Methods, compounds, compositions, formulations and uses for addressing mitochondrial disease resulting from nuclear DNA mutations

Abstract

The present disclosure provides methods, compounds, compositions, formulations or medicaments and related uses for treating, preventing, inhibiting, ameliorating or delaying the onset of a mitochondrial disease resulting from nuclear DNA mutations (e.g., mitochondrial DNA (mtDNA) depletion syndrome) in a subject. The methods comprise administering to the subject compounds, compositions, formulations or medicaments disclosed herein to thereby produce the aforementioned therapeutically beneficial effect(s).

Description

Atty. Dkt. No.091151-1691 METHODS, COMPOUNDS, COMPOSITIONS, FORMULATIONS AND USES FOR ADDRESSING MITOCHONDRIAL DISEASE RESULTING FROM NUCLEAR DNA MUTATIONS CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. Provisional Patent Application No.63 / 673,237, filed July 19, 2024, the content of which is incorporated herein by reference in its entirety for any and all purposes. TECHNICAL FIELD

[0002] The present application relates generally to methods, compounds, compositions / formulations / medicaments and related uses for treating, preventing, inhibiting, ameliorating or delaying the onset of mitochondrial disease resulting from nuclear DNA mutations (e.g., mitochondrial DNA (mtDNA) depletion syndrome) in a subject. The methods, compounds, compositions / formulations / medicaments and related uses all relate to the administration to the subject of a compound specifically identified herein. INTRODUCTION

[0003] The following description is provided to assist the understanding of the reader. None of the information provided or references cited herein is admitted as being prior art to the compounds, compositions, products, uses, and / or methods disclosed herein.

[0004] Mitochondria are the powerhouses of the cell. Their primary role is to perform oxidative phosphorylation to thereby generate adenosine triphosphate (ATP), a predominate source of energy for the cell. Mitochondria (an organelle of the cell) contain a set of DNA (the mtDNA) that encode a few dozen proteins, mostly subunits of the oxidative phosphorylation complexes involved in cellular respiration and ATP production. Almost all proteins (>99%) found within mitochondria are encoded by the cell’s nuclear, chromosomal DNA (nDNA).

[0005] The replication of mtDNA is critical to support mitochondrial function. All of the proteins necessary for mtDNA replication, the mtDNA Replisome, are encoded by nDNA. Among this family of enzymes are the POLG and TWNK genes, and their downstream protein products polymerase gamma and twinkle, respectively. Mutations in POLG and TWNK genes can lead to mutated forms of polymerase gamma and twinkle enzymes. TheseAtty. Dkt. No.091151-1691 mutations hinder mtDNA replicative / proofreading processes, and can result in mtDNA depletion, mtDNA deletions, and / or mtDNA mutations.

[0006] Mitochondrial DNA (mtDNA) depletion(s) and / or mitochondrial DNA (mtDNA) mutation(s) are associated with various diseases such as mitochondrial encephalopathy, lactic acidosis and stroke-like episodes (MELAS), myoclonic epilepsy and ragged red fibers (MERRF), and neuropathy, ataxia, and retinitis pigmentosa (NARP) syndrome.

[0007] Mitochondrial DNA depletion syndrome (MDDS) refers to any autosomal disorder caused by a significant drop in mitochondrial DNA in affected tissues. MDDS appears to be strongly correlated with deficiencies in mtDNA replication and nucleotide pool regulation. MDDS often manifests itself shortly after birth, a time when energy metabolism of a mammal transitions from glycolysis to oxidative phosphorylation. Symptoms of MDDS can be myopathic, hepatopathic, and / or encephalomyopathic. In some cases, MDDS is associated with neurodegenerative disease, such as Kearns–Sayre syndrome, Alzheimer’s disease, Parkinson’s disease (PD), sporadic Parkinson’s disease (sPD), or Alpers syndrome.

[0008] There is a need for effective therapeutics / treatments to address mitochondrial disease resulting from nuclear DNA mutations, including those that cause mitochondrial DNA (mtDNA) depletion(s), and / or increased mtDNA mutations / deletions, that contribute to mitochondrial DNA depletion syndrome (MDDS). SUMMARY

[0009] In one aspect, the present technology provides a method for treating, preventing, inhibiting, ameliorating or delaying the onset of a mitochondrial disease resulting from nuclear DNA mutations in a subject comprising administering to the subject in need thereof a therapeutically effective amount of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, wherein Compound 1 is:.Atty. Dkt. No.091151-1691

[0010] In some embodiments, the nuclear DNA mutations result in mitochondrial DNA (mtDNA) depletion(s) and / or mitochondrial DNA (mtDNA) rearrangement(s) in the subject. In some embodiments, the subject suffers from mitochondrial DNA (mtDNA) depletion syndrome. In some embodiments, the mitochondrial disease results from at least one mutation of the POLG gene. In some embodiments, mitochondrial disease results from at least one mutation of the TWNK (twinkle) gene. In some embodiments, the mitochondrial DNA (mtDNA) depletion(s) codes / code for at least one mutation of a complex I respiratory chain subunit. In some embodiments, the mitochondrial DNA (mtDNA) rearrangement(s) codes / code for at least one mutation of a complex I respiratory chain subunit.

[0011] In some embodiments, the subject has been diagnosed as having a mitochondrial disease that is neurodegenerative. In some embodiments, the mitochondrial disease is Kearns–Sayre syndrome or Alzheimer’s disease. In some embodiments, the mitochondrial disease is Parkinson’s disease (PD) or sporadic Parkinson’s disease (sPD). In some embodiments, the mitochondrial disease is Alpers syndrome.

[0012] In some embodiments, the subject is a mammal. In some embodiments, the subject is human.

[0013] In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 6 weeks or more. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, administered daily for 24 weeks or more. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 48 weeks or more. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 52 weeks or more.

[0014] In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered orally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered subcutaneously. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered topically. In some embodiments, Compound 1, orAtty. Dkt. No.091151-1691 pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intranasally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered systemically. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intravenously. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intraperitoneally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intradermally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intraocularly. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered ophthalmically. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intrathecally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intracerebroventricularly. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered iontophoretically. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered transmucosally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intravitreally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered or intramuscularly.

[0015] In some embodiments, the method further comprises separately, sequentially, or simultaneously administering at least one additional therapeutic agent to the subject. In some embodiments, the additional therapeutic agent comprises levodopa, optionally in combination with carbidopa (LODOSYN^).

[0016] In some embodiments, the combination of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, optionally in combination with the at least one additional therapeutic agent, has a synergistic effect in the treating, preventing, inhibiting, ameliorating or delaying the onset of the mitochondrialAtty. Dkt. No.091151-1691 disease. In some embodiments, administration of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, optionally in combination with the at least one additional therapeutic agent, prolongs survival of the subject, as compared with an untreated subject or an untreated control group of subjects.

[0017] In another aspect, the present technology provides for use of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, in the preparation of a composition, formulation or medicament for treating, preventing, inhibiting, ameliorating or delaying the onset of a mitochondrial disease resulting from nuclear DNA mutations in a subject, wherein Compound 1 is:.

[0018] In some embodiments of the use, the nuclear DNA mutations result in mitochondrial DNA (mtDNA) depletion(s) and / or mitochondrial DNA (mtDNA) rearrangement(s) in the subject. In some embodiments of the use, the subject suffers from mitochondrial DNA (mtDNA) depletion syndrome. In some embodiments of the use, the mitochondrial disease results from at least one mutation of the POLG gene. In some embodiments of the use, the mitochondrial disease results from at least one mutation of the TWNK (twinkle) gene. In some embodiments of the use, the mitochondrial DNA (mtDNA) depletion(s) codes / code for at least one mutation complex I respiratory chain subunit. In some embodiments of the use, the mitochondrial DNA (mtDNA) rearrangement(s) codes / code for at least one mutation of the complex I respiratory chain subunit.

[0019] In some embodiments of the use, the subject has been diagnosed as having a mitochondrial disease that is neurodegenerative. In some embodiments of the use, the mitochondrial disease is Kearns–Sayre syndrome or Alzheimer’s disease. In some embodiments of the use, the mitochondrial disease is Parkinson’s disease (PD) or sporadic Parkinson’s disease (sPD). In some embodiments, the mitochondrial disease is Alpers syndrome.

[0020] In some embodiments of the use, the subject is a mammal. In some embodiments of the use, the subject is human.Atty. Dkt. No.091151-1691

[0021] In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 6 weeks or more. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 24 weeks or more. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 48 weeks or more. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 52 weeks or more.

[0022] In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration orally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration subcutaneously. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration topically. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intranasally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration systemically. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intravenously. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intraperitoneally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administrationAtty. Dkt. No.091151-1691 intradermally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intraocularly. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration ophthalmically. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intrathecally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intracerebroventricularly. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration iontophoretically. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration transmucosally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intravitreally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intramuscularly.

[0023] In some embodiments of the use, administration of the composition, formulation or medicament further comprises separately, sequentially, or simultaneously administering an additional therapeutic agent to the subject. In some embodiments of the use, the additional therapeutic agent comprises levodopa, optionally in combination with carbidopa (LODOSYN^).

[0024] In some embodiments of the use, administering the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, optionally in combination with at least one additional therapeutic agent, has a synergistic effect in the treating, preventing, inhibiting,Atty. Dkt. No.091151-1691 ameliorating or delaying the onset of the mitochondrial disease. In some embodiments of the use, administering the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, optionally in combination with at least one additional therapeutic agent, prolongs survival of the subject, as compared with an untreated subject or an untreated control group of subjects.

[0025] In another aspect, the present technology provides a composition comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, for use in treating, preventing, inhibiting, ameliorating or delaying the onset of a mitochondrial disease resulting from nuclear DNA mutations in a subject, wherein.

[0026] In some embodiments of the foregoing composition, the nuclear DNA mutations result in mitochondrial DNA (mtDNA) depletion(s) and / or mitochondrial DNA (mtDNA) rearrangement(s) in the subject. In some embodiments of the foregoing composition, the subject suffers from mitochondrial DNA (mtDNA) depletion syndrome. In some embodiments of the foregoing composition, the mitochondrial disease results from at least one mutation of the POLG gene. In some embodiments of the foregoing composition, the mitochondrial disease results from at least one mutation of the TWNK (twinkle) gene. In some embodiments of the foregoing composition, the mitochondrial DNA (mtDNA) depletion(s) codes / code for at least one mutation of a complex I respiratory chain subunit. In some embodiments of the foregoing composition, the mitochondrial DNA (mtDNA) rearrangement(s) codes / code for at least one mutation of a complex I respiratory chain subunit.

[0027] In some embodiments of the foregoing composition, the subject has been diagnosed as having a mitochondrial disease that is neurodegenerative. In some embodiments of the foregoing composition, the mitochondrial disease is Kearns–Sayre syndrome or Alzheimer’s disease. In some embodiments of the foregoing composition, the mitochondrial disease is Parkinson’s disease (PD) or sporadic Parkinson’s disease (sPD). In some embodiments, the mitochondrial disease is Alpers syndrome.Atty. Dkt. No.091151-1691

[0028] In some embodiments of the foregoing composition, the subject is a mammal. In some embodiments of the foregoing composition, the subject is human.

[0029] In some embodiments of the foregoing composition, the composition is administered daily for 6 weeks or more. In some embodiments of the foregoing composition, the composition is administered daily for 24 weeks or more. In some embodiments of the foregoing composition, the composition is administered daily for 48 weeks or more. In some embodiments of the foregoing composition, the composition is administered daily for 52 weeks or more.

[0030] In some embodiments of the foregoing composition, the composition is formulated for administration orally or subcutaneously. In some embodiments of the foregoing composition, the composition is formulated for administration topically, intranasally, systemically, intravenously, intraperitoneally, intradermally, intraocularly, ophthalmically, intrathecally, intracerebroventricularly, iontophoretically, transmucosally, intravitreally, or intramuscularly.

[0031] In another aspect, the present technology provides for a method for treating a subject suffering from mitochondrial DNA (mtDNA) depletion syndrome, the method comprising: administering to the subject a therapeutically effective amount of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, wherein Compound 1 is:.

[0032] In some embodiments of the foregoing method, the subject has at least one mutation of the POLG gene. In some embodiments of the foregoing method, the subject has at least one mutation of the TWNK (twinkle) gene. In some embodiments of the foregoing method, the subject is a mammal. In some embodiments of the foregoing method, the mammalian subject is a human.Atty. Dkt. No.091151-1691

[0033] In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered orally to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered subcutaneously to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered topically to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intranasally to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered systemically to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intravenously to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intraperitoneally to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intradermally to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intraocularly to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered ophthalmically to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intrathecally to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intracerebroventricularly to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered iontophoretically to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered transmucosally to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administeredAtty. Dkt. No.091151-1691 intravitreally to the subject. In some embodiments of the foregoing method, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intramuscularly to the subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Fig.1 is a bar graph illustration of data obtained for an in vitro assay using HepG2 cells treated with dideoxycytosine triphosphate (ddC) to assess ATP production in the presence and absence of ddC.

[0035] Fig.2 is a bar graph illustration of data obtained for an in vitro assay using HepG2 cells treated alone or in combination with each of dideoxycytosine triphosphate (ddC) and Compound 1 wherein viable cell density is determined.

[0036] Fig.3 is a bar graph illustration of data obtained for an in vitro assay using ddC- treated HepG2 cells that were then administered vehicle treatment (no RSL3), RSL3 alone, or RSL3 in combination with Compound 1 to assess ATP production under the conditions examined.

[0037] Fig.4 is a bar graph illustration of data obtained for an in vitro assay using HepG2 cells that were co-treated with ddC and ethidium bromide (EtBr) and administered either vehicle treatment (no RSL3), RSL3 alone, or RSL3 with Compound 1 to assess ATP production under conditions examined.

[0038] Fig.5A is a chart showing mouse plasma PK profile after 5 days treatment with Compound 1 at (i) 60 mg / kg, 5% Kolliphor^ ELP (K-ELP) in PBS, (ii) 60 mg / kg, 15% K- ELP in PBS, and (iii) 180 mg / kg, 15% K-ELP in PBS.

[0039] Figs.5B-5C are charts showing mouse tissue (Fig.5B – heart; Fig.5C – brain) exposure profile after 5 days of treatment with Compound 1 at: (i) 60 mg / kg, 5% Kolliphor^ ELP (K-ELP) in PBS, (ii) 60 mg / kg, 15% K-ELP in PBS, and (iii) 180 mg / kg, 15% K-ELP in PBS. DETAILED DESCRIPTION

[0040] It is to be appreciated that certain aspects, modes, embodiments, variations and features of the present disclosure are described below in various levels of detail in order to provide a substantial understanding of the present technology. The definitions of certainAtty. Dkt. No.091151-1691 terms as used in this specification are provided below. Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this present technology belongs.

[0041] In practicing the present technology, many conventional techniques in molecular biology, protein biochemistry, cell biology, immunology, microbiology and recombinant DNA are used. These techniques are well-known and are explained in, e.g., Current Protocols in Molecular Biology, Vols. I-III, Ausubel, Ed. (1997); Sambrook et al., Molecular Cloning: A Laboratory Manual, Second Ed. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989); DNA Cloning: A Practical Approach, Vols. I and II, Glover, Ed. (1985); Oligonucleotide Synthesis, Gait, Ed. (1984); Nucleic Acid Hybridization, Hames & Higgins, Eds. (1985); Transcription and Translation, Hames & Higgins, Eds. (1984); Animal Cell Culture, Freshney, Ed. (1986); Immobilized Cells and Enzymes (IRL Press, 1986); Perbal, A Practical Guide to Molecular Cloning; the series, Meth. Enzymol., (Academic Press, Inc., 1984); Gene Transfer Vectors for Mammalian Cells, Miller & Calos, Eds. (Cold Spring Harbor Laboratory, N Y, 1987); and Meth. Enzymol., Vols.154 and 155, Wu & Grossman, and Wu, Eds., respectively. I. Definitions

[0042] Definitions of specific functional groups and chemical terms are described in more detail below. The chemical elements are identified in accordance with the Periodic Table of the Elements, GAS version, Handbook of Chemistry and Physics, 7Sh Ed., inside cover. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Thomas Sorrell, Organic Chemistry, University Science Books, Sausalito, 1999; Smith and March, March's Advanced Organic Chemistry, 5th Edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; and Carruthers, Some Modern Methods of Organic Synthesis, 3rd Edition, Cambridge University Press, Cambridge, 1987.

[0043] It is to be appreciated that certain aspects, modes, embodiments, variations and features of the technology are described below in various levels of detail in order to provide a substantial understanding of the present disclosure. The definitions of certain terms as used in this specification are provided below. Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this technology belongs.Atty. Dkt. No.091151-1691

[0044] As used in this specification and the appended embodiments, the singular forms “a,” “an” and “the” include plural references unless the content clearly dictates otherwise. For example, reference to “a cell” includes a combination of two or more cells, and the like.

[0045] As used herein, “administering” or the “administration” of an agent (i.e., a therapeutic agent) or compound / drug product (including a composition (i.e., a formulation or medicament)) to a subject includes any route of introducing or delivering to a subject a compound / drug product to perform its intended function. Administration may be carried out by any suitable route, such as oral administration. Administration can be carried out subcutaneously. Administration can be carried out intravenously. Administration can be carried out intraocularly. Administration can be carried out systemically. Alternatively, administration may be carried out topically, intranasally, intraperitoneally, intradermally, ophthalmically, intrathecally, intracerebroventricularly, iontophoretically, transmucosally, intravitreally, or intramuscularly. Administration includes self-administration, the administration by another or administration by use of a device (e.g., an infusion pump).

[0046] As used herein, to “ameliorate” or “ameliorating” a disease, disorder or condition refers to results that, in a statistical sample or specific subject, make the occurrence of the disease, disorder or condition (or a sign, symptom or condition thereof) better or more tolerable in a sample or subject administered a therapeutic agent relative to a control sample, control subject or group of control subjects.

[0047] As used herein the terms “carrier” or “pharmaceutically acceptable carrier” refer to a diluent, adjuvant, excipient, or vehicle with which a compound / drug product / composition (including a formulation or medicament) is administered or formulated for administration. Non-limiting examples of such pharmaceutically acceptable carriers include liquids, such as water, saline, oils and solids, such as gum acacia, gelatin, starch paste, talc, keratin, colloidal silica, silica particles (nanoparticles or microparticles) urea, and the like. In addition, auxiliary, stabilizing, thickening, lubricating, flavoring, and coloring agents may be used. Other examples of suitable pharmaceutical carriers are described in Remington’s Pharmaceutical Sciences by E.W. Martin, herein incorporated by reference in its entirety.

[0048] As used herein, the phrase “delaying the onset of” refers to, in a statistical sample, postponing, hindering the occurrence of a disease, disorder or condition, or causing one or more signs or symptoms of a disease, disorder or condition to occur more slowly thanAtty. Dkt. No.091151-1691 normal, in a sample or subject administered a therapeutic agent or agents relative to a control sample, control subject or group of control subjects.

[0049] As used herein, the term “effective amount” refers to a quantity of a compound / composition / drug product sufficient to achieve a desired therapeutic and / or prophylactic effect, e.g., an amount that treats, prevents, inhibits, ameliorates, or delays the onset of the disease, disorder or condition, or the physiological signs or symptoms of the disease, disorder, or condition. In the context of therapeutic or prophylactic applications, in some embodiments, the amount of a compound / composition / drug product administered to the subject will depend on the type and severity of the disease and on the characteristics of the individual, such as general health, age, sex, body weight and tolerance to drugs. In some embodiments, it will also depend on the degree, severity and type of disease. The skilled artisan will be able to determine appropriate dosages depending on these and other factors. The compounds / compositions / drug products can also be administered in combination with one or more additional therapeutic compounds / agents (a so called “co-administration” where, for example, the additional or other therapeutic agent(s) could be administered simultaneously, sequentially or by separate administration).

[0050] As used herein, the term "hydrate" refers to a compound which is associated (e.g., complexed) with water. The number of the water molecules contained in a hydrate of a compound may be (or may not be) in a definite ratio to the number of the compound molecules in the hydrate.

[0051] As used herein, “inhibit” or “inhibiting” refers to the reduction in a sign, symptom, or condition (e.g., risk factor) associated with a disease or disorder associated with α- synucleinopathies. In one embodiment, inhibit or inhibiting refers to the reduction by at least a statistically significant amount compared to a control (or control subject). In one embodiment, inhibit or inhibiting refers to a reduction by at least 5 percent compared to control (or control subject). In various individual embodiments, inhibit or inhibiting refers to a reduction by at least 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 33, 40, 50, 60, 67, 70, 75, 80, 90, 95, or 99 percent compared to a control sample, control subject or group of control subjects.

[0052] As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a therapeutic compound that can be prepared with relatively nontoxic acids or bases, depending on the particular substituents found on the compounds described herein. When compoundsAtty. Dkt. No.091151-1691 contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, magnesium salt, or a similar salt. When compounds contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Salts derived from pharmaceutically acceptable inorganic bases include ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, and zinc salts, and the like. Salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary and tertiary amines, including substituted amines, cyclic amines, naturally-occurring amines and the like, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2- diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N- methylmorpholine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperadine, polyamine resins, procaine, purines, theobromine, triethylamine (NEt3), trimethylamine, tripropylamine, tromethamine and the like, such as where the salt includes the protonated form of the organic base (e.g., [HNEt3]+). Salts derived from pharmaceutically acceptable inorganic acids include salts of boric, carbonic, hydrohalic (hydrobromic, hydrochloric, hydrofluoric or hydroiodic), nitric, phosphoric, sulfamic and sulfuric acids. Salts derived from pharmaceutically acceptable organic acids include salts of aliphatic hydroxyl acids (e.g., citric, gluconic, glycolic, lactic, lactobionic, malic, and tartaric acids), aliphatic monocarboxylic acids (e.g., acetic, butyric, formic, propionic and trifluoroacetic acids), amino acids (e.g., aspartic and glutamic acids), aromatic carboxylic acids (e.g., benzoic, p- chlorobenzoic, diphenylacetic, gentisic, hippuric, and triphenylacetic acids), aromatic hydroxyl acids (e.g., o-hydroxybenzoic, p-hydroxybenzoic, 1-hydroxynaphthalene-2- carboxylic and 3-hydroxynaphthalene-2-carboxylic acids), ascorbic, dicarboxylic acids (e.g., fumaric, maleic, oxalic and succinic acids), glucuronic, mandelic, mucic, nicotinic, orotic, pamoic, pantothenic, sulfonic acids (e.g., benzenesulfonic, camphorsulfonic, edisylic, ethanesulfonic, isethionic, methanesulfonic, naphthalenesulfonic, naphthalene-1,5-disulfonic, naphthalene-2,6-disulfonic, p-toluenesulfonic acids (PTSA)), xinafoic acid, and the like. In some embodiments, the pharmaceutically acceptable counterion is selected from the group consisting of acetate, benzoate, besylate, bromide, camphorsulfonate, chloride, chlorotheophyllinate, citrate, ethanedisulfonate, fumarate, gluceptate, gluconate, glucoronate,Atty. Dkt. No.091151-1691 hippurate, iodide, isethionate, lactate, lactobionate, laurylsulfate, malate, maleate, mesylate, methylsulfate, naphthoate, sapsylate, nitrate, octadecanoate, oleate, oxalate, pamoate, phosphate, polygalacturonate, succinate, sulfate, sulfosalicylate, tartrate, tosylate, and trifluoroacetate. In some embodiments, the salt is a tartrate salt, a fumarate salt, a citrate salt, a benzoate salt, a succinate salt, a suberate salt, a lactate salt, an oxalate salt, a phthalate salt, a methanesulfonate salt, a benzenesulfonate salt, a maleate salt, a trifluoroacetate salt, a hydrochloride salt, or a tosylate salt. Also included are salts of amino acids such as arginate and the like, and salts of organic acids such as glucuronic or galactunoric acids and the like (see, e.g., Berge et al, Journal of Pharmaceutical Science 66: 1-19 (1977)). Certain specific compounds may contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts or exist in zwitterionic form. These salts may be prepared by methods known to those skilled in the art. Any other pharmaceutically acceptable carriers known to those of skill in the art are suitable for use with the present technology.

[0053] As used herein, “prevention” or “preventing” of a disease, disorder, or condition refers to results that, in a statistical sample, exhibit a reduction in the occurrence of the disease, disorder, or condition in a sample or subject administered a therapeutic agent or agents relative to a control sample, control subject or group of control subjects. Such prevention is sometimes referred to as a prophylactic treatment.

[0054] As used herein, the term “separate” therapeutic use refers to an administration of at least two active ingredients (e.g., therapeutic agents) at the same time or at substantially the same time by different routes.

[0055] As used herein, the term “sequential” therapeutic use refers to administration of at least two active ingredients (e.g., therapeutic agents) at different times, the administration route being identical or different. More particularly, sequential use refers to the whole administration of one of the active ingredients (e.g., therapeutic agents) before administration of the other or others commences. It is thus possible to administer one of the active ingredients over several minutes, hours, or days before administering the other active ingredient or ingredients. There is no simultaneous treatment in this definition.Atty. Dkt. No.091151-1691

[0056] As used herein, the term “simultaneous” therapeutic use refers to the administration of at least two active ingredients (e.g., therapeutic agents) by the same route and at the same time or at substantially the same time.

[0057] As used herein, the term "solvate" refers to forms of the compound that are associated with a solvent, possibly by a solvolysis reaction. This physical association may include hydrogen bonding. Conventional solvents include water, methanol, ethanol, isopropanol, acetic acid, ethyl acetate, acetone, hexane(s), dimethylsulfoxide (DMSO), tetrahydrofuran (THF), diethyl ether, and the like.

[0058] As used herein, a “subject” refers to a living animal. In various embodiments, a subject is a mammal. In various embodiments, a subject is a non-human mammal, including, without limitation, a mouse, rat, hamster, guinea pig, rabbit, sheep, goat, cat, dog, pig, minipig, horse, cow, or non-human primate. In certain embodiments, the subject is a human.

[0059] It is also to be appreciated that the various modes of treatment or prevention of medical conditions as described herein, in some embodiments, are intended to mean “substantial,” which includes total but also less than total treatment or prevention, and wherein some biologically or medically relevant result is achieved.

[0060] As used herein, a “synergistic therapeutic effect” refers to a greater-than-additive therapeutic effect which is produced by a combination of at least two agents, and which exceeds that which would otherwise result from the individual administration of the agents.

[0061] As used herein, the term "tautomer" refers to compounds that are interchangeable forms of a particular compound structure, and that vary in the displacement of hydrogen atoms and electrons. Thus, two structures may be in equilibrium through the movement of π electrons and an atom (usually H). For example, enols and ketones are tautomers because they are rapidly interconverted by treatment with either acid or base. Tautomeric forms may be relevant to the attainment of the optimal chemical reactivity and biological activity of a compound of interest.

[0062] As used herein, the terms “treating” or “treatment” refer to therapeutic treatment, wherein the object is to reduce, alleviate or slow down (lessen) a pre-existing disease, disorder or condition, or its related signs, or symptoms. By way of example, but not by way of limitation, a subject is successfully “treated” for a disease if, after receiving an effectiveAtty. Dkt. No.091151-1691 amount of the compound / composition / drug product or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, the subject shows observable and / or measurable reduction in or absence of one or more signs, or symptoms associated with the disease, disorder or condition. It is also to be appreciated that the various modes of treatment of medical conditions as described are intended to mean “substantial,” which includes total alleviation of conditions, signs or symptoms of the disease, disorder or condition, as well as “partial,” where some biologically or medically relevant result is achieved. II. Pharmaceutical Compositions, Routes of Administration, and Dosing

[0063] The methods, uses and compositions / formulations / medicaments of the present application utilize a therapeutically effective amount of Compound 1 or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof, wherein Compound 1 is:. Compound 1, or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof can be formulated into a drug product suitable for administration to a subject in need thereof. Such drug product can be referred to as a composition, formulation or medicament depending on its usage. Any mixture prepared by mixing Compound 1, or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof with a solvent and / or other compound(s) is a composition or formulation and may, or may not, be intended for administration to a subject. A medicament is generally considered a composition or formulation specifically prepared for administration to a subject to address a disease, disorder, or condition (e.g., a mitochondrial disease resulting from nuclear DNA mutations). In some embodiments, one or more of the hydrogen atoms of Compound 1 as used in a method, use, medicament or formulation as disclosed herein, can be substituted with a deuterium atom without any significant deviation from its efficacy or therapeutic profile.

[0064] For purposes of brevity, whenever there is a reference to Compound 1, this reference is intended to encompass any pharmaceutically acceptable salts, stereoisomers,Atty. Dkt. No.091151-1691 mixtures of stereoisomers, tautomers, hydrates, and / or solvates of Compound 1, as well as any modifications associated with substitution of a deuterium atom for a hydrogen atom in the Compound.

[0065] Compound 1 may be administered alone or in combination with other therapeutic agent(s) to address the needs of subjects suffering from a mitochondrial disease resulting from nuclear DNA mutations. In order to be administered to a subject in need thereof, Compound 1 will generally need to be formulated (for individual administration or in a combined formulation) for the intended route of administration. In some embodiments, the same route of administration can be used to deliver Compound 1 and optionally one or more other (i.e., additional) therapeutic agents. In some embodiments, a different route of administration can be used to deliver each of Compound 1 and optionally one or more other (i.e., additional) therapeutic agents.

[0066] In some embodiments, the therapeutic agent(s) can be formulated with little or no excipient or carrier. In some embodiments, the therapeutic agent(s) can be formulated such that the majority of the formulation is excipient or carrier. In brief, one of skill in the art will tailor the composition / formulation / medicament to have a suitable amount of excipient or carrier based on the needs / condition of the subject, the kind and extent of the disease to be treated; the properties of the therapeutic agent or agents to be delivered and the selected mode of administration of the particular therapeutic agent or agents.

[0067] In certain embodiments, a pharmaceutical composition (e.g., a medicament) may further comprise at least one additional therapeutic agent other than Compound 1, or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof. The at least one other / additional therapeutic agent can be an agent useful in the treatment of a mitochondrial disease resulting from nuclear DNA mutations or could, for example, be administered to ameliorate the side effect of the administration of Compounds 1 or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof. Thus, in some embodiments, pharmaceutical compositions can be prepared, for example, by combining Compound 1 and, optionally, one or more additional therapeutical agents or otherwise merely administering the other / additional therapeutic agent(s) in combination with the administration of Compound 1Atty. Dkt. No.091151-1691 or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof.

[0068] Pharmaceutical compositions may contain an effective amount of one or more of the therapeutic agent or agents as described herein and may optionally be disbursed (e.g. dissolved, suspended or otherwise) in a pharmaceutically acceptable carrier. The components of the pharmaceutical composition(s) may also be capable of being commingled with Compound 1 of the present application, and with each other, in a manner such that there is no interaction that would substantially impair the desired pharmaceutical efficiency.

[0069] As stated above, an “effective amount” refers to any amount of a particular therapeutic agent that is sufficient to achieve a desired biological effect. Combined with the teachings provided herein, by choosing among the various therapeutic compound(s) and weighing factors such as potency, relative bioavailability, patient body weight, target tissue uptake, severity of adverse side-effects and mode of administration, an effective prophylactic (i.e. preventative) or therapeutic treatment regimen can be planned which does not cause substantial unwanted toxicity and yet is effective to address the particular condition, disorder or disease of a particular subject in a therapeutic way. The effective amount of a therapeutic agent for any particular indication can vary depending on such factors as the disease, disorder or condition being treated, the particular compound or compounds being administered, the size of the subject, the age of the subject, the overall health of the subject and / or the severity of the disease, disorder or condition. The effective amount may be determined during pre- clinical trials and / or clinical trials by methods familiar to physicians and clinicians. A maximum dose may be used, that is, the highest safe dose according to some medical judgment. Multiple doses per day may be contemplated to achieve appropriate systemic levels of compounds. Appropriate systemic levels can be determined by, for example, measurement of the patient’s peak or sustained plasma level of the drug. “Dose” and “dosage” are used interchangeably herein. A dose may be administered by oneself, by another or by way of a device (e.g., a pump).

[0070] For any therapeutic compound described herein the therapeutically effective amount can, for example, be initially determined from animal models. A therapeutically effective dose can also be determined from human data for compounds which have been tested in humans and for compounds which are known to exhibit similar pharmacological activities, such as other related active agents. Higher doses may be required for parenteral administration. The applied dose can be adjusted based on the relative bioavailability andAtty. Dkt. No.091151-1691 potency of the administered compound. Adjusting the dose to achieve maximal efficacy based on the methods described above and other methods as are well-known in the art is well within the capabilities of the ordinarily skilled artisan.

[0071] Therapeutic compounds (alone or as formulated in a pharmaceutical composition / medicament) for use in therapy or prevention can be tested in suitable animal model systems. Suitable animal model systems include, but are not limited to, rats, mice, chicken, cows, monkeys, rabbits, pigs, minipigs and the like, prior to testing in human subjects. In vivo testing of any animal model system known in the art can be used prior to administration to human subjects. In some embodiments, dosing can be tested directly in humans.

[0072] Dosage, toxicity and therapeutic efficacy of any therapeutic agents or compositions (e.g., formulations or medicaments comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof, other / additional therapeutic agents, or mixtures thereof can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50 / ED50. Compounds that exhibit high therapeutic indices are advantageous. While compounds that exhibit toxic side effects may be used, in such cases it may be prudent to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to uninfected cells and, thereby, reduce side effects.

[0073] An exemplary treatment regime can, for example, entail administration once per day, twice per day, thrice per day, once a week, or once a month. In therapeutic applications, a relatively high dosage at relatively short intervals is sometimes required until progression of the disease is delayed, reduced or terminated, or until the subject shows partial or complete amelioration of symptoms of disease. Thereafter, the patient can be administered a prophylactic regimen.

[0074] For use in therapy, an effective amount of the therapeutic compound (alone or as formulated) can be administered to a subject by any mode that delivers the compound to the desired surface. Administering a pharmaceutical composition may be accomplished by any means known to the skilled artisan. Routes of administration include but are not limited toAtty. Dkt. No.091151-1691 oral, topical, intranasal, systemic, intravenous, subcutaneous, intraperitoneal, intradermal, intraocular, ophthalmical, intrathecal, intracerebroventricular, iontophoretical, transmucosal, intravitreal, or intramuscular administration. Administration includes self-administration, administration by another and administration by a device (e.g., a pump).

[0075] A therapeutic compound / agent disclosed herein (e.g., Compound 1, or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof, optionally with one or more additional therapeutic agents, can be delivered to the subject in a formulation or medicament (i.e., a pharmaceutical composition). Formulations and medicaments can be prepared by, for example, dissolving or suspending a therapeutic compound / agent disclosed herein (e.g., Compound 1) in water, a solvent, a pharmaceutically acceptable carrier, salt, (e.g., NaCl or sodium phosphate), buffering agents, preservatives, compatible carriers, adjuvants, and optionally other therapeutically acceptable ingredients.

[0076] The pharmaceutical compositions (e.g. a formulation or medicament) can include a carrier (e.g., a pharmaceutically acceptable carrier), which can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thiomerasol, and the like. Glutathione and other antioxidants can be included to prevent oxidation. In many cases, it will be advantageous to include isotonic agents, for example, sugars (e.g., trehalose), polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate or gelatin.

[0077] Solutions or suspensions (e.g., a formulation or medicament) used for parenteral, intradermal, subcutaneous or intraocular application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pHAtty. Dkt. No.091151-1691 can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic. For convenience of the patient or treating physician, the dosing formulation can be provided alone or in a kit containing all necessary equipment (e.g., vials of drug, vials of diluent, syringes and needles) for a treatment course (e.g., 1, 2, 3, 4, 5, 6, 7 days or more of treatment).

[0078] The therapeutic compounds / agents or pharmaceutical compositions, when it is desirable to deliver them systemically, may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion (for example by IV injection or via a pump to meter the administration over a defined time). Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. Pharmaceutical compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and / or dispersing agents. Additionally, suspensions of the therapeutic compounds (e.g., Compound 1, or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof, optionally in combination with one or more additional therapeutic agents) may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.

[0079] Systemic formulations include those designed for administration by injection, e.g., subcutaneous, intravenous, intramuscular, intrathecal, or intraperitoneal injection, as well as those designed for transdermal, transmucosal oral or pulmonary administration.

[0080] For intravenous and other parenteral routes of administration, a compound (e.g., Compound 1, or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof, optionally in combination with one or more additional therapeutic agents) can be formulated as a lyophilized preparation, as a lyophilized preparation of liposome-intercalated or lipid-encapsulated therapeutic compound(s), as a lipid complex in aqueous suspension, or as a salt complex. Lyophilized formulations are generallyAtty. Dkt. No.091151-1691 reconstituted in suitable aqueous solution, e.g., in sterile water or saline, shortly prior to administration.

[0081] Pharmaceutical compositions (e.g., a formulation or medicament) suitable for injection can include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). A composition for administration by injection will generally be sterile and should be fluid to the extent that easy syringability exists. It should be stable under the conditions of manufacture and storage and may be preserved against the contaminating action of microorganisms such as bacteria and fungi.

[0082] Sterile injectable solutions (e.g., a formulation or medicament) can be prepared by incorporating the therapeutic compound(s) (e.g., Compound 1, or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof, optionally in combination with one or more additional therapeutic agents) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the therapeutic compound(s) into a sterile vehicle, that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, typical methods of preparation include vacuum drying and freeze drying, which can yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[0083] For oral administration, the compounds can be formulated readily by combining the therapeutic compound(s) (e.g., Compound 1, or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof, optionally in combination with one or more additional therapeutic agents) with pharmaceutically acceptable carriers well known in the art. Such carriers enable the therapeutic compound(s) to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a subject to be treated. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid,Atty. Dkt. No.091151-1691 Primogel^, or corn starch; a lubricant such as magnesium stearate or sterates; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

[0084] Pharmaceutical preparations for oral use can be obtained as solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate. Optionally the oral formulations may also be formulated in saline or buffers, e.g., EDTA for neutralizing internal acid conditions or may be administered without any carriers.

[0085] Also specifically contemplated are oral dosage forms of the above that may be chemically modified so that oral delivery of the derivative is efficacious. Generally, the chemical modification contemplated is the attachment of at least one moiety to the therapeutic agent(s), ingredient(s), and / or excipient(s), where said moiety permits (a) inhibition of acid hydrolysis; and (b) uptake into the blood stream from the stomach or intestine. Also desired is the increase in overall stability of the therapeutic agent(s), ingredient(s), and / or excipient(s) and increase in circulation time in the body. Examples of such moieties include: polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone and polyproline. Abuchowski and Davis, “Soluble Polymer-Enzyme Adducts”, In: Enzymes as Drugs, Hocenberg and Roberts, eds., Wiley-Interscience, New York, N.Y., pp.367-383 (1981); Newmark et al., J Appl Biochem 4:185-9 (1982). Other polymers that could be used are poly-1,3-dioxolane and poly-1,3,6-tioxocane. For pharmaceutical usage, as indicated above, polyethylene glycol (PEG) moieties of various molecular weights are suitable.

[0086] For the formulation of the therapeutic agent(s), ingredient(s), and / or excipient(s), the location of release may be the stomach, the small intestine (the duodenum, the jejunum, or the ileum), or the large intestine. One skilled in the art has available formulations which will not dissolve in the stomach yet will release the material in the duodenum or elsewhere in the intestine. Preferably, the release will avoid the deleterious effects of the stomachAtty. Dkt. No.091151-1691 environment, either by protection of a therapeutic compound / agent or by release of the biologically active material beyond the stomach environment, such as in the intestine.

[0087] A coating or mixture of coatings can also be used on tablets, which are not intended for protection against the stomach. This can include sugar coatings, or coatings which make the tablet easier to swallow. Capsules may consist of a hard shell (such as gelatin) for delivery of dry therapeutic agent (e.g., powder); for liquid forms, a soft gelatin shell may be used. The shell material of cachets could be thick starch or other edible paper. For pills, lozenges, molded tablets or tablet triturates, moist massing techniques can be used.

[0088] The therapeutic compound(s) / agent(s) refers to Compound 1, or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, hydrates, and / or solvates thereof and optionally one or more additional therapeutic agents that can be administered in a combination to address a mitochondrial disease resulting from nuclear DNA mutations. The formulation can comprise fine multi-particulates in the form of granules or pellets of particle size about, e.g., 1-2 mm. The formulation of the material for capsule administration could also be as a powder, lightly compressed plugs or even as tablets. The therapeutic compound(s) / agent(s) or pharmaceutical composition(s) could be prepared by compression.

[0089] Colorants and flavoring agents may all be included. For example, the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) may be formulated and then further contained within an edible product, such as a refrigerated beverage containing colorants and flavoring agents.

[0090] One may dilute or increase the volume of the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) with an inert material. These diluents could include carbohydrates, especially mannitol, lactose, anhydrous lactose, cellulose, sucrose, modified dextrans and starch. Certain inorganic salts may also be used as fillers including calcium triphosphate, magnesium carbonate and sodium chloride. Some commercially available diluents are Fast-Flo^, Emdex^, STARCH 1500^, Emcompress^ and Avicel^.

[0091] Disintegrants may be included in the formulation of the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) into a solid dosage form. Materials used as disintegrates include but are not limited to starch, including the commercial disintegrant based on starch, Explotab. Sodium starch glycolate, Amberlite^, sodium carboxymethylcellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acidAtty. Dkt. No.091151-1691 carboxymethyl cellulose, natural sponge and bentonite may all be used. Another form of the disintegrants are the insoluble cationic exchange resins. Powdered gums may be used as disintegrants and as binders and these can include powdered gums such as agar, karaya gum or tragacanth. Alginic acid and its sodium salt are also useful as disintegrants.

[0092] Binders may be used to hold the therapeutic agent(s) together to form a hard tablet and include materials from natural products such as acacia, tragacanth, starch and gelatin. Others include methyl cellulose (MC), ethyl cellulose (EC) and carboxymethyl cellulose (CMC). Polyvinyl pyrrolidone (PVP) and hydroxypropylmethyl cellulose (HPMC) could both be used in alcoholic solutions to granulate the therapeutic agent(s).

[0093] An anti-frictional agent may be included in the formulation of the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) to prevent sticking during the formulation process. Lubricants may be used as a layer between the therapeutic and the die wall, and these can include but are not limited to; stearic acid including its magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils and waxes. Soluble lubricants may also be used such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycol (PEG) of various molecular weights, Carbowax™ 4000 and 6000.

[0094] Glidants that might improve the flow properties of the drug during formulation and to aid rearrangement during compression might be added. The glidants may include starch, talc, pyrogenic silica and hydrated silicoaluminate.

[0095] To aid dissolution of the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) into the aqueous environment, a surfactant might be added as a wetting agent. Surfactants may include anionic detergents such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate. Cationic detergents which can be used and can include benzalkonium chloride and benzethonium chloride. Potential non-ionic detergents that could be included in the formulation or medicament as surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50 and 60, glycerol monostearate, polysorbate 40, 60, 65 and 80, sucrose fatty acid ester, methyl cellulose and carboxymethyl cellulose. These surfactants could be present in the formulation or medicament disclosed herein or derivative either alone or as a mixture in different ratios.

[0096] Pharmaceutical compositions which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixtureAtty. Dkt. No.091151-1691 with filler such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the therapeutic compound(s) may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. Microspheres formulated for oral administration may also be used. Such microspheres have been well defined in the art. All formulations for oral administration should be in dosages suitable for such administration.

[0097] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0098] For topical administration, the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) may be formulated as solutions, gels, ointments, creams, suspensions, etc. as are well-known in the art. Solutions, gels, ointments, creams or suspensions may be administered topically. The compounds may also be formulated in rectal or vaginal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.

[0099] For administration by inhalation, therapeutic compound(s) / agent(s) or pharmaceutical composition(s) for use according to the present application may be conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. Such methods include those described in U.S. Pat. No.6,468,798. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. For example, capsules and cartridges of e.g., gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the therapeutic compound / agent and a suitable powder base such as lactose or starch. Alternatively, the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

[0100] Nasal delivery of a therapeutic compound(s) / agent(s) or pharmaceutical composition(s) is also contemplated. Nasal delivery allows the passage of therapeutic compound(s) / agent(s) or pharmaceutical composition(s) to the blood stream directly after administering the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) to theAtty. Dkt. No.091151-1691 nose, without the necessity for deposition of the product in the lung. Formulations for nasal delivery include those with dextran or cyclodextran.

[0101] For nasal administration, one type of useful device is a small, hard bottle to which a metered dose sprayer is attached. In some embodiments, the metered dose is delivered by drawing a pharmaceutical composition (in solution form) into a chamber of defined volume, which chamber has an aperture dimensioned to aerosolize and aerosol formulation by forming a spray when a liquid in the chamber is compressed. The chamber is compressed to administer the therapeutic compound(s) / agent(s) or pharmaceutical composition(s). In a specific embodiment, the chamber is a piston arrangement. Such devices are commercially available.

[0102] Alternatively, a plastic squeeze bottle with an aperture or opening dimensioned to aerosolize an aerosol formulation by forming a spray when squeezed can be used. The opening is usually found in the top of the bottle, and the top is generally tapered to partially fit in the nasal passages for efficient administration of the aerosol formulation. Preferably, the nasal inhaler will provide a metered amount of the aerosol formulation, for administration of a measured dose of the therapeutic compound(s) / agent(s) or pharmaceutical composition(s).

[0103] Also contemplated herein is pulmonary delivery of the compounds disclosed herein. The therapeutic compound(s) / agent(s) or pharmaceutical composition(s)is / are delivered to the lungs of a mammal while inhaling and traverses across the lung epithelial lining to the blood stream. Other reports of inhaled molecules include Adjei et al., Pharm Res 7:565-569 (1990); Adjei et al., Int J Pharmaceutics 63:135-144 (1990) (leuprolide acetate); Braquet et al., J Cardiovasc Pharmacol 13(suppl.5):143-146 (1989) (endothelin-1); Hubbard et al., Annal Int Med 3:206-212 (1989) (α1-antitrypsin); Smith et al., 1989, J Clin Invest 84:1145- 1146 (a-1-proteinase); Oswein et al., 1990, “Aerosolization of Proteins”, Proceedings of Symposium on Respiratory Drug Delivery II, Keystone, Colorado, March, (recombinant human growth hormone); Debs et al., 1988, J Immunol 140:3482-3488 (interferon-gamma and tumor necrosis factor alpha) and Platz et al., U.S. Pat. No.5,284,656 (granulocyte colony stimulating factor; incorporated by reference). A method and composition for pulmonaryAtty. Dkt. No.091151-1691 delivery of drugs for systemic effect is described in U.S. Pat. No.5,451,569 (incorporated by reference), issued Sep.19, 1995, to Wong et al.

[0104] Contemplated for use in the practice of this technology are a wide range of mechanical devices designed for pulmonary delivery of therapeutic products, including but not limited to nebulizers, metered dose inhalers, and powder inhalers, all of which are familiar to those skilled in the art.

[0105] Some specific examples of commercially available devices suitable for the practice of this technology are the Ultravent™ nebulizer, manufactured by Mallinckrodt, Inc., St. Louis, Mo.; the Acorn II^ nebulizer, manufactured by Marquest Medical Products, Englewood, Colo.; the Ventolin^ metered dose inhaler, manufactured by Glaxo Inc., Research Triangle Park, North Carolina; and the Spinhaler^ powder inhaler, manufactured by Fisons Corp., Bedford, Mass.

[0106] All such devices require the use of formulations suitable for the dispensing of the therapeutic compound(s) / agent(s) or pharmaceutical composition(s). Typically, each formulation is specific to the type of device employed and may involve the use of an appropriate propellant material, in addition to the usual diluents, adjuvants, and / or carriers useful in therapy. Also, the use of liposomes, microcapsules, microspheres, nanoparticles, nanospheres, inclusion complexes, or other types of carriers is contemplated.

[0107] Formulations suitable for use with a nebulizer, either jet or ultrasonic, can, for example, comprise therapeutic compound(s) / agent(s) or pharmaceutical composition(s) dissolved in water at a concentration of about 0.01 to 50 mg of biologically active compound per mL of solution. The formulation may also include a buffer and optionally a simple sugar (e.g., for inhibitor stabilization and regulation of osmotic pressure). The nebulizer formulation may also contain a surfactant, to reduce or prevent surface induced aggregation of the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) disclosed herein caused by atomization of the solution in forming the aerosol.

[0108] Formulations for use with a metered-dose inhaler device may generally comprise a finely divided powder comprising the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) disclosed herein suspended in a propellant with the aid of a surfactant. The propellant may be any conventional material employed for this purpose, such as a chlorofluorocarbon, a hydrochlorofluorocarbon, a hydrofluorocarbon, or a hydrocarbon, including trichlorofluoromethane, dichlorodifluoromethane, dichlorotetrafluoroethanol, andAtty. Dkt. No.091151-1691 1,1,1,2-tetrafluoroethane, or combinations thereof. Suitable surfactants include sorbitan trioleate and soya lecithin. Oleic acid may also be useful as a surfactant.

[0109] Formulations for dispensing from a powder inhaler device may comprise a finely divided dry powder containing the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) and may also include a bulking agent, such as lactose, sorbitol, sucrose, or mannitol in amounts which facilitate dispersal of the powder from the device, e.g., 50 to 90% by weight of the formulation. The compound(s) / therapeutic agent(s) / pharmaceutical composition(s) can advantageously be prepared in particulate or nanoparticulate form with an average particle size of less than 10 micrometers (μm), most preferably 0.5 to 5 μm, for most effective delivery to the deep lung.

[0110] For ophthalmic or intraocular indications, any suitable mode of delivering the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) to the eye or regions near the eye can be used. For ophthalmic formulations generally, see Mitra (ed.), Ophthalmic Drug Delivery Systems, Marcel Dekker, Inc., New York, N.Y. (1993) and also Havener, W. H., Ocular Pharmacology, C.V. Mosby Co., St. Louis (1983). Nonlimiting examples of pharmaceutical compositions suitable for administration in or near the eye include, but are not limited to, ocular inserts, minitablets, and topical formulations such as eye drops, ointments, and in situ gels. In one embodiment, a contact lens is coated with a pharmaceutical composition (or contains a pharmaceutical composition encapsulated therein) comprising a therapeutic compound / agent. In some embodiments, a single dose can comprise from between 0.1 ng to 5000 μg, 1 ng to 500 μg, or 10 ng to 100 μg of the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) administered to the eye.

[0111] Eye drops can comprise a sterile liquid formulation that can be administered directly to the eye. In some embodiments, eye drops comprise at least one therapeutic agent (and possibly several) and may further comprise one or more preservatives. In some embodiments, the optimum pH for eye drops equals that of tear fluid and is about 7.4, the pH may be within any range that is not harmful to the eye of the subject. For eye drops, the therapeutic compound(s) / agent(s) can be present in the drop solution from about 0.1% to about 5% (w / v or v / v depending on the physical nature (i.e. solid or liquid) of the active ingredient). In some embodiments, the therapeutic compound / agent can be present in the drop solution from about 1% to about 3% (w / v or v / v, as appropriate). In some embodiments, the therapeutic compound / agent can be present in the drop solution from about 0.2% to about 1.5% (w / v or v / v, as appropriate). In some embodiments, the therapeuticAtty. Dkt. No.091151-1691 compound / agent can be present in the drop solution from about 0.1% to about 1.0% (w / v or v / v, as appropriate).

[0112] In situ gels are viscous liquids, showing the ability to undergo sol-to-gel transitions when influenced by external factors, such as appropriate pH, temperature, pressure, and / or the presence of electrolytes. This property causes slowing of drug drainage from the eyeball surface and increase of the active ingredient bioavailability. Polymers commonly used in in situ gel formulations include, but are not limited to, gellan gum, poloxamer, silicone containing formulations, silica-based formulations, and cellulose acetate phthalate. In some embodiments, the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) is / are formulated into an in-situ gel (as the formulation / medicament).

[0113] For topical ophthalmic administration, therapeutic compound(s) / agent(s) or pharmaceutical composition(s) is / are may be formulated as solutions, gels, ointments, creams, suspensions, etc. as are well-known in the art. Ointments are semisolid dosage forms for external use such as topical use for the eye or skin. In some embodiments, ointments comprise a solid or semisolid hydrocarbon base of melting or softening point close to human core temperature. In some embodiments, an ointment applied to the eye decomposes into small drops, which stay for a longer time period in conjunctival sac, thus increasing bioavailability.

[0114] Ocular inserts are solid or semisolid dosage forms without disadvantages of traditional ophthalmic drug forms. They are less susceptible to defense mechanisms like outflow through nasolacrimal duct, show the ability to stay in conjunctival sac for a longer period, and can be more stable than conventional dosage forms. They also offer advantages such as accurate dosing of one or more therapeutic compound(s) / agent(s) or pharmaceutical composition(s), slow release of one or more therapeutic compound(s) / agent(s) with constant speed and limiting of one or more therapeutic compounds’ / agents’ systemic absorption. In some embodiments, an ocular insert comprises one or more therapeutic compound(s) / agent(s) and one or more polymeric materials. The polymeric materials can include, but are not limited to, methylcellulose and its derivatives (e.g., hydroxypropyl methylcellulose (HPMC)), ethylcellulose, polyvinylpyrrolidone (PVP K-90), polyvinyl alcohol, chitosan, carboxymethyl chitosan, gelatin, and various mixtures of the aforementioned polymers. An ocular insert canAtty. Dkt. No.091151-1691 comprise silica. An ocular insert can comprise liposomes, nanoparticles or microparticles of degradable or biodegradable polymer (as described in more detail below).

[0115] Minitablets are biodegradable, solid drug forms, that transit into gels after application to the conjunctival sac, thereby extending the period of contact between active ingredient (i.e., the therapeutic compound(s) / agent(s)) and the eyeball surface, which in turn increases a therapeutic compounds’ / agents’ bioavailability. The advantages of minitablets include easy application to conjunctival sac, resistance to defense mechanisms like tearing or outflow through nasolacrimal duct, longer contact with the cornea caused by presence of mucoadhesive polymers, and gradual release of the active ingredient from the formulation in the place of application due to the swelling of the outer carrier layers. Minitablets can comprise one or more therapeutic compound(s) / agent(s) and one or more polymers. Nonlimiting examples of polymers suitable for use in in a minitablet formulation include cellulose derivatives, like hydroxypropyl methylcellulose (HPMC), hydroxyethyl cellulose (HEC), sodium carboxymethyl cellulose, ethyl cellulose, acrylates (e.g., polyacrylic acid and its cross-linked forms), Carbopol^ or carbomer, chitosan, and starch (e.g., drum-dried waxy maize starch). In some embodiments, minitablets further comprise one or more excipients. Nonlimiting examples of excipients include mannitol and magnesium stearate.

[0116] The ophthalmic or intraocular formulations and medicaments may contain non-toxic auxiliary substances such as antibacterial components which are generally non-injurious in use, for example, thimerosal, benzalkonium chloride, methyl and propyl paraben, benzyldodecinium bromide, benzyl alcohol, or phenylethanol; buffering ingredients such as sodium chloride, sodium borate, sodium acetate, sodium citrate, or gluconate buffers; and other conventional ingredients such as sorbitan monolaurate, triethanolamine, polyoxyethylene sorbitan monopalmitylate, ethylenediamine tetraacetic acid (EDTA), and the like.

[0117] In some embodiments, the viscosity of the ocular formulation comprising one or more therapeutic compound(s) / agent(s) is increased to improve contact with the cornea and bioavailability in the eye. Viscosity can be increased by the addition of hydrophilic polymers of high molecular weight which do not diffuse through biological membranes and which form three-dimensional networks in the water. Nonlimiting examples of such polymersAtty. Dkt. No.091151-1691 include polyvinyl alcohol, poloxamers, hyaluronic acid, carbomers, and polysaccharides, cellulose derivatives, gellan gum, and xanthan gum.

[0118] In addition to the formulations described above, therapeutic compound(s) / agent(s) or pharmaceutical composition(s) may also be formulated as a depot preparation. Such long acting formulations may be formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

[0119] In some embodiments, the therapeutic compound(s) / agent(s) or pharmaceutical composition(s) is / are administered as a depot formulation wherein the active therapeutic agent(s) is / are encapsulated by, or disposed within, silica-based microparticles. Such a formulation may be a controlled-release, delayed-release or extended release formulation (terms are defined below). Such controlled-release, delayed release or extended release formulation may comprise particles, such as microparticles or nanoparticles.

[0120] The pharmaceutical compositions also may comprise suitable solid or gel-phase carriers or excipients. Examples of such carriers or excipients include but are not limited to calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, silica / silicone and polymers such as polyethylene glycols.

[0121] Suitable liquid or solid pharmaceutical preparation forms (e.g., a formulation or medicament) can, for example, be aqueous or saline solutions for inhalation, microencapsulated, encochleated, coated onto microscopic gold particles, contained in liposomes, nebulized, aerosols, pellets for implantation into the skin, or dried onto a sharp object to be scratched into the skin. The pharmaceutical compositions / formulations may also include granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops or preparations with protracted release of therapeutic compound(s), in whose preparation excipients and additives and / or auxiliaries such as disintegrants, binders, coating agents, swelling agents, lubricants, flavorings, sweeteners, or solubilizers are customarily used as described above. The pharmaceutical compositions can be suitable for use in a variety of drug delivery systems. For a brief review of methods for drug delivery, see Langer R, Science 249:1527-33 (1990).

[0122] The therapeutic compound(s) / agent(s) or pharmaceutical composition(s) may be provided in particles. Particles as used herein means nanoparticles or microparticles (or in some instances larger particles) which can consist in whole or in part of the therapeuticAtty. Dkt. No.091151-1691 compound(s) / agent(s) as described herein. The particles may contain the therapeutic compound(s) / agent(s) in a core surrounded by a coating, including, but not limited to, an enteric coating. The therapeutic compound(s) / agent(s) also may be dispersed throughout the particles. The therapeutic compound(s) / agent(s) also may be adsorbed into the particles. The particles may be of any order release kinetics, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, and any combination thereof, etc. The particle may include, in addition to any therapeutic compound(s) / agent(s), any of those materials routinely used in the art of pharmacy and medicine, including, but not limited to, erodible, non-erodible, biodegradable, or nonbiodegradable material or combinations thereof. The particles may be microcapsules which contain the therapeutic compound(s) / agent(s) in a solution or in a semi-solid state. The particles may be of virtually any shape.

[0123] Both non-biodegradable and biodegradable polymeric materials can be used in the manufacture of particles for delivering the therapeutic compound(s) / agent(s). Such polymers may be natural or synthetic polymers. The polymer can be selected based on the period of time over which release is desired. Bioadhesive polymers of particular interest include bioerodible hydrogels described in Sawhney H S et al. (1993) Macromolecules 26:581-7, the teachings of which are incorporated herein. These include polyhyaluronic acids, casein, gelatin, glutin, polyanhydrides, polyacrylic acid, alginate, chitosan, polyethylene glycols (PEGs), polyvinylalcohols (PVAs), poly(methyl methacrylates), poly(ethyl methacrylates), poly(butylmethacrylate), poly(isobutyl methacrylate), poly-lactic acid (PLA), poly(lactic -co- glycolic) acid (PLGA), poly(hexylmethacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate) and poly(ε-caprolactone) or mixtures of two or more of the foregoing. The biodegradable polymeric materials may be substantially pure single polymer or mixes of two or more polymers wherein the materials comprise mixtures of single monomers, block co-polymers or a mixture thereof.

[0124] Therapeutic compound(s) / agent(s) or mixtures of two or more therapeutic compound(s) / agent(s) can be formulated in a carrier system. The carrier can be a colloidal system. The carrier or colloidal system can be a liposome, a phospholipid bilayer vehicle. In one embodiment, therapeutic compound(s) / agent(s) or mixtures of two or more therapeutic compound(s) / agent(s) can be encapsulated in a liposome while maintaining integrity of the therapeutic compound(s) / agent(s). One skilled in the art would appreciate that there are aAtty. Dkt. No.091151-1691 variety of methods to prepare liposomes. (See Lichtenberg, et al., Methods Biochem. Anal., 33:337-462 (1988); Anselem, et al., Liposome Technology, CRC Press (1993)). Liposomal formulations can delay clearance and increase cellular uptake (See Reddy, Ann. Pharmacother., 34(7-8):915-923 (2000)). For example, a therapeutic agent can also be loaded into a particle prepared from pharmaceutically acceptable ingredients including, but not limited to, soluble, insoluble, permeable, impermeable, biodegradable or gastroretentive polymers or liposomes. Such particles include, but are not limited to, nanoparticles, biodegradable nanoparticles, microparticles, biodegradable microparticles, nanospheres, biodegradable nanospheres, microspheres, biodegradable microspheres, capsules, emulsions, liposomes, micelles and viral vector systems.

[0125] The carrier can also be a polymer, e.g., a biodegradable, biocompatible polymer matrix. In one embodiment, the therapeutic compound(s) / agent(s) or mixtures of two or more therapeutic compound(s) / agent(s) can be embedded in the polymer matrix, while maintaining integrity of the composition. The polymer can be a microparticle or nanoparticle that encapsulates therapeutic compound(s) / agent(s). The polymer may be natural, such as polypeptides, proteins or polysaccharides, or synthetic, such as poly α-hydroxy acids. Examples include carriers made of, e.g., collagen, fibronectin, elastin, cellulose acetate, cellulose nitrate, polysaccharide, fibrin, gelatin, and combinations thereof. In some embodiments, the polymer is poly-lactic acid (PLA), poly lactic / glycolic acid (PLGA) or a mixture thereof. The polymeric matrices can be prepared and isolated in a variety of forms and sizes, including microspheres and nanospheres. Polymer formulations can lead to prolonged duration of therapeutic effect. (See Reddy, Ann. Pharmacother., 34(7-8):915-923 (2000)). A polymer formulation for human growth hormone (hGH) has been used in clinical trials. (See Kozarich and Rich, Chemical Biology, 2:548-552 (1998)).

[0126] Examples of polymer microsphere sustained release formulations are described in PCT publication WO 99 / 15154 (Tracy, et al.), U.S. Pat. Nos.5,674,534 and 5,716,644 (both to Zale, et al.), PCT publication WO 96 / 40073 (Zale, et al.), and PCT publication WO 00 / 38651 (Shah, et al.). U.S. Pat. Nos.5,674,534 and 5,716,644 and PCT publication WOAtty. Dkt. No.091151-1691 96 / 40073 describe a polymeric matrix containing particles of erythropoietin that are stabilized against aggregation with a salt.

[0127] In some embodiments, the nanoparticles or microparticles can be silica-based or silane-based (See for example: WO2002 / 080977 entitled: “Biodegradable carrier and method for preparation thereof”).

[0128] In some embodiments, the therapeutic compound(s) / agent(s) or mixtures of two or more therapeutic compound(s) / agent(s) can be prepared with carriers that will protect the therapeutic compound(s) / agent(s) or other therapeutic agent(s) or mixtures thereof against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Such formulations can be prepared using known techniques. The materials can also be obtained commercially, e.g., from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to specific cells with monoclonal antibodies to cell-specific antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No.4,522,811.

[0129] The therapeutic compound(s) / agent(s) or mixtures of two or more therapeutic compound(s) / agent(s) may be contained in controlled release systems. The term “controlled-release” is intended to refer to any drug-containing formulation in which the manner and profile of drug release from the formulation are controlled. This refers to immediate as well as non-immediate release formulations, with non-immediate release formulations including but not limited to sustained release and delayed release formulations. The term “sustained-release” (also referred to as “extended-release”) is used in its conventional sense to refer to a drug formulation that provides for gradual release of a drug over an extended period of time, and that preferably, although not necessarily, results in substantially constant blood levels of a drug over an extended time period. The term “delayed-release” is used in its conventional sense to refer to a drug formulation in which there is a time delay between administration of the formulation and the release of the drug therefrom to thereby make it available to the subject. “Delayed release” may or may notAtty. Dkt. No.091151-1691 involve gradual release of drug over an extended period of time, and thus may or may not be “sustained release.”

[0130] Use of a long-term controlled-release or sustained-release implant or depot formulation may be particularly suitable for treatment of chronic conditions. The term “implant” and “depot formulation” is intended to include a single composition (such as a mesh) or composition comprising multiple components (e.g., a fibrous mesh constructed from several individual pieces of mesh material) or a plurality of individual compositions where the plurality remains localized and provides the long-term sustained-release of active pharmaceutical ingredient(s) occurring from the aggregate of the one or plurality of compositions. “Long-term” release, as used herein, means that the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for at least 2 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for at least 7 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for at least 14 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for at least 30 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for at least 60 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient for at least 90 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for at least 180 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for at least one year. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for 15-30 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for 30-60 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for 60-90 days. In some embodiments, the implant orAtty. Dkt. No.091151-1691 depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for 90-120 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for 120-180 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active pharmaceutical ingredient(s) for up to one year. In some embodiments, the long-term sustained-release implants or depot formulation are well-known to those of ordinary skill in the art and include some of the release systems described above. In some embodiments, such implants or depot formulation can be administered surgically. In some embodiments, such implants or depot formulation can be administered topically or by injection. III. Compositions, Formulations, and Medicaments

[0131] Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, can be used, alone or in combination, with one or more other therapeutic agents to address the needs of subjects suffering from a mitochondrial disease resulting from nuclear DNA mutations. In order to be administered to a subject in need thereof, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and / or other therapeutic agent(s) will generally need to be formulated for the suitable route of administration. For example, if Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and / or other therapeutic agent(s) is / are to be administered to the subject by injection, it / they will typically be formulated into an injectable liquid or liquid suspension. This could, for example, be accomplished by dissolving or suspending the therapeutic agent(s) in a suitable diluent, adjuvant, excipient, vehicle or pharmaceutically acceptable carrier as described previously herein (See the section above entitled: Pharmaceutical Compositions, Routes of Administration, and Dosing). In some embodiments, the diluent, adjuvant, excipient, vehicle or pharmaceutically acceptable carrier can be water, saline or a buffered aqueous solution. In some embodiments, the diluent, adjuvant, excipient, vehicle or pharmaceutically acceptable carrier can comprise an oil such as sesame oil or other lipophilic excipient. Suitable methods, reagents and compositions for formulating Compound 1 and / or other therapeutic agent(s) into a suitable medicament are discussed above.Atty. Dkt. No.091151-1691

[0132] Similarly, if Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and / or other therapeutic agent(s) can be to be administered to the subject in oral form, the selected active ingredient(s) can be formulated into a pill, tablet, capsule or other vehicle for such administration as discussed above in the section entitled: “Pharmaceutical Compositions, Routes of Administration, and Dosing” or as otherwise known to those of ordinary skill in the art. Suitable methods, reagents and compositions for formulating Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and / or other therapeutic agent(s) into a suitable orally administrable medicament are discussed above.

[0133] Similarly, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and / or other therapeutic agent(s) can be formulated for ocular administration, buccal administration, topical administration, nasal administration or any other of the modes of administration previously discussed herein or that are known to those of ordinary skill in the art. Suitable methods, reagents and compositions for formulating Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and / or other therapeutic agent(s) into a suitable ocular, buccal, topical, or nasal administrable medicament are discussed above.

[0134] In brief, any of the formulations (which can also be referred to as a medicament or composition when formulated for administration to a subject having a certain affliction or medical condition that requires medical attention) described in the section above entitled: “Pharmaceutical Compositions, Routes of Administration, and Dosing” can be applied to produce a composition (i.e. a formulation or medicament) suitable for administration to a subject in need thereof. Thus, in some embodiments, this application is directed to compositions, formulations and medicaments suitable for administration to a subject suffering from, or believed to be suffering from, a mitochondrial disease resulting from nuclear DNA mutations.

[0135] In some embodiments, the composition, formulation, or medicament is administered subcutaneously. In some embodiments, the composition, formulation, or medicament is administered orally. In some embodiments, the composition, formulation, or medicament is administered topically, intranasally, systemically, intravenously, intraperitoneally, intradermally, intraocularly, ophthalmically, intrathecally, intracerebroventricularly, iontophoretically, transmucosally, intravitreally, or intramuscularly.Atty. Dkt. No.091151-1691

[0136] In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and / or other therapeutic agent(s) can be administered in formulations prepared using a pharmaceutically acceptable salt form. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and / or other therapeutic agent(s) can be administered in formulations prepared using a hydrate form. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and / or other therapeutic agent(s) can be administered in formulations prepared using a solvated form. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and / or other therapeutic agent(s) can be administered in formulations prepared using a tautomeric form. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and / or other therapeutic agent(s) can be administered in formulations prepared using a stereoisomer of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and / or the other therapeutic agent(s).

[0137] Thus, in one aspect, the present technology provides for a composition comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, for use in treating, preventing, inhibiting, ameliorating or delaying the onset of a mitochondrial disease resulting from nuclear DNA mutations in a subject, wherein.

[0138] In some embodiments of the foregoing composition, the nuclear DNA mutations result in mitochondrial DNA (mtDNA) depletion(s) and / or mitochondrial DNA (mtDNA) rearrangement(s) in the subject. In some embodiments of the foregoing composition, the subject suffers from mitochondrial DNA (mtDNA) depletion syndrome. In some embodiments of the foregoing composition, the mitochondrial disease results from at least one mutation of the POLG gene. In some embodiments of the foregoing composition, the mitochondrial disease results from at least one mutation of the TWNK (twinkle) gene. In some embodiments of the foregoing composition, the mitochondrial DNA (mtDNA)Atty. Dkt. No.091151-1691 depletion(s) codes / code for at least one mutation of a complex I respiratory chain subunit. In some embodiments of the foregoing composition, the mitochondrial DNA (mtDNA) rearrangement(s) codes / code for at least one mutation of a complex I respiratory chain subunit.

[0139] In some embodiments of the foregoing composition, the subject has been diagnosed as having a mitochondrial disease that is neurodegenerative. In some embodiments of the foregoing composition, the mitochondrial disease is Kearns–Sayre syndrome or Alzheimer’s disease. In some embodiments of the foregoing composition, the mitochondrial disease is Parkinson’s disease (PD) or sporadic Parkinson’s disease (sPD). In some embodiments, the mitochondrial disease is Alpers syndrome.

[0140] In some embodiments of the foregoing composition, the subject is a mammal. In some embodiments of the foregoing composition, the subject is human.

[0141] In some embodiments of the foregoing composition, the composition is administered daily for 6 weeks or more. In some embodiments of the foregoing composition, the composition is administered daily for 24 weeks or more. In some embodiments of the foregoing composition, the composition is administered daily for 48 weeks or more. In some embodiments of the foregoing composition, the composition is administered daily for 52 weeks or more.

[0142] In some embodiments of the foregoing composition, the composition is formulated for administration orally. In some embodiments of the foregoing composition, the composition is formulated for administration subcutaneously. In some embodiments of the foregoing composition, the composition is formulated for administration topically. In some embodiments of the foregoing composition, the composition is formulated for administration intranasally. In some embodiments of the foregoing composition, the composition is formulated for administration systemically. In some embodiments of the foregoing composition, the composition is formulated for administration intravenously. In some embodiments of the foregoing composition, the composition is formulated for administration intraperitoneally. In some embodiments of the foregoing composition, the composition is formulated for administration intradermally. In some embodiments of the foregoing composition, the composition is formulated for administration intraocularly. In some embodiments of the foregoing composition, the composition is formulated for administrationAtty. Dkt. No.091151-1691 ophthalmically. In some embodiments of the foregoing composition, the composition is formulated for administration intrathecally. In some embodiments of the foregoing composition, the composition is formulated for administration intracerebroventricularly. In some embodiments of the foregoing composition, the composition is formulated for administration iontophoretically. In some embodiments of the foregoing composition, the composition is formulated for administration transmucosally. In some embodiments of the foregoing composition, the composition is formulated for administration intravitreally. In some embodiments of the foregoing composition, the composition is formulated for administration intramuscularly. IV. Therapeutic Methods and Related Uses of the Disclosed Compounds

[0143] In one aspect, the present technology provides a method for treating, preventing, inhibiting, ameliorating or delaying the onset of a mitochondrial disease resulting from nuclear DNA mutations in a subject comprising administering to the subject in need thereof a therapeutically effective amount of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, wherein Compound 1 is:.

[0144] In some embodiments, the nuclear DNA mutations result in mitochondrial DNA (mtDNA) depletion(s) and / or mitochondrial DNA (mtDNA) rearrangement(s) in the subject. In some embodiments, the subject suffers from mitochondrial DNA (mtDNA) depletion syndrome. In some embodiments, the mitochondrial disease results from at least one mutation of the POLG gene. In some embodiments, mitochondrial disease results from at least one mutation of the TWNK (twinkle) gene. In some embodiments, the mitochondrial DNA (mtDNA) depletion(s) codes / code for at least one mutation of a complex I respiratory chain subunit. In some embodiments, the mitochondrial DNA (mtDNA) rearrangement(s) codes / code for at least one mutation of a complex I respiratory chain subunit.

[0145] In some embodiments, the subject has been diagnosed as having a mitochondrial disease that is neurodegenerative. In some embodiments, the mitochondrial disease is Kearns–Sayre syndrome or Alzheimer’s disease. In some embodiments, the mitochondrialAtty. Dkt. No.091151-1691 disease is Parkinson’s disease (PD) or sporadic Parkinson’s disease (sPD). In some embodiments, the mitochondrial disease is Alpers syndrome.

[0146] In some embodiments, the subject is a mammal. In some embodiments, the subject is human.

[0147] In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 6 weeks or more. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, administered daily for 24 weeks or more. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 48 weeks or more. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 52 weeks or more.

[0148] In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered orally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered subcutaneously. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered topically. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intranasally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered systemically. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intravenously. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intraperitoneally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intradermally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intraocularly. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered ophthalmically. In some embodiments, Compound 1, or pharmaceuticallyAtty. Dkt. No.091151-1691 acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intrathecally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intracerebroventricularly, iontophoretically. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered transmucosally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intravitreally. In some embodiments, Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered intramuscularly.

[0149] In some embodiments, the method further comprises separately, sequentially, or simultaneously administering at least one additional therapeutic agent to the subject. In some embodiments, the additional therapeutic agent comprises levodopa, optionally in combination with carbidopa (LODOSYN^). In some embodiments, the combination of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and an additional therapeutic agent has a synergistic effect in the treating, preventing, inhibiting, ameliorating or delaying the onset of the mitochondrial disease. In some embodiments, administration of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof and optionally including administration of the additional therapeutic agent, prolongs survival of the subject, as compared with an untreated subject or an untreated control group of subjects.

[0150] In another aspect, the present technology provides for use of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, in the preparation of a composition, formulation or medicament for treating, preventing, inhibiting, ameliorating or delaying the onset of a mitochondrial disease resulting from nuclear DNA mutations in a subject, wherein Compound 1 is:.

[0151] In some embodiments of the use, the nuclear DNA mutations result in mitochondrial DNA (mtDNA) depletion(s) and / or mitochondrial DNA (mtDNA) rearrangement(s) in the subject. In some embodiments of the use, the subject suffers from mitochondrial DNAAtty. Dkt. No.091151-1691 (mtDNA) depletion syndrome. In some embodiments of the use, the mitochondrial disease results from at least one mutation of the POLG gene. In some embodiments of the use, the mitochondrial disease results from at least one mutation of the TWNK (twinkle) gene. In some embodiments of the use, the mitochondrial DNA (mtDNA) depletion(s) codes / code for at least one mutation of the complex I respiratory chain subunit. In some embodiments of the use, the mitochondrial DNA (mtDNA) rearrangement(s) codes / code for at least one mutation of the complex I respiratory chain subunit.

[0152] In some embodiments of the use, the subject has been diagnosed as having a mitochondrial disease that is neurodegenerative. In some embodiments of the use, the mitochondrial disease is Kearns–Sayre syndrome or Alzheimer’s disease. In some embodiments of the use, the mitochondrial disease is Parkinson’s disease (PD) or sporadic Parkinson’s disease (sPD). In some embodiments, the mitochondrial disease is Alpers syndrome.

[0153] In some embodiments of the use, the subject is a mammal. In some embodiments of the use, the subject is human.

[0154] In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 6 weeks or more. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 24 weeks or more. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 48 weeks or more. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered daily for 52 weeks or more.

[0155] In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration orally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration subcutaneously. In some embodiments of the use,Atty. Dkt. No.091151-1691 the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration topically. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intranasally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration systemically. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intravenously. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intraperitoneally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intradermally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intraocularly. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration ophthalmically. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intrathecally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intracerebroventricularly. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration iontophoretically. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration transmucosally. In some embodiments of the use, the composition, formulation or medicament comprisingAtty. Dkt. No.091151-1691 Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intravitreally. In some embodiments of the use, the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is formulated for administration intramuscularly.

[0156] In some embodiments of the use, administration of the composition, formulation or medicament further comprises separately, sequentially, or simultaneously administering at least one additional therapeutic agent to the subject. In some embodiments of the use, the additional therapeutic agent comprises levodopa, optionally in combination with carbidopa (LODOSYN^). In some embodiments of the use, the combination of the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, and an additional therapeutic has a synergistic effect in the treating, preventing, inhibiting, ameliorating or delaying the onset of the mitochondrial disease. In some embodiments of the use, administration of the composition, formulation or medicament comprising Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof and optionally including administration of the additional therapeutic agent, prolongs survival of the subject, as compared with an untreated subject or an untreated control group of subjects.

[0157] In another aspect, the present technology provides for a method for treating a subject suffering from mitochondrial DNA (mtDNA) depletion syndrome, the method comprising: administering to the subject a therapeutically effective amount of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, wherein Compound 1 is:.

[0158] In some embodiments of the foregoing method, the subject has at least one mutation of the POLG gene. In some embodiments of the foregoing method, the subject has at least one mutation of the TWNK (twinkle) gene. In some embodiments of the foregoing method,Atty. Dkt. No.091151-1691 the subject is a mammal. In some embodiments of the foregoing method, the mammalian subject is a human. EXAMPLES

[0159] The present technology is further illustrated by the following examples, which should not be construed as limiting in any way. Example 1: In Vitro Assays 1. Methods:

[0160] Cell viability assessment (Figs.1-2)

[0161] HepG2 cells were obtained from American Type Culture Collection (ATCC; Product Number: HB-8065) and grown in Complete Media (Eagle’s Minimum Essential Medium supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin) at 37 degrees Centigrade with 5% CO2. To assess cell viability after extended culturing, 3 x 106HepG2 cells were seeded in T75 flasks and cultured for eight days with andwithout 400 μM dideoxycytosine triphosphate (ddC) and with and without 1 μM Compound 1. Media, including any drug treatments (as noted in the Figures), was replaced every 48 hours through simple exchange. For Fig.1, to confirm that ddC treatment indeed diminished ATP production, 10,000 cells per well with and without ddC treatment were seeded on a 96-well plate. Cells were incubated overnight and ATP production was measured using the CellTiter- Glo 2.0 cell viability assay the following day per manufacturer’s instructions(Promega). For Fig.2, viable cell density and percentage cell viability were measured at the end of eight days. From each condition (T75 flask), 10 μL of sample was measured using trypan blue staining with the Countess 3 Automated Cell Counter (ThermoFisher Scientific AMQAX2000). Relative light units (RLUs) were measured using the luminescence setting on a BioTek plate reader. Three samples were measured for each condition and their respective counts were averaged. Data were graphed using GraphPad Prism 10 and the data is presented in Figs.1-2.

[0162] RSL3 glutathione-depletion stress assays (Figs.3-4)

[0163] To deplete cellular mtDNA by ddC alone, HepG2 cells were grown in T75 flasks in complete media with 400 μM ddC for 8-12 days (Fig.3). To deplete cellular mtDNA using ddC + ethidium bromide (EtBr), HepG2 cells were grown in T75 flasks for 8-12 days withAtty. Dkt. No.091151-1691 100 μM ddC with 400 ng / mL of EtBr (Fig.4). Cells were harvested from flasks using 0.25% tryspin. Cells were seeded in a 6-well, tissue culture-treated plate and incubated at 37 degrees Centigrade with 5% CO2overnight. The following day, supernatant was gently removed from cells by aspirating, and cells were treated with 1 μM RSL3 in complete media in the presence and absence of 1 μM Compound 1 (Figs.3 and 4). Cells were incubated for 4 hours. ATP production was measured with the Cell Titer-Glo Cell Viability Assay 2.0 per manufacturer’s instructions (Promega). Relative light units (RLUs) were measured using the luminescence setting on a BioTek plate reader. Data were graphed using GraphPad Prism 10 and are presented in Figs.3-4. 2. Results & Discussion:

[0164] In the following, two commonly used models to simulate POLG disease (resulting from mtDNA depletion) were used; ddC treatment or ddC coupled with EtBr (See Michael V. Novotny, Weiling Xu, Anny Mulya, Allison J. Janocha, Serpil C. Erzurum, bioRxiv 2023.07.28.551015; doi: doi.org / 10.1101 / 2023.07.28.551015, now published in MethodsX doi: 10.1016 / j.mex.2023.102497; Zsurka G, Trombly G, Schöler S, Blei D, Kunz WS. Functional Assessment of Mitochondrial DNA Maintenance by Depletion and Repopulation Using 2',3'-Dideoxycytidine in Cultured Cells. Methods Mol Biol.2023;2615:229-240. doi: 10.1007 / 978-1-0716-2922-2_17. PMID: 36807796; and Lund KC, Peterson LL, Wallace KB. Absence of a universal mechanism of mitochondrial toxicity by nucleoside analogs. Antimicrob Agents Chemother.2007 Jul;51(7):2531-9. doi: 10.1128 / AAC.00039-07. Epub 2007 Apr 30. PMID: 17470651; PMCID: PMC1913246). Based on these literature models, both chemical depletion strategies were used to account for any differences between the models and to confirm any potential results from Compound 1 treatment. In these models, cells were chemically treated to deplete mitochondrial DNA (mtDNA) with either 400 μM dideoxycytosine triphosphate (ddC) or 100 μM ddC in combination with 400 ng / mL ethidium bromide (EtBr). In some experiments, the glutathione-inhibitor RSL3 was used in chemically treated cells to add additional stress to the model.

[0165] With reference to Fig.1, ATP production is reduced in HepG2 cells treated with 400 μM ddC as compared to vehicle-treated control cells. Percent viability was measured and compared in these groups and no significant difference was observed, suggesting an impairment in bioenergetics (growth) rather than increased death. The decrease in ATP production is consistent with deceased OXPHOS, which would be expected if mtDNA isAtty. Dkt. No.091151-1691 effectively depleted in the HepG2 cells. These data support the suitability of the model for testing the effect of a test compound (i.e., Compound 1) on modulating DNA depletion.

[0166] With reference to Fig.2, viable cell density was assessed in both vehicle- and ddC- treated HepG2 cells (dideoxycytosine triphosphate (a chain terminator of polymerase extension)). Each condition was co-cultured for eight days with either vehicle control or with Compound 1 present. Viable cell density (VCD) refers to the number of cells per mL of culture and is a metric to determine health of a cell culture. Addition of Compound 1 to vehicle-treated HepG2 cells appears to result in an increase in viable cell density, suggesting it has a beneficial effect to VCD in otherwise healthy cells. As predicted, ddC-treated HepG cells exhibited decreased VCD compared to vehicle-treated cells, exhibited presumably due to depletion of mtDNA, which would impair bioenergetics and inhibit oxidative phosphorylation (OXPHOS). Percent viability was also compared in these groups (not shown) and no significant difference was observed. This supports the premise that bioenergetics are impaired and that the decreased VCD observed in ddC-treated cells is due to slower growth rather than increased cell death. Addition of Compound 1 in the presence of ddC restores VCD to the same level as vehicle-treated cells. Without wishing to be bound by theory, this suggests that Compound 1 is either protective against the effects of mtDNA depletion or is able to improve bioenergetics in mtDNA-depleted cells.

[0167] With reference to Fig.3, the effect of Compound 1 to ameliorate the effects of a glutathione stressor (RSL3) were evaluated in the ddC mtDNA-depletion model. In this experiment, all cells were treated with ddC for one week to deplete mtDNA. Cells were then assayed and either administered no further treatment (untreated), treated with RSL3 only (RSL3), or administered RSL3 with Compound 1 (1 μM). In ddC-treated cells, RSL3 alone reduced ATP production significantly. The amount of ATP produced when the cells were treated with (ddC) RSL3 and Compound 1 is virtually identical to the untreated control (vehicle). This demonstrates that Compound 1 is protective of the ATP production and ameliorates the deleterious effects of the addition of RSL3 in this assay.

[0168] With reference to Fig.4, the effect of the glutathione-stressor RSL3 were assessed in the 100 μM ddC with 400 ng / mL EtBr model of mtDNA depletion. In this model, cells were either treated with vehicle (no RSL3), RSL3 alone, or RSL3 co-treated with Compound 1. As shown, RSL3 alone decreased ATP compared to vehicle control. In contrast, co- treatment of RSL3-stressed cells with Compound 1 appeared to restore ATP levels toAtty. Dkt. No.091151-1691 baseline (no RSL3 control) levels. This demonstrates that Compound 1 is effective in restoring OXPHOS (ATP production) in this model of mtDNA depletion. 3. Summary:

[0169] Compound 1 is protective against mtDNA depletion regardless of mtDNA depletion method. Compound 1 increases bioenergetics in mtDNA-depleted cells as reflected by increased VCD. Compound 1 ameliorates glutathione stress in mtDNA-depleted cells by restoring ATP production. Accordingly, these results demonstrate that Compound 1 is useful in methods for treating, preventing, inhibiting, ameliorating, or delaying the onset of mitochondrial disease resulting from nuclear DNA mutations in a subject in need thereof. Example 2: Compound 1 Pharmacokinetics and Tissue Exposure Following Five Daily Subcutaneous Doses to Male C57BL / 6 Mice

[0170] This example demonstrates the pharmacokinetics and dose proportionality of exposure in plasma and tissues (heart and brain) in mice dosed with; (i) 60 mg / kg Compound 1 or (ii) 180 mg / kg Compound 1 and (iii) provides a comparison of 60 mg / kg Compound 1 using 15% Kolliphor^ ELP to 60 mg / kg Compound 1 using 5% Kolliphor^ ELP.

[0171] Study design details: • Animals were allowed to acclimate to the test facility for at least two days prior to the beginning of the study. • Study animals were male C57BL / 6 mice weighing approximately 18-25 grams each. • Fasting was not required for the study. • Body weights were recorded prior to dose administration. The volume of each dose delivered (mL / kg) was based on each individual animal’s body weight. • This was a multiple dose serial PK dosed subcutaneously once daily over 5 days. Compound 1 was administered to two groups: Group 1 at 60 mg / kg and Group 2 at 180 mg / kg (6 and 18 mg / mL, 15% Kolliphor^ ELP in PBS, 10 mL / kg; N = 3 / group). Serial pharmacokinetic (PK) timepoints were collected following the 5thand final dose. Mice were taken down following their 5thand final dose for blood and tissue collection. • All dose syringes were weighed prior to and following dosing to gravimetrically determine the amount of formulation administered. • All animals were observed at dosing and each scheduled collection.Atty. Dkt. No.091151-1691 • The same design was used for Group 3 mice administered 60 mg / kg using 5% Kolliphor^ ELP subcutaneously (SC) once daily over 5 days, except that 3 mice were taken down at each of the four PK timepoints (n=12). Table 1: Study DesignTable 2: Formulation Design

[0172] Sample collection details: • Serial blood samples were collected via tail vein snip. • Blood samples were collected into tubes with anticoagulant. Tubes were stored on wet ice until processed to plasma by centrifugation (3500 rpm at 5℃ for 10 minutes) within 20 minutes of collection. Sample aliquots of 20 µL were transferred into individualAtty. Dkt. No.091151-1691 uniquely labeled matrix tubes and stored at nominal -80℃ until transferred to analytical chemistry for analysis. • Immediately following each terminal blood collection for all groups, animals were sacrificed and perfused with cold phosphate buffered saline (PBS). Heart and brain tissues were collected. o Perfusion methodology: The chest was opened lengthwise using scissors to expose the thoracic organs. A scalpel was used to cut a small hole into the left ventricle just large enough to insert a cannula tip. The cannula was attached to an appropriately sized tubing and syringe prefilled with cold PBS and primed to remove any air. The tip of the cannula was inserted into the left ventricle and directed into the ascending aorta. Hemostats were used to help keep the cannula in place during perfusion. A second hole was made in the right atrium to allow for the escape of fluid during the return circulation. The syringe attached to the cannula was manually pushed or placed into a pump programmed to infuse at 10 mL / min. Animals were perfused for a minimum of 3 minutes or more if needed (until the exiting fluid was clear). Table 3: Sample Collection DesignTable 4: Sample Collection DesignAtty. Dkt. No.091151-1691

[0173] Analyses: Groups 1 and 2 • N = 3 / group; • Collections relative to the final dose: o Plasma: 60 mg / kg at 2 and 24 hr.; 180 mg / kg at predose, 2, 8, and 24 hours (hrs.) o Heart and Brain: 24 hrs. (terminal, PBS-perfused); • RGA 2 bioanalysis. Group 3 • N = 3 / timepoint; • Collections relative to the final dose: o Plasma: 0.5, 2, 8, and 24 hrs. (terminal) o Heart and Brain: 0.5, 2, 8, and 24 hrs. (terminal, PBS-perfused) • RGA 2 bioanalysis.Atty. Dkt. No.091151-1691 Table 5: Sample AnalysesAtty. Dkt. No.091151-1691

[0174] Results - Mouse plasma PK profile after 5 days (Fig.5A): • 60 and 180 mg / kg SC was well tolerated over 5 days (10 mL / kg QD, 15% Kolliphor^ ELP in PBS); • 5% and 15% Kolliphor^ ELP produced similar Compound 1 plasma concentrations at 60 mg / kg; • Tmax ~0.5-2 hr.; • Steady-state achieved within 5 days; • Compound 1 plasma exposure was approximately proportional to dose. Table 6: Compound 1 Mouse Plasma Concentrations (Groups 1 and 2)Atty. Dkt. No.091151-1691 Table 7: Compound 1 Mouse Plasma Concentrations (Group 3)Table 8: Plasma PKNC = Not calculated

[0175] Results - Mouse tissue exposure profile after 5 days (Figs.5B-5C): • Similar Compound 1 concentrations found in heart and brain; • C24 (trough) exposure was approximately proportional to dose; • 180 mg / kg QD maintained heart and brain tissue concentrations > 1 µM after 24 hours – which should be a therapeutically effective tissue concentration. • 60 mg / kg QD maintained heart and brain tissue concentrations > 100 nM after 24 hours – which may be a therapeutically effective tissue concentration. • Compound 1 was well tolerated by the subjects under the conditions administered.Atty. Dkt. No.091151-1691 Table 9: Compound 1 Concentration in Mouse Tissue (Groups 1 and 2) Compound 1 Concentration in C57BL / 6 Mouse TissueTable 10: Compound 1 Concentration in Mouse Tissue (Group 3)Atty. Dkt. No.091151-1691 Table 11: Tissue ExposureEQUIVALENTS

[0176] The present technology is not to be limited in terms of the particular embodiments described in this application, which are intended as single illustrations of individual aspects of the present technology. Many modifications and variations of this present technology can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and apparatuses within the scope of the present technology, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. The present technology is to be limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled. It is to be understood that this present technology is not limited to particular methods, reagents, compounds compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0177] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0178] As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a nonlimiting example, each range discussed herein can be readily broken down into a lower third, middle third and upperAtty. Dkt. No.091151-1691 third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like, include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 cells refers to groups having 1, 2, or 3 cells. Similarly, a group having 1-5 cells refers to groups having 1, 2, 3, 4, or 5 cells, and so forth.

[0179] All patents, patent applications, provisional applications, and publications referred to or cited herein are incorporated by reference in their entirety, including all figures and tables, to the extent they are not inconsistent with the explicit teachings of this specification.

[0180] Other embodiments are set forth within the following claims.

Claims

Atty. Dkt. No.091151-1691 CLAIMS What is claimed is:

1. A method for treating, preventing, inhibiting, ameliorating or delaying the onset of a mitochondrial disease resulting from nuclear DNA mutations in a subject comprising administering to the subject in need thereof a therapeutically effective amount of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, wherein Compound 1 is:.

2. The method of claim 1, wherein the nuclear DNA mutations result in mitochondrial DNA (mtDNA) depletion(s) and / or mitochondrial DNA (mtDNA) rearrangement(s) in the subject.

3. The method of claims 1 or 2, wherein the subject suffers from mitochondrial DNA (mtDNA) depletion syndrome.

4. The method of claims 1, 2, or 3, wherein the mitochondrial disease results from at least one mutation of the POLG gene.

5. The method of claims 1, 2, or 3, wherein the mitochondrial disease results from at least one mutation of the TWNK (twinkle) gene.

6. The method of claims 2 or 3, wherein the mtDNA depletion(s) codes / code for at least one mutation of a complex I respiratory chain subunit.

7. The method of claim 2, wherein the mtDNA rearrangement(s) codes / code for at least one mutation of a complex I respiratory chain subunit.

8. The method of any one of claims 1 to 7, wherein the subject has been diagnosed as having a mitochondrial disease that is neurodegenerative.

9. The method of claim 8, wherein the mitochondrial disease is Kearns–Sayre syndrome or Alzheimer’s disease.Atty. Dkt. No.091151-1691 10. The method of claim 8, wherein the mitochondrial disease is Parkinson’s disease (PD), sporadic Parkinson’s disease (sPD), or Alpers syndrome.

11. The method of any one of claims 1 to 10, wherein the subject is a mammal.

12. The method of claim 11, wherein the mammalian subject is a human.

13. The method of any one of claims 1 to 12, wherein Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered orally or subcutaneously to the subject.

14. The method of any one of claims 1 to 12, wherein Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered topically, intranasally, systemically, intravenously, intraperitoneally, intradermally, intraocularly, ophthalmically, intrathecally, intracerebroventricularly, iontophoretically, transmucosally, intravitreally, or intramuscularly to the subject.

15. The method of any one of claims 1 to 14, further comprising separately, sequentially, or simultaneously administering at least one additional therapeutic agent to the subject.

16. The method of claim 15, wherein the additional therapeutic agent comprises levodopa, optionally in combination with carbidopa (LODOSYN^).

17. The method of claim 15 or claim 16, wherein the combination of administering Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, optionally in combination with at least one additional therapeutic agent, to the subject has a synergistic effect in the treating, preventing, inhibiting, ameliorating or delaying the onset of the mitochondrial disease.

18. The method of any one of claims 1 to 17, wherein administration of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, optionally in combination with at least one additional therapeutic agent, prolongs survival of the subject, as compared with an untreated subject or an untreated control group of subjects.

19. Use of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, in the preparation of a composition, formulation or medicament for treating, preventing, inhibiting, ameliorating or delaying the onset ofAtty. Dkt. No.091151-1691 a mitochondrial disease resulting from nuclear DNA mutations in a subject, wherein Compound 1 is:.

20. The use of claim 19, wherein the nuclear DNA mutations result in mtDNA depletion(s) and / or mitochondrial DNA (mtDNA) rearrangement(s) in the subject.

21. The use of claims 19 or 20, wherein the subject suffers from mitochondrial DNA (mtDNA) depletion syndrome.

22. The use of claims 19, 20, or 21, wherein the mitochondrial disease results from at least one mutation of the POLG gene.

23. The use of claims 19, 20, or 21, wherein the mitochondrial disease results from at least one mutation of the TWNK (twinkle) gene.

24. The use of claims 20 or 21, wherein the mtDNA depletion(s) codes / code for at least one mutation of a complex I respiratory chain subunit.

25. The use of claim 20, wherein the mtDNA rearrangement(s) codes / code for at least one mutation of a complex I respiratory chain subunit.

26. The use of any one of claims 19 to 25, wherein the subject has been diagnosed as having a mitochondrial disease that is neurodegenerative.

27. The use of claim 26, wherein the mitochondrial disease is Kearns–Sayre syndrome or Alzheimer’s disease.

28. The use of claim 26, wherein the mitochondrial disease is Parkinson’s disease (PD), sporadic Parkinson’s disease (sPD), or Alpers syndrome.

29. The use of any one of claims 19 to 28, wherein the subject is a mammal.

30. The use of claim 29, wherein the mammalian subject is a human.

31. The use of any one of claims 19 to 30, wherein the composition, formulation or medicament is formulated for oral or subcutaneous administration.Atty. Dkt. No.091151-1691 32. The use of any one of claims 19 to 30, wherein the composition, formulation or medicament is formulated for topical, intranasal, systemic, intravenous, intraperitoneal, intradermal, intraocular, ophthalmic, intrathecal, intracerebroventricular, iontophoretic, transmucosal, intravitreal, or intramuscular administration.

33. The use of any one of claims 19 to 32, further comprising separately, sequentially, or simultaneously administering an additional therapeutic agent to the subject.

34. The use of claim 33, wherein the additional therapeutic agent comprises levodopa, optionally in combination with carbidopa (LODOSYN^).

35. The use of claim 33 or claim 34, wherein administering the composition, formulation or medicament, optionally in combination with at least one additional therapeutic agent, to the subject has a synergistic effect in the treating, preventing, inhibiting, ameliorating or delaying the onset of the mitochondrial disease.

36. The method of any one of claims 19 to 35, wherein administering the composition, formulation or medicament, optionally in combination with at least one additional therapeutic agent, to the subject prolongs survival of the subject, as compared with an untreated subject or an untreated control group of subjects.

37. A composition comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, for use in treating, preventing, inhibiting, ameliorating or delaying the onset of a mitochondrial disease resulting from nuclear DNA mutations in a subject, wherein Compound 1 is:.

38. The composition of claim 37 comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the nuclear DNA mutations result in mitochondrial DNA (mtDNA) depletion(s) and / or mitochondrial DNA (mtDNA) rearrangement(s) in the subject.Atty. Dkt. No.091151-1691 39. The composition of claims 37 or 38 comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the subject suffers from mitochondrial DNA (mtDNA) depletion syndrome.

40. The composition of claims 37, 38, or 39 comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the mitochondrial disease results from at least one mutation of the POLG gene.

41. The composition of claims 37, 38, or 39 comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the mitochondrial disease results from at least one mutation of the TWNK (twinkle) gene.

42. The composition of claims 38 or 39 comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the mtDNA depletion(s) codes / code for at least one mutation of a complex I respiratory chain subunit.

43. The composition of claim 38 comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the mtDNA rearrangement(s) codes / code for at least one mutation of a complex I respiratory chain subunit.

44. The composition of any one of claims 37 to 43, comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the subject has been diagnosed as having a mitochondrial disease that is neurodegenerative.

45. The composition of claim 44 comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the mitochondrial disease is Kearns–Sayre syndrome or Alzheimer’s disease.

46. The composition of claim 44 comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the mitochondrial disease is Parkinson’s disease (PD), sporadic Parkinson’s disease (sPD), or Alpers syndrome.Atty. Dkt. No.091151-1691 47. The composition of any one of claims 37 to 46, comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the subject is a mammal.

48. The composition of claim 47 comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the mammalian subject is a human.

49. The composition of any one of claims 37 to 48 comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the composition is administered orally or subcutaneously to the subject.

50. The composition of any one of claims 37 to 48 comprising Compound 1, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, wherein the composition is administered topically, intranasally, systemically, intravenously, intraperitoneally, intradermally, intraocularly, ophthalmically, intrathecally, intracerebroventricularly, iontophoretically, transmucosally, intravitreally, or intramuscularly to the subject.

51. A method for treating a subject suffering from mitochondrial DNA (mtDNA) depletion syndrome, the method comprising: administering to the subject a therapeutically effective amount of Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, wherein Compound 1 is:.

52. The method of claim 51, wherein the subject has at least one mutation of the POLG gene.

53. The method of claim 51, wherein the subject has at least one mutation of the TWNK (twinkle) gene.

54. The method of any one of claims 51 to 53, wherein the subject is a mammal.

55. The method of claim 54, wherein the mammalian subject is a human.Atty. Dkt. No.091151-1691 56. The method of any one of claims 51 to 55, wherein Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered orally or subcutaneously to the subject.

57. The method of any one of claims 51 to 55, wherein Compound 1, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, and / or solvates thereof, is administered topically, intranasally, systemically, intravenously, intraperitoneally, intradermally, intraocularly, ophthalmically, intrathecally, intracerebroventricularly, iontophoretically, transmucosally, intravitreally, or intramuscularly to the subject.

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