WO2021242758A1

WO2021242758A1 - Improved nitro-fatty acid oral dose regimens

Info

Publication number: WO2021242758A1
Application number: PCT/US2021/034058
Authority: WO
Inventors: Francisco SALVA; William W. Leong; Theodore Danoff; Robert N. Willette
Original assignee: Imara Inc.
Priority date: 2020-05-26
Filing date: 2021-05-25
Publication date: 2021-12-02

Abstract

The present disclosure provides methods of treating a variety of diseases, wherein the method comprises administering to a human subject an oral daily dose in an amount of at least 450 mg to 2,000 mg of compounds and compositions comprising esters and acids of nitro-containing fatty acids and esters. The present disclosure also provides pharmaceutical compositions and oral dosage units comprising acids and esters of nitro-containing fatty acids.

Description

IMPROVED NITRO-FATTY ACID ORAL DOSE REGIMENS CROSS-REFERENCE [0001] This application claims the benefit of U.S. Provisional Application No.63/030,107, filed May 26, 2020, which is incorporated herein by reference in its entirety. BACKGROUND [0002] CXA-10 (10-nitro-9(E)-octadec-9-enoic acid) is a specific isomer of nitro-oleic acid (OA- NO2) with electrophilic properties that facilitate rapid and reversible protein adduction reactions with cysteine and, to a lesser extent, histidine residues (Baker et al., J. Biol. Chem. 282(42):31085-93), but not DNA bases. Of particular relevance to its potential as a therapeutic agent, CXA-10 attaches covalently, but reversibly, to key signaling proteins that are involved in metabolic and inflammatory processes, thereby modulating their activity, leading to cellular and tissue-protective anti-oxidant, anti-inflammatory and anti-fibrotic effects. [0003] The major actions of CXA-10 are to (i) selectively adduct key cysteine residues (Cys273 and Cys288) of the Nrf2 inhibitor, Keap 1, causing release and stabilization of the Nrf2 transcription factor, leading to upregulation of expression of antioxidant and detoxifying genes and production of effector proteins; (ii) selectively adduct Cys38 of the p65 subunit of NF-KB, disrupting the TLR4 signaling complex to prevent the elaboration of pro-inflammatory mediators such as cytokines and chemokines, pro-fibrotic agents, and adhesion molecules; and (iii) selectively de-repress HSF1-driven expression of heat shock proteins, increasing the concentration of stress-protecting protein chaperones. A fourth pharmacological action of CXA- 10 is inhibition of xanthine oxidoreductase, which is one of the major enzymes involved in the production of reactive oxygen species (ROS), agents that cause much of the damage associated with oxidative stress pathology. At a systems level, CXA-10 up-regulates the major anti- inflammatory and reparative pathway of the body, Nrf2, while inhibiting the major pro- inflammatory pathway, NF-KB; upregulates production of cell-protective chaperone proteins; and reduces production of reactive oxygen species. [0004] The absorption, distribution, metabolism, and elimination (ADME) of CXA-10 after oral administration is both complex and unusual for a small molecule drug, presenting pharmacokinetic (“PK”) and pharmacodynamic (“PD”) challenges, especially following oral administration. In particular, the parallel routes of distribution, including circulation both as the free fatty acid and esterified within triglycerides within lipoprotein particles, sequestration in adipose tissues, and difficulties in quantitative detection in plasma, render standard dose-finding protocols poorly suited to determining an effective dosage regimen. [0005] As a consequence, the first repeat dose oral trial of CXA-10 was conducted in obese subjects, in the hope of providing a higher dynamic range of inflammatory biomarkers than would be found in nonobese healthy normal subjects. This study (CXA-10-202; NCT02460146) demonstrated that CXA-10 decreased levels of biomarkers associated with altered inflammatory and metabolic stress at a 150 mg daily oral dose, but not at 25 mg per day or 450 mg per day. See Garner et al., “Pharmacokinetic and pharmacodynamic effects of oral CXA-10, a nitro fatty acid, after single and multiple ascending doses in healthy and obese subjects,” Clin. Transl. Sci. 12:667-676 (2019) and US Pat. No.10,537,541, incorporated herein by reference in their entireties. [0006] There exists a need for CXA-10 oral dosage regimens that are effective to treat patients with chronic inflammatory diseases, and in particular, effective to treat patients in whom the ADME of CXA-10 may differ from that observed in obese normal subjects. SUMMARY [0007] The CXA-10-202 repeat oral dose phase 1 trial in obese healthy normal subjects demonstrated that CXA-10 exhibited classical hormesis: plotting biomarker response as a function of dose produced an inverted “U” shaped curve: the exposures achieved at both low doses and high doses had no beneficial effect. After 14 days of daily dosing, only the oral dose of 150 mg led to reduced serum levels of certain inflammatory biomarkers; neither the 25 mg dose nor 450 mg dose reduced levels of these biomarkers. Accordingly, the daily oral doses selected for the phase 2 FIRSTx trial (NCT03422510), a study of oral CXA-10 in primary focal segmental glomerulosclerosis (FSGS), were 75 mg, 75 mg with possible increase to 150 mg, 150 mg, and 150 mg with possible increase to 300 mg. Initial anecdotal data on reduction in proteinuria — a clinical measure of impaired renal function — suggested that the higher doses were more effective in reducing proteinuria in this patient population, and further suggested that clinically effective doses could even exceed the upper threshold of biomarker effect seen in the CXA-10-202 trial. [0008] Both the CXA-10-202 trial and earlier studies had suggested that oral doses of CXA-10 at or above 300 mg per day could cause gastrointestinal side effects, and that lack of tolerability might independently limit the maximum daily oral doses. A new phase 2 trial disclosed herein shows that such side effects are self-limiting and/or can be mitigated, permitting higher doses to be administered with therapeutic effect. [0009] Accordingly, the present disclosure provides methods of treating a variety of diseases, wherein the method comprises administering to a human subject an oral daily dose in an amount of at least 450 mg to 2,000 mg of CXA-10 and related compounds and compositions comprising esters and acids of nitro-containing fatty acids. The present disclosure also provides pharmaceutical compositions and oral dosage units comprising esters and acids of nitro- containing fatty acids [0010] Accordingly, in a first aspect, the present disclosure provides a method of treating a disease, comprising administering to a human subject with the disease an oral daily dose of a compound of formula (I),

or a salt, a stereoisomer, a deuterated analog, or a fluorinated analog thereof, in an amount of at least 450 milligrams and less than 2,000 milligrams per day, wherein: X is selected from H and

; a is from 0-30; b is from 0-30; R¹ is selected from H, alkyl, haloalkyl, cycloalkyl, aryl, and heteroaryl, -C(O)-R², gluconate, glycoside, glucuronide, tocopherols, and PEG groups; R² is selected from alkyl, haloalkyl, cycloalkyl, aryl, and heteroaryl; and at least one pharmaceutically acceptable carrier or excipient. DETAILED DESCRIPTION [0011] In some embodiments of the method, the daily oral dose is 450 mg to 1,800 mg. [0012] In some embodiments of the method, the daily oral dose is 450 mg or 600 mg. [0013] In some embodiments of the method, the daily oral dose is 450 mg. [0014] In some embodiments of the method, the daily oral dose is 600 mg. [0015] In some embodiments of the method, the daily oral dose is administered once a day. [0016] In some embodiments of the method, the daily oral dose is administered twice a day. [0017] In some embodiments of the method, the daily oral dose is administered for at least 7 days. [0018] In some embodiments of the method, X is

. [0019] In some embodiments of the method, R¹ is H, methyl, ethyl, propyl, isopropyl, butyl, sec- butyl, tert-butyl, or pentyl. [0020] In some embodiments of the method, R¹ is H. [0021] In some embodiments of the method, the compound of formula (I) is nitro-oleic acid. [0022] In some embodiments of the method, the compound of formula (I) is selected from:

[0023] In some embodiments of the method, the compound of Formula I is 10-nitro-9(E)- octadec-9-enoic acid (CXA-10), or a salt, a stereoisomer, a deuterated analog, or a fluorinated analog thereof. [0024] In some embodiments of the method, the compound of Formula I is 9-nitro-9(E)-octadec- 9-enoic acid (CXA-9), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof. [0025] In some embodiments of the method, the disease is selected from solid organ fibrosis, inflammatory disease, cardiovascular disease, renal disease, kidney failure, ischemic kidney injury, acute kidney injury (AKI), chronic kidney injury (CKI), chronic kidney disease (CKD), obesity associated chronic kidney disease, diabetic nephropathy, kidney fibrosis, progressive kidney disease (renal fibrosis), focal segmental glomerular sclerosis (FSGS), primary FSGS, secondary FSGS, sickle cell nephropathy, glomerulonephritis, nephrotic syndrome, nonalcoholic steatohepatitis (NASH), fatty liver disease, non-fatty liver disease (NFLD), pulmonary arterial hypertension (PAH), pulmonary fibrosis, psoriasis, allergic airway disease, obesity, anti- adipogenic disease, type II diabetes, lithium induced diabetes insipidus, sickle cell disease, sickle cell crisis, idiopathic pulmonary fibrosis (IPF), interstitial lung diseases (ILDs), scleroderma, inflammatory gastrointestinal disease, colitis, inflammatory bowel disease, neurodegenerative disease, amyotrophic lateral sclerosis (ALS), metabolic syndrome, neuropathy, Charcot-Marie- Tooth disease, and mitochondrial related disease. [0026] In some embodiments of the method, the human subject has sickle cell disease. [0027] In some embodiments of the method, the human subject has sickle cell crisis. [0028] In some embodiments of the method, the compound of Formula I is formulated in a pharmaceutical composition. [0029] In some embodiments of the method, the pharmaceutical composition comprises a triglyceride. [0030] In some embodiments of the method, the triglyceride is a medium chain triglyceride. [0031] ] In some embodiments of the method, the medium chain triglyceride is selected from hexanoic acid, octanoic acid, decanoic acid, and dodecanoic acid. [0032] In some embodiments of the method, the pharmaceutical composition comprises miglyol. [0033] In some embodiments of the method, the pharmaceutical composition further comprises an antioxidant. [0034] In some embodiments of the method, the antioxidant is butylated hydroxytoluene (BHT). [0035] In some embodiments of the method, the pharmaceutical composition further comprises a thickening agent. [0036] In some embodiments of the method, the thickening agent is fumed silica. [0037] In some embodiments of the method, the pharmaceutical composition is in an oral unit dosage form. [0038] In some embodiments of the method, the oral unit dosage form is a capsule. [0039] In some embodiments of the method, the capsule comprises hydroxypropyl methylcellulose (HMPC). [0040] In some embodiments of the method, the human subject is between 18 and 60 years of age. [0041] In some embodiments of the method, the human subject is male. [0042] In some embodiments of the method, the human subject is female. [0043] In some embodiments of the method, the human subject has a body mass index (BMI) between 18.0 and 32.0 kg/m². [0044] 3 In some embodiments of the method, the daily oral dose is administered to the human subject with a meal. [0045] In some embodiments of the method, the daily oral dose is administered to the human subject within 30 minutes of a meal. [0046] In some embodiments of the method, wherein the daily oral dose is administered to the human subject within 60 minutes of a meal. [0047] In some embodiments of the method, wherein the daily oral dose is administered to the human subject without a meal. [0048] In some embodiments of the method, the compound of formula (I) is 10-nitro-9(E)- octadec-9-enoic acid (CXA-10), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily dose is 450 mg; and the human subject has sickle cell disease. [0049] In some embodiments of the method, the compound of formula (I) is 10-nitro-9(E)- octadec-9-enoic acid (CXA-10), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily does is 600 mg; and the human subject has sickle cell disease. [0050] In some embodiments of the method, the compound of formula (I) is 10-nitro-9(E)- octadec-9-enoic acid (CXA-10), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof; the oral daily does is 450 mg; and the human subject has sickle cell crisis. [0051] In some embodiments of the method, the compound of formula (I) is 10-nitro-9(E)- octadec-9-enoic acid (CXA-10), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily does is 600 mg; and the human subject has sickle cell crisis. [0052] In some embodiments of the method, the compound of formula (I) is 9-nitro-9(E)- octadec-9-enoic acid (CXA-9), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily dose is 450 mg; and the human subject has sickle cell disease. [0053] In some embodiments of the method, the compound of formula (I) is 9-nitro-9(E)- octadec-9-enoic acid (CXA-9), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily does is 600 mg; and the human subject has sickle cell disease. [0054] In some embodiments of the method, the compound of formula (I) is 9-nitro-9(E)- octadec-9-enoic acid (CXA-9), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily does is 450 mg; and the human patient has sickle cell crisis. [0055] In some embodiments of the method, the compound of formula (I) is 9-nitro-9(E)- octadec-9-enoic acid (CXA-9), or a salt, stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily does is 600 mg; and the human patient has sickle cell crisis. [0056] In a second aspect, the present disclosure provides a pharmaceutical composition comprising a compound of Formula I,

or a salt, a stereoisomer, a deuterated analog, or a fluorinated analog thereof, wherein: X is selected from H and

; a is from 0-30; b is from 0-30; R¹ is selected from H, alkyl, haloalkyl, cycloalkyl, aryl, and heteroaryl, -C(O)-R², gluconate, glycoside, glucuronide, tocopherols, and PEG groups; R² is selected from alkyl, haloalkyl, cycloalkyl, aryl, and heteroaryl; and at least one pharmaceutically acceptable carrier or excipient. [0057] In some embodiments of the pharmaceutical composition, X is

[0058] In some embodiments of the pharmaceutical composition, R¹ is H, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, or pentyl. [0059] In some embodiments of the pharmaceutical composition, R¹ is H. [0060] In some embodiments of the pharmaceutical composition, the compound of formula (I) is oleic acid. [0061] In some embodiments of the pharmaceutical composition, the compound of formula (I) is nitro-oleic acid. [0062] In some embodiments of the pharmaceutical composition, the compound of formula (I) is selected from:

[0063] In some embodiments of the pharmaceutical composition, the compound of formula (I) is 10-nitro-9(E)-octadec-9-enoic acid (CXA-10), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof. [0064] In some embodiments of the pharmaceutical composition, the compound of formula (I) is 9-nitro-9(E)-octadec-9-enoic acid (CXA-9), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof. [0065] In some embodiments, the pharmaceutical composition further comprises a triglyceride. [0066] In some embodiments of the pharmaceutical composition, the triglyceride is a medium chain triglyceride. [0067] In some embodiments of the pharmaceutical composition, the medium chain triglyceride is selected from hexanoic acid, octanoic acid, decanoic acid, and dodecanoic acid. [0068] In some embodiments, the pharmaceutical composition comprises miglyol. [0069] In some embodiments, the pharmaceutical composition further comprises an antioxidant. [0070] In some embodiments of the pharmaceutical composition, the antioxidant is butylated hydroxytoluene (BHT). [0071] In some embodiments, the pharmaceutical composition further comprises a thickening agent. [0072] In some embodiments of the pharmaceutical composition, the thickening agent is fumed silica. [0073] In a third aspect, the present disclosure provides an oral unit dosage form, comprising: a capsule shell; and the pharmaceutical composition as described herein. [0074] In some embodiments of the oral dosage unit, the capsule shell comprises hydroxypropyl methylcellulose (HMPC). [0075] In some embodiments of the oral dosage unit, the capsule contains 25 mg to 300 mg of an active pharmaceutical ingredient (API). [0076] In some embodiments of the oral dosage unit, the capsule contains 150 mg of an active pharmaceutical ingredient (API). [0077] In some embodiments of the oral dosage unit, the API is a compound of formula (I). [0078] In some embodiments of the oral dosage unit, the API is 10-nitro-9(E)-octadec-9-enoic acid or 9-nitro-9(E)-octadec-9-enoic acid. [0079] In some embodiments of the oral dosage unit, the API is 10-nitro-9(E)-octadec-9-enoic acid. [0080] In some embodiments of the oral dosage unit, the API is 9-nitro-9(E)-octadec-9-enoic acid. DETAILED DESCRIPTION Definitions [0081] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. [0082] It is understood that the definitions provided herein are not intended to be mutually exclusive. Accordingly, some chemical moieties may fall within the definition of more than one term. [0083] As used herein, a dash ( " —") that is not between two letters or symbols refers to a point of bonding or attachment for a substituent. For example, -NH2 is attached through the nitrogen atom. [0084] As used herein, “alkyl”, as used herein, refers to an unbranched or branched saturated hydrocarbon chain. In some embodiments, alkyl as used herein has 1 to 50 carbon atoms ((C₁- C₅₀)alkyl), 1 to 20 carbon atoms ((C₁-C₂₀)alkyl), 1 to 12 carbon atoms ((C₁-C₁₂)alkyl), 1 to 8 carbon atoms ((C₁-C₈)alky), 1 to 6 carbon atoms ((C₁-C₆)alkyl), or l to 4 carbon atoms ((C₁- C₄)alkyl). Examples of alkyl groups may, for example, include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, 2-pentyl, isopentyl, neopentyl, n-hexyl, 2- hexyl, 3-hexyl, and 3-methyl pentyl. When an alkyl residue having a specific number of carbons is named, all geometric isomers having that number of carbons may be encompassed. Thus, for example, “but, can include n-butyl, sec-butyl, isobutyl and t-butyl, and “propel” can include n- propyl and isopropyl. [0085] As used herein, “haloalkyl”, as used herein, refers to an alkyl group substituted with one or more halo, which may be selected independently. Thus, haloalkyl may include alkyl substituted with one or more halo independently selected from the group consisting of fluoro, chloro, iodo, and bromo. Haloalkyl may include, for example, ---CH₂F, ---CHF₂, ---CF₃, ---CH₂C1, CHCl₂, —CC1₃, —CH₂CHFC1, —CHFCH₃, —CH₂Br, and —CH₂CHFCH₂CH₂Br. [0086] As used herein, “cyeloalkvl”, as used herein, refers to a monocyclic or polycyclic saturated hydrocarbon. In some embodiments, cycloalkyl has 3 to 50 carbon atoms ((C₃- C₅₀)cycloalkyl), 3 to 20 carbon atoms ((C₃-C₂₀)cycloalkyl), 3 to 12 carbon atoms ((C₃-C₁₂)cycloalkyl), 3 to 8 carbon atoms ((C₃-C₈)cycloalkyl), 3 to 6 carbon atoms ((C₃-C₆)cycloalkyl), or 3 to 5 carbon atoms ((C₃-C₅)cycloaikyl). Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, octahydropentalenyl, octahydro-1H- indene, decahydronaphthalene, cubane, bicyclo[3.1.0]hexane, and bicyclo[1.1.1]pentane. [0087] As used herein, “aryl”, as used herein, refers to a monocyclic or polycyclic group having at least one hydrocarbon aromatic ring, wherein all of the ring atoms of the at least one hydrocarbon aromatic ring are carbon. Wherein an includes a polyeyelic system, no aromatic ring heteroatoms are present. Aryl may include groups with a single aromatic ring (e.g., phenyl) and multiple fused aromatic rings (e.g., naphthyl, anthryl). Aryl may further include groups with one or more aromatic hydrocarbon rings fused to one or more non-aromatic hydrocarbon rings (e.g., fluorenyl; 2,3-di hydro-1H-indene, 1,2,3,4- tetrahydronaphthalene). In certain embodiments, aryl includes groups with an aromatic hydrocarbon ring fused to a non-aromatic ring, wherein the non-aromatic ring comprises at least one ring heteroatom independently selected from the group consisting of N, O, and S. For example, in some embodiments, aryl includes groups with a phenyl ring fused to a non-aromatic ring, wherein the non-aromatic ring comprises at least one ring heteroatom independently selected from the group consisting of N, O, and S (e.g., chromane; thiochromane; 2, 3-dihydrobenzofura.n; indoline). In some embodiments, aryl as used herein has from 6 to 14 carbon atoms ((C₆-C₁₄)aryl), or 6 to 10 carbon atoms ((C₆- C₁₀)aryl). Where the aryl includes fused rings, the aryl may connect to one or more substituents or moieties of the formulae described herein through any atom of the fused ring for which valency permits. [0088] As used herein, “heteroaryl”, as used herein, refers to a monocyclic or polycyclic group comprising at least one aromatic ring, wherein the aromatic ring comprises at least one ring heteroatom independently selected from the group consisting of N, O, and S. The heteroaryl group may comprise 5, 6, 7, 8, 9, 10, 11, 12, or more ring atoms, where ring atoms refer to the sum of carbon and heteroatoms in the one or more rings (e.g., be a 5-membered, 6-membered, 7- membered, 8-membered, 9-membered, 10-membered, 11-membered, or 12-membered heteroaryl). In some embodiments, heteroaryl includes groups with an aromatic ring that comprises at least one ring heteroatom independently selected from the group consisting of N, O, and S, (e.g., pyridinyl, pyrazinyl, furanyl, thiophenyl). In certain embodiments, heteroaryl includes polycyclic groups with an aromatic ring comprising at least one ring heteroatom, fused to a non-aromatic hydrocarbon ring (e.g., 5, 6, 7, 8-tetrahydroquinolinyl; 4, 5, 6, 7- tetrahydroisobenzofuranyl.). In some embodiments, heteroaryl includes polycyclic groups with an aromatic ring comprising at least one ring heteroatom fused to an aromatic hydrocarbon ring (e.g., quinolinyl, quinoxalinyl, benzothiazolyl). In still further embodiments, heteroaryl includes polycyclic groups with two fused aromatic rings, wherein each ring comprises at least one ring heteroatom (e.g., naphthyridinyl). Heteroaryl may include groups comprising 1 to 5 ring heteroatoms, l to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 or 2 ring heteroatoms, or 1 ring heteroatom, wherein each ring heteroatom is independently selected from the group consisting of N, O, and S. In one example, a heteroaryl has 3 to 8 ring carbon atoms, with 1 to 3 ring heteroatoms independently selected from N, O, and S. Examples of heteroaryl groups include pyridyl, pyridazinyl, pyrimidinyl, benzothiazolyl, and pyrazolyl. [0089] As used herein, “halo” or “halogen” includes bromo, chloro, fluoro, and iodo. [0090] As used herein, the term “substituted” used herein means a group wherein at least one hydrogen atom is replaced by a bond to a non-hydrogen atom such as, but not limited to: a halogen atom such as F, Cl, Br, and I; an oxygen atom in a hydroxyl group; and a nitrogen atom in an amino group. [0091] As used herein, the term “substituted alkyl” refers to an alkyl group as defined herein wherein one or more carbon atoms in the alkyl chain have been optionally replaced with a heteroatom such as 0-, N-, S-, -S(O)n- (where n is 0 to 2), -NR- (where R is hydrogen or alkyl) and having from 1 to 5 substituents selected from the group consisting of alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO- aryl, -SO-heteroaryl, -SO₂-alkyl, -SO₂-aryl, SO₂-heteroaryl, and —NR’R”, wherein R’ and R” may be the same or different and are chosen from hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic. [0092] As used herein, the terms “heteroatom-containing alkyl” and “heteroalkyl” refer to an alkyl substituent in which at least one carbon atom is replaced with a heteroatom, as described in further detail infra. If not otherwise indicated, the terms “alkyl” and “lower alkyl” include linear, branched, cyclic, unsubstituted, substituted, and/or heteroatom-containing alkyl or lower alkyl, respectively. [0093] As used herein, the term “alkylene” refers to a di-radical alkyl group. Unless otherwise indicated, such groups include saturated hydrocarbon chains containing from 1 to 24 carbon atoms, which may be substituted or unsubstituted, may contain one or more alicyclic groups, and may be heteroatom-containing. “Lower alkylene” refers to alkylene linkages containing from 1 to 6 carbon atoms. Examples include, methylene (—CH₂—), ethylene (—CH₂CH₂—),propylene (—CH₂CH₂CH₂—), 2-methylpropylene (—CH₂—CH(CH₃) —CH2—), hexylene (—(CH₂)₆—) and the like. [0094] As used herein, the term “alkynyl” refers to straight or branched monovalent hydrocarbyl groups having from 2 to 6 carbon atoms and preferably 2 to 3 carbon atoms and having at least 1 and preferably from 1 to 2 sites of acetylenic (C≡C—) unsaturation. Examples of such alkynyl groups include, but are not limited to, acetylenyl (C≡CH), and propargyl (CH₂C≡CH). [0095] As used herein, the terms “alkenylene,” “alkynylene,” “arylene,” “arylalkylene,” and “alkylarylene” refer to di-radical alkenyl, alkynyl, aryl, arylalkyl, and alkylaryl groups, respectively. [0096] As used herein, the term “amido” refers to a “C-amido” group represented by — C(=O)NR^XR^Y and an “N-amido” group represented by —NR^XC(=O)R^Y, wherein R^X and R^Y are independently selected from the group consisting of hydrogen, alkyl, aryl, haloalkyl, or heteroaryl, and each of which may be optionally substituted. [0097] As used herein, the term “amidino” refers to R^X C(=NR^Y )NR^Z wherein R^X ,R^Y andR^Z are independently selected from the group consisting of hydrogen, alkyl, aryl, haloalkyl, or heteroaryl, and each of which may be optionally substituted. [0098] As used herein, the term “amino” refers to the group -NRR’ wherein R and R’ are independently hydrogen or nonhydrogen substituents, with nonhydrogen substituents including, for example, alkyl, aryl, alkenyl, arylalkyl, and substituted and/or heteroatom-containing variants thereof. [0099] As used herein, the term “aryl”, unless otherwise specified, refers to an aromatic substituent generally, although not necessarily, containing 5 to 30 carbon atoms and containing a single aromatic ring or multiple aromatic rings that are fused together, directly linked, or indirectly linked (such that the different aromatic rings are bound to a common group such as a methylene or ethylene moiety). Aryl groups may, for example, contain 5 to 20 carbon atoms, and as a further example, aryl groups may contain 5 to 12 carbon atoms. For example, aryl groups may contain one aromatic ring or two or more fused or linked aromatic rings (i.e., biaryl, aryl- substituted aryl, etc.). Examples include phenyl, naphthyl, biphenyl, diphenylether, diphenylamine, benzophenone, and the like. [0100] As used herein, the term “substituted aryl” refers to an aryl moiety substituted with one or more substituent groups in which at least one carbon atom is replaced with a heteroatom, as will be described in further detail infra. Aryl is intended to include stable cyclic, heterocyclic, polycyclic, and polyheterocyclic unsaturated C₃-C₁₄ moieties, exemplified but not limited to phenyl, biphenyl, naphthyl, pyridyl, furyl, thiophenyl, imidazoyl, pyrimidinyl, and oxazoyl; which may further be substituted with one to five members selected from the group consisting of hydroxy, C₁-C₈ alkoxy, C₁-C₈ branched or straight-chain alkyl, acyloxy, carbamoyl, amino, N- acylamino, halogen, trifluoromethyl, cyano, and carboxyl (see i.e., Katritzky, Handbook of Heterocyclic Chemistry). If not otherwise indicated, the term “aryl” includes unsubstituted, substituted, and/or heteroatom-containing aromatic substituents. [0101] As used herein, the term “arylalkyl” refers to an alkyl group with an aryl substituent, and the term “alkylaryl” refers to an aryl group with an alkyl substituent, wherein “alkyl” and “aryl” are as defined above. In general, arylalkyl and alkylaryl groups herein contain 6 to 30 carbon atoms. Arylalkyl and alkylaryl groups may, for example, but are not limited to, contain 6 to 20 carbon atoms, and as a further example, such groups may contain 6 to 12 carbon atoms. [0102] As used herein, the term “azido” refers to group —N₃. [0103] As used herein, the term “carboxyl,” “carboxy” or “carboxylate” refers to — CO₂H or salts thereof. [0104] As used herein, the term “carbamoyl” refers to the group NH₂CO—. [0105] As used herein, the terms “cycloalkyl” and “bicycloalkyl” refers to cyclic alkyl groups of from 3 to 10 carbon atoms having single or multiple cyclic rings including fused, bridged, and spiro ring systems. Examples of suitable cycloalkyl groups include, but are not limited to, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl and the like. Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantanyl, and the like. [0106] As used herein, the terms “substituted cycloalkyl” and “substituted bicycloalkyl” refer to cycloalkyl groups having from 1 to 5 substituents, or from 1 to 3 substituents. selected from alkyl, substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, bicycloalkyl, substituted bicycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -SO-alkyl, -SO- substituted alkyl, -SO-aryl, -SO-heteroaryl, -SO₂-alkyl, SO₂-substituted alkyl, -SO₂-aryl and - SO₂-heteroaryl. [0107] As used herein, the groups

and

are used interchangeably and refer to a cyclohexyl group. [0108] As used herein, the term “cyano” and “carbonitrile” refer to the group —CN. [0109] As used herein, the term “formyl” refers to the group —C(O)H. [0110] As used herein, the term “guanidino” refers to the group —NHC(=NH)NH₂. [0111] As used herein, the terms “halo” and “halogen” are used in the conventional sense to refer to a chloro, bromo, fluoro or iodo substituent. [0112] As used herein, the terms “hydroxy” and “hydroxyl” refer to the group -OH. [0113] As used herein, the term “heteroalkyl” refers to an alkyl substituent in which one or more of the carbon atoms and any attached hydrogen atoms are independently replaced with the same or different heteroatomic group. For example, 1, 2, or 3 carbon atoms may be independently replaced with the same or different heteroatomic substituent. [0114] As used herein, the term “heteroaryl” refers to an aromatic group of from 4 to 10 carbon atoms and 1 to 4 heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur within the ring. Such heteroaryl groups can have a single ring (i.e., pyridinyl or furyl) or multiple condensed rings (i.e., indolizinyl or benzothienyl) wherein the condensed rings may or may not be aromatic and/or contain a heteroatom provided that the point of attachment is through an atom of the aromatic heteroaryl group. In one embodiment, the nitrogen and/or the sulfur ring atom(s) of the heteroaryl group are optionally oxidized to provide for the N oxide (N→O), sulfinyl, or sulfonyl moieties. Preferred heteroaryls include 5 or 6 membered heteroaryls such as pyridinyl, pyrrolyl, indolyl, thiophenyl, and furanyl. [0115] As used herein, the term “substituted heteroaryl” refers to heteroaryl groups that are substituted with from 1 to 5, preferably 1 to 3, or more preferably 1 to 2 substituents selected from the group consisting of the same group of substituents defined for substituted aryl. [0116] As used herein, the terms “heterocycle,” “heterocyclic” and “heterocycloalkyl” refer to a saturated or partially saturated, but not aromatic, group having from 2 to 10 ring carbon atoms and from 1 to 4 ring heteroatoms selected from the group consisting of nitrogen, sulfur, or oxygen. Cycloalkyl or heterocycloalkyl refers to a group having x number of ring carbon atoms excluding the ring heteroatoms. Heterocycle encompasses single ring or multiple condensed rings, including fused, bridged and spiro ring systems. In fused ring systems, one or more the rings can be cycloalkyl, aryl or heteroaryl provided that the point of attachment is through the non-aromatic ring. In one embodiment, the nitrogen and/or sulfur atom(s) of the heterocyclic group are optionally oxidized to provide for the N-oxide, sulfinyl, sulfonyl moieties. In another embodiment, heterocycles can be saturated or partially unsaturated, monocyclic or bicyclic, bridged, or fused. [0117] As used herein, the terms “substituted heterocyclic” or “substituted heterocycloalkyl” or “substituted heterocyclyl” refers to heterocyclyl groups that are substituted with from 1 to 5 or preferably 1 to 3 of the same substituents as defined for substituted cycloalkyl. Examples of heterocycle and heteroaryl include, but are not limited to, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, dihydroindole, dexahydroindole, dihydropyridine, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, imidazolinone, piperidine, piperazine, indoline, phthalimide, 1,2,3,4-tetrahydroisoquinoline, 4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene, benzo[b]thiophene, morpholinyl, thiomorpholinyl (also referred to as thiamorpholinyl), 1,1-dioxothiomorpholinyl, piperidinyl, pyrrolidine, and tetrahydrofuranyl. [0118] As used herein, the term “nitro” refers to the group —NO₂. [0119] As used herein, the term “oxo” refers to the group (=O) or (O). [0120] As used herein, the term “oxetanyl” refers to the group of a four-membered saturated cycloalkane ring with three carbon atoms and one oxygen atom. [0121] As used herein, the term “sulfinyl” refers to the divalent group —SO—. [0122] As used herein, the term “sulfonyl” refers to the group —SO₂R, where R may be alkyl, haloalkyl, cycloalkyl, heterocycloalkyl, heteroaryl, aryl, arylalkyl, or alkylaryl. Examples include, but are not limited to, methylsulfonyl, ethylsulfonyl, phenylsulfonyl, and pyridylsulfonyl. [0123] As used herein, the term “tetrahydrofuranyl” refers to the group of a five-membered saturated cycloalkane ring with four carbon atoms and one oxygen atom. [0124] As used herein, the term “thiocyanate” refers to the group —SCN. [0125] As used herein, the term “isothiocyante” refers to the group —NCS. [0126] As used herein, the term “thiol” or “mercapto” refers to the group —SH. [0127] As used herein, the term “thione” refers to the group =S. [0128] As used herein, the term “substituted” refers that any one or more hydrogen atoms on the designated atom or group is replaced with one or more substituents other than hydrogen, provided that the designated atom’s normal bonding valence is not exceeded. The one or more substituents include, but are not limited to, alkyl alkenyl, alkynyl, alkoxy, acyl, amino, amido, amidino, aryl, azido, carbamoyl, carboxyl, carboxyl ester, cyano, guanidino, halo, haloalkyl, heteroalkyl, heteroaryl, heterocycloalkyl, hydroxy, hydrazino, imino, oxo, nitro, alky sulfinyl, sulfonic acid, alkylsulfonyl, thiocyanate, thiol, thione, or combinations thereof. By way of example, there may be one, two, three, four, five, or six substituents. [0129] As used herein, the term “isomers” refers to compounds comprising the same numbers and types of atoms, groups or components, but with different structural arrangement and connectivity of the atoms. [0130] As used herein, the term “tautomer” refers to one of two or more structural isomers which readily convert from one isomeric form to another and which exist in equilibrium. [0131] As used herein, “stereoisomer” refers to a compound made up of the sam.e atoms bonded by the same bonds but having different three-dimensional structures, which are not interchangeable. The present invention contemplates various stereoisomers and mixtures thereof and includes “enantiomers”, which refers to two stereoisomers whose molecules are non- superimposeabie mirror images of one another. [0132] Individual enantiomers and diastereomers of compounds of the present disclosure can be prepared synthetically from commercially available starting materials that contain asymmetric or stereogenic centers, or by preparation of racemic mixtures followed by resolution methods well known to those of ordinary skill in the art. These methods of resolution are exemplified by (1) attachment of a mixture of enantiomers to a chiral auxiliary, separation of the resulting mixture of diastereomers by recrystallization or chromatography and liberation of the optically pure product from the auxiliary, (2) salt formation employing an optically active resolving agent, (3) direct separation of the mixture of optical enantiomers on chiral liquid chromatographic columns, or (4) kinetic resolution using stereoselective chemical or enzymatic reagents. Racemic mixtures also can be resolved into their respective enantiomers by well-known methods, such as chiral- phase gas chromatography or crystallizing the compound in a chiral solvent. Stereoselective syntheses, a chemical or enzymatic reaction in which a single reactant forms an unequal mixture of stereoisomers during the creation of a new stereocenter or during the transformation of a pre- existing one, are well known in the art. Stereoselective syntheses encompass both enantio- and diastereoselective transformations. See, for example, Carreira and Kvaerno, Classics in Stereoselective Synthesis, Wiley-VCH: Weinheim, 2009. [0133] Geometric isomers, resulting from the arrangement of substituents around a carbon- carbon double bond or arrangement of substituents around a cycloalkyl or heterocyclic ring, can also exist in the compounds of the present disclosure. The symbol = denotes a bond that may be a single, double or triple bond as described herein. Substituents around a carbon-carbon double bond are designated as being in the “Z” or “E” configuration, where the terms “Z” and “E” are used in accordance with IUPAC standards. Unless otherwise specified, structures depicting double bonds encompass both the “E” and “Z” isomers. [0134] Substituents around a carbon-carbon double bond alternatively can be referred to as “cis” or “trans,” where “cis” represents substituents on the same side of the double bond and “trans” represents substituent on opposite sides of the double bond. The arrangement of substituents around a carbocyclic ring can also be designated as “cis” or “trans.” The term “cis” represents substituents on the same side of the plane of the ring and the term “trans” represents substituents on opposite sides of the plane of the ring. Mixtures of compound wherein the substituents are disposed on both the same and opposite sides of the plane of the ring are designated “cis/trans.” [0135] The present disclosure also encompasses isotopically-labeled compounds which are identical to those compounds recited herein, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature (“isotopologues”). Examples of isotopes that can be incorporated into compounds described herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as ²H (“D”), ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵s,¹⁸F, and ³⁶Cl, respectively. For example, a compound described herein can have one or more H atoms replaced with deuterium. [0136] Certain isotopically-labeled compounds, such as those labeled with ³H and ¹⁴C, can be useful in compound and/or substrate tissue distribution assays. Tritiated (³H) and carbon-14 (¹⁴C) isotopes can be particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium can afford certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and hence can be preferred in some circumstances. Isotopically-labeled compounds can generally be prepared by following procedures analogous to those disclosed herein, for example, in the Examples section, by substituting an isotopically-labeled reagent for a non- isotopically-labeled reagent. [0137] As used herein, the term “pharmaceutically acceptable salt” refers to a salt which is acceptable for administration to a subject. It is understood that such salts, with counter ions, will have acceptable mammalian safety for a given dosage regime. Such salts can also be derived from pharmaceutically acceptable inorganic or organic bases and from pharmaceutically acceptable inorganic or organic acids, and may comprise organic and inorganic counter ions. The neutral forms of the compounds described herein may be converted to the corresponding salt forms by contacting the compound with a base or acid and isolating the resulting salts. [0138] As used herein, singular articles such as “a,” “an” and “the” and similar referents in the context of describing the elements are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, including the upper and lower bounds of the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (i.e., “such as”) provided herein, is intended merely to better illuminate the embodiments and does not pose a limitation on the scope of the claims unless otherwise stated. [0139] In some embodiments, where the use of the term “about” is before a quantitative value, the present disclosure also includes the specific quantitative value itself, unless specifically stated otherwise. As used herein, the term “about” refers to a ±10% variation from the nominal value unless otherwise indicated or implied. Where a percentage is provided with respect to an amount of a component or material in a composition, the percentage should be understood to be a percentage based on weight, unless otherwise stated or understood from the context. [0140] Where a molecular weight is provided and not an absolute value, for example, of a polymer, then the molecular weight should be understood to be an average molecule weight, unless otherwise stated or understood from the context. [0141] As used herein, the terms “active agent,” “drug,” “pharmacologically active agent,” “active pharmaceutical ingredient” are used interchangeably to refer to a compound or composition which, when administered to a subject, induces a desired pharmacologic or physiologic effect by local or systemic action or both. [0142] As used herein, the term “promoiety” refers to a form of protecting group that, when used to mask a functional group within an active agent, converts the active agent into a prodrug. In some embodiments, the promoiety will be attached to the drug via bond(s) that are cleaved by enzymatic or non-enzymatic means in vivo. In some embodiments, the promoiety is attached to an amine, a hydroxyl, or a carboxylic acid group of the subject compounds. In some embodiments, the promoiety is an acyl or substituted acyl group. In certain cases, the promoiety is an alkyl or substituted alkyl group. [0143] As used herein, the term “prodrug” refers to compounds that are transformed in vivo to provide a compound or pharmaceutically acceptable salt, hydrate or solvate of the compound described herein. The transformation can occur by various mechanisms (i.e., esterase, amidase, phosphatase, oxidative and/or reductive metabolism) in various locations I (i.e., in the intestinal lumen or upon transit into the intestine, blood, or liver). [0144] The terms “individual,” “host,” and “subject,” are used interchangeably herein, and refer to an animal, including, but not limited to, human and non-human primates, including simians and humans; rodents, including rats and mice; bovines; equines; ovines; felines; canines; and the like. “Mammal” means a member or members of any mammalian species, and includes, by way of example, canines, felines, equines, bovines, ovines, rodentia, etc. and primates, i.e., non- human primates, and humans. Non-human animal models, i.e., mammals, non-human primates, murines, lagomorpha, etc. may be used for experimental investigations. As used herein, “patient” refers to a human subject. [0145] As used herein, the terms “treating,” “treatment,” and the like, refer to obtaining a desired pharmacologic and/or physiologic effect, such as reduction of proteinuria in an inflammatory renal disease. The effect may be therapeutic in terms of a partial or complete cure for a disease and/or adverse effect attributable to the disease. “Treatment,” as used herein, covers any treatment of a disease in a mammal, particularly in a human, and includes: (a) preventing the disease or a symptom of a disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it (i.e., including diseases that may be associated with or caused by a primary disease); (b) inhibiting the disease, i.e., arresting its development; and (c) relieving the disease, i.e., causing regression of the disease (i.e., reduction in of tumor burden). [0146] [0147] Examples of salts include, but are not limited to: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, oxalate, palmoate, pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, undecanoate, and the like. [0148] Other examples of salts include anions of the compounds of the present disclosure compounded with a suitable cation such as N⁺, NH₄ ⁺, and NW₄ ⁺ (where W can be a C₁-C₈ alkyl group), and the like. For therapeutic use, salts of the compounds of the present disclosure can be pharmaceutically acceptable. However, salts of acids and bases that are non-pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound. [0149] Compounds included in the present compositions that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. The acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, including but not limited to, malate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate (i.e., 1,1’-methylene-bis-(2- hydroxy-3-naphthoate )) salts. [0150] Compounds included in the present compositions that are acidic in nature are capable of forming base salts with various pharmacologically acceptable cations. Examples of such salts include alkali metal or alkaline earth metal salts and, particularly, calcium, magnesium, sodium, lithium, zinc, potassium, and iron salts. [0151] Compounds included in the present compositions that include a basic or acidic moiety can also form pharmaceutically acceptable salts with various amino acids. The compounds of the disclosure can contain both acidic and basic groups; for example, one amino and one carboxylic acid group. In such a case, the compound can exist as an acid addition salt, a zwitterion, or a base salt. [0152] As used herein, the phrase “therapeutically effective amount” refers to the amount of a compound that, when administered to a mammal or other subject for treating a disease, condition, or disorder, is sufficient to affect such treatment for the disease, condition, or disorder. The “therapeutically effective amount” will vary depending on the compound, the disease and its severity and the age, weight, etc., of the subject to be treated. [0153] As used herein, the term “unit dosage form” refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of a compound (i.e., an aminopyrimidine compound, as described herein) calculated in an amount sufficient to produce the desired effect in association with a pharmaceutically acceptable diluent, carrier or vehicle. The specifications for unit dosage forms depend on the particular compound employed and the effect to be achieved, and the pharmacodynamics associated with each compound in the host. [0154] As used herein, the terms “pharmaceutically acceptable excipient,” “pharmaceutically acceptable diluent,” “pharmaceutically acceptable carrier,” and “pharmaceutically acceptable adjuvant” refer to an excipient, diluent, carrier, and adjuvant that are useful in preparing a pharmaceutical composition that are generally safe, non-toxic and neither biologically nor otherwise undesirable, and include an excipient, diluent, carrier, and adjuvant that are acceptable for veterinary use as well as human pharmaceutical use. The phrase “A pharmaceutically acceptable excipient, diluent, carrier and adjuvant” as used in the specification and claims includes both one and more than one such excipient, diluent, carrier, and adjuvant. [0155] As used herein, the term “pharmaceutical composition” is meant to encompass a composition suitable for administration to a subject, such as a mammal, especially a human. In general a “pharmaceutical composition” is sterile, and preferably free of contaminants that are capable of eliciting an undesirable response within the subject (i.e., the compound(s) in the pharmaceutical composition is pharmaceutical grade). Pharmaceutical compositions can be designed for administration to subjects or patients in need thereof via a number of different routes of administration including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, intracheal, intramuscular, subcutaneous, and the like. [0156] As used herein, the phrases “of the formula” and “of the structure” is not intended to be limiting and are used in the same way that the term “comprising” is commonly used. The term “independently selected from” is used herein to indicate that the recited elements, i.e., R groups or the like, can be identical or different. [0157] the phrase “optionally substituted” means that a non-hydrogen substituent may or may not be present on a given atom, and, thus, the description includes structures wherein a non- hydrogen substituent is present and structures wherein a non-hydrogen substituent is not present. [0158] The above definitions are not intended to include impermissible substitution patterns (i.e., methyl substituted with 5 fluorines or heteroaryl groups having two adjacent oxygen ring atoms). Such impermissible substitution patterns are well known to the skilled artisan. When used to modify a chemical group, the term “substituted” may describe other chemical groups defined herein. For example, the term “substituted aryl” includes, but is not limited to, “alkylaryl.” Unless specified otherwise, where a group is described as optionally substituted, any substituents of the group are themselves unsubstituted. [0159] Generally, reference to or depiction of a certain element such as hydrogen or H is meant to include all isotopes of that element. For example, if an R group is defined to include hydrogen or H, it also includes deuterium and tritium. Compounds comprising radioisotopes such as tritium, ¹⁴C, ³²P and ³⁵S are thus within the scope of the present technology. Procedures for inserting such labels into the compounds of the present technology will be readily apparent to those skilled in the art based on the disclosure herein. [0160] Unless the specific stereochemistry is expressly indicated, all chiral, diastereomeric, and racemic forms of a compound are intended. Thus, compounds described herein include enriched or resolved optical isomers at any or all asymmetric atoms as are apparent from the depictions. Racemic mixtures of R-enantiomer and S-enantiomer, and enantio-enriched stereomeric mixtures comprising of R- and S-enantiomers, as well as the individual optical isomers can be isolated or synthesized so as to be substantially free of their enantiomeric or diastereomeric partners, and these stereoisomers are all within the scope of the present technology. [0161] The compounds described herein may exist as solvates, especially hydrates, and unless otherwise specified, all such solvates and hydrates are intended. Hydrates may form during manufacture of the compounds or compositions comprising the compounds, or hydrates may form over time due to the hygroscopic nature of the compounds. Compounds of the present technology may exist as organic solvates as well, including DMF, ether, and alcohol solvates, among others. The identification and preparation of any particular solvate is within the skill of the ordinary artisan of synthetic organic or medicinal chemistry. [0162] or prevent everyday life functions but do not constitute a substantial disruption. [0163] As described herein, the text refers to various embodiments of the present compounds, compositions, and methods. The various embodiments described are meant to provide a variety of illustrative examples and should not be construed as descriptions of alternative species. Rather, it should be noted that the descriptions of various embodiments provided herein may be of overlapping scope. The embodiments discussed herein are merely illustrative and are not meant to limit the scope of the present technology. Method of Treating a Disease [0164] In a first aspect, the present disclosure provides a method of treating a disease, comprising administering to a human subject with a disease an oral daily dose of a compound of Formula I,

or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof, in an amount of at least 450 milligrams and less than 2,000 milligrams per day, wherein: X is selected from H and

; a is from 0-30; b is from 0-30; R¹ is selected from H, alkyl, haloalkyl, cycloalkyl, aryl, and heteroaryl, -C(O)-R², gluconate, glycoside, glucuronide, tocopherols, and PEG groups; R² is selected from alkyl, haloalkyl, cycloalkyl, aryl, and heteroaryl; and at least one pharmaceutically acceptable carrier or excipient. [0165] In some embodiments, the daily oral dose is about 400 mg to about 1,500 mg. In some embodiments, the daily oral dose is about 400 mg to about 500 mg, about 400 mg to about 600 mg, about 400 mg to about 700 mg, about 400 mg to about 800 mg, about 400 mg to about 900 mg, about 400 mg to about 1,000 mg, about 400 mg to about 1,100 mg, about 400 mg to about 1,200 mg, about 400 mg to about 1,300 mg, about 400 mg to about 1,400 mg, about 400 mg to about 1,500 mg, about 500 mg to about 600 mg, about 500 mg to about 700 mg, about 500 mg to about 800 mg, about 500 mg to about 900 mg, about 500 mg to about 1,000 mg, about 500 mg to about 1,100 mg, about 500 mg to about 1,200 mg, about 500 mg to about 1,300 mg, about 500 mg to about 1,400 mg, about 500 mg to about 1,500 mg, about 600 mg to about 700 mg, about 600 mg to about 800 mg, about 600 mg to about 900 mg, about 600 mg to about 1,000 mg, about 600 mg to about 1,100 mg, about 600 mg to about 1,200 mg, about 600 mg to about 1,300 mg, about 600 mg to about 1,400 mg, about 600 mg to about 1,500 mg, about 700 mg to about 800 mg, about 700 mg to about 900 mg, about 700 mg to about 1,000 mg, about 700 mg to about 1,100 mg, about 700 mg to about 1,200 mg, about 700 mg to about 1,300 mg, about 700 mg to about 1,400 mg, about 700 mg to about 1,500 mg, about 800 mg to about 900 mg, about 800 mg to about 1,000 mg, about 800 mg to about 1,100 mg, about 800 mg to about 1,200 mg, about 800 mg to about 1,300 mg, about 800 mg to about 1,400 mg, about 800 mg to about 1,500 mg, about 900 mg to about 1,000 mg, about 900 mg to about 1,100 mg, about 900 mg to about 1,200 mg, about 900 mg to about 1,300 mg, about 900 mg to about 1,400 mg, about 900 mg to about 1,500 mg, about 1,000 mg to about 1,100 mg, about 1,000 mg to about 1,200 mg, about 1,000 mg to about 1,300 mg, about 1,000 mg to about 1,400 mg, about 1,000 mg to about 1,500 mg, about 1,100 mg to about 1,200 mg, about 1,100 mg to about 1,300 mg, about 1,100 mg to about 1,400 mg, about 1,100 mg to about 1,500 mg, about 1,200 mg to about 1,300 mg, about 1,200 mg to about 1,400 mg, about 1,200 mg to about 1,500 mg, about 1,300 mg to about 1,400 mg, about 1,300 mg to about 1,500 mg, or about 1,400 mg to about 1,500 mg. In some embodiments, the daily oral dose is about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1,000 mg, about 1,100 mg, about 1,200 mg, about 1,300 mg, about 1,400 mg, or about 1,500 mg. In some embodiments, the daily oral dose is at least about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1,000 mg, about 1,100 mg, about 1,200 mg, about 1,300 mg, or about 1,400 mg. In some embodiments, the daily oral dose is at most about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1,000 mg, about 1,100 mg, about 1,200 mg, about 1,300 mg, about 1,400 mg, or about 1,500 mg. [0166] In some embodiments, the daily oral dose is about 1,500 mg to about 2,000 mg. In some embodiments, the daily oral dose is about 1,500 mg to about 1,600 mg, about 1,500 mg to about 1,700 mg, about 1,500 mg to about 1,800 mg, about 1,500 mg to about 1,900 mg, about 1,500 mg to about 2,000 mg, about 1,600 mg to about 1,700 mg, about 1,600 mg to about 1,800 mg, about 1,600 mg to about 1,900 mg, about 1,600 mg to about 2,000 mg, about 1,700 mg to about 1,800 mg, about 1,700 mg to about 1,900 mg, about 1,700 mg to about 2,000 mg, about 1,800 mg to about 1,900 mg, about 1,800 mg to about 2,000 mg, or about 1,900 mg to about 2,000 mg. In some embodiments, the daily oral dose is about 1,500 mg, about 1,600 mg, about 1,700 mg, about 1,800 mg, about 1,900 mg, or about 2,000 mg. In some embodiments, the daily oral dose is at least about 1,500 mg, about 1,600 mg, about 1,700 mg, about 1,800 mg, or about 1,900 mg. In some embodiments, the daily oral dose is at most about 1,600 mg, about 1,700 mg, about 1,800 mg, about 1,900 mg, or about 2,000 mg. [0167] In some embodiments, the daily oral dose is 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 825 mg, 850 mg, 875 mg, 900 mg, 925 mg, 950 mg, 975 mg, 1,000 mg, 1,100 mg, 1,200 mg, 1,300 mg, 1,400 mg, 1,500 mg, 1,600 mg, 1,700 mg, 1,800 mg, 1,900 mg, or 2,000 mg. [0168] In some embodiments, the daily oral dose is 450 mg or 600 mg. [0169] In some embodiments, the daily oral dose is 450 mg. [0170] In some embodiments, the daily oral dose is 600 mg. [0171] In some embodiments, the daily oral dose is administered as a single undivided dose once a day. In some embodiments, the daily oral dose is administered in divided subdoses, typically equally divided subdoses, twice a day. In some embodiments, the daily oral dose is administered in divided subdoses, typically equally divided subdoses, three times a day. [0172] In some embodiments, the daily oral dose is administered once a day. [0173] In some embodiments, the daily oral dose is administered twice a day. [0174] In some embodiments, the daily oral dose is administered three times a day. [0175] In some embodiments, the daily oral dose is administered for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, or at least 14 days. In some embodiments, the daily oral dose is administered for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, or 6 months. In some embodiments, the daily oral dose is administered for at least 1 year. [0176] In some embodiments of the compound of formula (I), is

. [0177] In some embodiments of the compound of formula (I), R^1- is H, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, or pentyl. [0178] In some embodiments of the compound of formula (I), R^1- is H. [0179] In some embodiments, the compound of formula (I) is nitro-oleic acid. [0180] In some embodiments, the compound of formula (I) is one of the structures of one of the compounds in Table 1.

[0181] In some embodiments, the compound of formula (I) is 10-nitro-9(E)-octadec-9-enoic acid (CXA-10), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof. [0182] In some embodiments, the compound of formula (I) is 9-nitro-9(E)-octadec-9-enoic acid (CXA-9), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof. [0183] In some embodiments, the disease is selected from solid organ fibrosis, inflammatory disease, cardiovascular disease, renal disease, kidney failure, ischemic kidney injury, acute kidney injury (AKI), chronic kidney injury (CKI), chronic kidney disease (CKD), obesity associated chronic kidney disease, diabetic nephropathy, kidney fibrosis, progressive kidney disease (renal fibrosis), focal segmental glomerular sclerosis (FSGS), primary FSGS, secondary FSGS, sickle cell nephropathy, glomerulonephritis, nephrotic syndrome, nonalcoholic steatohepatitis (NASH), fatty liver disease, non-fatty liver disease (NFLD), pulmonary arterial hypertension (PAH), pulmonary fibrosis, psoriasis, allergic airway disease, obesity, anti- adipogenic disease, type II diabetes, lithium induced diabetes insipidus, sickle cell disease, sickle cell crisis, idiopathic pulmonary fibrosis (IPF), interstitial lung diseases (ILDs), scleroderma, inflammatory gastrointestinal disease, colitis, inflammatory bowel disease, neurodegenerative disease, amyotrophic lateral sclerosis (ALS), metabolic syndrome, neuropathy, Charcot-Marie- Tooth disease, and mitochondrial related disease. [0184] In some embodiments, the human subject has sickle cell disease. [0185] In some embodiments, the human subject has sickle cell crisis. [0186] In some embodiments of the method, the compound of formula I is formulated in a pharmaceutical composition. [0187] In some embodiments of the method, the human subject is between 18 and 60 years of age. In another embodiment, the human subject is male. In another embodiment, the human subject is female. [0188] In some embodiments of the method, the human subject has a body mass index (BMI) between 18.0 and 32.0 kg/m². [0189] In some embodiments of the method, the oral daily dose is administered to the human subject with a meal. In another embodiment, the oral daily dose is administered to the human subject within 30 minutes of a meal. In another embodiment, the oral daily dose is administered to the human subject within 60 minutes of a meal. [0190] In some embodiments, the oral daily dose is administered to the human subject without a meal. [0191] In some embodiments of the method, the compound of formula (I) is 10-nitro-9(E)- octadec-9-enoic acid (CXA-10), or a salt, a stereoisomer, a deuterated analog, or a fluorinated analog thereof; the oral daily dose is 450 mg; and the human subject has sickle cell disease. [0192] In some embodiments of the method, the compound of formula (I) is 10-nitro-9(E)- octadec-9-enoic acid (CXA-10), or a salt, a stereoisomer, a deuterated analog, or a fluorinated analog thereof; the oral daily does is 600 mg; and the human subject has sickle cell disease. [0193] In some embodiments of the method, the compound of formula (I) is 10-nitro-9(E)- octadec-9-enoic acid (CXA-10), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily does is 450 mg; and the human subject has sickle cell crisis. [0194] In some embodiments of the method, the compound of formula (I) is 10-nitro-9(E)- octadec-9-enoic acid (CXA-10), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily does is 600 mg; and the human subject has sickle cell crisis. [0195] In some embodiments of the method, the compound of formula (I) is 9-nitro-9(E)- octadec-9-enoic acid (CXA-9), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily dose is 450 mg; and the human subject has sickle cell disease. [0196] In some embodiments of the method, the compound of formula (I) is 9-nitro-9(E)- octadec-9-enoic acid (CXA-9), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily does is 600 mg; and the human subject has sickle cell disease. [0197] In some embodiments of the method, the compound of formula (I) is 9-nitro-9(E)- octadec-9-enoic acid (CXA-9), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily does is 450 mg; and the human patient has sickle cell crisis. [0198] In some embodiments of the method, the compound of formula (I) is 9-nitro-9(E)- octadec-9-enoic acid (CXA-9), or a salt, a stereoisomer, a deuterated analog, and a fluorinated analog thereof; the oral daily does is 600 mg; and the human patient has sickle cell crisis. Pharmaceutical Composition [0199] In a second aspect, the present disclosure provides for a pharmaceutical composition comprising a compound of formula (I),

or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof, wherein: X is selected from H and

; a is from 0-30; b is from 0-30; R¹ is selected from H, alkyl, haloalkyl, cycloalkyl, aryl, and heteroaryl, -C(0)-R², gluconate, glycoside, glucuronide, tocopherols, and PEG groups; R² is selected from alkyl, haloalkyl, cycloalkyl, aryl, and heteroaryl; and at least one pharmaceutically acceptable carrier or excipient. [0200] In some embodiments of the pharmaceutical composition, X is

. [0201] In some embodiments of the pharmaceutical composition, R¹ is H, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, or pentyl. [0202] In some embodiments of the pharmaceutical composition, R¹ is H. [0203] In some embodiments of the pharmaceutical composition, the compound of formula (I) is oleic acid. [0204] In some embodiments of the pharmaceutical composition, the compound of formula (I) is nitro-oleic acid. [0205] In some embodiments of the pharmaceutical composition, the compound of formula (I) is represented by the structure of one of the compounds in Table 1. [0206] In some embodiments, the pharmaceutical composition comprises triglyceride. In another embodiment, the triglyceride is a medium chain triglyceride. [0207] In some embodiments of the method, the medium chain triglyceride is selected from hexanoic acid, octanoic acid, decanoic acid, and dodecanoic acid. [0208] In some embodiments, the pharmaceutical composition comprises Miglyol. In some embodiments, Miglyol comprises octanoic acid, decanoic acid, and propylene glycol. [0209] In some embodiments, the pharmaceutical composition further comprises an antioxidant. In some embodiments, the antioxidant is glycine, α-tocopherol or ascorbate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), or a combination thereof. [0210] In some embodiments of the pharmaceutical composition, the antioxidant is butylated hydroxytoluene (BHT). [0211] In some embodiments, the pharmaceutical composition further comprises a thickening agent. In another embodiment, the thickening agent is fumed silica. [0212] In some embodiments, the pharmaceutical composition is formulated for oral administration. [0213] In some embodiments, the pharmaceutical composition is formulated as a capsule. In another embodiment, the pharmaceutical composition is formulated as a tablet. Unit Oral Dosage [0214] In a third aspect, the present disclosure provides for an oral unit dosage form, comprising a capsule shell; and any of the pharmaceutical composition described herein. [0215] In some embodiments of the oral dosage unit, the capsule shell comprises hydroxypropyl methylcellulose (HMPC). [0216] In some embodiments of the oral dosage unit, the capsule contains 25 mg to 300 mg of an active pharmaceutical ingredient (API). In another embodiment, the capsule contains 150 mg of an active pharmaceutical ingredient (API). [0217] In some embodiments of the oral dosage unit, the API is a compound of formula (I). In another embodiment, the API is 10-nitro-9(E)-octadec-9-enoic acid or 9-nitro-9(E)-octadec-9- enoic acid. [0218] In some embodiments of the oral dosage unit, the API is 10-nitro-9(E)-octadec-9-enoic acid (CXA-10). [0219] In some embodiments of the oral dosage unit, the API is 9-nitro-9(E)-octadec-9-enoic acid (CXA-9). EXAMPLES [0220] The following examples are offered to illustrate the present technology and are not to be construed in any way as limiting the scope of the present technology. Any methods that are functionally equivalent are within the scope of the present technology. Various modifications of the present technology in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications fall within the scope of the appended claims Abbreviations

Example 1 - High-Dose Tolerance and Safety of CXA-10 [0221] Both the CXA-10-202 clinical trial and earlier studies had suggested that oral doses of CXA-10 at or above 300 mg per day could cause gastrointestinal side effects, and that lack of tolerability might independently limit the maximum daily oral doses. The phase 2 trial described below, CXA-10-1007, shows that such side effects are self-limiting and/or can be mitigated, permitting higher doses to be administered with therapeutic effect. [0222] The CXA-10-1007 clinical trial is a randomized, double-blind, placebo-controlled study investigating the safety, tolerability, and pharmacokinetics of steady-state administration of CXA-10 at doses of 300 mg/day or higher. [0223] The purpose of the current repeated-dose study is to explore the safety and gastrointestinal (GI) tolerability of high doses of CXA-10 when administered with food to healthy volunteers. [0224] The study is conducted in 2 cohorts. In Cohort 1, CXA-10 are administered 300 mg twice daily (BID) with food (containing up to approximately 30% fat) for 7 days. New subjects are enrolled in Cohort 2 to receive CXA-10 at 300 or 450 mg once daily (QD) in the morning with food (containing up to approximately 30% fat) for 7 days to assess the overall safety and GI tolerability. Overall Study Design [0225] This is a single-center, randomized, double blind, placebo-controlled study in healthy subjects. Up to two cohorts of approximately 12 or 16 subjects each are enrolled as follows:

BID = twice daily; QD = once daily. Note that the planned dosing regimens maybe be subject to change. [0226] The Screening Visit is performed within 21 days of first dose. For both cohorts, eligible subjects are admitted to the clinical study unit on Day -1. Cohort 1 and Cohort 2 are run in a staggered fashion such that dosing in Cohort 1 begins at least 3 days prior to the first does in Cohort 2. Cohort 1 [0227] On Days 1 to 7, subjects are given the randomized study drugs, namely CXA-10 at 300 mg BID or placebo. The CXA-10 or placebo dose is administered with breakfast and dinner (both meals containing up to approximately 30% fat). CXA-10 or placebo is administered BID on Days 1 to 6. The final dose is administered on the morning of Day 7. Subjects is discharged on Day 8 after the Day 724-hour postdose PK sample has been collected. Subjects return for an additional postdose PK sample on Day 10 (± 1 day). A follow-up visit takes place on Day 14 (± 1 day) for the final PK sample and safety assessments. [0228] If the 300 mg BID dose is poorly tolerated by individual subjects, the evening dose is skipped as needed. Cohort 2 [0229] On Days 1 to 7, subjects are given the randomized study drug (CXA-10300 mg or 450 mg QD or placebo). The CXA-10 or placebo dose is administered QD in the morning with breakfast (containing up to approximately 30% fat). Subjects are discharged on Day 8 after the Day 724-hour post-dose PK sample has been collected. Subjects are given an additional post- dose PK sample on Day 10 ((±1 day). A follow up visit is to take place on Day 14 ((±1 day) for the final PK sample and safety assessments. [0230] Cohort 2 does not commence dosing until Cohort 1 has completed at least 3 days of dosing. If tolerability in Cohort 1 is deemed acceptable by the principle investigator and medical monitor, then subjects of Cohort 2 are given the planned CXA-10 regimen of 300 or 450 mg WD. If tolerability in Cohort 1 is deemed poor by the principal investigator and medical monitor, then subjects of Cohort 2 are given 300 mg QD but may amend the 450 mg regimen to 150 mg QD for 2 days with dose escalated to 450 mg QD for Days 3 to 7. [0231] If a subject is withdrawn, efforts are made to perform safety assessments for an Early Termination visit, if possible. Selection of Doses [0232] The doses planned in this study are 300 or 450 mg QD and 300 mg BID. As stated in the section above, these planned doses may be amended based on emerging tolerability data. If dose changes are warranted based on principal investigator and medical monitor discretion, the resulting daily doses are lower than the planned dosing regimen. [0233] In the current study, the tolerability of the 450 mg dose in the fed state is confirmed. A 300 mg dose is also being studied in a subset of subjects to compare safety, PK, and PD with the higher doses. [0234] The current study also investigates higher exposures by administering the 600 mg dose as a divided 300 mg BID regimen. CXA-10 at 300 mg QD in the fed state has been shown to be well-tolerated in patients with primary sickle cell disease. Primary Objective [0235] The primary objective of this study is to investigate the safety and tolerability of CXA-10 administered at doses of 300 mg/day and higher for 7 days. Secondary Objectives [0236] The secondary objectives of this study are: (i) to investigate the pharmacokinetic (PK) profile of CXA-10 and its major metabolites (8, 9 alkene and 10-nitrostearate), (ii) to investigate the PK profile of triglycerides which have incorporated CXA-10; and (iii) to investigate the pharmacodynamic (PD) effects of CXA-10 on select biomarkers in the plasma and peripheral blood mononuclear cells (PBMCs). Safety Endpoint [0237] The safety endpoints of this study include: (i) adverse events (AEs) including gastrointestinal symptoms (ii) vital signs including blood pressure (BP), heart rate (HR), 12-lead ECGs, and physical examination; and (iii) clinical laboratory values, such as hematology, biochemistry, and urinalysis. Pharmacokinetics (PK)Endpoints [0238] PK endpoints of this study include: (i) Plasma-concentration-time profile of CXA-10 and major metabolites relative to Day 1 and Day 7 dosing; (ii) CXA-10 parent and metabolite(s): Cmax, tmax, t1/2, AUC0-24, AUC0-t, AUC0-∞, CL/F, (iii) Vd/F, λz„ as data permit. (iv) Accumulation ratio: Cmaxday 7/CmaxDayl and AUCDay7/AUCDay1 (v) Other PK parameters may be calculated, as appropriate (vi) If a validated bioanalytical assay is available: plasma concentration time profile of CXA-10 incorporated into triglycerides relative to Day 1 and Day 7 dosing. Cmax, tmax, t1/2, AUC0-24, AUC0-t, AUC0-∞, CL/F, Vd/F, and accumulation ratios will be calculated as data permit. Pharmacodynamics (PD) Endpoints [0239] PD endpoints of this study include: (i) Transcriptomic panel endpoints in PBMCs: genes for nuclear factor E2 related factor 2 (Nrf2) activation and heat shock protein (HSP) activation; (ii) Plasma fibroblast growth factor 21 and 19 (FGF21, FGF19); and (iii) Ex vivo assay of CXA-10 activity markers. Investigational Products Drug Product [0240] Capsules containing 150 mg CXA-10 (or matching placebo) are provided for this study. [0241] The drug product is a size `1’ white, opaque hydroxypropylmethyl cellulose capsule filled with a solution of CXA-10 (25% or 50% weight/weight) in medium chain triglyceride oil with 0.05% of butylated hydroxytoluene (BHT) as an antioxidant. Fumed silica is optionally added as a thickening agent. Each capsule contains 300 mg of formulation (equivalent to 150 mg CXA- 10). The filled capsules are packaged in white high-density polyethylene containers (18 capsules per bottle) with child resistant closures. [0242] The placebo is a size ‘1’ white, opaque hydroxypropylmethyl cellulose capsule containing the same inactive ingredients as the active drug product to a total fill weight of 300 mg, but no CXA-10. The filled capsules are packaged in white high-density polyethylene containers (18 capsules per bottle) with child resistant closures. Blinding [0243] This is a double-blind study. The pharmacist (or designee) at the study site prepares medication before use according to instructions provided separately. Subjects are randomized to receive study medications according to a randomization list provided to the site. The pharmacist is required to maintain the blind, and the pharmacist does not otherwise participate in this study. [0244] The study statistician provides the randomization code in advance of the study. Only the pharmacist and a statistician have access to this code. The sponsor and all staff involved with the conduct of the study at the clinic are blinded. The laboratories involved with analysis of PK and laboratory-based PD samples, as well as a separate statistician, are unblinded to the data. [0245] Other study personnel who are not involved with direct subject care are also optionally unblinded to treatment assignments; however, these personnel need to be documented at the site and must maintain the confidentiality of the blind. [0246] A copy of subject treatment assignments is retained in a secure file by the study pharmacist. In the event of an adverse event or a medical emergency when knowledge of the treatment assignment will influence the subject’s care, the decision to unblind a subject’s treatment assignment are made by the Investigator. The Investigator must document the event with information regarding the reasons for unblinding and notify the Complexa Medical Monitor, Chief Medical Officer or designee of the decision as soon as reasonably possible. Study Treatment Administration [0247] Cohort 1: Study medication is planned for BID administration (approximately 10 hours apart) in the fed state on Days 1 to 6, and the final dose on the morning of Day 7. All subjects are instructed to receive two capsules on each dosing occasion: 1. CXA-10300 mg: Two 150-mg capsules 2. Placebo: Two matching placebo capsules. [0248] The morning dose is administered within 30 minutes of consuming a standard breakfast (containing up to approximately 30% of calories from fat). The evening dose is administered within 30 minutes of completely consuming a standard dinner (containing up to approximately 30% of calories from fat). Subjects are3 encouraged to complete the entire meal, but they are not required to complete the entire meal in order to receive their dose of study drug. Each dose is administered orally with approximately 240 mL of room temperature water. Based on individual subject tolerability and if deemed appropriate by the principal investigator and medical monitor, the evening dose is optionally skipped such that the subject only receives the morning dose. [0249] Cohort 2: Study medication is planned for QD administration in the morning in the fed state on Days 1 to 7. All subjects in this cohort are to receive 3 capsules each day: 1. CXA-10450 mg QD: Three 150-mg capsules 2. CXA-10300 mg QD: Two 150-mg capsules + 1 matching placebo capsule 3. Placebo: Three matching placebo capsules. [0250] All doses are planned for administration within 30 minutes of subject’s completing their standard breakfast (containing up to approximately 30% of calories from fat). Each dose is administered orally with approximately 240 mL of room temperature water. [0251] If there is early indication of poor tolerability by subjects in Cohort 1, the principal investigator and medical monitor may decide to amend the Cohort 2 regimen such that subjects receive 150 mg QD on Days 1 and 2 followed by 450 mg QD on Days 3 to 7. The relevant subjects are still given 3 capsules on Days 1 and 2 to maintain the blind (One 150-mg capsule + 2 placebo capsules). Treatment of Investigational Product Overdose [0252] There is currently no information available on the medical effects of an overdose of oral CXA-10, although a single dose of 900 mg and a total daily dose of 1800 mg (900 mg twice within 6 hours) was administered with the most notable effect of Grade 1 to Grade 2 diarrhea. [0253] If an overdose is suspected, the subject is maintained under close medical observation and be intensively monitored for potential GI and cardiovascular events. Diarrhea has been the most obvious and consistent side effect thus far seen when dosed in the fasted state, and an overdose could possibly result in profound and persistent diarrhea. Subjects may experience vasovagal signs and symptoms, dehydration and electrolyte abnormalities should such an event occur due to an overdose. Medical intervention and subsequent care should be instituted as appropriate to the signs and symptoms of any AE. Medical personnel should be trained in advanced care life support with access to a medical defibrillation device. Please refer to the IB for full details on toxicities observed in animals and AEs observed in completed clinical studies. Concomitant Medications [0254] Subjects are refrained from the use of over-the-counter or prescription medications (including herbal medications) within 14 days or 5 half-lives (whichever is longer) prior to dosing until completion of the follow-up visit. By exception, use of acetaminophen is allowed. In addition, concomitant medications are optionally used to treat AEs based on principal investigator judgment. Study Population [0255] Subjects eligible for enrollment in the study must meet all the following criteria: Inclusion Criteria ^ Males or females, of any race, between 18 and 60 years of age, inclusive. ^ The following applies to female subjects: i. Non-pregnant, non-lactating, female of childbearing potential who agrees to use a reliable method of contraception (as defined in Section 6.4), or ii. Female is of non-childbearing potential defined as surgically sterile (hysterectomy or bilateral tubal ligation) or post-menopausal (amenorrhea for a minimum of 12 consecutive months with follicle stimulating hormone [FSH] > 40 U/L). ^ Body mass index (BMI) between 18.0 and 32.0 kg/m2, inclusive. ^ In good health, determined by no clinically significant findings from medical history, physical examination, 12-lead ECG (Screening only), vital sign measurements, and clinical laboratory at Screening and Check-in as assessed by the Investigator (or designee). ^ Able to comprehend and willing to sign an informed consent form (ICF) and to abide by the study restrictions and study duration. Exclusion Criteria ^ Significant history or clinical manifestation of any metabolic, allergic, dermatological, hepatic, renal, hematological, pulmonary, cardiovascular, GI, neurological, respiratory, endocrine, immunological, or psychiatric disorder, as determined by the Investigator (or designee). The subject has any clinical history of cardiovascular events, including clinically significant arrhythmias, fainting, palpitations, personal or family history of congenital prolonged QT syndromes, or sudden unexpected death due to a cardiac reason. ^ History of significant hypersensitivity, intolerance, or allergy to any drug compound, food, or other substance, unless approved by the Investigator (or designee). ^ History of stomach or intestinal surgery or resection (including cholecystectomy) that would potentially alter absorption and/or excretion of orally administered drugs (uncomplicated appendectomy and hernia repair will be allowed). ^ The subject has a history of any primary malignancy, including a history of melanoma or suspicious undiagnosed skin lesions, with the exception of basal cell or squamous cell carcinomas of the skin curatively treated and with no evidence of disease for at least 5 years. ^ The subject has serum triglyceride, very low-density lipoprotein, or low-density lipoprotein concentrations > 1.5 times the upper limit of normal at Screening. May be confirmed by repeated fasted laboratory evaluation. ^ The subject has a supine ECG at Screening with QTcF interval > 450 msec or any other abnormal ECG finding at Screening considered clinically significant by the Investigator. If the QTcF measurement is > 450 msec, the ECG may be repeated 2 additional times and a triplicate average will be used to determine eligibility. ^ The subject has elevated blood pressure defined by an average blood pressure > 145/95 mmHg calculated from the last 2 of the triplicate supine blood pressure measurements at Screening unless deemed acceptable by the Investigator (or designee). ^ History of alcoholism or drug/chemical abuse within 2 years prior to Check-in. ^ Alcohol consumption of > 21 units per week. One unit of alcohol equals 12 oz (360 mL) of beer, 11/2 oz (45 mL) of liquor, or 5 oz (150 mL) of wine. ^ Positive alcohol breath test result or positive urine drug screen at Screening or Check-in. ^ Positive hepatitis panel and/or positive human immunodeficiency virus test. ^ The subject has received a live attenuated vaccine within 6 weeks prior to Day 1 or plans to receive a live attenuated vaccine during the study period. ^ Participation in a clinical study involving administration of an investigational drug (new chemical entity) in the past 30 days, or 5 half-lives (if known) of the investigational product, whichever is longer, prior to dosing. ^ Use of over-the-counter or prescription medications (including herbal medications) within 14 days or 5 half-lives (whichever is longer) prior to dosing until completion of the follow-up visit. By exception, use of acetaminophen will be allowed. ^ Smokers (or other nicotine users) who cannot refrain from nicotine use while confined to the clinical unit on Days -1 to 8 of the study. ^ Receipt of blood products within 2 months prior to Check-in. ^ Donation of blood from 3 months prior to Screening, plasma from 2 weeks prior to Screening, or platelets from 6 weeks prior to Screening. ^ Poor peripheral venous access. ^ Have previously participated in any study investigating CXA-10. ^ Subjects who, in the opinion of the Investigator (or designee), should not participate in this study Pregnancy [0256] All female participants must have a negative pregnancy test (serum β-hCG) at screening. [0257] Females of childbearing potential and females with a history of bilateral tubal ligation (in the absence of documented menopause) must have a negative serum pregnancy test at all subsequent study visits. Negative serum pregnancy test results must be confirmed before the first dose of study medication is administered. [0258] Female subjects who have had a hysterectomy or are postmenopausal (as confirmed by FSH > 40U/L at Screening [Visit 1]) do not require urine pregnancy testing after Screening Visit 1. [0259] Reports of pregnancies in female subjects (or in female partners of male subjects) will be collected after the start of dosing and until the final follow-up visit on Day 14 ± 1 day. [0260] Female subjects found to be pregnant are withdrawn from the study. [0261] The Investigator collects pregnancy information on any female subject who becomes pregnant while participating in this study. The Investigator records pregnancy information on the appropriate form and submit it to the Medical Monitor within 24 hours of learning of the pregnancy. The subject is followed to determine the outcome of the pregnancy. Generally, follow-up is no longer than 6 to 8 weeks following the estimated delivery date. Any premature termination of the pregnancy is reported. [0262] All pregnancies are reported from the site and documented according to the same procedures as SAE reporting. While a pregnancy is be considered as an SAE, any pregnancy complication or elective termination of a pregnancy for medical reasons is recorded as an AE or SAE. [0263] A spontaneous abortion is always considered to be an SAE and is reported as such. Furthermore, any SAE occurring as a result of a post-study pregnancy and which is considered reasonably related to the investigational product by the Investigator, is reported to the Sponsor. While the Investigator is not obligated to actively seek this information in former study participants, he or she may learn of an SAE through spontaneous reporting. Subjects are asked to contact the clinic in these situations. Study Assessments and Procedures [0264] Details for study assessments are provided. The timing of planned study assessments, including safety and PL assessments may be altered during the course of the study based on newly available data to ensure appropriate monitoring, but the number of assessments is not increased, unless required to care for any emerging clinical events at the discretion of the Investigator. The change in timing for any planned study assessments must be approved and documented by the Sponsor, but this does not constitute a protocol amendment. The addition of any new study assessments to the protocol would constitute an amendment, unless required for appropriate medical care due to an AE at the discretion of the principal investigator. [0265] In addition to the protocol-specified procedures, at any time during the study appropriate medical evaluations and interventions should be implemented, as necessary, based on the clinical presentation of the subject according to standard of care. Changes to the medical conditions, history and medications and all other medical interventions are documented in the electronic Case Report Forms (eCRFs). Adverse Events [0266] The condition of each subject is monitored from the time of signing the ICF through the final follow-up visit. Subjects are observed for any signs or symptoms and asked about their condition by open questioning, such as “How have you been feeling since you were last asked?”, at least once each day while resident at the study site and at each study visit. Subjects are also be encouraged to spontaneously report AEs occurring at any other time during the study. [0267] All non-serious AEs, whether reported by the subject voluntarily or upon questioning, or noted on physical examination, are recorded from initiation of study drug until study completion. Serious AEs are recorded from the time the subject signs the ICF until study completion. The nature, time of onset, duration, and severity are documented, together with an Investigator’s (or designee’s) opinion of the relationship to study drug. [0268] Adverse events recorded during the course of the study are followed up, where possible, until resolution or until the unresolved AEs are judged by the Investigator (or designee) to have stabilized. This is completed at the Investigator’s (or designee’s) discretion. [0269] In addition to standard AE assessments, subjects are asked to complete a GI Symptom Questionnaire at the time points indicated. Clinical Laboratory Evaluations [0270] Blood and urine samples are collected for clinical laboratory evaluations according to Table 3. Subjects are asked to provide urine samples for drugs of abuse screen and are asked to undergo an alcohol breath test at the times indicated. An Investigator performs a clinical assessment of all clinical laboratory data. Any clinically significant laboratory abnormality should be recorded as an AE.

Vital Signs [0271] Supine blood pressure, supine pulse rate, and oral body temperature are assessed at the times indicated in the Schedule of Assessments. Vital signs are optionally taken at other times if judged to be clinically appropriate or if the ongoing review of the data suggests a more detailed assessment of vital signs is required. [0272] Subjects must be supine and at rest for at least 5 minutes before blood pressure and pulse rate measurements. All measurements are performed singly and repeated if outside the relevant clinical reference range, except for blood pressure at Screening and Day -1, which are performed in triplicate. Any clinically significant vital sign abnormality should be reported as an AE. 12-lead Electrocardiogram [0273] Resting 12-lead ECGs are recorded after the subject has been supine and at rest in a quiet environment, free from distractions, for at least 5 minutes at the times indicated in the Schedule of Assessments. In addition to heart rate, the following intervals are recorded: PR, RR, QRS, QT, and QTcF. The Investigator (or designee) makes an assessment of any abnormal finding as clinically significant or not clinically significant. Any clinically significant ECG abnormality should be reported as an AE. [0274] Single 12-lead ECGs is repeated twice (for a total of 3 ECGs) if the Investigator deems a repeat is indicated to assess prolonged QT intervals. [0275] Additional 12-lead ECGs is optionally performed at other times if judged to be clinically appropriate or if the ongoing review of the data suggests a more detailed assessment of ECGs is required. Physical Examination [0276] A full physical examination or limited physical examination is performed at the timepoints specified in Table 2. The limited physical examination includes a review of the following body systems: cardiovascular, respiratory, dermatologic, and gastrointestinal. Pharmacokinetics [0277] Blood samples for PK (approximately 6 mL each) are collected over the course of the study. The PK blood sample collection times is recorded eCRF. The time of study medication administration is also be recorded in eCRF. [0278] MicroConstatns bioanalytical assay PK blood sample processing must begin within 4 minutes of collection. Blood samples are collected, processed, stored, and shipped according to the instructions provided in the SPM. AIT Bioscience Assay [0279] Blood samples for PK (approximately 5 mL each) are collected relative to Day 7 dose. The PK blood sample collection times are recorded in the eCRF. The time of study medication administration are also recorded in the eCRF. Additional PK samples may be collected in any subject who experiences QTcF prolongation. Pharmacodynamics [0280] Blood (approximately 10 mL per time point) is collected for transcriptomic panel endpoints. The panel will include genes for Nrf2 activation and HSP activation. Blood is processed for PBMCs as detailed in the SPM. [0281] Blood (approximately 5 mL per time point) is collected for FGF21 and FGF19). Blood is processed for plasma as detailed in the SPM. [0282] Blood (approximately 10 mL per time point) is collected for and ex vivo assay of CXA- 10 activity markers. Blood is processed for plasma as detailed in the SPM. [0283] Based on emerging data, additional pharmacodynamic (PD) parameters are optionally examined in these samples. Lifestyle and Dietary Restrictions [0284] Subjects are fasted from all food and drink (except water) for at least 8 hours prior to each morning dose. On dosing days: ^ Cohorts 1 and 2 must eat a standard breakfast (containing up to approximately 30% fat) within approximately 30 minutes prior to each morning dose; ^ Cohort 1 only approximately 10 hours after the morning dose, subjects are given dinner (containing up to approximately 30% fat) within approximately 30 minutes prior to the evening dosing ^ Subjects will refrain from consuming water until approximately 2 hours postdose, excludng the amount of water consumed at dosing, and are fasted until approximately 4 hours postdose. At all other times during the study, subjects may consume water ad libitum. [0285] During all clinic days, subjects areprovided breakfast, lunch and dinner if appropriate. [0286] On Day -1, Day 1, Day 6, and Day 7, the meals and snacks must be low in fructose (fructose may interfere with some of the PD markers). [0287] Consumption of alcohol is not permitted from 72 hours prior to Check-in until Discharge. Subjects are not permitted to use tobacco- or nicotine-containing products from Check-in until Discharge. [0288] Subjects are abstained from strenuous exercise for 48 hours prior to each blood collection for clinical laboratory tests. Strenuous exercise includes strength (weightlifting, barre, etc.) and speed-strength (boot camp, cross fit, etc.) activities. Exercise such as walking or a slow jog is permitted as long as it is within the subject’s typical regimen. [0289] Because reproductive toxicology studies have not yet been conducted, the risks of CXA- 10 on the fetus are unknown. [0290] Male subjects are required to use condoms during the study until final follow-up visit if they engage in sexual intercourse with a female of childbearing potential. Alternatively, the male subject must practice abstinence or have a medical history that includes vasectomy. [0291] Females of childbearing potential are required to use double-barrier methods of contraception; condoms plus the use of caps (with spermicide) or plus the use of intrauterine device (IUD) are acceptable during the study until final follow-up visit if they engage in sexual intercourse with a man. Alternatively, the female of childbearing potential must: ^ Abstain from sexual intercourse; ^ Have a male partner who is sterile prior to the female subject’s entry into the study and is the sole sexual partner for that female subject; and ^ Have a same-sex partner and abstains from bisexual activities where there is any risk of pregnancy Data Analysis and Statistical Considerations [0292] Sample size of at least 12 subjects per cohort (at least 9 active and 3 placebo) is based on feasibility and is deemed suitable for assessing the safety, tolerability, PK, and PD of CXA 10. [0293] Safety population for analysis includes all subjects who receive at least one dose of study medication. [0294] PK population for analysis includes all subjects who receive at least one dose of study medication and have at least 1 quantifiable CXA-10 concentration. [0295] PD population for analysis includes all subjects who receive at least one dose of study medication and have at least 1 evaluable PD assessment. [0296] In general, categorical variables are summarized by the count (N) and percentage of subjects (%). Continuous variables are summarized by the number of non-missing observations (N), mean, standard deviation, median, minimum, and maximum values. Summary tables are provided by time point and by dose group and overall (i.e., pooled dose groups). When appropriate, estimates are provided with two-sided 95% confidence intervals. All study data are displayed in the data listings. [0297] Safety analyses are performed on data from all subjects in the Safety Population. AEs, clinical laboratory evaluations, and other safety measures (e.g., vital signs) are listed and summarized. No formal statistical analysis of safety data is planned. All available data are reviewed throughout the study, as the data become available. Pharmacokinetic Analysis [0298] Concentration time data for CXA-10 and its metabolites (8, 9 alkene and 10-nitrostearate) (MicroConstants bioanalytical assay) are evaluated using standard non-compartmental analysis (NCA) methods. If feasible, PK parameters at all doses include Cmax, Tmax, t¹/₂, AUC0-last, AUC0-inf, CL/F, Vd/F, and λz. Other PK parameters are calculated, as appropriate. Accumulation ratio is determined based on Cmax and AUC on Days 1 and 7. [0299] If the concentration time data are available from the AIT Biosciences bioanalytical method, then a similar PK analysis approach is conducted as outlined above. Depending on the availability of the AIT Biosciences bioanalytical, this analysis is optionally conducted at a later date. [0300] If concentration time data for CXA-10 and/or its metabolites in triglycerides are available, then a similar PK analysis approach is conducted as outlined above. Depending on the availability of a bioanalytical assay to measure CXA-10 in triglycerides, this analysis is optionally conducted at a later date. [0301] Derived PK parameters and plasma concentrations are listed and summarized descriptively by dose. Descriptive statistics (n, arithmetic mean, standard deviation, minimum, median, maximum, geometric mean, and %CV for the geometric mean) are calculated for all pharmacokinetic parameters. Pharmacodynamic Analysis [0302] PD data is listed and summarized descriptively in tabular or graphical format as appropriate. Change from baseline is optionally calculated and summarized, as appropriate. Time Period, Frequency, and Method of Detecting AEs and SAEs [0303] At each visit, AEs/SAEs of the subjects are evaluated. All AEs must be recorded irrespective of whether they are considered drug-related. [0304] Any AEs/SAEs already documented at a previous assessment and designated as ongoing, should be reviewed at subsequent visits as necessary. If these have resolved, this should be documented. Changes in intensity or frequency of AEs/SAEs should be recorded as separate events (i.e., a new record started). Recording AEs and SAEs [0305] All clinical events, including either observed or volunteered problems, complaints or symptoms are recorded on the AE page(s) of the eCRF. The need to capture this information is not dependent upon whether the clinical event is associated with study medication. Adverse clinical events resulting from concurrent illnesses or reactions to concurrent medications are also recorded. In order to avoid vague, ambiguous, or colloquial expressions, the AE should be recorded in standard medical terminology rather than the subject’s own words. [0306] Each adverse clinical event is evaluated for duration, whether it was continuous or intermittent, intensity, and relationship to the study medication or other causes. Start and stop dates, relationship to study medication, medical management, outcome, and alternative causality of event are recorded in the AE section of the eCRF. [0307] For AEs other than diarrhea, the severity of an adverse event (AE and SAE) is scored according to the following scale: Mild: Awareness of sign or symptom, but easily tolerated Moderate: Discomfort enough to cause interference with usual activity Severe: Incapacitating with inability to work or perform usual activity [0308] An AE that is assessed as severe should not be confused with a SAE. [0309] For AEs of diarrhea, the severity of diarrhea is graded according to the Common Terminology Criteria for Adverse Events (CTCAE) version 4.03: Grade 1: Increase of < 4 stools per day over baseline; mild increase in ostomy output compared with baseline Grade 2: Increase of 4 to 6 stools per day over baseline; moderate increase in ostomy output compared with baseline Grade 3: Increase > 7 stools per day over baseline; incontinence; hospitalization indicated; severe increase in ostomy output compared with baseline; limiting self-care activities of daily living Grade 4: Life-threatening consequences; urgent intervention indicated Grade 5: Death [0310] AEs are assessed at each visit. At the end of the study, subjects with ongoing AEs considered at least possibly related to study medication are followed until resolution or for a total 30 days as warranted by the nature of the AE. [0311] After the initial AE/SAE report, the Investigator is required to proactively follow each subject. For clinically important AEs and all SAEs, the Investigator provides further information on the subject’s condition to the Medical Monitor as described in Section 12.6. [0312] Any SAE occurring in a subject receiving treatment or any SAE that the Investigator becomes aware of post-treatment during the follow up period, must be reported by the Investigator to the Medical Monitor or designee within 24 hours even if the SAE does not appear to be drug-related. This should be done by following the SAE reporting instructions in the SPM. Additionally, it may be necessary for the Sponsor to directly communicate with the Investigator if additional information is required. Example 2 - High-Dose of CXA-10 on HbF in Cultured K562 Cells [0001] This study assesses the effects of different dosages of CXA-10 on the percentage of cultured K562 erythroleukemic cells positive for fetal hemoglobin (HbF). K562 cells are cultured at 37°C in IMDM® to which the required concentrations of test items or DMSO (negative control) are added. Plated cells are maintained at 37°C for 3 days at the end of which the presence of HbF within cells is detected by flow cytometry. The treatment groups are consistent with Table 3.

BID = twice daily; QD = once daily. Note that the planned dosing regimens maybe be subject to change. [0313] Treatment with CXA-10 produces statistically significant increases in HbF positive cells. Compared to vehicle treated cells, CXA-10 elicits statistically significant increases in HbF cells to between 50% and 70% at 300-450 mg per day, and 70% to 85% at 450-900mg per day, with a peak increase to 85.6% at 900mg. [0314] For reference, the above experiment is repeated with hydroxyurea (HU), a known inducer of HbF. In comparison, HU elicits concentration dependent and statistically significant increases in HbF-positive cells at concentrations of 10 μM and above (but not at 1 and 3 μM), with the highest percentage of HbF-positive cells observed (92.6%) at the 100 μM concentration, which is 10 times the concentration of CXA-10 requires to elicit a similar response. Consistent results are observed in a similar experiment in which a broader range of concentrations was examined. Example 3 - High-Dose of CXA-10 on HbF Production in CD34+ Derived Red Blood Cells from SCD Subjects [0315] This study assesses the effects of CXA-10 and HU on the production of HbF in red blood cells (RBCs) of 5 subjects with SCD. Blood derived CD34+ cells from 5 SCD subjects undergoing transfusions are cultured for 5 days under continuous exposure to various doses of CXA-10 (see Table 4) or 30 μM HU and both the percentage of HbF positive cells and the amount of HbF within the cells are measured.

BID = twice daily; QD = once daily. Note that the planned dosing regimens maybe be subject to change. [0316] CXA-10 significantly increases the percentage of HbF positive CD36+ cells relative to control treated cells, from a mean of 15-20% in controls to a mean of 20-30%, and the amount of HbF within these cells, from a mean MFI of 100-150%. [0317] Hydroxyurea elicits greater than 80% cell death in cultures from 2 of the 5 subjects such that no assessments are made for them. For the remaining 3 subjects, HU does not significantly increase the percentage of HbF positive CD36+ cells (mean of 23.9%) relative to control-treated cells, but significantly increases the amount of HbF expressed from a mean MFI of 7,484 in controls to 19,383 (258%). [0318] Example 4 – Effect of Hydroxyurea vs. CXA-10 on HbF Positive and Sickled Red Blood Cells in Berkeley Sickle Cell Transgenic Mice [0319] This study assesses the effects of chronic dosing (30 day) of CXA-10 and HU on HbF and cell sickling in a mouse model of sickle cell disease. Berkeley sickle cell transgenic mice (Hbatm1PazHbbtm1Tow Tg(HBA-HBBs)41Paz/J) are divided into groups of 7 to 8 animals and are dosed once daily or twice daily, by gavage, for 30 days with either vehicle (PEG:water), 5 mg/kg/day CXA-10, 7.25 mg/kg/day CXA-10, 10 mg/kg/day CXA-10, 15 mg/kg/day CXA-10, or 100 mg/kg/day of HU. This mouse genotype mimics the genetic, hematologic and histopathologic features that are found in humans afflicted with sickle cell anemia, including irreversibly sickled RBCs, anemia and multi-organ pathology. Blood is collected from treated animals on Day 30 for limited routine hematology as well as derivation of the percent of sickled and HbF positive RBCs and total bilirubin. At termination the spleen is removed and weighed. [0320] After 30 days of treatment, both CXA-10 and HU results in statistically significant decreases in the percentage of sickled RBCs and increases in the percentage of HbF positive RBCs relative to controls. In addition, both compounds lead to statistically significant decreases in total bilirubin, total leucocyte count, and spleen weight. They reduce neutrophil levels and leukotytosis. These changes are not associated with any apparent alterations in RBC count, hemoglobin concentration or hematocrit. [0321] Administration of 5 mg/kg/day CXA-10, 7.25 mg/kg/day CXA-10, 10 mg/kg/day CXA- 10, 15 mg/kg/day CXA-10, for 30 days are well tolerated with no related deaths or abnormal clinical signs. Administration of 100 mg/kg HU is also well tolerated. Conclusion [0322] Repeated administration of high oral doses of CXA-10 (450 mg to 1800 mg per day) are well tolerated when CXA-10 is administered with food. Initial gastrointestinal effects are self- limited and diminish upon repeat oral dosing. Thus, oral doses of 450-1800 mg per day are safe with no material adverse events or serious adverse events. EQUIVALENTS AND INCORPORATION BY REFERENCE [0323] While the invention has been particularly shown and described with reference to a preferred embodiment and various alternate embodiments, it will be understood by persons skilled in the relevant art that various changes in form and details can be made therein without departing from the spirit and scope of the invention. [0324] All references, issued patents and patent applications cited within the body of the instant specification are hereby incorporated by reference in their entirety, for all purposes.

Claims

CLAIMS 1. A method of treating a disease, comprising: administering to a human subject with a disease an oral daily dose of a compound of Formula I,

or a salt, stereoisomer, deuterated analog, or fluorinated analog thereof, in an amount of at least 450 milligrams and less than 2,000 milligrams per day, wherein: X is selected from H and

a is from 0-30; b is from 0-30; R¹ is selected from H, alkyl, haloalkyl, cycloalkyl, aryl, and heteroaryl, -C(0)-R², gluconate, glycoside, glucuronide, tocopherols, and PEG groups; R² is selected from alkyl, haloalkyl, cycloalkyl, aryl, and heteroaryl.

2. The method of claim 1, wherein the daily oral dose is 450 mg to 1,800 mg.

3. The method of claim 1 or 2, wherein the daily oral dose is 450 mg or 600 mg.

4. The method of any one of claims 1 to 3, wherein the daily oral dose is 450 mg.

5. The method of any one of claims 1 to 3, wherein the daily oral dose is 600 mg.

6. The method of any one of claims 1 to 5, wherein the daily oral dose is administered in an undivided dose once a day.

7. The method of any one of claims 1 to 5, wherein the daily oral dose is administered in divided subdoses twice a day.

8. The method of any one of claims 1 to 5, wherein the daily oral dose is administered in divided subdoses three times a day.

9. The method of any one of claims 1 to 7, wherein the daily oral dose is administered for at least 7 days.

10. The method of any one of claims 1 to 9, wherein X is

11. The method of any one of claims 1 to 10, wherein R¹ is H, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, or pentyl.

12. The method of any one of claims 1 to 11, wherein R¹ is H.

13. The method of any one of claims 1 to 12, wherein the compound of Formula I is nitro-oleic acid.

14. The method of any one of claims 1 to 13, wherein the compound of Formula I is selected from:

15. The method of any one of claims 1 to 14, wherein the compound of Formula I is 10-nitro- 9(E)-octadec-9-enoic acid (CXA-10), or a salt, stereoisomer, deuterated analog, or fluorinated analog thereof.

16. The method of any one of claims 1 to 14, wherein the compound of Formula I is 9-nitro-9(E)- octadec-9-enoic acid (CXA-9), or a salt, stereoisomer, deuterated analog, or fluorinated analog thereof.

17. The method of any one of claims 1 to 16, wherein the disease is selected from solid organ fibrosis, inflammatory disease, cardiovascular disease, renal disease, kidney failure, ischemic kidney injury, acute kidney injury (AKI), chronic kidney injury (CKI), chronic kidney disease (CKD), obesity associated chronic kidney disease, diabetic nephropathy, kidney fibrosis, progressive kidney disease (renal fibrosis), focal segmental glomerular sclerosis (FSGS), primary FSGS, secondary FSGS, sickle cell nephropathy, glomerulonephritis, nephrotic syndrome, non-alcoholic steatohepatitis (NASH), fatty liver disease, non-fatty liver disease (NFLD), pulmonary arterial hypertension (PAH), pulmonary fibrosis, psoriasis, allergic airway disease, obesity, anti-adipogenic disease, type II diabetes, lithium induced diabetes insipidus, sickle cell disease, sickle cell crisis, idiopathic pulmonary fibrosis (IPF), interstitial lung diseases (ILDs), scleroderma, inflammatory gastrointestinal disease, colitis, inflammatory bowel disease, neurodegenerative disease, amyotrophic lateral sclerosis (ALS), metabolic syndrome, neuropathy, Charcot-Marie-Tooth disease, and mitochondrial related disease.

18. The method of any one of claims 1 to 17, wherein the human subject has sickle cell disease.

19. The method of any one of claims 1 to 17, wherein the human subject has sickle cell crisis.

20. The method of any one of claims 1 to 19, wherein the compound of Formula I is formulated in a pharmaceutical composition, wherein the pharmaceutical composition comprises the compound of Formula I and at least one pharmaceutically acceptable carrier or excipient.

21. The method of claim 20, wherein the pharmaceutically acceptable carrier or excipient comprises a triglyceride.

22. The method of claim 21, wherein the triglyceride is a medium chain triglyceride.

23. The method of claim 22, wherein the medium chain triglyceride is selected from hexanoic acid, octanoic acid, decanoic acid, and dodecanoic acid.

24. The method of any one of claims 20 to 23, wherein the pharmaceutical composition comprises Miglyol.

25. The method of any one of claims 20 to 24, wherein the pharmaceutical composition further comprises an antioxidant.

26. The method of claim 25, wherein the antioxidant is butylated hydroxytoluene (BHT).

27. The method of any one of claims 23 to 26, wherein the pharmaceutical composition further comprises a thickening agent.

28. The method of claim 27, wherein the thickening agent is fumed silica.

29. The method of any one of claims 20 to 28, wherein the pharmaceutical composition is in an oral unit dosage form.

30. The method of claim 29, wherein the oral unit dosage form is a capsule.

31. The method of claim 30, wherein the capsule comprises hydroxypropyl methylcellulose (HMPC).

32. The method of any one of claims 1 to 31, wherein the human subject is between 18 and 60 years of age.

33. The method of any one of claims 1 to 32, wherein the human subject is male.

34. The method of any one of claims 1 to 33, wherein the human subject is female.

35. The method of any one of claims 1 to 34, wherein the human subject has a body mass index (BMI) between 18.0 and 32.0 kg/m².

36. The method of any one of claims 1 to 35, wherein the daily oral dose is administered to the human subject with a meal.

37. The method of any one of claims 1 to 35, wherein the daily oral dose is administered to the human subject within 30 minutes of a meal.

38. The method of any one of claims 1 to 35, wherein the daily oral dose is administered to the human subject within 60 minutes of a meal.

39. The method of any one of claims 1 to 35, wherein the daily oral dose is administered to the human subject without a meal.

40. The method of any one of claims 1 to 39, wherein: the compound of Formula I is 10-nitro-9(E)-octadec-9-enoic acid (CXA-10), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof; the oral daily dose is 450 mg; and the human subject has sickle cell disease.

41. The method of any one of claims 1 to 40, wherein: the compound of Formula I is 10-nitro-9(E)-octadec-9-enoic acid (CXA-10), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof; the oral daily does is 600 mg; and the human subject has sickle cell disease.

42. The method of any one of claims 1 to 39, wherein: the compound of Formula I is 10-nitro-9(E)-octadec-9-enoic acid (CXA-10), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof; the oral daily does is 450 mg; and the human subject has sickle cell crisis.

43. The method of any one of claims 1 to 39, wherein: the compound of Formula I is 10-nitro-9(E)-octadec-9-enoic acid (CXA-10), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof; the oral daily does is 600 mg; and the human subject has sickle cell crisis.

44. The method of any one of claims 1 to 39, wherein: the compound of Formula I is 9-nitro-9(E)-octadec-9-enoic acid (CXA-9), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof; the oral daily dose is 450 mg; and the human subject has sickle cell disease.

45. The method of any one of claims 1 to 39, wherein: the compound of Formula I is 9-nitro-9(E)-octadec-9-enoic acid (CXA-9), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof; the oral daily does is 600 mg; and the human subject has sickle cell disease.

46. The method of any one of claims 1 to 39, wherein: the compound of Formula I is 9-nitro-9(E)-octadec-9-enoic acid (CXA-9), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof; the oral daily does is 450 mg; and the human patient has sickle cell crisis.

47. The method of any one of claims 1 to 39, wherein: the compound of Formula I is 9-nitro-9(E)-octadec-9-enoic acid (CXA-9), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof; the oral daily does is 600 mg; and the human patient has sickle cell crisis.

48. A pharmaceutical composition comprising a compound of Formula I,

a is from 0-30; b is from 0-30; R¹ is selected from H, alkyl, haloalkyl, cycloalkyl, aryl, and heteroaryl, -C(O)-R², gluconate, glycoside, glucuronide, tocopherols, and PEG groups; R² is selected from alkyl, haloalkyl, cycloalkyl, aryl, and heteroaryl; and at least one pharmaceutically acceptable carrier or excipient.

49. The pharmaceutical composition of claim 48, wherein X is

50. The pharmaceutical composition of claim 48 or 49, wherein R¹ is H, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, or pentyl.

51. The pharmaceutical composition of any one of claims 48 to 50, wherein R¹ is H.

52. The pharmaceutical composition of any one of claims 48 to 51, wherein the compound of Formula I is oleic acid.

53. The pharmaceutical composition of any one of claims 48 to 52, wherein the compound of Formula is nitro-oleic acid.

54. The pharmaceutical composition of any one of claims 48 to 53, wherein the compound of Formula I is selected from:

55. The pharmaceutical composition of any one of claims 48 to 54, wherein the compound of Formula I is 10-nitro-9(E)-octadec-9-enoic acid (CXA-10), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof.

56. The pharmaceutical composition of any one of claims 48 to 54, wherein the compound of Formula I is 9-nitro-9(E)-octadec-9-enoic acid (CXA-9), or a salt, stereoisomer, deuterated analog, and fluorinated analog thereof.

57. The pharmaceutical composition of any one of claims 48 to 56, wherein the pharmaceutical composition further comprises a triglyceride.

58. The pharmaceutical composition of claim 57, wherein the triglyceride is a medium chain triglyceride.

59. The pharmaceutical composition of claim 58, wherein the medium chain triglyceride is selected from hexanoic acid, octanoic acid, decanoic acid, and dodecanoic acid.

60. The pharmaceutical composition of any one of claims 48 to 59, wherein the pharmaceutical composition comprises miglyol.

61. The pharmaceutical composition of any one of claims 48 to 60, wherein the pharmaceutical composition further comprises an antioxidant.

62. The pharmaceutical composition of claim 61, wherein the antioxidant is butylated hydroxytoluene (BHT).

63. The pharmaceutical composition of any one of claims 48 to 62, wherein the pharmaceutical composition further comprises a thickening agent.

64. The pharmaceutical composition of claim 63, wherein the thickening agent is fumed silica.

65. An oral unit dosage form, comprising: a capsule shell; and the pharmaceutical composition of any one of claims 48 to 64.

66. The oral unit dosage form of claim 65, wherein the capsule shell comprises hydroxypropyl methylcellulose (HMPC).

67. The oral unit dosage form of claim 65 or 66, wherein the capsule contains 25 mg to 300 mg of an active pharmaceutical ingredient (API).

68. The oral unit dosage form of claim 67, wherein the capsule contains 150 mg of an active pharmaceutical ingredient (API).

69. The oral unit dosage form of claim 67 or 68, wherein the API is a compound of Formula I.

70. The oral unit dosage form of any one of claims 67 to 69, wherein the API is 10-nitro-9(E)- octadec-9-enoic acid or 9-nitro-9(E)-octadec-9-enoic acid.

71. The oral unit dosage form of any one of claims 67 to 69, wherein the API is 10-nitro-9(E)- octadec-9-enoic acid.

72. The oral unit dosage form of any one of claims 67 to 69, wherein the API is 9-nitro-9(E)- octadec-9-enoic acid.