Compositions for improving cell viability, mitochondrial function and frataxin expression, and methods of use thereof

Abstract

The present disclosure provides methods and compositions for improving cell viability, mitochondrial function and / or frataxin expression or treating Friedreich's ataxia (FRDA) comprising two or more of monomethyl fumarate (MMF) or a MMF prodrug such as dimethyl fumarate (DMF), nicotinamide (NAM), idebenone (IDB) and nitisinone (NTS).

Description

[0001] Docket: M1643.70002WG00

[0002] COMPOSITIONS FOR IMPROVING CELL VIABILITY, MITOCHONDRIAL FUNCTION AND FRATAXIN EXPRESSION, AND METHODS OF USE THEREOF

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS

[0004] This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application No. 63 / 695,268, filed on September 16, 2024, and U.S. Provisional Application No. 63 / 814,778 filed on May 30, 2025, the entire contents of each of which are incorporated herein by reference.

[0005] BACKGROUND

[0006] Friedreich’s Ataxia (FRDA) is a monogenic recessive inherited disease caused by a deficiency in frataxin which leads to mitochondrial dysfunction and a progressive loss of function in patients characterized by weakness and heart failure. Friedreich’s Ataxia affects over six thousand patients in the USA per NIH estimates. There is only one currently approved drug, omaveloxolone (Omav), marketed as Skyclarys. Omav acts through Nrf-2 activation and has no effect on frataxin levels. It has shown, at most, a benefit equivalent to a year’s worth of deterioration, but it is accompanied by many side effects including derangement in liver function tests, gastro-intestinal toxicity and changes in brain natriuretic peptide, a sensitive marker of heart failure. Thus, there is a need for improved therapies to treat FRDA.

[0007] SUMMARY

[0008] Described herein are compositions comprising two or more of monomethyl fumarate (MMF) (or a MMF prodrug, such as dimethyl fumarate, DMF), nicotinamide (NAM), idebenone (IDB), and nitisinone (NTS), as well as methods of using these compositions, such as for the treatment of Friedreich’s Ataxia (FRDA). These compositions and methods produce greater improvements in cell viability, mitochondrial function, frataxin expression, NAD(P)H quinone dehydrogenase 1 expression, thioredoxin reductase 1 expression, and / or Nrf-2 signaling activation relative to embodiments comprising only one of these agents. Moreover, in some embodiments, these compositions and methods produce greater improvements in cell viability, mitochondrial function, frataxin expression, NAD(P)H quinone dehydrogenase 1 expression, thioredoxin reductase 1 expression, and / or Nrf-2 signaling activation relative to omaveloxolone.

[0009] 12523362.1 In some aspects, the disclosure relates to a method for increasing frataxin levels in a cell, increasing NAD(P)H quinone dehydrogenase 1 levels in a cell, increasing thioredoxin reductase 1 expression in a cell, increasing Nrf-2 signaling activation in a cell, increasing viability of a cell, and / or increasing mitochondrial function of a cell.

[0010] In some embodiments, the method comprises contacting the cell with an effective amount of a composition comprising two or more of: a) dimethyl fumarate (DMF) or monomethyl fumarate (MMF); b) nicotinamide (NAM); c) idebenone (IDB); and d) nitisinone (NTS).

[0011] In some embodiments, the composition comprises three or more of DMF or MMF, NAM, IDB, and NTS, optionally wherein the composition comprises each of DMF or MMF, NAM, IDB, and NTS.

[0012] In some embodiments, the method increases frataxin levels in the cell to a greater extent than the frataxin levels of a control cell contacted with a composition comprising two or fewer of DMF or MMF, NAM, IDB, and NTS, such as a composition consisting essentially of DMF or MMF and NAM, DMF or MMF and IDB, NAM and IDB, DMF or MMF and NTS, NAM and NTS, or IDB and NTS. In some embodiments, the method increases frataxin mRNA and / or protein levels in the cell by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or at least 50%. In some embodiments, the method increases frataxin levels in the cell to an equal or greater extent than the sum of frataxin levels in control cells contacted with each of DMF or MMF, NAM, IDB, or NTS alone.

[0013] In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 levels in the cell. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 mRNA and / or protein levels in the cell by at least 50%, at least 75%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, at least 550%, or at least 600%. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 levels in the cell to an equal or greater extent than the sum of NAD(P)H quinone dehydrogenase 1 levels in control cells contacted with each of MMF or NAM alone.

[0014] In some embodiments, the method increases thioredoxin reductase 1 levels in the cell. In some embodiments, the method increases thioredoxin reductase 1 mRNA and / or protein levels in the cell by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, at least 225%, at least 250%, at least 270%, or at least 300%. In some

[0015] 12523362.1 embodiments, the method increases thioredoxin reductase 1 levels in the cell to an equal or greater extent than the sum of thioredoxin reductase 1 levels in control cells contacted with each of MMF or NAM alone.

[0016] In some embodiments, the method increases viability of the cell.

[0017] In some embodiments, the cell is contacted with DMF at a concentration of 5-15 pM, 10- 20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95- 105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170- 180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250- 260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM. In some embodiments, the cell is contacted with DMF at a concentration of about lOpM, about 30 pM, or about 90 pM.

[0018] In some embodiments, the cell is contacted with MMF at a concentration of 5-15 pM, 10- 20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95- 105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170- 180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250- 260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM. In some embodiments, the cell is contacted with MMF at a concentration of about 40 pM, about 50 pM, or about 60 pM.

[0019] In some embodiments, the cell is contacted with NAM at a concentration of 0.5-10 mM, 0.5-1 mM, 1-1.5 mM, 1.5-2 mM, 2-2.5 mM, 2.5-3 mM, 3-3.5 mM, 3.5-4 mM, 4-4.5 mM, 4.5-5 mM, 5-5.5 mM, 5.5-6 mM, 6-6.5 mM, 6.5-7 mM, 7-7.5 mM, 7.5-8 mM, 8-8.5 mM, 8.5-9 mM, 9- 9.5 mM, 9.5-10 mM, 10-10.5 mM, 10.5-11 mM, 11-11.5 mM, 11.5 mM-12 mM, 12-12.5 mM, 12.5-13 mM, 13-13.5 mM, 13.5-14 mM, 14-14.5 mM, or 14.5-15 mM. In some embodiments, the cell is contacted with NAM at a concentration of about 1 mM, about 3 mM, about 5mM, or about 9 mM.

[0020] In some embodiments, the method comprises administering ImM NAM and 30 pM DMF to the subject. In some embodiments, the method comprises administering ImM NAM and 90 pM DMF to the subject. In some embodiments, the method comprises administering 3mM NAM and 30 pM DMF to the subject. In some embodiments, the method comprises administering 3mM NAM and 90 pM DMF to the subject.

[0021] In some embodiments, the composition comprises DMF or MMF in an amount that is less than 480 mg. In some embodiments, the composition comprises NAM in an amount that is

[0022] 12523362.1 less than 5 g. In some embodiments, the composition comprises IDB in an amount that is less than 450 mg. In some embodiments, the composition comprises NTS in an amount that is less than 70 mg.

[0023] In some embodiments, the method comprises contacting the cell with the composition twice daily.

[0024] In some embodiments, the method comprises contacting the cell with an effective amount of a composition comprising: a) dimethyl fumarate (DMF) or monomethyl fumarate (MMF); and b) nicotinamide (NAM); wherein the composition comprises less than 480 mg DMF or MMF and less than 5 g NAM.

[0025] In some embodiments, the composition further comprises idebenone (IDB) and / or nitisinone (NTS). In some embodiments, the composition comprises less than 450 mg IDB. In some embodiments, the composition comprises less than 70 mg.

[0026] In some embodiments, the method increases frataxin mRNA and / or protein levels in the cell by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, or least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150%. In some embodiments, the method increases frataxin levels in the cell to an equal or greater extent than the sum of frataxin levels in control cells contacted with each of DMF or MMF or NAM alone.

[0027] In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 levels in the cell. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 mRNA and / or protein levels in the cell by at least 50%, at least 75%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, at least 550%, or at least 600%. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 levels in the cell to an equal or greater extent than the sum of NAD(P)H quinone dehydrogenase 1 levels in control cells contacted with each of MMF or NAM alone.

[0028] In some embodiments, the method increases thioredoxin reductase 1 levels in the cell. In some embodiments, the method increases thioredoxin reductase 1 mRNA and / or protein levels in the cell by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, at least 225%, at least 250%, at least 270%, or at least 300%. In some embodiments, the method increases thioredoxin reductase 1 levels in the cell to an equal or

[0029] 12523362.1 greater extent than the sum of thioredoxin reductase 1 levels in control cells contacted with each of MMF or NAM alone.

[0030] In some embodiments, the method increases viability of the cell.

[0031] In some aspects, the disclosure provides a method for treating Friedreich’s Ataxia, the method comprising administering to a subject in need thereof an effective amount of a composition comprising two or more of: a) dimethyl fumarate (DMF) or monomethyl fumarate (MMF); b) nicotinamide (NAM); c) idebenone (IDB); and d) nitisinone (NTS).

[0032] In some embodiments, the composition comprises three or more of DMF or MMF, NAM, IDB, and NTS, optionally wherein the composition comprises each of DMF or MMF, NAM, IDB, and NTS.

[0033] In some embodiments, the method increases frataxin levels in cells of the subject to a greater extent than the frataxin levels of cells of a subject administered a composition comprising two or fewer of DMF or MMF, NAM, IDB, and NTS, such as a composition consisting essentially of DMF or MMF and NAM, DMF or MMF and IDB, or NAM and IDB. In some embodiments, the method increases frataxin mRNA and / or protein levels in cells of the subject by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150%. In some embodiments, the method increases frataxin levels in cells of the subject to an equal or greater extent than the sum of frataxin levels in cells of a subjects administered DMF or MMF, NAM, or IDB alone.

[0034] In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 levels in cells of the subject. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 mRNA and / or protein levels in cells of the subject by at least 50%, at least 75%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, at least 550%, or at least 600%. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 levels in cells of the subject to an equal or greater extent than the sum of NAD(P)H quinone dehydrogenase 1 levels in control cells contacted with each of MMF or NAM alone.

[0035] In some embodiments, the method increases thioredoxin reductase 1 levels in cells of the subject. In some embodiments, the method increases thioredoxin reductase 1 mRNA and / or protein levels in cells of the subject by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, at least 225%, at least 250%, at least 270%, or at least

[0036] 12523362.1 300%. In some embodiments, the method increases thioredoxin reductase 1 levels in cells of the subject to an equal or greater extent than the sum of thioredoxin reductase 1 levels in control cells contacted with each of MMF or NAM alone.

[0037] In some embodiments, the method increases viability of cells in the subject.

[0038] In some embodiments, the composition is administered to the subject orally.

[0039] In some embodiments, the composition comprises DMF in an amount sufficient that cells of the subject are exposed to DMF at a concentration of 5-15 pM, 10-20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM. In some embodiments, the composition comprises DMF in an amount sufficient that cells of the subject are exposed to DMF at a concentration of about lOpM, about 30 pM, or about 90 pM. In some embodiments, the composition comprises DMF or MMF in an amount that is less than 480 mg.

[0040] In some embodiments, the composition comprises MMF in an amount sufficient that cells of the subject are exposed to MMF at a concentration of 5-15 pM, 10-20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM. In some embodiments, the composition comprises MMF in an amount sufficient that cells of the subject are exposed to MMF at a concentration of about 40 pM, about 50 pM, or about 60 pM.

[0041] In some embodiments, the composition comprises NAM in an amount sufficient that cells of the subject are exposed to NAM at a concentration of 0.5-10 mM, 0.5-1 mM, 1-1.5 mM, 1.5-2 mM, 2-2.5 mM, 2.5-3 mM, 3-3.5 mM, 3.5-4 mM, 4-4.5 mM, 4.5-5 mM, 5-5.5 mM, 5.5-6 mM, 6-6.5 mM, 6.5-7 mM, 7-7.5 mM, 7.5-8 mM, 8-8.5 mM, 8.5-9 mM, 9-9.5 mM, 9.5-10 mM, 10-10.5 mM, 10.5-11 mM, 11-11.5 mM, 11.5 mM-12 mM, 12-12.5 mM, 12.5-13 mM, 13-13.5 mM, 13.5-14 mM, 14-14.5 mM, or 14.5-15 mM. In some embodiments, the composition comprises NAM in an amount sufficient that cells of the subject are exposed to NAM at a concentration of about 1 mM, about 3 mM, or about 9 mM.

[0042] 12523362.1 In some embodiments, the composition comprises DMF or MMF in an amount that is less than 480 mg. In some embodiments, the composition comprises NAM in an amount that is less than 5 g. In some embodiments, the composition comprises IDB in an amount less than 450 mg. In some embodiments, the composition comprises NTS in an amount less than 70 mg.

[0043] In some embodiments, the method comprises administering a first composition comprising two or more of DMF or MMF, NAM, IDB, and NTS; and administering a second composition comprising one or more of DMF or MMF, NAM, IDB, and NTS; wherein the first composition and the second composition differ; and wherein the first composition and the second composition are administered simultaneously. In some embodiments, the method comprises administering a composition comprising DMF or MMF, NAM, IBD, and / or NTS twice daily.

[0044] In some embodiments, after treatment with the method, the subject has a lower International Cooperative Ataxia Rating Scale (ICARS) or Functional Assessment Rating Scale (FARS) score than before treatment with the method.

[0045] In some embodiments, the method causes fewer side effects in the subject than treatment with omaveloxolone.

[0046] In some embodiments, the method comprises administering a composition comprising DMF or MMF, NAM, IBD, and / or NTS twice daily.

[0047] In some aspects, the disclosure relates to a method for reducing oxidative stress in a cell, the method comprising contacting the cell an effective amount of a composition comprising two or more of: a) dimethyl fumarate (DMF) or monomethyl fumarate (MMF); b) nicotinamide (NAM); c) idebenone (IDB); and d) nitisinone (NTS). In some embodiments, the method reduces the level of reactive oxygen species (ROS) in the cell.

[0048] In some embodiments, the cell is contacted with DMF at a concentration of 5-15 pM, 10- 20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM. In some embodiments, the cell is contacted with DMF at a concentration of about lOpM, about 30 pM, or about 90 pM.

[0049] In some embodiments, the cell is contacted with MMF at a concentration of 5-15 pM, 10- 20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-

[0050] 12523362.1 105 pM, 100-110 pM, 110-120 pM, 120-130 pM. 130-140 pM. 140-150 pM, 150-160 pM, 170- 180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250- 260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM. In some embodiments, the cell is contacted with MMF at a concentration of about 40 pM, about 50 pM, or about 60 pM.

[0051] In some embodiments, the cell is contacted with NAM at a concentration of 0.5-1 mM, 1-1.5 mM, 1.5-2 mM, 2-2.5 mM, 2.5-3 mM, 3-3.5 mM, 3.5-4 mM, 4-4.5 mM, 4.5-5 mM, 5-5.5 mM, 5.5-6 mM, 6-6.5 mM, 6.5-7 mM, 7-7.5 mM, 7.5-8 mM, 8-8.5 mM, 8.5-9 mM, 9-9.5 mM, 9.5-10 mM, 10-10.5 mM, 10.5-11 mM, 11-11.5 mM, 11.5 mM-12 mM, 12-12.5 mM, 12.5-13 mM, 13-13.5 mM, 13.5-14 mM, 14-14.5 mM, or 14.5-15 mM. In some embodiments, the cell is contacted with NAM at a concentration of about 1 mM, about 3 mM, or about 9 mM.

[0052] In some embodiments, the cell is contacted with 1 mM NAM and 30 pM DMF. In some embodiments, the cell is contacted with 1 mM NAM and 90 pM DMF. In some embodiments, the cell is contacted with 3 mM NAM and 30 pM DMF. In some embodiments, the cell is contacted with 3 mM NAM and 90 pM DMF. In some embodiments, the composition comprises 5 mM NAM and 50 pM MMF.

[0053] BRIEF DESCRIPTION OF THE DRAWINGS

[0054] FIG. 1 shows a comparison of percentage ATP generated in treated cells relative to control cells. Treating with 90 pM DMF and 1 mM NAM showed greater improvement in ATP generation compared to either treatment alone.

[0055] FIGs. 2A-2D show the average frataxin mRNA levels upon treatment with DMF and NAM, alone and in combination. FIG. 2A shows increased frataxin mRNA levels upon treatment of cells with 30 pM DMF and ImM NAM, compared to control (vehicle) and an equivalent amount of each compound alone. FIG. 2B shows increased frataxin mRNA levels upon treatment of cells with 30 pM DMF and 3 mM NAM, compared to control (vehicle) and an equivalent amount of each compound alone. FIG. 2C shows increased frataxin mRNA levels upon treatment of cells with 90 pM DMF and 1 mM NAM, compared to control (vehicle) and an equivalent amount of each compound alone. FIG. 2D shows increased frataxin mRNA levels upon treatment of cells with 90 pM DMF and 3 mM NAM, compared to control (vehicle) and an equivalent amount of each compound alone.

[0056] 12523362.1 FIGs. 3A-3D show the absolute percent increase of frataxin mRNA (copies / ug mRNA) with low and high NAM and DMF concentrations. The low NAM concentration is 1 mM; the high NAM concentration is 3 mM. The low DMF concentration is 30 pM; the high DMF concentration is 90 pM. FIG. 3A shows increased frataxin mRNA levels upon treatment of cells with 30 pM DMF and 1 mM NAM, compared to control (vehicle) and an equivalent amount of each compound alone. FIG. 3B shows increased frataxin mRNA levels upon treatment of cells with 90 pM DMF and 1 mM NAM, compared to control (vehicle) and an equivalent amount of each compound alone. FIG. 3C shows increased frataxin mRNA levels upon treatment of cells with 30 pM DMF and 3 mM NAM, compared to control (vehicle) and an equivalent amount of each compound alone. FIG. 3D shows increased frataxin mRNA levels upon treatment of cells with 90 pM DMF and 3 mM NAM, compared to control (vehicle) and an equivalent amount of each compound alone.

[0057] FIGs. 4A-4C show the average fold change in expression of frataxin (FXN) mRNA (FIG. 4A), NAD(P)H quinone dehydrogenase 1 (NQO1) mRNA (FIG. 4B), and thioredoxin reductase 1 (TXNRD1) mRNA (FIG. 4C) upon treatment with 50 pM MMF and 5 mM NAM, alone and in combination (N5M50). Expression is compared to treatment with control (DMSO) and treatment with 50 nM omaveloxolone. FIG. 4A shows increased frataxin mRNA levels upon treatment of cells with 50 pM MMF and 5 mM NAM, compared to control (DMSO), 50 nM omaveloxolone, and an equivalent amount of each compound alone. FIG. 4B shows increased NAD(P)H quinone dehydrogenase 1 mRNA levels upon treatment of cells with 50 pM MMF and 5 mM NAM, compared to control (DMSO) and an equivalent amount of each compound alone. FIG. 4C shows increased thioredoxin reductase 1 mRNA levels upon treatment of cells with 50 pM MMF and 5 mM NAM, compared to control (DMSO), 50 nM omaveloxolone, and an equivalent amount of each compound alone.

[0058] FIG. 5 shows the average percent cell viability upon exposure to 200 pM H2O2 and treatment with 50 pM MMF and 5 mM NAM, alone and in combination (N5M50). Expression is compared to treatment with control (DMSO) and treatment with 50 nM omaveloxolone. Highest viability was seen upon treatment of cells with 50 pM MMF and 5 mM NAM.

[0059] DETAILED DESCRIPTION

[0060] The present disclosure reports the discovery that compositions comprising two or more of dimethyl fumarate (DMF) (or another monomethyl fumarate prodrug), monomethyl fumarate

[0061] 12523362.1 (MMF), nicotinamide (NAM), idebenone (IDB), and nitisinone (NTS) can produce greater improvements in cell viability, mitochondrial function, frataxin expression, NAD(P)H quinone dehydrogenase 1 expression, and / or thioredoxin reductase 1 expression than each those agents can alone. This surprising discovery suggests that combinations of these agents will be more effective in treating FRDA, than each agent alone.

[0062] I. Compositions

[0063] In some aspects, the disclosure relates to compositions comprising two or more of dimethyl fumarate (DMF) (or another monomethyl fumarate prodrug, such as diroximel fumarate), monomethyl fumarate (MMF), nicotinamide (NAM), idebenone (IDB), and nitisinone (NTS). The compositions described herein may comprise any combination of DMF (or another MMF prodrug, such as diroximel fumarate), MMF, NAM, IDB, and NTS. Exemplary compositions provided herein comprise the components as depicted in Table 1.

[0064] Table 1: Exemplary components of compositions described herein.

[0065] In some embodiments, a composition comprises MMF or DMF (or another MMF prodrug, such as diroximel fumarate) and each of NAM, IDB, and NTS.

[0066] In some embodiments, a composition further comprises a pharmaceutically acceptable vehicle or pharmaceutically acceptable excipient. Exemplary pharmaceutical vehicles and pharmaceutically acceptable excipients are known to those having ordinary skill in the art.

[0067] In some embodiments, the disclosure relates to a composition consisting essentially of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and NAM, DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and IDB, NAM and IDB, DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and NTS, NAM and NTS, or IDB

[0068] 12523362.1 and NTS. In some embodiments, a composition consisting essentially of a combination of the agents listed above further comprises additional non-therapeutic agents. In some embodiments, additional non-therapeutic agents comprise excipients, diluents, and the like.

[0069] In some embodiments, a composition described herein, when contacted with a cell, is capable of: activating Nrf-2 signaling in the cell; increasing frataxin levels in the cell; increasing NAD(P)H quinone dehydrogenase 1 levels in the cell; increasing thioredoxin reductase 1 levels in the cell; and / or decreasing levels of reactive oxygen species (ROS) in the cell. In some embodiments, a composition described herein, when contacted with a cell, is capable of activating Nrf-2 signaling in the cell. In some embodiments, a composition described herein, when contacted with a cell, is capable of increasing frataxin levels in the cell. In some embodiments, a composition described herein, when contacted with a cell, is capable of increasing NAD(P)H quinone dehydrogenase 1 levels in the cell. In some embodiments, a composition described herein, when contacted with a cell, is capable of increasing thioredoxin reductase 1 levels in the cell. In some embodiments, a composition described herein, when contacted with a cell, is capable of decreasing levels of reactive oxygen species (ROS) in the cell. In some embodiments, a composition described herein, when contacted with a cell, is capable of: activating Nrf-2 in the cell; increasing frataxin levels in the cell; and decreasing levels of reactive oxygen species (ROS) in the cell. In some embodiments, a composition described herein, when contacted with a cell, is capable of: activating Nrf-2 signaling in the cell; increasing frataxin levels in the cell; increasing NAD(P)H quinone dehydrogenase 1 levels in the cell; increasing thioredoxin reductase 1 levels in the cell; and decreasing levels of reactive oxygen species (ROS) in the cell.

[0070] Dimethyl Fumarate

[0071] In some embodiments, a composition comprises dimethyl fumarate (DMF) (or another

[0072] MMF prodrug, such as diroximel fumarate). DMF is a compound having the structure shown below:

[0073] 12523362.1

[0074] In some embodiments, a composition comprises about 160 milligrams (mg), about 240 mg, or about 480 mg DMF (or another MMF prodrug, such as diroximel fumarate). In some embodiments, a composition comprises 100-200 mg, 125-225 mg, 150-250 mg, 175-275 mg, 200-300 mg, 225-325 mg, 250-350 mg, 275-375mg, 300-400mg, 325-425 mg, 350-450 mg, 375-475 mg, or 400-500 mg DMF (or another MMF prodrug, such as diroximel fumarate).

[0075] In some embodiments, a composition provided herein comprises DMF (or another MMF prodrug, such as diroximel fumarate) in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of about 10 pM, about 30 pM, or about 90 pM. In some embodiments, a composition provided herein comprises DMF (or another MMF prodrug, such as diroximel fumarate) in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 5-15 pM, 10-20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM.

[0076] Monomethyl Fumarate

[0077] In some embodiments, a composition comprises monomethyl fumarate (MMF). MMF is a metabolite of DMF, and has the structure shown below:

[0078] 12523362.1

[0079] In some embodiments, a composition comprises about 95 mg, 125 mg, about 190 mg, or about 380 mg MMF. In some embodiments, a composition comprises 50-150 mg, 75-175 mg, 100-200 mg, 125-225 mg, 150-250 mg, 175-275 mg, 200-300 mg, 225-325 mg, 250-350 mg, 275-375mg, 300-400 mg, 325-425 mg, 350-450 mg, 375-475 mg, 400-500, 425-525 mg, 450- 550 mg, 475-575 mg, or 500-600 mg MMF.

[0080] In some embodiments, a composition provided herein comprises MMF in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to MMF at a concentration of about 40 pM, about 50 pM, or about 60 pM. In some embodiments, a composition provided herein comprises MMF in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to MMF at a concentration of 5-15 pM, 10-20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM.

[0081] Nicotinamide

[0082] In some embodiments, a composition comprises nicotinamide (NAM). NAM is a chemical compound having the structure shown below:

[0083] 12523362.1 In some embodiments, a composition comprises about 1 gram (g), 1.5 g, about 1.75 g, about 2 g, about 2.25 g, about 2.5 g, about 2.75 g, about 3 g, about 3.25 g, about 3.5 g, about 3.75 g, about 4 g, about 4.25 g, about 4.5 g, about 4.75 g, about 5 g, about 5.25 g, about 5.5 g, about 5.75 g, about 6 g, about 6.25 g, or about 6.5 g NAM. In some embodiments, a composition comprises 1-2 g, 1.25-2.25 g, 1.5-2.5 g, 1.75-2.75 g, 2-3 g, 2.25-3.25 g, 2.5g-3.5 g, 2.75-3.75 g, 3-4 g, 3.25-4.25 g, 3.5-4.5 g, 3.75-4.75 g, 4-5 g, 4.25-5.25 g, 4.5-5.5 g, 4.75-5.75 g, 5-6 g, 5.25- 6.25 g, or 5.5-6.5 g NAM.

[0084] In some embodiments, a composition provided herein comprises NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of about 1 mM, about 3 mM, about 5 mM, or about 9 mM. In some embodiments, a composition provided herein comprises NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of 0.5-10 mM, 0.5-1 mM, 1- 1.5 mM, 1.5-2 mM, 2-2.5 mM, 2.5-3 mM, 3-3.5 mM, 3.5-4 mM, 4-4.5 mM, 4.5-5 mM, 5-5.5 mM, 5.5-6 mM, 6-6.5 mM, 6.5-7 mM, 7-7.5 mM, 7.5-8 mM, 8-8.5 mM, 8.5-9 mM, 9-9.5 mM, 9.5-10 mM, 10-10.5 mM, 10.5-11 mM, 11-11.5 mM, 11.5 mM-12 mM, 12-12.5 mM, 12.5-13 mM, 13-13.5 mM, 13.5-14 mM, 14-14.5 mM, or 14.5-15 mM.

[0085] Idebenone

[0086] In some embodiments, a composition comprises idebenone (IDB). IDB is a chemical compound having the structure shown in below:

[0087] In some embodiments, a composition comprises about 1.167 g, about 1.75 g, or about 3.5 g IDB. In some embodiments, a composition comprises 1-2 g, 1.25-2.25 g, 1.5-2.5 g, 1.75-2.75

[0088] 12523362.1 g, 2-3 g, 2.25-3.25 g, 2.5g-3.5 g, 2.75-3.75 g, 3-4 g, 3.25-4.25 g, 3.5-4.5 g, 3.75-4.75 g, 4-5 g, 4.25-5.25 g, 4.5-5.5 g, 4.75-5.75 g, 5-6 g, 5.25-6.25 g, or 5.5-6.5 g IDB.

[0089] Nitisinone

[0090] In some embodiments, a composition comprises nitisinone (NTS). NTS is a chemical compound having the structure shown below:

[0091] In some embodiments, a composition comprises about 35 mg, about 70 mg, or about 140 mg NTS. In some embodiments, a composition comprises 25-125 mg, 50-150 mg, 75-175 mg, 100-200 mg, 125-225 mg, 150-250 mg, 175-275 mg, 200-300 mg, 225-325 mg, 250-350 mg, 275-375mg, 300-400 mg, 325-425 mg, 350-450 mg, 375-475 mg, 400-500, 425-525 mg, 450- 550 mg, 475-575 mg, or 500-600 mg NTS.

[0092] Excmplary Combinations of Dimethyl Fumarate and Nicotinamide

[0093] In some embodiments, the disclosure relates to a composition comprising or consisting essentially of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM.

[0094] In some embodiments, a composition provided herein that comprises or consists essentially of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM comprises:

[0095] DMF (or another MMF prodrug, such as diroximel fumarate) in an amount of about 160 milligrams (mg), about 240 mg, or about 480 mg DMF (or another MMF prodrug, such as diroximel fumarate). In some embodiments, a composition comprises 100-200 mg, 125-225 mg, 150-250 mg, 175-275 mg, 200-300 mg, 225-325 mg, 250-350 mg, 275-375 mg, 300-400 mg,

[0096] 12523362.1 325-425 mg, 350-450 mg, 375-475 mg, or 400-500 mg DMF (or another MMF prodrug, such as diroximel fumarate); and / or

[0097] NAM in an amount of about 1 gram (g), 1.5 g, about 1.75 g, about 2 g, about 2.25 g, about 2.5 g, about 2.75 g, about 3 g, about 3.25 g, about 3.5 g, about 3.75 g, about 4 g, about

[0098] 4.25 g, about 4.5 g, about 4.75 g, about 5 g, about 5.25 g, about 5.5 g, about 5.75 g, about 6 g, about 6.25 g, or about 6.5 g NAM. In some embodiments, a composition comprises 1-2 g, 1.25-

[0099] 2.25 g, 1.5-2.5 g, 1.75-2.75 g, 2-3 g, 2.25-3.25 g, 2.5g-3.5 g, 2.75-3.75 g, 3-4 g, 3.25-4.25 g, 3.5- 4.5 g, 3.75-4.75 g, 4-5 g, 4.25-5.25 g, 4.5-5.5 g, 4.75-5.75 g, 5-6 g, 5.25-6.25 g, or 5.5-6.5 g NAM.

[0100] In some embodiments, a composition provided herein that comprises or consists essentially of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM comprises:

[0101] DMF (or another MMF prodrug, such as diroximel fumarate) in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 5-15 pM, 10-20 pM, or about 10 pM; and

[0102] NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of 0.5-1 mM, 1-1.5 mM, 1.5-2 mM, or about 1 mM.

[0103] In some embodiments, a composition provided herein that comprises or consists essentially of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM comprises:

[0104] DMF (or another MMF prodrug, such as diroximel fumarate) in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 5-15 pM, 10-20 pM, or about 10 pM; and

[0105] NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of 2-2.5 mM, 2.5-3 mM, 3-3.5 mM, 3.5-4 mM, 4-4.5 mM, 4.5-5 mM, 5-5.5 mM, 5.5-6 mM, 6-6.5 mM, 6.5-7 mM, or about 3 mM.

[0106] 12523362.1 In some embodiments, a composition provided herein that comprises or consists essentially of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM comprises:

[0107] DMF (or another MMF prodrug, such as diroximel fumarate) in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 5-15 pM, 10-20 pM, or about 10 pM; and

[0108] NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of 7-7.5 mM,

[0109] 7.5-8 mM, 8-8.5 mM, 8.5-9 mM, 9-9.5 mM, 9.5-10 mM, 10-10.5 mM, 10.5-11 mM, 11-11.5 mM, 11.5 mM-12 mM, 12-12.5 mM, 12.5-13 mM, 13-13.5 mM, 13.5-14 mM, 14-14.5 mM,

[0110] 14.5-15 mM, or about 9 mM.

[0111] In some embodiments, a composition provided herein that comprises or consists essentially of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM comprises:

[0112] DMF (or another MMF prodrug, such as diroximel fumarate) in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, or about 30 pM; and

[0113] NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of 0.5-1 mM, 1-1.5 mM, 1.5-2 mM, or about 1 mM.

[0114] In some embodiments, a composition provided herein that comprises or consists essentially of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM comprises:

[0115] DMF (or another MMF prodrug, such as diroximel fumarate) in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to DMF at a concentration of 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, or about 30 pM; and

[0116] NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of 2-2.5 mM,

[0117] 12523362.1 2.5-3 mM, 3-3.5 mM, 3.5-4 mM, 4-4.5 mM, 4.5-5 mM, 5-5.5 mM, 5.5-6 mM, 6-6.5 mM, 6.5-7 mM, or about 3 mM.

[0118] In some embodiments, a composition provided herein that comprises or consists essentially of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM comprises:

[0119] DMF (or another MMF prodrug, such as diroximel fumarate) in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, or about 30 pM; and

[0120] NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of 7-7.5 mM,

[0121] 7.5-8 mM, 8-8.5 mM, 8.5-9 mM, 9-9.5 mM, 9.5-10 mM, 10-10.5 mM, 10.5-11 mM, 11-11.5 mM, 11.5 mM-12 mM, 12-12.5 mM, 12.5-13 mM, 13-13.5 mM, 13.5-14 mM, 14-14.5 mM,

[0122] 14.5-15 mM, or about 9 mM.

[0123] In some embodiments, a composition provided herein that comprises or consists essentially of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM comprises:

[0124] DMF (or another MMF prodrug, such as diroximel fumarate) in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM, or about 90 pM; and

[0125] NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of 0.5-1 mM, 1-1.5 mM, 1.5-2 mM, or about 1 mM.

[0126] In some embodiments, a composition provided herein that comprises or consists essentially of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM comprises:

[0127] 12523362.1 DMF (or another MMF prodrug, such as diroximel fumarate) in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM, or about 90 pM; and

[0128] NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of 2-2.5 mM,

[0129] 2.5-3 mM, 3-3.5 mM, 3.5-4 mM, 4-4.5 mM, 4.5-5 mM, 5-5.5 mM, 5.5-6 mM, 6-6.5 mM, 6.5-7 mM, or about 3 mM.

[0130] In some embodiments, a composition provided herein that comprises or consists essentially of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM comprises:

[0131] DMF (or another MMF prodrug, such as diroximel fumarate) in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM, or about 90 pM; and

[0132] NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of 7-7.5 mM,

[0133] 7.5-8 mM, 8-8.5 mM, 8.5-9 mM, 9-9.5 mM, 9.5-10 mM, 10-10.5 mM, 10.5-11 mM, 11-11.5 mM, 11.5 mM-12 mM, 12-12.5 mM, 12.5-13 mM, 13-13.5 mM, 13.5-14 mM, 14-14.5 mM,

[0134] 14.5-15 mM, or about 9 mM.

[0135] In some embodiments, a composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 10 pM and NAM at a concentration of 1 mM. In some embodiments, a composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 30 pM and NAM at a concentration of 1 mM. In some embodiments, a composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 90 pM and NAM at a concentration of 1 mM. In some

[0136] 12523362.1 embodiments, a composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 270 pM and NAM at a concentration of 1 mM. In some embodiments, a composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 10 pM and NAM at a concentration of 3 mM. In some embodiments, a composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 30 pM and NAM at a concentration of 3 mM. In some embodiments, a composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 90 pM and NAM at a concentration of 3 mM. In some embodiments, a composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 270 pM and NAM at a concentration of 3 mM. In some embodiments, a composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 10 pM and NAM at a concentration of 9 mM. In some embodiments, a composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 30 pM and NAM at a concentration of 9 mM. In some embodiments, a composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 90 pM and NAM at a concentration of 9 mM. In some embodiments, a composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) at a concentration of 270 pM and NAM at a concentration of 9 mM.

[0137] Exemplary Combinations of Monomethyl Fumarate and Nicotinamide In some embodiments, the disclosure relates to a composition comprising or consisting essentially of MMF and NAM.

[0138] In some embodiments, a composition provided herein that comprises or consists essentially of MMF and NAM comprises:

[0139] MMF in an amount of about 160 milligrams (mg), about 240 mg, or about 480 mg MMF. In some embodiments, a composition comprises 100-200 mg, 125-225 mg, 150-250 mg, 175- 275 mg, 200-300 mg, 225-325 mg, 250-350 mg, 275-375 mg, 300-400 mg, 325-425 mg, 350- 450 mg, 375-475 mg, or 400-500 mg MMF; and / or

[0140] NAM in an amount of about 1 gram (g), 1.5 g, about 1.75 g, about 2 g, about 2.25 g, about 2.5 g, about 2.75 g, about 3 g, about 3.25 g, about 3.5 g, about 3.75 g, about 4 g, about

[0141] 4.25 g, about 4.5 g, about 4.75 g, about 5 g, about 5.25 g, about 5.5 g, about 5.75 g, about 6 g, about 6.25 g, or about 6.5 g NAM. In some embodiments, a composition comprises 1-2 g, 1.25-

[0142] 2.25 g, 1.5-2.5 g, 1.75-2.75 g, 2-3 g, 2.25-3.25 g, 2.5g-3.5 g, 2.75-3.75 g, 3-4 g, 3.25-4.25 g, 3.5-

[0143] 12523362.1 4.5 g, 3.75-4.75 g, 4-5 g, 4.25-5.25 g, 4.5-5.5 g, 4.75-5.75 g, 5-6 g, 5.25-6.25 g, or 5.5-6.5 g NAM.

[0144] In some embodiments, a composition provided herein that comprises or consists essentially of MMF and NAM comprises:

[0145] MMF in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to MMF at a concentration of 5-15 pM, 10-20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM; and

[0146] NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of 0.5-10 mM, 0.5-1 mM, 1-1.5 mM, 1.5-2 mM, 2-2.5 mM, 2.5-3 mM, 3-3.5 mM, 3.5-4 mM, 4-4.5 mM, 4.5-5 mM, 5-5.5 mM, 5.5-6 mM, 6-6.5 mM, 6.5-7 mM, 7-7.5 mM, 7.5-8 mM, 8-8.5 mM, 8.5-9 mM, 9-9.5 mM, 9.5-10 mM, 10-10.5 mM, 10.5-11 mM, 11-11.5 mM, 11.5 mM-12 mM, 12-12.5 mM, 12.5-13 mM, 13-13.5 mM, 13.5-14 mM, 14-14.5 mM, or 14.5-15 mM.

[0147] In some embodiments, a composition provided herein that comprises or consists essentially of MMF and NAM comprises:

[0148] MMF in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to MMF at a concentration of 20-80 pM, 40-60 pM, or about 50 pM; and

[0149] NAM in an amount sufficient that, when the composition is administered to a subject (e.g., a human subject), cells of the subject are exposed to NAM at a concentration of 0.5-10 mM, 2-8 mM, 3-7 mM, 4-6 mM, or about 5 mM.

[0150] In some embodiments, a composition comprises MMF at a concentration of 50 pM and NAM at a concentration of 5 mM.

[0151] II. Methods of Increasing Frataxin Levels, NAD(P)H Quinone Dehydrogenase 1 Levels, and / or Thioredoxin Reductase 1 Levels

[0152] In some aspects, the present disclosure provides methods of increasing frataxin levels in a cell. In some aspects, the present disclosure provides methods of increasing NAD(P)H

[0153] 12523362.1 quinone dehydrogenase 1 levels in a cell. In some aspects, the present disclosure provides methods of increasing thioredoxin reductase 1 levels in a cell. In some aspects, the present disclosure provides a method of increasing frataxin levels, NAD(P)H quinone dehydrogenase 1 levels, and thioredoxin reductase 1 levels in a cell.

[0154] In some embodiments, the methods comprising a step of contacting the cell with an effective amount of a composition described herein. In some embodiments, a composition comprises the components depicted in Table 1.

[0155] In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM (e.g., a composition described above in “Exemplary Combinations of Dimethyl

[0156] Fumarate and Nicotinamide’ ’) In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of MMF and NAM (e.g., a composition described above in “Exemplary Combinations of Monomethyl Fumarate and Nicotinamide”). In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of NAM and IDB. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of DMF (or another MMF prodrug, such as diroximel fumarate), NAM, and IDB. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of MMF, NAM, and IDB. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of DMF (or another MMF prodrug, such as diroximel fumarate), NAM, and NTS. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of MMF, NAM, and NTS. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of DMF (or another MMF prodrug, such as diroximel fumarate), NAM, IDB, and NTS. In some embodiments, the method comprises contacting the cell with an effective amount of a composition comprising MMF, NAM, IDB, and NTS.

[0157] In some embodiments, increasing frataxin levels means increasing expression of the frataxin gene. In some embodiments, increasing frataxin levels in a cell means increasing frataxin mRNA levels present in a cell. In some embodiments, the method increases frataxin mRNA levels in the cell by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least

[0158] 12523362.1 150%, or at least 200%. In some embodiments, the method increases frataxin mRNA levels in the cell by 10-15%, 10-20%, 10-25%, 10-30%, 10-35%, 10-40%, 10-45%, 10-50%, 10-60%, 10- 70%, 10-80%, 10-90%, 10-100%, 10-110%, 10-120%, 10-130%, 10-140%, 10-150%, 10-200%, 15-20%, 15-25%, 15-30%, 15-35%, 15-40%, 15-45%, 15-50%, 15-60%, 15-70%, 15-80%, 15- 90%, 15-100%, 15-110%, 15-120%, 15-130%, 15-140%, 15-150%, 15-200%, 20-25%, 20-30%, 20-35%, 20-40%, 20-45%, 20-50%, 20-60%, 20-70%, 20-80%, 20-90%, 20-100%, 20-110%, 20-120%, 20-130%, 20-140%, 20-150%, 20-200%, 25-30%, 25-35%, 25-40%, 25-45%, 25- 50%, 25-60%, 25-70%, 25-80%, 25-90%, 25-100%, 25-110%, 25-120%, 25-130%, 25-140%, 25-150%, 10-200%, 30-35%, 30-40%, 30-45%, 30-50%, 30-60%, 30-70%, 30-80%, 30-90%, 30-100%, 30-110%, 30-120%, 30-130%, 30-140%, 30-150%, 30-200%, 50-60%, 50-70%, 50- 80%, 50-90%, 50-100%, 50-110%, 50-120%, 50-130%, 50-140%, 50-150%, or 50-200%. In some embodiments, the method increases frataxin mRNA levels in the cell by about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 110%, about 120%, about 130%, about 140%, or about 150%.

[0159] In some embodiments, increasing frataxin levels means increasing levels of frataxin protein in a cell. In some embodiments, the method increases frataxin protein levels in the cell by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, or at least 200%. In some embodiments, the method increases frataxin protein levels in the cell by 10-15%, 10-20%, 10-25%, 10-30%, 10-35%, 10-40%, 10-45%, 10-50%, 10-60%, 10-70%, 10-80%, 10-90%, 10- 100%, 10-110%, 10-120%, 10-130%, 10-140%, 10-150%, 10-200%, 15-20%, 15-25%, 15-30%, 15-35%, 15-40%, 15-45%, 15-50%, 15-60%, 15-70%, 15-80%, 15-90%, 15-100%, 15-110%, 15-120%, 15-130%, 15-140%, 15-150%, 15-200%, 20-25%, 20-30%, 20-35%, 20-40%, 20- 45%, 20-50%, 20-60%, 20-70%, 20-80%, 20-90%, 20-100%, 20-110%, 20-120%, 20-130%, 20- 140%, 20-150%, 20-200%, 25-30%, 25-35%, 25-40%, 25-45%, 25-50%, 25-60%, 25-70%, 25- 80%, 25-90%, 25-100%, 25-110%, 25-120%, 25-130%, 25-140%, 25-150%, 10-200%, 30-35%, 30-40%, 30-45%, 30-50%, 30-60%, 30-70%, 30-80%, 30-90%, 30-100%, 30-110%, 30-120%, 30-130%, 30-140%, 30-150%, 30-200%, 50-60%, 50-70%, 50-80%, 50-90%, 50-100%, 50- 110%, 50-120%, 50-130%, 50-140%, 50-150%, or 50-200%. In some embodiments, the method increases frataxin protein levels in the cell by about 10%, about 15%, about 20%, about 25%,

[0160] 12523362.1 about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 110%, about 120%, about 130%, about 140%, or about 150%.

[0161] In some embodiments, the method increases frataxin mRNA and / or protein levels in the cell to the level of frataxin mRNA and / or protein observed in an asymptomatic FRDA carrier. In some embodiments, the method increases frataxin mRNA and / or protein levels in the cell to the level of a frataxin mRNA and / or protein observed in healthy controls.

[0162] In some embodiments, the method increases frataxin levels in the cell to a greater extent than the frataxin levels of a control cell contacted with DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and NAM, DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and IDB, NAM and IDB, NTS and IDB, NTS and NAM, or NTS and DMF (or another MMF prodrug, such as diroximel fumarate) or MMF.

[0163] In some embodiments, the method increases frataxin levels in the cell to an equal or greater extent than the sum of frataxin levels in control cells contacted with each of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, or IDB alone. In some embodiments, the sum of frataxin levels as described above comprises the sum of 1) the level of frataxin in a cell contacted with DMF (or another MMF prodrug, such as diroximel fumarate) or MMF only, 2) the level of frataxin in a cell contacted with NAM only, and 3) the level of frataxin in a cell contacted with IDB only. In some embodiments, the method increases frataxin levels in the cell to an equal or greater extent than the sum of frataxin levels in control cells contacted with each of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, IDB, or NTS alone. In some embodiments, the sum of frataxin levels as described above comprises the sum of 1) the level of frataxin in a cell contacted with DMF (or another MMF prodrug, such as diroximel fumarate) or MMF only, 2) the level of frataxin in a cell contacted with NAM only, 3) the level of frataxin in a cell contacted with IDB only, and 4) the level of frataxin in a cell contacted with NTS only.

[0164] In some embodiments, the method has an additive effect on frataxin levels relative to administration of each of the agents alone. In some embodiments, the method has a synergistic effect on frataxin levels relative to administration of each of the agents alone. In some embodiments, the method has a potentiating effect on frataxin levels relative to administration of each of the agents alone. In some embodiments, a potentiating effect is an effect where the combination of the agents administered together, at a less than maximal dose of each agent, has a greater effect on frataxin levels than the effect of the maximum amount of any agent alone.

[0165] 12523362.1 In some embodiments, increasing NAD(P)H quinone dehydrogenase 1 levels means increasing expression of the NAD(P)H quinone dehydrogenase 1 gene. In some embodiments, increasing NAD(P)H quinone dehydrogenase 1 levels in a cell means increasing NAD(P)H quinone dehydrogenase 1 mRNA levels present in a cell. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 mRNA levels in the cell by at least 50%, at least 75%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, at least 550%, or at least 600%. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 mRNA levels in the cell by 50-600%, 75-600%, 100-600%, 150-600%, 200-600%, 250-600%, 300-600%, 350- 600%, 400-600%, 450-600%, 500-600%, 550-600%, 50-550%, 50-500%, 50-450%, 50-400%, 50-350%, 50-300%, 50-250%, 50-200%, 50-150%, 100-500%, or 200-400%. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 mRNA levels in the cell by about 50%, about 75%, about 100%, about 150%, about 200%, about 250%, about 300%, about 350%, about 400%, about 450%, about 500%, about 550%, or about 600%.

[0166] In some embodiments, increasing NAD(P)H quinone dehydrogenase 1 levels means increasing levels of NAD(P)H quinone dehydrogenase 1 protein in a cell. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 protein levels in the cell by at least 50%, at least 75%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, at least 550%, or at least 600%. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 protein levels in the cell by 50-600%, 75-600%, 100-600%, 150-600%, 200-600%, 250-600%, 300-600%, 350- 600%, 400-600%, 450-600%, 500-600%, 550-600%, 50-550%, 50-500%, 50-450%, 50-400%, 50-350%, 50-300%, 50-250%, 50-200%, 50-150%, 100-500%, or 200-400%. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 protein levels in the cell by about 50%, about 75%, about 100%, about 150%, about 200%, about 250%, about 300%, about 350%, about 400%, about 450%, about 500%, about 550%, or about 600%.

[0167] In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 mRNA and / or protein levels in the cell to the level of NAD(P)H quinone dehydrogenase 1 mRNA and / or protein observed in an asymptomatic FRDA carrier. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 mRNA and / or protein levels in the cell to the level of a NAD(P)H quinone dehydrogenase 1 mRNA and / or protein observed in healthy controls.

[0168] 12523362.1 In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 levels in the cell to a greater extent than the NAD(P)H quinone dehydrogenase 1 levels of a control cell contacted with DMF (or another MMF prodrug, such as diroximel fumarate), MMF, IDB, NAM, or NTS.

[0169] In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 levels in the cell to an equal or greater extent than the sum of NAD(P)H quinone dehydrogenase 1 levels in control cells contacted with each of MMF or NAM alone. In some embodiments, the sum of NAD(P)H quinone dehydrogenase 1 levels as described above comprises the sum of 1) the level of NAD(P)H quinone dehydrogenase 1 in a cell contacted with DMF (or another MMF prodrug, such as diroximel fumarate) or MMF only, 2) the level of NAD(P)H quinone dehydrogenase 1 in a cell contacted with NAM only, and 3) the level of NAD(P)H quinone dehydrogenase 1 in a cell contacted with IDB only. In some embodiments, the method increases NAD(P)H quinone dehydrogenase 1 levels in the cell to an equal or greater extent than the sum of NAD(P)H quinone dehydrogenase 1 levels in control cells contacted with each of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, IDB, or NTS alone. In some embodiments, the sum of NAD(P)H quinone dehydrogenase 1 levels as described above comprises the sum of 1) the level of NAD(P)H quinone dehydrogenase 1 in a cell contacted with DMF (or another MMF prodrug, such as diroximel fumarate) or MMF only, 2) the level of NAD(P)H quinone dehydrogenase 1 in a cell contacted with NAM only, 3) the level of NAD(P)H quinone dehydrogenase 1 in a cell contacted with IDB only, and 4) the level of NAD(P)H quinone dehydrogenase 1 in a cell contacted with NTS only.

[0170] In some embodiments, the method has an additive effect on NAD(P)H quinone dehydrogenase 1 levels relative to administration of each of the agents alone. In some embodiments, the method has a synergistic effect on NAD(P)H quinone dehydrogenase 1 levels relative to administration of each of the agents alone. In some embodiments, the method has a potentiating effect on NAD(P)H quinone dehydrogenase 1 levels relative to administration of each of the agents alone. In some embodiments, a potentiating effect is an effect where the combination of the agents administered together, at a less than maximal dose of each agent, has a greater effect on NAD(P)H quinone dehydrogenase 1 levels than the effect of the maximum amount of any agent alone.

[0171] In some embodiments, increasing thioredoxin reductase 1 levels means increasing expression of the thioredoxin reductase 1 gene. In some embodiments, increasing thioredoxin

[0172] 12523362.1 reductase 1 levels in a cell means increasing thioredoxin reductase 1 mRNA levels present in a cell. In some embodiments, the method increases thioredoxin reductase 1 mRNA levels in the cell at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, at least 225%, at least 250%, at least 270%, or at least 300%. In some embodiments, the method increases thioredoxin reductase 1 mRNA levels in the cell by 50-300%, 75-300%, 100- 300%, 150-300%, 200-300%, 250-300%, or 75-250%, 75-200%, 75-150%, 75-100%, or 100- 200%. In some embodiments, the method increases thioredoxin reductase 1 mRNA levels in the cell by about 50%, about 75%, about 100%, about 150%, about 200%, about 250%, or about 300%.

[0173] In some embodiments, increasing thioredoxin reductase 1 levels means increasing levels of thioredoxin reductase 1 protein in a cell. In some embodiments, the method increases thioredoxin reductase 1 protein levels in the cell at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, at least 225%, at least 250%, at least 270%, or at least 300%. In some embodiments, the method increases thioredoxin reductase 1 protein levels in the cell by 50-300%, 75-300%, 100-300%, 150-300%, 200-300%, 250-300%, or 75-250%, 75-200%, 75-150%, 75-100%, or 100-200%. In some embodiments, the method increases thioredoxin reductase 1 protein levels in the cell by about 50%, about 75%, about 100%, about 150%, about 200%, about 250%, or about 300%.

[0174] In some embodiments, the method increases thioredoxin reductase 1 mRNA and / or protein levels in the cell to the level of thioredoxin reductase 1 mRNA and / or protein observed in an asymptomatic FRDA carrier. In some embodiments, the method increases thioredoxin reductase 1 mRNA and / or protein levels in the cell to the level of a thioredoxin reductase 1 mRNA and / or protein observed in healthy controls.

[0175] In some embodiments, the method increases thioredoxin reductase 1 levels in the cell to a greater extent than the thioredoxin reductase 1 levels of a control cell contacted with DMF (or another MMF prodrug, such as diroximel fumarate), MMF, IDB, NAM, or NTS.

[0176] In some embodiments, the method increases thioredoxin reductase 1 levels in the cell to an equal or greater extent than the sum of thioredoxin reductase 1 levels in control cells contacted with each of MMF or NAM alone. In some embodiments, the sum of thioredoxin reductase 1 levels as described above comprises the sum of 1) the level of thioredoxin reductase 1 in a cell contacted with DMF (or another MMF prodrug, such as diroximel fumarate) or MMF only, 2) the level of thioredoxin reductase 1 in a cell contacted with NAM only, and 3) the level

[0177] 12523362.1 of thioredoxin reductase 1 in a cell contacted with IDB only. In some embodiments, the method increases thioredoxin reductase 1 levels in the cell to an equal or greater extent than the sum of thioredoxin reductase 1 levels in control cells contacted with each of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, IDB, or NTS alone. In some embodiments, the sum of thioredoxin reductase 1 levels as described above comprises the sum of 1) the level of thioredoxin reductase 1 in a cell contacted with DMF (or another MMF prodrug, such as diroximel fumarate) or MMF only, 2) the level of thioredoxin reductase 1 in a cell contacted with NAM only, 3) the level of thioredoxin reductase 1 in a cell contacted with IDB only, and 4) the level of thioredoxin reductase 1 in a cell contacted with NTS only.

[0178] In some embodiments, the method has an additive effect on thioredoxin reductase 1 levels relative to administration of each of the agents alone. In some embodiments, the method has a synergistic effect on thioredoxin reductase 1 levels relative to administration of each of the agents alone. In some embodiments, the method has a potentiating effect on thioredoxin reductase 1 relative to administration of each of the agents alone. In some embodiments, a potentiating effect is an effect where the combination of the agents administered together, at a less than maximal dose of each agent, has a greater effect on thioredoxin reductase 1 levels than the effect of the maximum amount of any agent alone.

[0179] III. Methods of Improving Cell Viability

[0180] In some aspects, the present disclosure provides methods of improving cell viability, the methods comprising a step of contacting the cell with an effective amount of a composition described herein. In some embodiments, a composition comprises the components depicted in Table 1.

[0181] In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM (e.g., a composition described above in “Exemplary Combinations of Dimethyl Fumarate and Nicotinamide”). In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of MMF and NAM (e.g., a composition described above in “Exemplary Combinations of Monomethyl Fumarate and Nicotinamide”). In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of NAM and IDB. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of DMF (or another MMF prodrug, such

[0182] 12523362.1 as diroximel fumarate), NAM, and IDB. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of MMF, NAM, and IDB. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of DMF (or another MMF prodrug, such as diroximel fumarate), NAM, and NTS. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of MMF, NAM, and NTS. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of DMF (or another MMF prodrug, such as diroximel fumarate), NAM, IDB, and NTS. In some embodiments, the method comprises contacting the cell with an effective amount of a composition comprising MMF, NAM, IDB, and NTS.

[0183] In some embodiments cell viability is improved relative to a control cell. In some embodiments, the control cell is a healthy cell. In some embodiments, the control cell is an untreated diseased cell. In some embodiments, a diseased cell is a cell having a decreased level of frataxin relative to a healthy control or having the mutation responsible for FRDA. In some embodiments, improving cell viability comprises decreasing cell death relative to an untreated diseased cell.

[0184] In some embodiments, improving cell viability comprises decreasing cell death after exposure to reactive oxygen species (ROS) relative to an untreated diseased cell.

[0185] In some embodiments, cell viability is improved by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or at least 150% relative to an untreated diseased cell.

[0186] In some embodiments, the method has an additive effect on cell viability relative to administration of each of the agents alone. In some embodiments, the method has a synergistic effect on cell viability relative to administration of each of the agents alone. In some embodiments, the method has a potentiating effect on cell viability relative to administration of each of the agents alone. In some embodiments, a potentiating effect is an effect where the combination of the agents administered together, at a less than maximal dose of each agent, has a greater effect on cell viability than the effect of the maximum amount of any agent alone.

[0187] IV. Methods of Improving Mitochondrial Function

[0188] In some aspects, the present disclosure provides methods of improving mitochondrial function, the methods comprising a step of contacting the cell with an effective amount of a

[0189] 12523362.1 composition described herein. In some embodiments, a composition comprises the components depicted in Table 1.

[0190] In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of DMF (or another MMF prodrug, such as diroximel fumarate) and NAM (e.g., a composition described above in “Exemplary Combinations of Dimethyl Fumarate and Nicotinamide”). In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of MMF and NAM (e.g., a composition described above in “Exemplary Combinations of Monomethyl Fumarate and Nicotinamide”). In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of NAM and IDB. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of DMF (or another MMF prodrug, such as diroximel fumarate), NAM, and IDB. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of MMF, NAM, and IDB. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of DMF (or another MMF prodrug, such as diroximel fumarate), NAM, and NTS. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of MMF, NAM, and NTS. In some embodiments, the method comprises contacting the cell with a composition comprising an effective amount of DMF (or another MMF prodrug, such as diroximel fumarate), NAM, IDB, and NTS. In some embodiments, the method comprises contacting the cell with an effective amount of a composition comprising MMF, NAM, IDB, and NTS.

[0191] In some embodiments, improved mitochondrial function comprises improved production of adenosine triphosphate (ATP). In some embodiments, production of cellular adenosine triphosphate is increased by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or at least 150% relative to an untreated diseased cell. Methods of measuring cellular adenosine triphosphate are known to those having ordinary skill in the art.

[0192] In some embodiments, the method has an additive effect on mitochondrial function relative to administration of each of the agents alone. In some embodiments, the method has a synergistic effect on mitochondrial function relative to administration of each of the agents alone. In some embodiments, the method has a potentiating effect on mitochondrial function relative to administration of each of the agents alone. In some embodiments, a potentiating effect

[0193] 12523362.1 is an effect where the combination of the agents administered together, at a less than maximal dose of each agent, has a greater effect on mitochondrial function than the effect of the maximum amount of any agent alone.

[0194] V. Methods of Treatment

[0195] In some aspects, the present disclosure provides methods of treating a subject in need thereof, the methods comprising a step of administering an effective amount of a composition described herein to the subject. In some embodiments, a composition comprises the components depicted in Table 1.

[0196] In some embodiments, the subject is a mammal. In some embodiments, the subject is a primate. In some embodiments, the subject is a human.

[0197] In some embodiments, a subject in need of treatment is a subject having, suspected of having, or at risk for Friedreich’s Ataxia. In some embodiments, the subject presents with one or more symptoms of Friedreich’s Ataxia. In some embodiments, the subject has been diagnosed with Friedreich’s Ataxia. In some embodiments, the subject has one or more genetic mutation associated with Friedreich’s Ataxia. Genetic mutations associated with Friedreich’s Ataxia are known to those having ordinary skill in the art.

[0198] In some embodiments, treating a subject comprises reducing one or more symptoms of Friedreich’s Ataxia. In some embodiments treating a subject comprises reducing one or more of: trouble walking, tiredness, loss of reflexes, slow or slurred speech, hearing loss, vision loss, and chest pain. In some embodiments, treating a subject comprises reducing two or more of: trouble walking, tiredness, loss of reflexes, slow or slurred speech, hearing loss, vision loss, and chest pain.

[0199] In some embodiments, treating a subject comprises preventing or delaying progression of Friedreich’s Ataxia. In some embodiments, preventing progression of FRDA means preventing onset of FRDA symptoms. In some embodiments, preventing progression of FRDA means preventing worsening of FRDA symptoms. In some embodiments, delaying progression means delaying progression relative to an untreated individual. In some embodiments, progression is delayed by at least 6 months, at least 1 year, at least 18 months, at least two years, at least three years, at least four years, at least five years, at least six years, at least seven years, at least eight years, at least nine years, or at least ten years.

[0200] 12523362.1 In some embodiments, the method of treatment increases frataxin levels in a cell(s) of the subject (as described above), increases NAD(P)H quinone dehydrogenase 1 levels in a cell(s) of the subject (as described above), increases thioredoxin reductase 1 levels in a cell(s) of the subject (as described above), increases viability of a cell(s) of the subject (as described above), and / or increases mitochondrial function in a cell(s) of the subject (as described above).

[0201] For example, in some embodiments, the method increases frataxin mRNA and / or protein levels in cells of the subject (e.g., a human subject) by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, or at least 200%. In some embodiments, the method increases frataxin mRNA and / or protein levels cells of the subject (e.g., a human subject) by 10-15%, 10- 20%, 10-25%, 10-30%, 10-35%, 10-40%, 10-45%, 10-50%, 10-60%, 10-70%, 10-80%, 10-90%, 10-100%, 10-110%, 10-120%, 10-130%, 10-140%, 10-150%, 10-200%, 15-20%, 15-25%, 15- 30%, 15-35%, 15-40%, 15-45%, 15-50%, 15-60%, 15-70%, 15-80%, 15-90%, 15-100%, 15- 110%, 15-120%, 15-130%, 15-140%, 15-150%, 15-200%, 20-25%, 20-30%, 20-35%, 20-40%, 20-45%, 20-50%, 20-60%, 20-70%, 20-80%, 20-90%, 20-100%, 20-110%, 20-120%, 20-130%, 20-140%, 20-150%, 20-200%, 25-30%, 25-35%, 25-40%, 25-45%, 25-50%, 25-60%, 25-70%, 25-80%, 25-90%, 25-100%, 25-110%, 25-120%, 25-130%, 25-140%, 25-150%, 10-200%, 30- 35%, 30-40%, 30-45%, 30-50%, 30-60%, 30-70%, 30-80%, 30-90%, 30-100%, 30-110%, 30- 120%, 30-130%, 30-140%, 30-150%, 30-200%, 50-60%, 50-70%, 50-80%, 50-90%, 50-100%, 50-110%, 50-120%, 50-130%, 50-140%, 50-150%, or 50-200%. In some embodiments, the method increases frataxin mRNA and / or protein levels in cells of the subject (e.g., a human subject) by about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 110%, about 120%, about 130%, about 140%, or about 150%.

[0202] In some embodiments, after treatment with a method or composition as described herein, the subject has a lower International Cooperative Ataxia Rating Scale (ICARS) or modified Functional Assessment Rating Scale (mFARS) score than before treatment with the method. In some embodiments, after treatment with a method or composition as described herein, the subject maintains the same International Cooperative Ataxia Rating Scale (ICARS) or modified Functional Assessment Rating Scale (mFARS) score for a longer time than an untreated individual with FRDA.

[0203] 12523362.1 In some embodiments, treating a subject with the compositions and methods as disclosed herein results in fewer side effects than treatment with omaveloxolone. In some embodiments, treating a subject with the compositions and methods described herein results in improved Liver Function Tests, Gastro-Intestinal Toxicity and changes in Brain Natriuretic Peptide compared with omaveloxolone.

[0204] In some embodiments, a composition described herein (e.g., a pharmaceutical composition) is administered orally. In some embodiments, the composition is administered as one or more tablets, capsules, or sachets.

[0205] In some embodiment, a composition described herein (e.g., a pharmaceutical composition) is administered intrathecally or intravenously.

[0206] In some embodiments, at least two of the DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, IDB, and / or NTS are administered as a single composition. In some embodiments, the DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and NAM are administered as a single composition. In some embodiments, the DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and IDB administered as a single composition. In some embodiments, the DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and NTS are administered as a single composition. In some embodiments, the DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, and IDB are administered as a single composition. In some embodiments, NAM and IDB are administered as a single composition. In some embodiments, at least two of MMF and NAM are administered as a single composition. In some embodiments, the DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, IDB, and NTS are administered as a single composition. In some embodiments, each the DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, IDB, and / or NTS are administered as a separate pharmaceutical composition.

[0207] In some embodiments, the pharmaceutical composition is administered daily. In some embodiments, the pharmaceutical composition is administered twice daily. In some embodiments, the pharmaceutical composition is administered three times daily. In some embodiments, the pharmaceutical composition is administered every other day.

[0208] In some embodiments, the composition comprises a lower dose of at least one of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, IDB, and NTS than would be prescribed in a non-combination treatment. In some embodiments, the dose of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF is less than 480 mg / day DMF

[0209] 12523362.1 or MMF. In some embodiments, the dose of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF is less than 240 mg provided twice daily. In some embodiments, the dose of NAM is less than 5 g / day NAM. In some embodiments, the dose of IDB is less than 450 mg / day. In some embodiments, the dose of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF is 480 mg / day or greater and the dose of NAM is less than 5 g / day. In some embodiments, the dose of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF is 240 mg / day or higher and the dose of IDB is less than 450 mg / day. In some embodiments, the dose of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF is 480 mg / day, the dose of NAM 5 g / day, and the dose of IDB is less than 450 mg / day. In some embodiments, the dose of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF is 240 mg / day, the dose of NAM is less than 5 g / day, and the dose of IDB is 450 mg / day or higher. In some embodiments, the dose of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF is 240 mg / day, the dose of NAM is less than 5 g / day, and the dose of IDB less than 450 mg / day. In some embodiments, the dose of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF is less than 240 mg / day and the dose of NAM is 5 g / day or higher. In some embodiments, the dose of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF is less than 240 mg / day and the dose of NAM is 5 g / day or higher, and the dose of IDB is 450 mg / day or higher. In some embodiments, the dose of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF is less than 240 mg / day and the dose of NAM is 5 g / day or higher, and the dose of IDB less than 450 mg / day. In some embodiments, the dose of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF is less than 240 mg / day and the dose of NAM is less than 5 g / day, and the dose of IDB is 450 mg / day or higher. In some embodiments, the dose of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF is less than 240 mg / day and the dose of NAM is less than 5 g / day, and the dose of IDB less than 450 mg / day.

[0210] In some embodiments, the composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) and NAM (e.g., a combination described above in “Exemplary Combinations of Dimethyl Fumarate and Nicotinamide”).

[0211] In some embodiments, the composition comprises MMF and NAM (e.g., a combination described above in “Exemplary Combinations of Monomethyl Fumarate and Nicotinamide”).

[0212] In some embodiments, the method comprises administering a first composition comprising two or more of DMF (or another MMF prodrug, such as diroximel fumarate) or

[0213] 12523362.1 MMF, NAM, IDB, and NTS; and administering a second composition comprising one or more of DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, IDB, and NTS, wherein the first and the second composition differ. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and NAM and the second composition comprises IDB and NTS. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, and IDB and the second composition comprises NTS. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, and NTS and the second composition comprises IDB. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and IDB and the second composition comprises NAM and NTS. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, IDB and NTS and the second composition comprises NAM. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and NTS and the second composition comprises IDB and NAM. In some embodiments, the first composition comprises NAM and IDB and the second composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and NTS. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and NTS and the second composition comprises IDB and NAM.

[0214] In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and NAM and the second composition comprises NAM, IDB and NTS. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, and IDB and the second composition comprises NAM and NTS. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, and IDB and the second composition comprises IDB and NTS. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, and NTS and the second composition comprises NAM and IDB. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF and IDB and the second composition comprises IDB, NAM, and NTS. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, IDB and NTS and the second composition comprises NAM. In some embodiments, the

[0215] 12523362.1 first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, and NTS and the second composition comprises IDB and NAM. In some embodiments, the first composition comprises NAM and IDB and the second composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, and NTS. In some embodiments, the first composition comprises DMF (or another MMF prodrug, such as diroximel fumarate) or MMF, NAM, and NTS and the second composition comprises IDB and NAM.

[0216] In some embodiments, the first composition and the second composition are administered simultaneously. In some embodiments, the first composition and the second composition are administered at different times.

[0217] In some embodiments, the combination of the first composition and the second composition has an additive effect. In some embodiments, the combination of the first composition and the second composition has a synergistic effect. In some embodiments, the combination of the first composition and the second composition has a potentiating effect. In some embodiments, a potentiating effect is an effect where the combination of the first composition and the second composition at a less than maximal dose has a greater effect than the effect of the maximum amount of either the first or the second composition alone.

[0218] EXAMPLES

[0219] Example 1. Experimental Design

[0220] The compounds DMF and NAM were evaluated individually and in combinations as a means to affect two measurements: mitochondrial function (measured as ATP production) (Example 2); and frataxin expression (measured as mRNA) (Example 3). Fibroblasts isolated from 4 human donors diagnosed with Friedrich’s Ataxia were used for the experiments described in these examples (2-3).

[0221] Example 2. Mitochondrial Function

[0222] CellTiter Gio (CTG)® assays were performed to test combinations of DMF and NAM for their ability to increase mitochondrial function.

[0223] Briefly, human fibroblasts were grown and cultured to 70% confluence, followed by incubation with the appropriate agent(s). Hydrogen peroxide (H2O2) was added between 20- and 24- hours of the onset of treatment. Treatment groups include monotherapy and combination therapy of dimethyl fumarate and / or nicotinamide. Post treatment, cells were

[0224] 12523362.1 harvested and processed for downstream analysis. Cells treated with 90pM DMF and ImM NAM showed increased ATP production after hydrogen peroxide treatment compared to monotherapy with either drug. Results are shown in FIG. 1.

[0225] Example 3. Frataxin Expression

[0226] Combinations of DMF and NAM were tested for their ability to increase frataxin expression.

[0227] Briefly, human fibroblasts were grown and cultured to 70% confluence, followed by incubation with the appropriate agent(s). Treatment groups include monotherapy and combination therapy of dimethyl fumarate and / or nicotinamide. Post treatment, cells were harvested and processed for downstream analysis. Combinations of DMF and NAM at respective concentrations of 30 pM and 1 mM, 30 pM and 3 mM, 90 pM and 1 mM, and 30 pM and 3 mM were found to have an additive, synergistic, or potentiating effect (z.e., increase frataxin expression) compared with DMF or NAM given alone. Results are shown in FIGs. 2A- 2D and 3A-3D and in the tables below. Table 2 shows a summary of data describing the cell lines used. Tables 3-6 show patient-level values of frataxin mRNA (copies / pg mRNA) in each fibroblast cell line as determined by ddPCR.

[0228] Table 2: Cell line characteristics

[0229] Table 3: Frataxin mRNA levels in cell line 156

[0230] 12523362.1 Table 4: Frataxin mRNA levels in cell line 179

[0231] Table 5: Frataxin mRNA levels in cell line 4675 Table 6: Frataxin mRNA levels in cell line 4807

[0232] Example 4. Frataxin Expression and Nrf-2 Pathway Activation

[0233] Combinations of MMF and NAM were tested for their ability to (1) increase frataxin expression and (2) activate the Nrf-2 pathway. Nrf-2 is a transcription factor known to activate expression of various genes, including NAD(P)H quinone dehydrogenase 1 and thioredoxin reductase 1. As such, expression of NAD(P)H quinone dehydrogenase 1 and thioredoxin reductase 1 was used to assess Nrf-2 pathway activation. Comparisons were made to a negative control (DMOS) and a positive control (omaveloxolone, a known Nrf-2 activator).

[0234] Briefly, GM 16223 cells (a Friedreich Ataxia lymphoblastoid cell line) were seeded in 6- well plates at a density of 1 x 106cells / mL and immediately treated with: dimethyl sulfoxide (DMSO; vehicle control), monomethyl fumarate (MMF; 50 pM), omaveloxolone (Omav; 50 nM), nicotinamide (NAM; 5 mM), or a combination of NAM (5 mM) + MMF (50 pM) (denoted as N5M50). Each treatment condition was conducted in biological triplicates (n = 3). Cells were

[0235] 12523362.1 incubated under standard culture conditions (37°C, 5% CO2) for 72 hours prior to downstream analysis.

[0236] Following the 72-hour treatment, total RNA was extracted using TRIzol reagent (Invitrogen) according to the manufacturer’s instructions. The mRNA expression levels of the target genes, frataxin (FXN), NAD(P)H quinone dehydrogenase 1 (NQO1), and thioredoxin reductase 1 (TXNRD1), were quantified using quantitative real-time PCR (qRT-PCR). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal control to normalize for variations in RNA input and reverse transcription efficiency.

[0237] Relative gene expression levels were quantified using the 2A-AACt method. Briefly, the cycle threshold (Ct) values for the target genes and the housekeeping gene were determined for each sample. The ACt value was calculated by subtracting the Ct value of the housekeeping gene from the Ct value of the target gene. The AACt value was calculated by subtracting the ACt value of the control group (DMSO-treated cells) from the ACt values of the treated groups. The fold change in gene expression was then calculated as 2A-AACt. Statistical analysis was performed using Prism software to determine the statistical significance of the observed differences between treatment groups and the control group. Data are presented as mean ± standard error of the mean (SEM). Statistical significance was determined using an unpaired t- test or one-way ANOVA with post-hoc analysis as appropriate, with a p-value 0.05 considered statistically significant.

[0238] Results are shown in FIGs. 4A-4C. The combination of 50 pM MMF and 5 mM NAM had synergistic effects relative to the equivalent amounts of each compound alone.

[0239] Example 5. Cell Viability Assay

[0240] Combinations of MMF and NAM were tested for their ability to improve cell viability following exposure to H2O2.

[0241] Briefly, GM16223 cells were seeded into white 96-well plates at a density of 3 x 104cells / well and immediately exposed to 200 pM H2O2 and treated with: dimethyl sulfoxide (DMSO; vehicle control), monomethyl fumarate (MMF; 50 pM), omaveloxolone (Omav; 50 nM), nicotinamide (NAM; 5 mM), or a combination of NAM (5 mM) + MMF (50 pM) (denoted as N5M50). Each treatment condition was conducted in biological triplicates (n = 3).

[0242] 24 hours after treatment, cell viability was assessed using the CellTiter-Glo Luminescent Cell Viability Assay (Cat. No. G9242, Promega). Briefly, 50 pL of CellTiter-Glo reagent was added directly to each well, followed by a 2-minute incubation on an orbital shaker to induce

[0243] 12523362.1 cell lysis. Luminescence was then measured using a plate reader. Cell viability was calculated as a percentage relative to the DMSO-treated control (set as 100%) and a blank (set as 0%).

[0244] Results are shown in FIG. 5. Of the conditions tested, the combination of 50 pM MMF and 5 mM NAM improved cell viability to the greatest extent.

[0245] OTHER EMBODIMENTS

[0246] All of the features disclosed in this specification may be combined in any combination. Each feature disclosed in this specification may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Thus, unless expressly stated otherwise, each feature disclosed is only an example of a generic series of equivalent or similar features.

[0247] From the above description, one skilled in the art can easily ascertain the essential characteristics of the present disclosure, and without departing from the spirit and scope thereof, can make various changes and modifications of the disclosure to adapt it to various usages and conditions. Thus, other embodiments are also within the claims.

[0248] EQUIVALENTS

[0249] While several inventive embodiments have been described and illustrated herein, those of ordinary skill in the art will readily envision a variety of other means and / or structures for performing the function and / or obtaining the results and / or one or more of the advantages described herein, and each of such variations and / or modifications is deemed to be within the scope of the inventive embodiments described herein. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary and that the actual parameters, dimensions, materials, and / or configurations will depend upon the specific application or applications for which the inventive teachings is / are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific inventive embodiments described herein. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims and equivalents thereto, inventive embodiments may be practiced otherwise than as specifically described and claimed. Inventive embodiments of the present disclosure are directed to each individual feature, system, article, material, kit, and / or method described herein. In addition, any

[0250] 12523362.1 combination of two or more such features, systems, articles, materials, kits, and / or methods, if such features, systems, articles, materials, kits, and / or methods are not mutually inconsistent, is included within the inventive scope of the present disclosure.

[0251] All definitions, as defined and used herein, should be understood to control over dictionary definitions, definitions in documents incorporated by reference, and / or ordinary meanings of the defined terms.

[0252] The indefinite articles “a” and “an,” as used herein in the specification and in the claims, unless clearly indicated to the contrary, should be understood to mean “at least one.”

[0253] The phrase “and / or,” as used herein in the specification and in the claims, should be understood to mean “either or both” of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with “and / or” should be construed in the same fashion, i.e., “one or more” of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the “and / or” clause, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, a reference to “A and / or B”, when used in conjunction with open-ended language such as “comprising” can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.

[0254] As used herein in the specification and in the claims, “or” should be understood to have the same meaning as “and / or” as defined above. For example, when separating items in a list, “or” or “and / or” shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as “only one of’ or “exactly one of,” or, when used in the claims, “consisting of,” will refer to the inclusion of exactly one element of a number or list of elements. In general, the term “or” as used herein shall only be interpreted as indicating exclusive alternatives (i.e., “one or the other but not both”) when preceded by terms of exclusivity, such as “either,” “one of,” “only one of,” or “exactly one of.” “Consisting essentially of,” when used in the claims, shall have its ordinary meaning as used in the field of patent law.

[0255] As used herein in the specification and in the claims, the phrase “at least one,” in reference to a list of one or more elements, should be understood to mean at least one element

[0256] 12523362.1 selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase “at least one” refers, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, “at least one of A and B” (or, equivalently, “at least one of A or B,” or, equivalently “at least one of A and / or B”) can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.

[0257] It should also be understood that, unless clearly indicated to the contrary, in any methods claimed herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited. In the claims, as well as in the specification above, all transitional phrases such as “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “holding,” “composed of,” and the like are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases “consisting of’ and “consisting essentially of’ shall be closed or semi-closed transitional phrases, respectively. It should be appreciated that embodiments described in this document using an open-ended transitional phrase (e.g., “comprising”) are also contemplated, in alternative embodiments, as “consisting of’ and “consisting essentially of’ the feature described by the open-ended transitional phrase. For example, if the disclosure describes “a composition comprising A and B”, the disclosure also contemplates the alternative embodiments “a composition consisting of A and B” and “a composition consisting essentially of A and B”.

[0258] 12523362.1

Claims

CLAIMSWhat is claimed is:

1. A method for increasing frataxin levels in a cell, increasing viability of a cell, and / or increasing mitochondrial function of a cell, the method comprising contacting the cell with an effective amount of a composition comprising two or more of: a. dimethyl fumarate (DMF) or monomethyl fumarate (MMF); b. nicotinamide (NAM); c. idebenone (IDB); and d. nitisinone (NTS).

2. The method of claim 1, wherein the composition comprises three or more of DMF or MMF, NAM, IDB, and NTS, optionally wherein the composition comprises each of DMF or MMF, NAM, IDB, and NTS.

3. The method of claim 2, wherein the method increases frataxin levels in the cell to a greater extent than the frataxin levels of a control cell contacted with a composition comprising two or fewer of DMF or MMF, NAM, IDB, and NTS, such as a composition consisting essentially of DMF or MMF and NAM, DMF or MMF and IDB, NAM and IDB, DMF or MMF and NTS, NAM and NTS, or IDB and NTS.

4. The method of any one of claims 1-3, wherein the method increases frataxin mRNA and / or protein levels in the cell by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150%.

5. The method of any one of claims 1-4, wherein the method increases frataxin levels in the cell to an equal or greater extent than the sum of frataxin levels in control cells contacted with each of DMF or MMF, NAM, IDB, or NTS alone.

6. The method of any one of claims 1-5, wherein the method increases NAD(P)H quinone dehydrogenase 1 levels in the cell.12523362.

17. The method of claim 6, wherein the method increases NAD(P)H quinone dehydrogenase 1 mRNA and / or protein levels in the cell by at least 50%, at least 75%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, at least 550%, or at least 600%.

8. The method of claim 6 or claim 7, wherein the method increases NAD(P)H quinone dehydrogenase 1 levels in the cell to an equal or greater extent than the sum of NAD(P)H quinone dehydrogenase 1 levels in control cells contacted with each of MMF or NAM alone.

9. The method of any one of claims 1-8, wherein the method increases thioredoxin reductase 1 levels in the cell.

10. The method of claim 9, wherein the method increases thioredoxin reductase 1 mRNA and / or protein levels in the cell by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, at least 225%, at least 250%, at least 270%, or at least 300%.

11. The method of claim 9 or claim 10, wherein the method increases thioredoxin reductase 1 levels in the cell to an equal or greater extent than the sum of thioredoxin reductase 1 levels in control cells contacted with each of MMF or NAM alone.

12. The method of any one of claims 1-11, wherein the method increases viability of the cell.

13. The method of any one of claims 1-12, wherein the cell is contacted with DMF at a concentration of 5-15 pM, 10-20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM.12523362.

114. The method of claim 13, wherein the cell is contacted with DMF at a concentration of about 10 pM, about 30 pM, or about 90 pM.

15. The method of any one of claims 1-14, wherein the cell is contacted with MMF at a concentration of 5-15 pM, 10-20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM. 130-140 pM. 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM.

16. The method of claim 15, wherein the cell is contacted with MMF at a concentration of about 40 pM, about 50 pM, or about 60 pM.

17. The method of any one of claims 1-16, wherein the cell is contacted with NAM at a concentration of 0.5-10 mM, 0.5-1 mM, 1-1.5 mM, 1.5-2 mM, 2-2.5 mM, 2.5-3 mM, 3-3.5 mM, 3.5-4 mM, 4-4.5 mM, 4.5-5 mM, 5-5.5 mM, 5.5-6 mM, 6-6.5 mM, 6.5-7 mM, 7-7.5 mM, 7.5-8 mM, 8-8.5 mM, 8.5-9 mM, 9-9.5 mM, 9.5-10 mM, 10-10.5 mM, 10.5-11 mM, 11-11.5 mM, 11.5 mM-12 mM, 12-12.5 mM, 12.5-13 mM, 13-13.5 mM, 13.5-14 mM, 14-14.5 mM, or 14.5-15 mM.

18. The method of claim 17, wherein the cell is contacted with NAM at a concentration of about 1 mM, about 3 mM, about 5mM, or about 9 mM.

19. The method of any one of claims 1-18, wherein the composition comprises DMF or MMF in an amount that is less than 480 mg.

20. The method of any one of claims 1-19, wherein the composition comprises NAM in an amount that is less than 5 g.

21. The method of any one of claims 1-20, wherein the composition comprises IDB in an amount that is less than 450 mg.12523362.

122. The method of any one of claims 1-21, wherein the composition comprises NTS in an amount that is less than 70 mg NTS.

23. The method of any one of claims 1-22, wherein the method comprises contacting the cell with the composition twice daily.

24. A method for increasing frataxin levels in a cell, increasing viability of a cell, and / or increasing mitochondrial function of a cell, the method comprising contacting the cell with an effective amount of a composition comprising: a. dimethyl fumarate (DMF) or monomethyl fumarate (MMF); and b. nicotinamide (NAM); wherein the composition comprises less than 480 mg DMF or MMF and less than 5 g NAM.

25. The method of claim 24, wherein the composition further comprises idebenone (IDB) and / or nitisinone (NTS).

26. The method of claim 25, wherein the composition comprises less than 450 mg IDB.

27. The method of claim 25 or claim 26, wherein the composition comprises less than 70 mg NTS.

28. The method of any one of claims 24-27, wherein the method increases frataxin mRNA and / or protein levels in the cell by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150%.

29. The method of any one of claims 24-28, wherein the method increases frataxin levels in the cell to an equal or greater extent than the sum of frataxin levels in control cells contacted with each of DMF or MMF or NAM alone.12523362.

130. The method of any one of claims 24-29, wherein the method increases NAD(P)H quinone dehydrogenase 1 levels in the cell.

31. The method of claim 30, wherein the method increases NAD(P)H quinone dehydrogenase 1 mRNA and / or protein levels in the cell by at least 50%, at least 75%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, at least 550%, or at least 600%.

32. The method of claim 30 or claim 31, wherein the method increases NAD(P)H quinone dehydrogenase 1 levels in the cell to an equal or greater extent than the sum of NAD(P)H quinone dehydrogenase 1 levels in control cells contacted with each of MMF or NAM alone.

33. The method of any one of claims 24-32, wherein the method increases thioredoxin reductase 1 levels in the cell.

34. The method of claim 33, wherein the method increases thioredoxin reductase 1 mRNA and / or protein levels in the cell by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, at least 225%, at least 250%, at least 270%, or at least 300%.

35. The method of claim 33 or claim 34, wherein the method increases thioredoxin reductase 1 levels in the cell to an equal or greater extent than the sum of thioredoxin reductase 1 levels in control cells contacted with each of MMF or NAM alone.

36. The method of any one of claims 24-35, wherein the method increases viability of the cell.

37. A method for treating Friedreich’s Ataxia, the method comprising administering to a subject in need thereof an effective amount of a composition comprising two or more of: a. dimethyl fumarate (DMF) or monomethyl fumarate (MMF); b. nicotinamide (NAM); c. idebenone (IDB); and d. nitisinone (NTS).12523362.

138. The method of claim 37, wherein the composition comprises three or more of DMF or MMF, NAM, IDB, and NTS, optionally wherein the composition comprises each of DMF or MMF, NAM, IDB, and NTS.

39. The method of claim 38, wherein the method increases frataxin levels in cells of the subject to a greater extent than the frataxin levels of cells of a subject administered a composition comprising two or fewer of DMF or MMF, NAM, IDB, and NTS, such as a composition consisting essentially of DMF or MMF and NAM, DMF or MMF and IDB, or NAM and IDB.

40. The method of any one of claims 37-39, wherein the method increases frataxin mRNA and / or protein levels in cells of the subject by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, or at least 150%.

41. The method of any one of claims 37-40, wherein the method increases frataxin levels in cells of the subject to an equal or greater extent than the sum of frataxin levels in cells of a subjects administered DMF or MMF, NAM, or IDB alone.

42. The method of any one of claims 37-41, wherein the method increases NAD(P)H quinone dehydrogenase 1 levels in cells of the subject.

43. The method of claim 42, wherein the method increases NAD(P)H quinone dehydrogenase 1 mRNA and / or protein levels in cells of the subject by at least 50%, at least 75%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, at least 350%, at least 400%, at least 450%, at least 500%, at least 550%, or at least 600%.

44. The method of claim 42 or claim 43, wherein the method increases NAD(P)H quinone dehydrogenase 1 levels in cells of the subject to an equal or greater extent than the sum of NAD(P)H quinone dehydrogenase 1 levels in control cells contacted with each of MMF or NAM alone.12523362.

145. The method of any one of claims 37-44, wherein the method increases thioredoxin reductase 1 levels in cells of the subject.

46. The method of claim 45, wherein the method increases thioredoxin reductase 1 mRNA and / or protein levels in cells of the subject by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, at least 225%, at least 250%, at least 270%, or at least 300%.

47. The method of claim 45 or claim 46, wherein the method increases thioredoxin reductase 1 levels in cells of the subject to an equal or greater extent than the sum of thioredoxin reductase 1 levels in control cells contacted with each of MMF or NAM alone.

48. The method of any one of claims 37-47, wherein the method increases viability of cells in the subject.

49. The method of any one of claims 37-48, wherein the composition is administered to the subject orally.

50. The method of any one of claims 37-49, wherein the composition comprises DMF in an amount sufficient that cells of the subject are exposed to DMF at a concentration of 5-15 pM, 10- 20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95- 105 pM, 100-110 pM, 110-120 pM, 120-130 pM. 130-140 pM. 140-150 pM, 150-160 pM, 170- 180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250- 260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM.

51. The method of claim 50, wherein the composition comprises DMF in an amount sufficient that cells of the subject are exposed to DMF at a concentration of about lOpM, about 30 pM, or about 90 pM.

52. The method of any one of claims 37-51, wherein the composition comprises MMF in an amount sufficient that cells of the subject are exposed to MMF at a concentration of 5-15 pM, 10-20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-6012523362.1pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM.

53. The method of claim 52, wherein the composition comprises MMF in an amount sufficient that cells of the subject are exposed to MMF at a concentration of about 40pM, about 50 pM, or about 60 pM.

54. The method of any one of claims 37-53, wherein the composition comprises DMF or MMF in an amount that is less than 480 mg.

55. The method of any one of claims 37-54, wherein the composition comprises NAM in an amount sufficient that cells of the subject are exposed to NAM at a concentration of 0.5-10 mM, 0.5-1 mM, 1-1.5 mM, 1.5-2 mM, 2-2.5 mM, 2.5-3 mM, 3-3.5 mM, 3.5-4 mM, 4-4.5 mM, 4.5-5 mM, 5-5.5 mM, 5.5-6 mM, 6-6.5 mM, 6.5-7 mM, 7-7.5 mM, 7.5-8 mM, 8-8.5 mM, 8.5-9 mM, 9- 9.5 mM, 9.5-10 mM, 10-10.5 mM, 10.5-11 mM, 11-11.5 mM, 11.5 mM-12 mM, 12-12.5 mM, 12.5-13 mM, 13-13.5 mM, 13.5-14 mM, 14-14.5 mM, or 14.5-15 mM.

56. The method of any one of claims 37-55, wherein the composition comprises NAM in an amount sufficient that cells of the subject are exposed to NAM at a concentration of about 1 mM, about 3 mM, about 5mM, or about 9 mM.

57. The method of any one of claims 37-56, wherein the composition comprises NAM in an amount that is less than 5 g.

58. The method of any one of claims 37-57, wherein the composition comprises IDB in an amount less than 450 mg.

59. The method of any one of claims 37-58, wherein the composition comprises NTS in an amount less than 70mg.

60. The method of any one of claims 37-59, comprising:12523362.1administering a first composition comprising two or more of DMF or MMF, NAM, IDB, and NTS; and administering a second composition comprising one or more of DMF or MMF, NAM, IDB, and NTS; wherein the first composition and the second composition differ; and wherein the first composition and the second composition are administered simultaneously.

61. The method of any one of claims 37-60, wherein the method comprises administering a composition comprising DMF or MMF, NAM, IBD, and / or NTS twice daily.

62. The method of any one of claims 37-61, wherein after treatment with the method, the subject has a lower International Cooperative Ataxia Rating Scale (ICARS) or Functional Assessment Rating Scale (FARS) score than before treatment with the method.

63. The method of any one of claims 37-62, wherein the method causes fewer side effects in the subject than treatment with omaveloxolone.

64. The method of any one of claims 37-63, wherein the method comprises administering a composition comprising DMF or MMF, NAM, IBD, and / or NTS twice daily.

65. A method for reducing oxidative stress in a cell, the method comprising contacting the cell an effective amount of a composition comprising two or more of: a. dimethyl fumarate (DMF) or monomethyl fumarate (MMF); b. nicotinamide (NAM); c. idebenone (IDB); and d. nitisinone (NTS).

66. The method of claim 65, wherein the method reduces the level of reactive oxygen species (ROS) in the cell.

67. The method of claim 65 or claim 66, wherein the cell is contacted with DMF at a concentration of 5-15 pM, 10-20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM,12523362.140-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM.

68. The method of claim 67, wherein the cell is contacted with DMF at a concentration of about 10 pM, about 30 pM, or about 90 pM.

69. The method of any one of claims 65-68, wherein the cell is contacted with MMF at a concentration of 5-15 pM, 10-20 pM, 15-25 pM, 20-30 pM, 25-35 pM, 30-40 pM, 35-45 pM, 40-50 pM, 45-55 pM, 50-60 pM, 55-65 pM, 60-70 pM, 65-75 pM, 70-80 pM, 75-85 pM, 80-90 pM, 85-95 pM, 90-100 pM, 95-105 pM, 100-110 pM, 110-120 pM, 120-130 pM, 130-140 pM, 140-150 pM, 150-160 pM, 170-180 pM, 190-200 pM, 200-210 pM, 210-220 pM, 220-230 pM, 230-240 pM, 240-250 pM, 250-260 pM, 260-270 pM, 270-280 pM, 280-290 pM, or 290-300 pM.

70. The method of claim 69, wherein the cell is contacted with MMF at a concentration of about 40 pM, about 50 pM, or about 60 pM.

71. The method of any one of claims 65-70, wherein the cell is contacted with NAM at a concentration of 0.5-10 mM, 0.5-1 mM, 1-1.5 mM, 1.5-2 mM, 2-2.5 mM, 2.5-3 mM, 3-3.5 mM, 3.5-4 mM, 4-4.5 mM, 4.5-5 mM, 5-5.5 mM, 5.5-6 mM, 6-6.5 mM, 6.5-7 mM, 7-7.5 mM, 7.5-8 mM, 8-8.5 mM, 8.5-9 mM, 9-9.5 mM, 9.5-10 mM, 10-10.5 mM, 10.5-11 mM, 11-11.5 mM, 11.5 mM-12 mM, 12-12.5 mM, 12.5-13 mM, 13-13.5 mM, 13.5-14 mM, 14-14.5 mM, or 14.5-15 mM.

72. The method of claim 71, wherein the cell is contacted with NAM at a concentration of about 1 mM, about 3 mM, or about 9 mM.

73. The method of any one of claims 1-72, wherein the composition comprises 1 mM NAM and 30 pM DMF.12523362.

174. The method of any one of claims 1-73, wherein the composition comprises 1 mM NAM and 90 pM DMF.

75. The method of any one of claims 1-73, wherein the composition comprises 3 mM NAM and 30 pM DMF.

76. The method of any one of claims 1-73, wherein the composition comprises 3 mM NAM and 90 pM DMF.

77. The method of any one of claims 1-73, wherein the composition comprises 5 mM NAM and 50 pM MMF.12523362.1

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