancer, Extrahepatic; Bladder Cancer; Bone Cancer; Osteosarcoma and Malignant Fibrous Histiocytoma; Brain Stem Glioma, Childhood; Brain Tumor, Adult; Brain Tumor, Brain Stem Glioma, Childhood; Brain Tumor, Central Nervous System Atypical Teratoid/Rhabdoid Tumor, Childhood; Central Nervous System Embryonal Tumors; Cerebellar Astrocytoma; Cerebral Astrocytoma/Malignant Glioma;

Craniopharyngioma; Ependymoblastoma; Ependymoma; Medulloblastoma; Medulloepithelioma; Pineal Parenchymal Tumors of intermediate Differentiation; Supratentorial Primitive Neuroectodermal Tumors and Pineob!astoma; Visual Pathway and Hypothalamic Glioma; Brain and Spinal Cord Tumors; Breast Cancer; Bronchial Tumors; Burkitt Lymphoma; Carcinoid Tumor; Carcinoid Tumor, Gastrointestinal; Central Nervous System Atypical Teratoid/Rhabdoid Tumor; Central Nervous System Embryonal Tumors; Central Nervous System Lymphoma; Cerebellar Astrocytoma; Cerebral Astrocytoma/Malignant Glioma, Childhood; Cervical Cancer; Chordoma, Childhood; Chronic Lymphocytic Leukemia; Chronic Myelogenous Leukemia; Chronic Myeloproliferative Disorders; Colon Cancer; Colorectal Cancer; Craniopharyngioma; Cutaneous T-Cei! Lymphoma; Esophageal Cancer; Ewing Family of Tumors; Extragonadai Germ Ceil Tumor; Extrahepatic Bile Duct Cancer; Eye Cancer, intraocular Melanoma; Eye Cancer, Retinoblastoma; Gallbladder Cancer; Gastric (Stomach) Cancer; Gastrointestinal Carcinoid Tumor; Gastrointestinal Stromal Tumor (GIST); Germ Cell Tumor, Extracranial; Germ Ceil Tumor, Extragonadai; Germ Ceil Tumor, Ovarian; Gestational Trophoblastic Tumor; Glioma; Glioma, Childhood Brain Stem; Glioma, Childhood Cerebral Astrocytoma; Glioma, Childhood Visual Pathway and Hypothalamic; Hairy Cell Leukemia; Head and Neck Cancer; Hepatocellular (Liver) Cancer; Histiocytosis, Langerhans Ceil; Hodgkin Lymphoma; Hypopharyngeai Cancer; Hypothalamic and Visual Pathway Glioma; intraocular Melanoma; Islet Cell Tumors; Kidney (Renal Cell) Cancer; Langerhans Ceil Histiocytosis; Laryngeal Cancer; Leukemia, Acute Lymphoblastic; Leukemia, Acute Myeloid; Leukemia, Chronic Lymphocytic; Leukemia, Chronic Myelogenous; Leukemia, Hairy Ceil; Lip and Oral Cavity Cancer; Liver Cancer; Lung Cancer, Non-Small Cell; Lung Cancer, Small Cell; Lymphoma, AIDS- Related; Lymphoma, Burkitt; Lymphoma, Cutaneous T-Ceii; Lymphoma, Hodgkin; Lymphoma, Non-Hodgkin; Lymphoma, Primary Central Nervous System; Macrog!obulinemia, Waldenstrom; Malignant Fibrous Histiocytoma of Bone and Osteosarcoma; Medulloblastoma; Melanoma; Melanoma, Intraocular (Eye); Merkel Cell Carcinoma; Mesothelioma; Metastatic Squamous Neck Cancer with Occult. Primary; Mouth Cancer; Multiple Endocrine Neoplasia Syndrome, (Childhood); Multiple Myeioma/Plasma Ceil Neoplasm; Mycosis Fungoides; Myelodyspiastic Syndromes; MyeiodyspSastic/MyeloproSiferative Diseases; Myelogenous Leukemia, Chronic; Myeloid Leukemia, Adult Acute; Myeloid Leukemia, Childhood Acute; Myeloma, Multiple; Myeloproliferative Disorders, Chronic; Nasal Cavity and Paranasal Sinus Cancer; Nasopharyngeal Cancer; Neuroblastoma; Non-Small Cell Lung Cancer; Oral Cancer; Oral Cavity Cancer; Oropharyngeal Cancer; Osteosarcoma and Malignant Fibrous Histiocytoma of Bone; Ovarian Cancer; Ovarian Epithelial Cancer; Ovarian Germ Cell Tumor; Ovarian Low Malignant Potential Tumor; Pancreatic Cancer; Pancreatic Cancer, Islet Cell Tumors; Papillomatosis; Parathyroid Cancer; Penile Cancer; Pharyngeal Cancer; Pheochromocytoma; Pineal Parenchymal Tumors of Intermediate Differentiation; Pineobiastorna and Supratentorial Primitive Neuroectodermal Tumors; Pituitary Tumor; Plasma Cell Neoplasm/Multiple Myeloma; Pieuropulmonary Blastoma; Primary Central Nervous System Lymphoma; Prostate Cancer; Rectal Cancer; Renal Cell (Kidney) Cancer; Renal Peivis and Ureter, Transitional Cell Cancer; Respiratory Tract Carcinoma involving the NUT Gene on Chromosome 15; Retinoblastoma; Rhabdomyosarcoma; Salivary Gland Cancer; Sarcoma, Ewing Family of Tumors; Sarcoma, Kaposi; Sarcoma, Soft Tissue; Sarcoma, Uterine; Sezary Syndrome; Skin Cancer (Nonmelanoma); Skin Cancer (Melanoma); Skin Carcinoma, Merkel Cell; Small Ceil Lung Cancer; Small Intestine Cancer; Soft Tissue Sarcoma; Squamous Cell Carcinoma, Squamous Neck Cancer with Occult. Primary, Metastatic; Stomach (Gastric) Cancer; Supratentorial Primitive Neuroectodermal Tumors; I -Cell Lymphoma, Cutaneous; Testicular Cancer; Throat Cancer; Thymoma and Thymic Carcinoma; Thyroid Cancer; Transitional Ceil Cancer of the Renal Pelvis and Ureter; Trophoblastic Tumor, Gestational; Urethral Cancer; Uterine Cancer, Endometrial; Uterine Sarcoma; Vaginal Cancer; Vulvar Cancer; Waldenstrom Macroglobullnemia; and Wilms Tumor

[0128] Further disclosed herein are methods for treating cancer in a human or mammal, comprising co-administering to a human or mammal, together with one or more chemotherapeutic agents or chemotherapeutic compounds, one or more of the disclosed HiF-la prolyl hydroxylase inhibitors.

[0129] Also disclosed herein are methods for treating a human or mammal diagnosed with cancer, co-administering to a human or mammal, together with one or more chemotherapeutic agent or chemotherapeutic compound one or more of the disclosed HiF-la prolyl hydroxylase inhibitors. [0130] Chemotherapeutic cancer agents that can be used in combination with the disclosed HiF-lct inhibitors include, but are not limited to, mitotic inhibitors (vinca alkaloids). These include vincristine, vinblastine, vindesine and Mavelbine™ (vinore!bine,5'-noranhydrobiastine) In yet other embodiments, chemotherapeutic cancer agents include topolsomerase i inhibitors, such as camptothecin compounds.

As used herein, "’camptothecin compounds" include Camptosar™ (irinotecan HCL), Hycamtin™ (topotecan HCL) and other compounds derived from camptothecin and its analogues. Another category of chemotherapeutic cancer agents that may be used in the methods and compositions disclosed herein are podophyliotoxin derivatives, such as etoposide, teniposide and mitopodozide. The present disclosure further encompasses other chemotherapeutic cancer agents known as alkylating agents, which alkylate the genetic material in tumor ceils. These include without limitation eisp!atin, cyclophosphamide, nitrogen mustard, trlmethyiene thiophosphoramide, carmustine, busuifan, chSorambucil, belustine, uracil mustard, ehlomaphazin, and dac.arhaz.ine. The disclosure encompasses antimetabolites as chemotherapeutic agents. Examples of these types of agents include cytosine arabinoside, fluorouracil, methotrexate, mercaptopurine, azathioprime, and procarbazine. An additional category of chemotherapeutic cancer agents that may be used in the methods and compositions disclosed herein include antibiotics. Examples include without limitation doxorubicin, bleomycin, dactmomyein, daunorubicin, mithramycin, mitomycin, mytomycin C, and daunomycin. There are numerous liposomal formulations commercially available for these compounds. The present disclosure further encompasses other chemotherapeutic cancer agents including without limitation anti tumor antibodies, dacarbazine, azacytidine, amsacrine, meiphaian, ifosfamide and mitoxantrone.

[0131] The disclosed HiF-la prolyl hydroxylase Inhibitors herein can be administered in combination with other anti-tumor agents, including cytotoxic/antineoplastic agents and anti -angiogenic agents. Cytotoxic/anti- neoplastic agents are defined as agents which attack and kill cancer cells. Some cytotoxic/anti-neopiastic agents are alkylating agents, which alkylate the genetic material in tumor ceils, e.g., cis-platin, cyclophosphamide, nitrogen mustard, trimethyiene thiophosphoramide, carmustine, busuifan, chlorambucil, belustme, uracil mustard, cbiomaphazin, and dacabazine. Other cytotoxic/anti-neopiastic agents are antimetabolites for tumor cells, e.g., cytosine arabinoside, fluorouracil, methotrexate, mercapfopuirine, azathioprime, and procarbazine. Other cytotoxic/anti-neoplastic agents are antibiotics, e.g., doxorubicin, bleomycin, dactinomycin, daunorubicin, mithramycin, mitomycin, mytomycin €, and daunomydn. There are numerous liposomal formulations commercially available for these compounds. Still other cytotoxic/anti-neoplastic agents are mitotic inhibitors (vinca alkaloids). These include vincristine, vinblastine and etoposide. Miscellaneous cytotoxic/anti-neoplastic agents include taxol and its derivatives, L-asparaginase, anti tumor antibodies, dacarbazine, azacytidine, amsacrine, meipbaian, VM-26, ifosfamide, mitoxantrone, and vindesine.

[0132] Anti-angiogenic agents are well known to those of skill in the art. Suitable anti- angiogenic agents for use in the disclosed methods and compositions include anti- VEGF antibodies, including humanized and chimeric antibodies, anti-VFGF aptamers and antisense oligonucleotides. Other known inhibitors of angiogenesis include angiostatin, endostatin, interferons, interleukin 1 (including a and b) interleukin 12, retinoic acid, and tissue inhibitors of metaSioproteinase-l and -2. (TIMP-I and -2). Small molecules, including topolsomerases such as razoxane, a topoisomerase II inhibitor with anti-angiogenic activity, can also be used.

[0133] Other anti-cancer agents that can be used in combination with the disclosed HIF-lo. inhibitors include, but are not limited to: acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin; altretamine; amhomydn; ametantrone acetate; aminoglutethimide; amsacrine; anastrozo!e; anthramycin; asparaginase; asperlin; azacitidine; azet.epa; az.otomyc.in; batimastat; henzodepa; bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate; brequinar sodium; bropirimine; busuifan; cactinomycin; caiusterone; caracemide; carbetimer; carboplatin; carmustine; carubicin hydrochloride; carzeiesin; cedefingol; chlorambucil; drolemydn; dspiatin; cladribine; crisnatol mesylate; cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicin hydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; docetaxei; doxorubicin; doxorubicin hydrochloride; droloxifene; droioxifene citrate; dromostanoione propionate; duazomycin; edatrexate; eflornithine hydrochloride; eSsamitrucin; enloplatin; enpromate; epipropidine; epirubicin hydrochloride; erbulozole; esorubicin hydrochloride; estramustine; estramustine phosphate sodium; etanidazoie; etoposide; etoposide phosphate; etoprine; fadrozoie hydrochloride; fazarabine; fenretinide; floxuridine; fiudarabine phosphate; fluorouracil; f!uorocitabine; fosquidone; fostrlecin sodium; gemcitabine; gemcitablne hydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide; iimofosine; interleukin ii (including recombinant interleukin II, or rlL2), interferon aiia-2a; interferon aifa-2b; interferon alfa-nl; interferon alfa-nB; interferon beta-i a; interferon gammad b; iproplatin; irinotecan hydrochloride; ianreotide acetate; !etrozole; ieuproiide acetate; liarozole hydrochloride; lometrexoi sodium; iomustine; losoxantrone hydrochloride; masoprocoi; maytansine; mechiorethamine hydrochloride; rnegestroi acetate; meiengestroi acetate; rneiphaian; menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogii!in; mitomaidn; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole; nogaiamycin; ormapiatin; oxisuran; paciitaxel; pegaspargase; peliomycin; pentamustine; pepiomycin sulfate; perfosfamide; pipobroman; piposuifan; piroxantrone hydrochloride; piicamycin; piomestane; porflmer sodium; porfiromycin; prednimustine; procarbazine hydrochloride; puromycin; puromycin hydrochloride; pyrazofurin; riboprine; rog!etimide; safingol; safingoi hydrochloride; semustine; simtrazene; sparfosate sodium; sparsornycin; spirogerrrtanium hydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin; su!ofenur; talisomydn; tecogalan sodium; tegafur; teioxantrone hydrochloride; temoporfin; teniposide; teroxirone; testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifene citrate; trestoione acetate; triciribine phosphate; trimetrexate; trimetrexate glucuronate; triptoreiin; tubulozole hydrochloride; uracil mustard; uredepa; vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate; vindesine; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate; vin!eurosine sulfate; vinorelbine tartrate; vinrosidlne sulfate; vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorublcin hydrochloride. Other anti-cancer drugs include, but are not limited to: 20-epi-l,25 dihydroxyvitamin D3; 5-etbynyluracil; abiraterone; adarubicin; acylfulvene; adecypenoi; adozelesin; aldesleukin; ALL-TK antagonists; aitretamine; ambamustine; amidox; amifostine; aminolevulinic acid; arnrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis inhibitors; antagonist D; antagonis G; antarelix; anti-dorsalizing morphogenetic protein-1; antiandrogen, prostatic carcinoma; antiestrogen; antineoplaston; antisense oligonucleotides; aphidicoiin glycinate; apoptosis gene modulators; apoptosis regulators; apurmic acid; ara-CDP-DL- PTBA; arginine deaminase; asuiacrlne; atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatin ill derivatives; balanol; batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactam derivatives; beta-alethine; betadamydn B; betulinic acid; bFGF inhibitor; bica!utamide; bisantrene; bisaziridinylspermine; bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine; caicipotrioi; ca!phostin C; camptothecin derivatives; canarypox IL-2; capecitabme; carboxamide-amino-triazole; carboxyamidotriazole; CaR.est M3; CAR.N 700; cartilage derived inhibitor; carzeiesin; casein kinase inhibitors (1COS); castanospermme; cecropin B; cetroreiix; chlorins; chloroquinoxaline sulfonamide; cicaprost; ds-porphyrin; cladribine; clomifene analogues; clotrimazole; collismycin A; co!lismydn B; combretastatin A4; combretastatin analogue; eonagenin; crambesddln 816; crisnatol; eryptophycin 8; cryptophydn A derivatives; curacin A; cydopentanthraquinones; cydoplatam; cypemydn; cytarabine ocfosfate; cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamii; diaziquone; didemnin B; didox; diethylnorspermine; dihydro-S- azacytidine; dihydrotaxol, 9-; dioxamycin; diphenyl spiromustine; docetaxel; docosanol; dolasetron; doxlfluridine; droloxlfene; dronabinol; duocarmydn SA; ebselen; ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur; epirubicln; epristeride; estramustine analogue; estrogen agonists; estrogen antagonists; etanidazoie; etoposide phosphate; exemestane; fadrozoie; fazarabine; fenretinide; filgrastim; finasteride; flavopiridoi; flezelastine; fiuasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex; formestane; fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate; gaiocitabine; ganireiix; gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsuifam; heregulin; hexamethyiene bisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene; idramantone; ilmofosine; ilomastat; imidazoacridones; imiquimod; irnmunostimuiant peptides; msu Si n-Uke growth factor-1 receptor inhibitor; interferon agonists; interferons; interleukins; iobenguane; iododoxorubiein; ipomeano!, 4-; iroplact; irsogladine; isobengazole; isohomoha!icondrin B; itasetron; jasplakinolide; kahaialide F; SameSiarin- N triacetate; ianreotide; leinamycin; lenograstim; lentinan sulfate; leptoistatin; letrozole; leukemia inhibiting factor; leukocyte alpha interferon; leupro!lde- estrogen÷progesterone; leuprore!in; levamisole; liarozole; linear polyamine analogue; lipophilic disaccharide peptide; lipophilic platinum compounds; lissoc!inamide 7; !ohapiatin; lombricine; lometrexoi; lonidamine; losoxantrone; lovastatin; loxoribine; lurtoteean; lutetium texaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A; marimastat; masoprocol; maspin; matriiysin inhibitors; matrix metalloproteinase inhibitors; menogarii; merbarone; metere!in; metbioninase; metoclopramide; !F inhibitor; mifepristone; miitefosine; mirimostim; mismatched double stranded RNA; mitoguazone; mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonal antibody, human chorionic gonadotrophin; monophosphoryl lipid A-rmyo bacterium cell wail sk; mopidamol; multiple drug resistance gene inhibitor; multiple tumor suppressor l-hased therapy; mustard anticancer agent; mycaperoxide B; mycobacterial cell wall extract; myriaporone; N-acety!dlnaline; -substituted benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatln; nemorubicin; neridronic acid; neutral endopeptidase; nilutamide; nisamycin; nitric oxide modulators; nitroxlde antioxidant; nitrullyn; 06- benzylguanlne; octreotide; okicenone; oligonucleotides; onapristone; ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin; osaterone; oxaliplatin; oxaunomycin; paclitaxel; paciitaxel analogues; paditaxel derivatives; pa!auamine; palmitoylrhizoxin; pamidronic acid; panaxytrioi; panomifene; parabactin; pazelliptine; pegaspargase; peidesine; pentosan polysulfate sodium; pentostatin; pentrozoie; perflubron; perfosfamide; periiiyl alcohol; phenazinomycin; pheny!acetate; phosphatase inhibitors; pidbanil; pilocarpine hydrochloride; pirarubicin; piritrexim; piacetin A; plac.et.in B; plasminogen activator inhibitor; platinum complex; platinum compounds; platinum-triamine complex; porfimer sodium; porfiromycin; prednisone; propyl bis-acridone; prostaglandin J2; proteasome inhibitors; protein A based immune modulator; protein kinase C inhibitor; protein kinase C inhibitors, microalgal; protein tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurins; pyrazo!oacridine; pyridoxyiated hemoglobin polyoxyethylene conjugate; raf antagonists; ra!titrexed; ramosetron; ras farnesyl protein transferase inhibitors; ras inhibitors; ras-GAP inhibitor; reteiiiptine demethy!ated; rhenium Re 186 etidronate; rhizoxin; ribozymes; Rii retinamide; rogletimide; rohitukine; romurtide; roquinimex; rubiginone Bl; ruboxyl; safingol; saintopin; SarCIMU; sarcophytol A; sargramosti ; Sdi 1 mimetics; semustine; senescence derived inhibitor 1; sense oligonucleotides; signal transduction inhibitors; signal transduction modulators; single chain antigen binding protein; sizofuran; sobuzoxane; sodium borocaptate; sodium pheny!acetate; solveroi; somatomedin binding protein; sonermin; sparfosic acid; spicamycin D; spiromustine; spienopentin; spongistatin 1; squalamine; stem ceil inhibitor; stem-cell division inhibitors; stipiamide; stromelysin inhibitors; sulfinosine; superactive vasoactive intestinal peptide antagonist; suradista; suramin; swainsonine; synthetic giycosaminoglycans; taiiimustine; tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur; te!!urapyry!ium; telomerase inhibitors; temoporfin; temozolo ide; teniposide; tetrachiorodecaoxide; tetrazomine; thaiibiastine; thiocora!ine; thrombopoietin; thrombopoietin mimetic; thymaifasin; thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocene bichloride; topsentin; toremifene; totipotent stem ceil factor; translation inhibitors; tretinoin; triacetyiuridine; triciribine; trimetrexate; triptoreiin; tropisetron; turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenital sinus-derived growth inhibitory factor; urokinase receptor antagonists; vapreotide; varioiin B; vector system, erythrocyte gene therapy; veiaresoi; veramine; verdins; verteporfin; vinore!bine; vinxaitine; vitaxin; vorozole; zanoterone; zenipiatin; zilascorb; and zinostatin stimaiamer. in one embodiment, the anti-cancer drug is 5-f!uorouraei!, taxoi, or ieucovorin

Methods Related to Treatment of Conditions Invoiving Microorganisms [0134] Disclosed is a method for prophylacticaliy treating a human or a mammal against infection by a microorganism, comprising administering to a human or mammal an effective amount of one or more of the disclosed HIF-Ia prolyl hydroxylase inhibitors.

[0135] Further disclosed is a method for decreasing the virulence of a microorganism when a human or a mammal is infected with a microorganism, comprising administering to a human or mammal an effective amount of one or more of the disclosed HIF-la prolyl hydroxylase inhibitors.

[0136] Yet further disclosed is a method for treating an infection in a human or mammal caused by a microorganism, comprising administering to a human or mammal an effective amount of one or more of the disclosed HIF -la prolyl hydroxylase inhibitors.

[0137] Still further disclosed is a method for treating a human or mammal diagnosed with an infection caused by a microorganism, comprising administering to a human or mammal an effective amount of one or more of the disclosed HIF-la prolyl hydroxylase inhibitors.

[0138] Also disclosed is a method for preventing transmission of a disease caused by a microorganism from a human or mammal to a human or mammal, comprising administering to a human or mammal an effective amoun t of one or more of the disclosed HIF-la prolyl hydroxylase inhibitors.

[0139] Still yet further disclosed is a method for preventing infection of a human or a mammal during a surgical procedure, comprising administering to a human or mammal an effective amount of one or more of the disclosed HIF-la prolyl hydroxylase inhibitors. [0140] The microorganism can be any benign or virulent microorganism, for example, bacteria, viruses, yeasts, fungi, or parasites. The following are non-limiting examples of microorganisms that can be affected by the disclosed H IF- let prolyl hydroxylase inhibitors. By the term "affected" is meant, the virulence of the microorganism is reduced, diminished or eliminated. The cause of the reduction, diminishment, or elimination of the virulence can be from the stabilization of H!F-1 and/or from the increased level of phagocytosis due to the administration of one or more of the disclosed H!F-lct prolyl hydroxylase inhibitors.

[0141] Acinetobacter caicoaceticus , Acinetobocter haemoiyticus , Aeromonas hydrophilic i, Agrobacterium iumefaciens , Badlius anthracis, Badl!us haiadurans, Bacillus suhtilis , Bocteroldes distasonis , Bacteroides eggerthii, Bacteroides fragiiis, Bacteroides ovolus , Bacteroides B452A homology group, Bacteroides spianchnicus , Bacteroides thetaiotaomicron , Bacteroides uniformi , Bacteroides vuigatus , Bordeteiia bronchiseptica , Bordeteiia parapertussis , Bordeteiia pertussis , Barre!ia burgdorferi , Branhameila catarrhalis, Brucella melitensls, Burkhoideria cepacia , Burkhoideria pseudomoliei , Campylobacter coil Campylobacterfetu , Campylobacter jejuni, Caulobacter crescentus, Citrobacter freundii, Clostridium difficile, Clostridium perftingens , Carynebacterium diphtherias, Corynebacterium giutamicum , Corynebacterium u!cerans, Edwardsieiia tarda, Enterobacter aerogenes, Erwinia chrysanthemi, Enterobacter cloacae , Enterococcus faecaiis, Enterococcus faecium, Escherichia coii, Frandseiia tularensis, Gardnerelia vaginalis, Haemophilus ducreyi, Haemophilus haemoiyticus , Haemophilus influenzae, Haemophilus parahaemolyticus, Haemophilus parainfluenzas, Helicobacter pylori, Klebsiella oxytoca, Klebsiella pneumoniae, Kiuyvera cryocrescens, Legionella pneumophila. Listeria innocua. Listeria monocytogenes, Listeria welshimeri, Methanosardna acetivorans, Methanosarcina mazes, Morganeiia morganii, Mycobacterium avium , Mycobacterium introceiiuiare, Mycobacterium ieprae, Mycobacterium tuberculosis, Mesorhizobium loti, Neisseria gonorrhoeas, Neisseria meningitidis, Pasteure!ia haemoiytica, Pasteurelia muitocida, Providencia alcalifaciens, Providencia rettgeri, Providencia stuartii, Proteus rnirabilis, Proteus vulgaris. Pseudomonas acidovorans, Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonasfluorescens, Pseudomonas putida , Raistonia soionacearum , Salmonella enterica subsp. enteridtidis , Salmonella enterica subsp. paratyphi, Salmonella enterica, subsp. typhlmurium , Salmonella enterica, subsp. typhi, Serratla marcescen , Shigella dysentenae, Shigella flexnerl, Shigella sonnei, Sinorhlzobium meiiioti , Staphylococcus aureus , Streptococcus criceti, Staphylococcu epldemmidis , Staphylococcus haemolyticus , Staphylococcus hominis , Staphylococcus hyicu , Staphylococcus intermedins , Stenotrophomonas rnaltophi!ia , Staphylococcus saccharoiyticus, Staphylococcus saprophyticus, Staphylococcus sciuri, Streptomyces avermltiiis , Streptomyces coelicoior, Streptococcus agalactiae , Streptococcus pneumoniae , Streptococcus pyogenes Suifohaibiobus soffiataricus. Thermotoga maritime , Vibrio choierae , Vibrio parahaemolyticus , Vogeseiia indigofera Xanthomonas axonopodis , Xanthamanas campestris , Yersinia enterocoiltica , Yersinia intermedia , Yersinia pestis, and Yersinia pseudotuhercuiosis.

Methods Re ating to Vaccination or Inoculation

[0142] Disclosed herein are methods for enhancing the effectiveness of a vaccine, comprising co-administering to a human or mammal a vaccine in combination with one or more H1F-Ia prolyl hydroxylase inhibitors. Non -limiting examples of vaccines are those for stimulating antibodies against hepatitis, influenza, measles, rubella, tetanus, polio, rabies, and the like. Therefore, the disclosed methods includes administering, or in the case of contacting cells in vitro, in vivo or ex vivo, the one or more HIF-Ia prolyl hydroxylase inhibitors and any co-administered compounds topically, buccaiiy, orally, Intradermally, subcutaneously, mucosally in the eye, vagina, rectum, and nose, intravenously, and intramuscularly.

[0143] Compounds having the structure of any one of Formulas (I), (I la) or (Mb), may be synthesized using standard synthetic techniques known to those of skill in the art. To this end, the reactions, processes and synthetic methods described herein are not limited to the specific conditions described in the following experimental section, but rather are intended as a guide to one with suitable skill in this field. For example, reactions may be carried out in any suitable solvent, or other reagents to perform the transformation^] necessary. Generally, suitable solvents are protic or aprotic solvents which are substantially non-reactive with the reactants, the intermediates or products at the temperatures at which the reactions are carried out (i.e., temperatures which may range from the freezing to boiling temperatures). A given reaction may be carried out in one solvent or a mixture of more than one solvent. Depending on the particular reaction, suitable solvents for a particular work-up following the reaction may be employed.

[0144] Unless otherwise indicated, conventional methods of mass spectroscopy (MS), liquid chromatography-mass spectroscopy (LCMS), NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques, and pharmacology are employed. Compounds are prepared using standard organic chemistry techniques such as those described in, for example, March's Advanced Organic Chemistry, 7th Edition, John Wiley and Sons, Inc (2013). Alternate reaction conditions for the synthetic transformations described herein may be employed such as variation of solvent, reaction temperature, reaction time, as well as different chemical reagents and other reaction conditions. As necessary, the use of appropriate protecting groups may be required. The incorporation and cleavage of such groups may be carried out using standard methods described in Peter G. M. Wuts and Theodora W. Green, Protecting Groups in Organic Synthesis, 4th Edition, Wiley-lnterscience. (2006). All starting materials and reagents are commercially available or readily prepared.

Methods for Making the Compounds of Formula (I)

[014S] Compounds of Formula (I) may be prepared from known or readily prepared starting materials, following methods known to one skilled in the art of organic synthesis. Methods useful for making the Compounds of Formula (I) are set forth in the Examples below and generalized in Schemes A and B below. Alternative synthetic pathways and analogous structures will be apparent to those skilled in the art of organic synthesis.

Scheme A: Preparation of Pyridazinones of formula lla

[0146] Commercially available 4,5-dibromopyridazin-B(2H)-one can be alkylated on the N-2 nitrogen by reaction with an alkyl halide (Z-W-X) in the presence of a base such as potassium carbonate in DMF, followed by reaction with sodium methoxide in methanol at room temperature to provide 5-bromo-4-methoxy-2-alkylpyridazin-3(2H)- ones. These intermediate halides can be cross-coupled with a reagent such as an alkyl tetrafluoroborate salt (Y-T-BF3 K⁺) in the presence of a catalytic palladium source such as Pd(OAc)₂ and a ligand such as 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (Xphos) and a base such as cesium carbonate at elevated temperatures.

Demethylation using standard methodologies, such as NaSCH3 in water at elevated temperatures, provides target pyridazinones of formula I la.

Scheme B: Preparation of Pyrimidinones of formula Mb

[0147] Commercially available 5-hydroxypyrimidin-4(3H)-one can be alkylated by reaction with an alkyl halide (Z-W-X) in the presence of a base such as potassium carbonate or triethylamine in DMF, followed by halogenation with a halogenating agent such as N-iodosuccinimide in DMF to provide 3-alkyl-5-hydroxy-6-halopyrimidin- 4(3H)-one intermediates. These can be cross-coupled with a reagent such as an alkyl tetrafluoroborate salt (R²BF3 K⁺) in the presence of catalytic palladium source such as Pd(OAc)2 and a ligand such as 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (Xphos) and a base such as cesium carbonate at elevated temperatures to provide target pyrimidinones of formula Mb.

EXAMPLES

PREPARATION OF COMPOUNDS

General Methods

[0148] Solvents, reagents, and intermediates that are commercially available were used as received. Reagents and intermediates that are not commercially available were prepared in the manner as described below. Flash column chromatography was performed using pre-packed normal phase silica from Biotage, Inc. or bulk silica from Fisher Scientific. Unless otherwise indicated, column chromatography was performed using a gradient elution of hexanes/ethyl acetate, from 100% hexanes to 100% ethyl acetate.

[0149] Example 1: Preparation of tert-butyi-4-((5-hydroxy-l-(4-methoxybenzyi)-6-oxo- l,6-dihydropyridazin-4-yl)methyl)piperazine-l-carboxylate

Step 1: Preparation of 4,5-dibromo-2-(4-methoxybenzyi)pyridazin-3(2H)-one

To a stirred solution of p-methoxybenzyl chloride (1.96 g, 12.5 mmol) in DMF (15 mL) was added K₂C0₃ (5.53 g, 40.0 m mol) at room temperature. After 5 min, 4,5- dibromopyridazin-3(2H)-one (2.54 g, 10.0 mmol) was added. The mixture was stirred at room temperature for 2 h and the reaction mixture was partitioned between water (20 mL) and EtOAc (100 mL). The organic layer was separated and washed with brine (2x20mL). The organic layer was separated, dried (Na₂S04), filtered and concentrated under reduced pressure. The crude residue was purified (silica gel; eluting with 15 %~25 % EtOAc in Petroleum Ether), to afford the title compound (2.70 g) as an off white solid. ^XH NMR (400 MHz, DMSO-d⁶) d 8.17 (s, 1H), 7.26 (d, J = 8.7 Hz, 2H), 6.89 (d, J = 8.6 Hz, 2H), 5.18 (s, 2H), 3.72 (s, 3H). LCMS (M+H) = 374.9. Step 2: Preparation of 5-bromo-4-methoxy-2-(4-methoxybenzyl)pyridazin-3(2H)-one

To a solution of 4,5-dibromo-2-(4-methoxybenzyl)pyridazin-3(2H)-one (1.12 g, 3.0 mmol) in 1,4-dioxane (15 mL) was added 30% NaOCH₃ in methanol (0.6 mL, 3.3 mmol). The mixture was stirred at room temperature for 1 h. The reaction mixture was partitioned between water (20 mL) and EtOAc (100 mL). The organic layer was separated and washed with brine (10 mL). The organic layer was separated, dried (Na₂S0₄), filtered and concentrated under reduced pressure to afford 5-bromo-4- methoxy-2-(4-methoxybenzyl)pyridazin-3(2H)-one (880 mg) as an off white solid. The crude product was used for the next step without purification. ¹H NMR (400 MHz, CDCI₃) d 7.79 (s, 1H), 7.37 (d, J = 8.7 Hz, 2H), 6.85 (d, J = 8.7 Hz, 2H), 5.21 (s, 2H), 4.25 (s, 3H), 3.78 (s, 3H). LCMS (M+H) = 325.0.

Step 3: Preparation oftert-butyi-4-((5-methoxy-l-(4-methoxybenzyi)-6-oxo-l,6- dihydropyridazin-4-yl)methyl)piperazine-l-carboxylate

To a stirred solution of 5-bromo-4-methoxy-2-(4-methoxybenzyl)pyridazin-3(2H)-one (325 mg, 1.0 mmol) in l,4-dioxane/H₂0 (10 mL/2 mL), was added potassium ((4-(tert- butoxycarbonyl)piperazin-l-yl)methyl)trifluoroborate (459.3 mg, 1.5 mmol), Cs₂C0₃ (977 mg, 3.0 mmol), Xphos (48 mg, 0.1 mmol ) and Pd(OAc)₂ (11 mg, 0.05 mmol). The mixture was stirred at 100°C for 16 h under nitrogen. The reaction mixture was partitioned between water (10 mL) and EtOAc (30 mL). The organic layer was separated and washed with brine (10 mL). The organic layer was separated, dried (Na₂S0 ), filtered and concentrated under reduced pressure. The crude residue was purified (silica gel; eluting with 50%~70% EtOAc in Petroleum Ether), to afford tert- butyl-4-((5-methoxy-l-(4-methoxybenzyl)-6-oxo-l,6-dihydropyridazin-4- yl)methyl)piperazine-l-carboxylate (300 mg) as an off-white solid. ^XH NMR (400 MHz, CDCI₃) 6 7.82 (s, 1H), 7.40 (d, J = 8.7 Hz, 2H), 6.86 (d, J = 8.7 Hz, 2H), 5.23 (s, 2H), 4.13 (s, 3H), 3.78 (s, 3H), 3.40 (m, 6H), 2.39 (m, 4H), 1.45 (s, 9H). LCMS (M+H) = 445.3.

Step 4: Preparation of tert-butyl-4-((5-hydroxy-l-(4-methoxybenzyl)-6-oxo-l,6- dihydropyridazin-4-yl)methyl)piperazine-l-carboxylate (Compound 1A)

Boc

Compound 1A

To a stirred solution of tert-butyl-4-((5-methoxy-l-(4-methoxybenzyl)-6-oxo-l,6- dihydropyridazin-4-yl)methyl)piperazine-l-carboxylate (180 mg, 0.4 mmol) in DMF (5 mL), was added (20%) CH₃SNa (70 mg, 2.0 mmol). The mixture was stirred at 140°C for 2 h, cooled to ambient temperature and then partitioned between water (10 mL) and EtOAc (30 mL). The organic layer was separated and washed with brine (2xl0mL). The organic layer was separated, dried (NazSC ), filtered and concentrated under reduced pressure. The crude residue was purified Pre-TLC eluted with MeOH: DCM=1:15 to afford Compound 1A (61 mg) as an off-white solid. ¹H NMR (400 MHz, CD₃OD) d 7.78 (s, 1H), 7.23 (d, J = 8.2 Hz, 2H), 6.63 (d, J = 7.6 Hz, 2H), 5.19 (s, 2H), 3.92 (s, 2H), 3.69 (s, 3H), 3.47 (m, 4H), 2.85 (m, 4H), 1.45 (s, 9H). LCMS (M+H) - 431.3.

The compounds in the table below were prepared in a manner as described for

Compound 1A in Example 1 above by substituting the appropriate alkylating agent in Step 1.

[0150] Example 2: Preparation of 3-(4-((4-(tert-butoxycarbonyl)piperazin-l-yl)methyl)- 5-hydroxy-6-oxopyridazin-l(6H)-yl)propanoic acid

Compound 1J Compound 1M

To a stirred solution of methyl 3-(4-((4-(tert-butoxycarbonyl)piperazin-l-yl)methyl)-5- hydroxy-6-oxopyridazin-l(6H)-yl)propanoate (39.7mg, 0.1 mmol) in THF (lmL) and methanol (lmL) was added lithium hydroxide solution (0.3mL, IN) and the mixture stirred for 16h at room temperature. The solvents were removed under reduced pressure and the residue purified by reverse phase HPLC. Pure fractions were lyophilized to afford Compound 6A as a white solid. 1H NMR (DMSO-d⁶) d 7.74 (s, 1H), 4.24 (t, J=7.1Hz, 2H), 3.38 (s, 2H), 3.30 (s, 4H), 2.69 (T, J=7.1Hz, 3H), 2.33 (s, 4H), 1.39 (s, 9H). LCMS (M+H) = 383.1.

The compounds in the table below were prepared in a manner as described for Compound 1M in Example 2 above, i.e. by hydrolysis of compound II to provide compound IN; compound IK to provide compound 10; and compound 1L to provide compound IP.

[0151] Example 3: Preparation oftert-butyi-4-((5-hydroxy-l-(4-methoxybenzyi)-6-oxo- l,6-dihydropyrimidin-4-yl)methyl)piperazine-l-carboxylate (Compound 2A)

Step 1. Preparation of 5-hydroxy-3-(4-methoxybenzyl)pyrimidin-4(3H)-one

To a stirred solution of 5-hydroxypyrimidin-4(3H)-one (2.00 g, 17.8 mmol) in DMF (15 mL) was added TEA (5.40 g, 53.5 mmol) and l-(chloromethyl)-4-methoxybenzene (3.35 g, 21.4 mmol). The mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated and purified (silica gel; eluting with 25 %~35 % EtOAc in Petroleum Ether) to afford 5-hydroxy-3-(4-methoxybenzyl)pyrimidin-4(3H)-one (710 mg) as an off white solid. ^XH NMR (400 MHz, DMSO-d⁶) d 9.62 (s, 1H), 8.19 (s, 1H), 7.43 (s, 1H), 7.31 (d, J = 8.7 Hz, 2H), 6.90 (d, J = 8.7 Hz, 2H), 5.04 (s, 2H), 3.72 (s, 3H). LCMS (M+H) = 233.1.

Step 2. Preparation of 5-hydroxy-6-iodo-3-(4-methoxybenzyl)pyrimidin-4(3H)-one

To a stirred solution of 5-hydroxy-3-(4-methoxybenzyl)pyrimidin-4(3H)-one (505 mg, 2.2 mmol) in DMF (10 mL) was added N-iodosuccinamide (587 mg, 2.6 mmol). The mixture was stirred at room temperature for 2 h. The reaction mixture was purified by a reverse column with 25% acetonitrile in water. The fractions were collected and concentrated to afford 5-hydroxy-6-iodo-3-(4-methoxybenzyl)pyrimidin-4(3H)-one (560 mg) as an off white solid. *H NMR (400 MHz, DMSO-d⁶) d 10.47 (s, 1H), 8.08 (s, 1H), 7.31 (d, J = 8.6 Hz, 2H), 6.90 (d, J = 8.6 Hz, 2H), 5.03 (s, 2H), 3.72 (s, 3H). LCMS (M+H) = 358.9.

Step 3. Preparation of tert-butyl-4-((5-hydroxy-l-(4-methoxybenzyl)-6-oxo-l,6- dihydropyrimidin-4-yl)methyl)piperazine-l-carboxylate (Compound 2A)

To a stirred solution of compound 5-hydroxy-6-iodo-3-(4-methoxybenzyl)pyrimidin- 4(3H)-one (185 mg, 0.52 mmol) in THF (5 mL) was added water (0.5 mL), potassium (4- tert-butoxycarbonylpiperazin-l-yl)methyltrifluoroborate (316 mg, 1.03 mmol), CS₂CO₃ (505 mg, 1.55 mmol), Pd(OAc)₂ (6 mg, 0.026 mmol) and XPhos (25 mg, 0.052 mmol). The mixture was stirred for 16 h at 70°C under nitrogen atmosphere. The mixture was concentrated, and the residue purified by reverse phase column chromatography with 45% acetonitrile in water. The fractions were collected and concentrated to afford Compound 2A (70 mg) as an off white solid. ¹H NMR (400 MHz, CD₃OD) d 8.04 (s, 1H), 7.32 (d, J = 8.6 Hz, 2H), 6.89 (d, J = 8.5 Hz, 2H), 5.11 (s, 2H), 3.77 (s, 3H), 3.62 (s, 2H), 3.45 (s, 4H), 2.63 - 2.44 (m, 4H), 1.45 (s, 9H). LCMS (M+H) = 431.2.

The compounds in the table below were prepared in a manner similar to that described for Compound 2A in Example 2 above by substituting the appropriate alkylating agent in Step 1.

NOTE: Carboxylic acids 2N and 2P were prepared in a manner similar to that described for the preparation of compound 1M.

The compounds in the table below were prepared in a manner similar to that described for Compound 2A in Example 2 above by substituting the appropriate coupling reagent in Step S.

(0152] Example 4: Preparation of methyl 3-(l-(4-chlorobenzyl)-5-hydroxy-6-oxo-l,6- dihydropyrimidin-4-yl)propanoate (Compound 2V)

Step 1. Preparation of methyl 3-(l-(4-chlorobenzyl)-5-hydroxy-6-oxo-l,6- dihydropyrimidin-4-yl)propenoate

To a stirred solution of 3-(4-chlorobenzyl)-5-hydroxy-6-iodopyrimidin-4(3H)-one (1.0 mmol) in DMF, in a dry 10 mL capacity Schlenk tube equipped with a magnetic stirrer bar was added methyl acrylate (1.1 mmol) and triethylamine (1.0 mL). Pd(PPh₃)₂CI₂ was added, and the reaction heated under stirring at 80°C for 6h. The mixture was then cooled, poured into water and extracted with DCM x3. The combined extracts were washed with brine then dried over Na₂SC>₄. Pure compound was isolated after column chromatography (97:3 DCM/CH₃OH). Step 2. Preparation of methyl 3-(l-(4-chlorobenzyl)-5-hydroxy-6-oxo-l,6- dihydropyrimidin-4-yl)propanoate

Compound 2V

To a solution of the methyl ester in ethyl acetate (lOmL) was added 10%Pd/C (50mg) and the mixture was stirred under a balloon of hydrogen gas for lh. The catalyst was removed by filtration through Celite and the filtrate evaporated and purified by reverse phase HPLC to afford the desired compound as a white solid. ¹H NMR (400 MHz, DMSO- d⁶) d 8. OB (s, 1H), 7.38-7.33 (m, 4H), 5.15 (s, 2H), 3.65 (s, 3H), 2.99 (t, J=7.6Hz, 2H), 2.67 (t, J=7.6Hz, 2H). LCMS (M+H) = 321.1.

[0153] Example 5: Preparation of methyl 4-((5-hydroxy-6-oxo-4-(piperazin-l- ylmethyl)pyrimidin-l(6H)-yl)methyl)benzoate TFA salt (Compound 2W)

Compound 2M Compound 2W

To a stirred solution of compound 2M (46 mg, 0.1 mmol) in dichloromethane (2 mL) was added TFA (0.5 mL) at room temperature. The mixture was stirred for 3h then concentrated and purified by HPLC to afford compound 3A (30 mg) as an off white solid after lyophilization. ^XH NMR (400 MHz, DMSO-d⁶) d 8.81 (s, 1H), 7.83 (d, J=8.0Hz, 2H), 7.77 (s, 1H), 7.38 (d, J = 8.0 Hz, 2H), 5.32 (s, 2H), 3.82 (s, 3H), 3.20 (m, 4H), 2.95 (m, 4H). LCMS (M+H) = 359.1.

IN VITRO ACTIVITY OF COMPOUNDS [0154] Example 6: PHD Enzyme Assay

HTRF assay to measure the inhibition of PHD1, PHD2 and PHD3 enzyme activity.

HIF-Ia peptides labeled with a Cy5 fluorophore (Cy5-DLEMLAPYIPMDDDFQL and Cy5- DLEMLA[hydroxy-P]YIPMDDDFQL) were prepared. Hydroxylation of the HIFl-a peptide at Proline 564 was measured using a primary antibody specific for hydroxylated HIF-la (Cell Signaling Technology, S4S4) and a Terbium-labeled secondary antibody (Cisbio,

61PARTAF), as follows: 4x concentrated test compounds were transferred to a 384 well low volume HTRF plate and pre-incubated with 4x concentrated PHD enzyme for 10 minutes at room temperature in buffer containing 50mM HEPES and 0.05% Triton X-1Q0 (lOnM final for PHD1, 5nM final for PHD2, and 5n final for PHD3). Control wells contained enzyme or buffer with vehicle 2x HIF-Ia peptide was prepared in assay buffer containing 50mM HEPES, 0.05% Triton X-1Q0, 2GuM Ferrous ammonium sulfate, 400uM Ascorbic Acid, and 4uM 2-OG and plated with compound and enzyme (24nM final peptide for PHD1 and PHD2, 5nM final peptide for PHDS). The reaction was incubated at room temperature for 30 minutes. An HTRF solution was prepared in 50mM HEPES with 0.05% Triton X containing 1:2000 anti-hydroxy HIF-la antibody, 1.5ng/wel! Terbium anti-rabbit antibody, and 2Q0uM Succinic Acid and plated at 1:2 to stop the enzymatic reaction. After a 1 hour incubation, the 384 well plate was read on a SpectraMax sD5 instrument using HTRF settings (The HTRF ratio is a ratio of acceptor emission at 665nm and donor emission at 616nm).

The compounds described herein were assayed as described above.

PHD IC50 values are provided for the compounds of the present invention in Table 2. With respect to PHD activity: "A" denotes and IC50 <0.5 uM, "B" denotes and IC50 of from 0.5uM - 5uM, and "C" denotes IC50 >5uM.

Table 2 : Activity of Representative Compounds

Compound No PHD1 PHD2 PHD3

2B C C B 2D B B A 2E C C C 2F C C C 2G C C C 2H C C C 21 C C C 2J C C B 2K C C C 2L C C C 2M C C B 2N C C B 20 C C C 2P C C C 2Q C C C 2R C C C 2S c c c 2T C C B 2U C C B 2 V C C B

[0155] This application claims the benefit of priority to U.S. Provisional Application No. 62/951,871, filed December 20, 2019, which application is hereby incorporated by reference in its entirety.

Claims

CLAIMS WHAT IS CLAIMED IS:

1. A compound of formula (I):

X is -N=CR^X- or -CR^X=N-;

R^x is H or halogen;

W is -CH₂-, -CH₂-CH₂-, -CH₂-CH₂-CH₂- or -CH₂-CH₂-CH₂-CH₂-;

Z is -COOH, -COOMe, -COOEt or

T is a bond, -CH₂- or -CH₂-CH₂-;

Y is -COOH, -COOMe, -COOEt or

2. A compound of claim 1, wherein R^x is H.

3. A compound of claim 1 or claim 2, wherein W is -CH2- .

4. A compound of any one of claims 1-3, wherein Z is

5. A compound of any one of claims 1-4, wherein R^z is halogen.

6. A compound of any one of claims 1-5, wherein n is 1 or 2.

7. A compound of any one of claims 1-6, wherein T is -CH2- .

8. A compound of any one of claims 1-7, wherein Y is

9. A compound of formula (lla):

R^x is H or halogen;

W is -CH2-, -CH2-CH2-, -CH2-CH2-CH2- or -CH2-CH2-CH2-CH2-;

Z is -COOH, -COOMe, -COOEt or

T is a bond or -CH2- ;

Y is -COOH, -COOMe, -COOEt or

10. A compound of formula (lib):

R^x is H or halogen;

W is -CH2-, -CH2-CH2-, -CH2-CH2-CH2- or -CH2-CH2-CH2-CH2-;

Z is -COOH, -COOMe, -COOEt or

T is a bond or -CH2- ;

Y is -COOH, -COOMe, -COOEt or

11. A compound having a structure listed in Table 1, or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof.

12. A pharmaceutical composition comprising the compound of any one of claims 1-11 or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof, and at least one pharmaceutically acceptable excipient.

13. Use of a compound of any one of claims 1-11 or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof in the manufacture of a medicament.

14. The use of claim 13, wherein the medicament is for the treatment of an inflammatory disease, cancer or an infection.

15. A method of inhibiting hypoxia-inducible factor 1-a (HIF-la) activity, comprising contacting the HIF-Ia with an effective amount of a compound of any one of claims 1- 11 or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof.

16. A method of inhibiting hypoxia-inducible factor 1-s (HIF-Ia) activity in a subject, comprising administering to the subject an effective amount of a compound of any one of claims 1-11 or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof.

17. A method of up regulating of phosphoglycerate kinase (PGK) activity, comprising contacting the PGK with an effective amount of a compound of any one of claims 1-11 or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof.

18. A method of increasing the immune response in a subject, comprising administering to the subject an effective amount of a compound of any one of claims 1-11 or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof.

19. The method of claim 18, wherein the subject has a medical condition causing a decreased cellular immunity.

20. The method of claim 19, wherein the medical condition is an inflammatory disease, cancer or an infection.

21. A method of treating a subject with an inflammatory epithelial disease, comprising administering to the subject an effective amount of a compound of any one of claims 1-11 or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof.

22. The method of claim 21, wherein the inflammatory epithelial disease is a disease affecting the Gl tract, the respiratory tract, mucosa, skin, the lining of major organs and endocrine glands or vascular tissue.

23. The method of claim 21, wherein the inflammatory epithelial disease is a disease affecting intestinal epithelial tissue.

24. The method of claim 21, wherein the inflammatory epithelial disease is Irritable Bowel Syndrome (IBS).

25. The method of claim 21, wherein the inflammatory epithelial disease is Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, diversion colitis, Behget's syndrome or indeterminate colitis.

26. A method of treating an infection in a subject, comprising administering to the subject an effective amount of a compound of any one of claims 1-11 or a pharmaceutically acceptable salt, solvate, hydrate, isomer, tautomer, racemate, or isotope thereof.

27. The method of claim 26, wherein the infection is caused by bacteria, a virus, a fungus, a yeast or a parasite.

28. The method of claim 26, wherein the infection is caused by a bacterium.

29. The method of claim 28, wherein the bacterium is Staphylococcus aureus, methiciliin resistant Staphylococcus aureus, Streptococcus pyogenes, Pseudomonas aeruginosa, or Adnetobacter baumannii.

30. The method of claim 26, wherein comprising administering an antimicrobial agent.