Methods and compositions for treating or reducing PMP22 genetic expression in PMP22-related neuropathy

Abstract

Provided herein are compositions and methods for reducing expression of PMP22 and for treating PMP22-related neuropathy in subjects in need thereof, by administering compositions comprising omaveloxolone.

Description

Attorney Docket No.: RRBS-007 / 01WO 353016-2013METHODS AND COMPOSITIONS FOR TREATING OR REDUCING PMP22 GENETIC EXPRESSION IN PMP22-RELATED NEUROPATHY CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63 / 738,362. filed December 23, 2024, which is incorporated herein in its entirety for all purposes.TECHNICAL FIELD

[0002] The disclosure relates to compositions and methods for reducing expression of PMP22 and for treating PMP22-related neuropathy in subjects in need thereof, by administering compositions comprising omaveloxolone.BACKGROUND

[0003] Pathogenic genetic variants in the PMP22 gene cause a spectrum of hereditary neuropathies including Charcot Marie Tooth disease, types 1A (CMT1A) and IE (CMT1E) as well as Hereditary Neuropathy with Liability to Pressure Palsies (HNPP). Duplications of PMP22 are the cause of CMT1 A, while deletions of PMP22 cause HNPP, and point mutations in PMP22 lead to CMT1E. CMT1 is most commonly caused by triploidy and subsequent overexpression of the PMP22 gene.

[0004] The PMP22 gene encodes an integral membrane protein that is a major component of myelin in the peripheral nervous system. PMP22-related neuropathy syndromes are inherited in an autosomal dominant pattern and affect males and females equally. They represent a relatively common cause of hereditary neuropathy, with variants of PMP22 being responsible for >50% cases of CMT. According to Orphanet, the prevalence of CMT1A is estimated to be around 1 / 7,000 worldwide. New therapeutic options are needed.SUMMARY

[0005] In some aspects, provided herein is a method of treating or preventing a disease or condition characterized by increased expression of PMP22, in a subject in need thereof, comprising administering an effective amount of a composition comprising omaveloxolone, or a pharmaceutically acceptable salt thereof. In some embodiments, the subject comprises one or more mutations in at least one allele of the PMP22 gene. In some embodiments, the one or more mutations comprise a duplication of the PMP22 gene, a duplication upstream of the131222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013PMP22 gene, or a point mutation.

[0006] In some embodiments, subject is determined to have the one or more mutations by analysis of the subject’s PMP22 genome sequence. In some embodiments, analysis of the subject’s PMP22 genome sequence is performed using a method selected from the group consisting of: PMP22 gene dosage analysis, sequence analysis of the entire PMP22 coding region, sequence analysis of select exons, mutation scanning of select exons, mutation scanning of the entire PMP22 coding region, or PMP22 RNA analysis.

[0007] In some embodiments, the disease or condition is Charcot Marie Tooth disease.

[0008] In some embodiments, the effective amount of the composition is about 0.5 mg / kg to about 20 mg / kg per day. In some embodiments, the effective amount of the composition is about 1 mg / kg to about 5 mg / kg per day. In some embodiments, the effective amount of the composition is about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg per day, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or about 10 mg / kg per day.

[0009] In some embodiments, the composition is administered orally. In some embodiments, the composition is administered once or twice per day. In some embodiments, the composition is administered for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 6 months, at least about 9 months, at least about 12 months, at least about 15 months, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years.

[0010] In some embodiments, the method results in reduction or improvement of at least one symptom of the disease or condition. In some embodiments, the method results in prevention of at least one symptom of the disease or condition.

[0011] In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0012] In some embodiments, the composition additionally comprises a pharmaceutically acceptable carrier.

[0013] In some aspects, provided herein is a method of decreasing the level of PMP22 transcript and / or protein in a cell, wherein the method comprises contacting a cell with an effective amount of a composition comprising omaveloxolone, or a pharmaceutically acceptable salt thereof.231222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013

[0014] In some embodiments, the cell comprises one or more mutations in one allele of the PMP22 gene. In some embodiments, the one or more mutations comprise a duplication of the PMP22 gene, a duplication upstream of the PMP22 gene, or a point mutation.

[0015] In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by about 10% to about 90%, compared to a cell treated with vehicle / control.

[0016] In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased to a level comparable to that of a cell that does not comprise a mutation in either allele of the PMP22 gene.

[0017] In some embodiments, the cell is a glia cell or a neuron.

[0018] In some embodiments, the contact occurs in vivo or ex vivo.

[0019] In another aspect, the present disclosure provides a composition comprising omaveloxolone, or a pharmaceutically acceptable salt thereof, for use in decreasing the level of PMP22 transcript and / or protein in a cell.

[0020] In another aspect, the present disclosure provides a composition comprising omaveloxolone, or a pharmaceutically acceptable salt thereof, for use in the treatment of a disease or condition. In some embodiments, the disease or condition is Charcot Marie Tooth disease.FIGURE DESCRIPTIONS

[0021] FIG. 1 shows gene expression of PMP22 in human iPSC-derived Schwann cells treated with omaveloxolone.

[0022] FIGS. 2A-2B show expression of PMP22 in human fibroblasts with a confirmed PMP22 duplication, following treatment with omaveloxolone or vehicle. FIG. 2A shows PMP22 transcript levels and FIG. 2B shows PMP22 protein levels.

[0023] FIG. 3 shows expression of PMP22 protein in human iPSC-derived Schwann cells treated with omaveloxolone or vehicle.DETAILED DESCRIPTION

[0024] The present disclosure provides compositions and methods of treating a disease or condition characterized by increased expression of PMP22 (e.g., a PMP22-related neuropathy) in a subject in need thereof, comprising administering an effective amount of a composition331222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013comprising omaveloxolone, or a pharmaceutically acceptable salt thereof. The disclosure also provides compositions and methods for reducing transcript and / or protein levels of PMP22 in a cell, by contacting the cell with an effective amount of a composition comprising omaveloxolone, or a pharmaceutically acceptable salt thereof.Definitions

[0025] Unless otherwise defined, all terms of art, notations, and other scientific terms or terminology' used herein are intended to have the meanings commonly understood by those of skill in the art to which this application pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art. Many of the techniques and procedures described or referenced herein are well understood and commonly employed using conventional methodology by those skilled in the art. All publications, patent applications, patents, GenBank or other accession numbers and other references mentioned herein are incorporated by reference in their entirety for all purposes.

[0026] The term “a” or “an” refers to one or more of that entity, i.e., can refer to a plural referent. As such, the terms “a” or “an”, and “one or more” are used interchangeably herein. In addition, reference to “an element” by the indefinite article “a” or “an” does not exclude the possibility that more than one of the elements is present, unless the context clearly requires that there is one and only one of the elements.

[0027] 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.

[0028] It is understood that aspects and embodiments of the disclosure described herein include431222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013“comprising,” “consisting,” and “consisting essentially of’ aspects and embodiments. As used herein, “comprising” is synonymous with “including,” “containing,” or “characterized by.” and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. As used herein, “consisting of’ excludes any elements, steps, or ingredients not specified in the claimed composition or method. As used herein, “consisting essentially of’ does not exclude materials or steps that do not materially affect the basic and novel characteristics of the claimed composition or method. Any recitation herein of the term “comprising”, particularly in a description of components of a composition or in a description of steps of a method, is understood to encompass those compositions and methods consisting essentially of and consisting of the recited components or step.

[0029] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the disclosure. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure.

[0030] Certain ranges are presented herein with numerical values being preceded by the term “approximately” or “about.” The terms “approximately” and “about” are used interchangeably and mean a quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length that varies by as much as 20%, preferably up to 10%. more preferably up to 5%, and more preferably still up to 1%. of a reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length, inclusive of the endpoints. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth, unless otherwise apparent from context that it is impossible to extend the boundary beyond certain points (e.g., below 0% or above 100% in some cases).

[0031] “Treating” or “treatment” as used herein covers the treatment of the disease, injury, or condition of interest, e.g., a PMP22 -related neuropathy, in a biological material, e.g., mammal, preferably a human, having the disease or condition of interest, and includes: (i) preventing or inhibiting the disease, injury, or condition from occurring in a biological material, e.g., mammal, in particular, when such mammal is predisposed to the condition but has not yet been 531222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013diagnosed as having it; (ii) reducing the severity or duration of the disease, injury or condition, e.g., when it occurs, e.g.. in a mammal predisposed to the condition; (iii) inhibiting the disease, injury, or condition, i.e., arresting its development; (iv) relieving the disease, injury, or condition, i.e., causing regression of the disease or condition; or (v) relieving the symptoms resulting from the disease, injury', or condition.

[0032] In certain embodiments, as used herein, the term “prevention’’ includes inhibiting or impeding the onset or progression of a disease, or reducing the amount of damage caused by a disease. As used herein, the terms “disease,” “disorder,” and “condition” may be used interchangeably.

[0033] The terms “increase” and “increased” are all used herein generally to mean an increase by a statistically significant amount. However, for avoidance of doubt, “increase” and “increased” mean an increase by at least 10% as compared to a reference level, for example an increase by at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least 1-fold, at least 2-fold, at least 3-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 50-fold, at least 100-fold, at least 200-fold, at least 500-fold, or at least 1000-fold, or any increase between 10% and 1000-fold as compared to a reference level.

[0034] The terms “decrease” and “decreased” are all used herein generally to mean a decrease by a statistically significant amount. However, for avoidance of doubt, “decrease” and “decreased” mean a decrease by at least 10% as compared to a reference level, for example a decrease by at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least 1-fold, at least 2-fold, at least 3-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 50-fold, at least 100-fold, at least 200-fold, at least 500-fold, or at least 1000-fold, or any decrease between 10% and 1000-fold as compared to a reference level.

[0035] The term “expression” as used herein refers to the level of a transcript and / or protein (e.g, a PMP22 transcript and / or a PMP22 protein). “Expression” may refer to the level of the transcript and / or protein within a cell. “Expression” may also refer to the level of the transcript and / or protein within a subject.

[0036] As used herein, a “subject” or an “individual” includes animals, such as human (e.g., human subjects) and non-human animals. In some embodiments, a “subject” or “individual” is a patient under the care of a physician. Thus, the subject can be a human patient or an individual 631222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013who has, is at risk of having, or is suspected of having a disease of interest (e.g., cancer) and / or one or more symptoms of the disease. The subject can also be an individual who is diagnosed with a risk of the condition of interest at the time of diagnosis or later. The term “non-human animals” includes all vertebrates, e.g., mammals, e.g., rodents, e.g., mice, e.g. non-human primates, and non- mammals, e.g., sheep, dogs, cows, chickens, amphibians, reptiles, etc.

[0037] “Administration,” “administering” and the like, refer to administration to a subject by a medical professional or by self-administration by the subject, as well as to indirect administration, which may be the act of prescribing a composition of the invention. Typically, an effective amount is administered, which amount can be determined by one of skill in the art. Any method of administration may be used. Administration to a subject can be achieved by, for example, oral administration, in liquid or solid form, e.g. in capsule or tablet form; intravascular injection; intramyocardial delivery; or other suitable forms of administration.

[0038] “Effective amount” as used herein refers to an amount of an agent effective in achieving a particular effect, e.g., reducing transcript and / or protein levels of PMP22. In certain embodiments, the reduction is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, or at least 70%, as compared to the amount prior to or without treatment. In the context of therapeutic treatment of a subject, an effective amount may be, e.g., an amount effective or sufficient to reduce one or more disease symptoms in the subject, e.g., a subject with a PMP22-related neuropathy. The amount of a compound or composition of the invention which constitutes an “effective amount” will vary depending on the compound or composition, the disease, injury or condition and its severity, the manner of administration, and the age of the subject to be treated, but can be determined routinely by one of ordinary skill in the art having regard to his own knowledge and to this disclosure.

[0039] A “composition” can comprise an active agent and a carrier, inert or active, e.g., a pharmaceutically acceptable carrier, diluent or excipient. A composition may be a pharmaceutical composition. In particular embodiments, the compositions are sterile, substantially free of endotoxins or non-toxic to recipients at the dosage or concentration employed.

[0040] “Pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and / or dosage forms which are. within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity7, irritation, allergic response, or other problem or complication,731222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013commensurate with a reasonable benefit / risk ratio.

[0041] As used herein, the term “pharmaceutically acceptable salt’' refers to any salt of an acidic or a basic group that may be present in a compound of the present invention (e.g, omaveloxolone). which salt is compatible with pharmaceutical administration.

[0042] As is known to those of skill in the art, “salts” of compounds may be derived from inorganic or organic acids and bases. Examples of acids include, but are not limited to, hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, ethanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic and benzenesulfonic acid. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds described herein and their pharmaceutically acceptable acid addition salts.

[0043] Examples of bases include, but are not limited to, alkali metal (e.g., sodium and potassium) hydroxides, alkaline earth metal (e.g., magnesium and calcium) hydroxides, ammonia, and compounds of formula NW4+, wherein W is Ci-4 alkyd, and the like.

[0044] Examples of salts include, but are not limited to, acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate. undecanoate, and the like. Other examples of salts include anions of the compounds of the present invention compounded with a suitable cation such as Na+, K+, Ca2+, NH4+, and NW4+(where W can be a Ci-4 alkyl group), and the like.

[0045] For therapeutic use, salts of the compounds of the present invention (e.g., omaveloxolone) are contemplated as being pharmaceutically acceptable. However, salts of acids and bases that are non-pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound.

[0046] As used herein, “pharmaceutically acceptable excipient” refers to a substance that aids the administration of an active agent to and / or absorption by a subject and can be included in the compositions of the present invention without causing a significant adverse toxicological 831222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013effect on the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, such as a phosphate buffered saline solution, emulsions (e.g., such as an oil / water or water / oil emulsions), lactated Ringer’s, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer’s solution), alcohols, oils, gelatins, carbohydrates, fatty acid esters, and colors, and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and / or aromatic substances and the like that do not deleteriously react with the compounds of the invention. For examples of excipients, see Martin, Remington’s Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA (1975).

[0047] As used herein, “solid dosage form” means a pharmaceutical dose(s) in solid form, e.g. , tablets, capsules, granules, powders, sachets, reconstitutable powders, dry powder inhalers and chewables.

[0048] As used herein, the term “mutation” refers to a point mutation, a gene fusion, a substitution, a gain-of-function mutation, a stop-gain mutation, an insertion mutation, a deletion mutation, a duplication mutation and / or a translocation. The mutation may be in one or more genes. The mutation may be naturally occurring. Alternatively, the mutation may be induced or engineered.

[0049] “Pharmaceutical compositions” include compositions of one or more inhibitors disclosed herein and one or more pharmaceutically acceptable carrier, excipient, or diluent.

[0050] “Dose” means a specified quantity of a pharmaceutical agent provided in a single administration, or in a specified time period. In certain embodiments, a dose can be administered in two or more boluses, tablets, or injections. In certain embodiments, a dose can be administered in two or more injections to minimize injection site reaction in an individual. Doses can be stated as the amount of pharmaceutical agent per hour, day, week or month. “Dosage amount” may be used interchangeably with “dose”.

[0051] “Dosage regimen” means a schedule according to which doses of a pharmaceutical agent are provided, e.g., daily, weekly, or other schedule capable of being developed by a person of ordinary skill in the art.

[0052] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of cell biology, molecular biology techniques), microbiology, biochemistry and immunology, which are within the scope of those of skill in the art. Such 931222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013techniques are explained fully in the literature, such as, "Molecular Cloning: A Laboratory Manual", second edition (Sambrook etal.. 1989); "Oligonucleotide Synthesis" (M. J. Gait, ed., 1984); "Animal Cell Culture" (R. I. Freshney, ed., 1987); "Methods in Enzymology" (Academic Press, Inc.); "Handbook of Experimental Immunology" (D. M. Weir & C. C. Blackwell, eds.); "Gene Transfer Vectors for Mammalian Cells" (J. M. Miller & M. P. Calos, eds., 1987); "Current Protocols in Molecular Biology’" (F. M. Ausubel etal., eds., 1987); "PCR: The Polymerase Chain Reaction", (Mullis et al., eds., 1994); and "Cunent Protocols in Immunology" (J. E. Coligan et al., eds., 1991), each of which is expressly’ incorporated by reference herein.

[0053] Several aspects of the invention are described below with reference to example applications for illustration. It should be understood that numerous specific details, relationships, and methods are set forth to provide a full understanding of the invention. One having ordinary skill in the relevant art, however, will readily recognize that the invention can be practiced without one or more of the specific details or with other methods. The present invention is not limited by the illustrated ordering of acts or events, as some acts may occur in different orders and / or concurrently with other acts or events. Furthermore, not all illustrated acts or events are required to implement a methodology in accordance with the present invention.PMP22 Genetic Variants and PMP22-Related Neuropathy

[0054] The disclosure provides compositions and methods of treating or preventing a disease or condition characterized by increased expression of PMP22 in a subject in need thereof, comprising administering an effective amount of a composition comprising omaveloxolone, or a pharmaceutically acceptable salt thereof. In some embodiments, increased expression of PMP22 comprises increased transcript levels of PMP22. In some embodiments, increased expression of PMP22 comprises increased protein levels of PMP22. The disclosure also provides compositions and methods for reducing transcript and / or protein levels of PMP22 in a cell, by contacting the cell with an effective amount of a composition comprising omaveloxolone.

[0055] The PMP22 gene encodes an integral membrane protein that is a major component of myelin in the peripheral nervous system. Myelin is a protective substance that covers nerves and promotes the efficient transmission of nen e impulses. Myelin is produced primarily by specialized cells called Schwann cells. Within Schwann cells, PMP22 contributes to development and maintenance of myelin as well as cell growth and differentiation.1031222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013

[0056] The human PMP22 gene has 12 transcripts. 10 of which encode a protein isoform of PMP22. In some embodiments, the increased transcript level of PMP22 comprises transcript NRJ04018.2, NRJ04017.2, NM_001281456.2, NM_000304.4, NMJ53321.3, XM_047436306.1, NM_001281455.2, NMJ53322.3, NM_001330143.2, XM_024450806.2, XM_054316516.1, and / or XM_054316517.1.

[0057] In some embodiments, the increased protein level of PMP22 comprises NP 001268385.1, NP_000295.1, NP_696996.1, XP_047292262.1, NP_001268384.1, NP 696997.1, NP_001317072.1, XP_024306574.1, XP 054172491.1, and / or XP_054172492.1.

[0058] In various embodiments, the subject comprises one or more mutations in at least one allele of the PMP22 gene (NCBI Gene ID: 5376). In some embodiments, the one or more mutations comprise a duplication of the PMP22 gene. In some embodiments, the one or more mutations comprise a triplication of the PMP22 gene. In some embodiments, the duplication or triplication of the PMP22 gene comprises a 1.4 Mb duplication or triplication on chromosome 17. In some embodiments, the duplication or triplication of the PMP22 gene comprises a 1.5 Mb duplication or triplication on chromosome 17. In some embodiments, the duplication or triplication on chromosome 17 is at the 17pl 1.2-12 locus. In some embodiments, the one or more mutations comprise one or more point mutations in the PMP22 gene. In some embodiments, the one or more point mutations in the PMP22 gene correspond to one or more amino acid substitutions in the PMP22 protein (Uniprot Accession No: Q6FH25). Non-limiting examples of point mutations include those described in Kovach et al. Am J. Human Genetics.1999. 64(6): 1580-1593; Saporta et al. Ann Neurol. 2011; 69(l):22-33.; and / or Liao et al. Sci Rep. 2017; 7(1): 15363, which are hereby incorporated by reference in their entirety. In some embodiments, the one or more mutations comprise a duplication of a region upstream of PMP22. Non-limiting examples of duplications include those described in Weterman et al. Eur J Hum Genet. 2010;18(4):421-428 and Zhang et al. Am J Hum Genet. 2010;86(6):892-903).

[0059] In some embodiments, the subject is determined to have the one or more mutations by analysis of the subject’s PMP22 genome sequence. In some embodiments, the analysis of the subject's PMP22 genome sequence is performed using a method selected from the group consisting of: PMP22 gene dosage analysis, sequence analysis of the entire PMP22 coding region, sequence analysis of select exons, mutation scanning of select exons, mutation scanning of the entire PMP22 coding region, or PMP22 RNA analysis. Details of suitable methods to detect PMP22 mutations in subjects are provided in the Genetic Testing Registry of the 1131222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013National Library of Medicine.

[0060] In some embodiments, the disease or condition associated with increased expression of PMP22 is a PMP2-related neuropathy syndrome. In some embodiments, the disease or condition associated with increased expression of PMP22 is Charcot Marie Tooth disease (CMT). In some embodiments, the disease or condition associated with increased expression of PMP22 is CMT t pe 1A (CMT1A). In some embodiments, the disease or condition associated with increased expression of PMP22 is CMT type IE (CMT1E). In some embodiments, the disease or condition associated with increased expression of PMP22 is Hereditary Neuropathy with Liability to Pressure Palsies (HNPP).

[0061] PMP22-related neuropathy syndromes are inherited in an autosomal dominant pattern and affect males and females equally. They represent a relatively common cause of hereditary neuropathy, with variants of PMP22 being responsible for >50% cases of CMT. According to Orphanet, the prevalence of CMT1A is estimated to be around 1 / 7,000 worldwide.Methods of Treatment

[0062] In one aspect, the present disclosure provides methods of treating or preventing a disease or condition characterized by increased expression of PMP22 in a subject in need thereof. In some embodiments, the method of treating or preventing a disease or condition characterized by increased expression of PMP22 in a subject in need thereof comprises administering an effective amount of a composition that reduces expression of PMP22 at the transcript and / or protein level. In some embodiments, the compound is omavel oxoIone, or a pharmaceutically acceptable salt thereof.

[0063] In some embodiments, the subject afflicted with a disease or condition associated with increased expression of PMP22 comprises one or more mutations in at least one allele of the PMP22 gene described herein. In some embodiments, the subject in need thereof is administered an effective amount of a composition comprising omaveloxolone, or a pharmaceutically acceptable salt thereof.

[0064] Omaveloxolone, also known as N-((4aS,6aR,6bS,8aR,12aS,14aR,14bS)-ll-cyano-2,2,6a,6b,9,9,12a-heptamethyl-10,14-dioxo-l,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,12a,I4,14a,I4b-octadecahydropicen-4a-yl)-2,2-difluoropropanamide or RTA-408, has the chemical structure of formula (I), as depicted below:1231222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013

[0065] The dose and dosage regimen may depend upon a variety of factors readily determined by a physician, such as the nature of the disease or condition, the characteristics of the subject, and the subject's history.

[0066] In some embodiments, the effective amount of the composition is about 0.5 mg / kg to about 20 mg / kg. In some embodiments, the effective amount of the composition is about 1 mg / kg to about 5 mg / kg. In some embodiments, the effective amount of the composition is about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or about 10 mg / kg.

[0067] In some embodiments, the effective amount of the composition is about 1 mg to about 1000 mg per day. In some embodiments, the effective amount of the composition is about 2 mg to about 500 mg per day. In some embodiments, the effective amount of the composition is 20 mg to about 200 mg per day. In some embodiments, the effective amount is about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg. about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg. about 180 mg, about 190 mg, or about 200 mg per day. In some embodiments, the effective amount of omaveloxolone is about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, or about 150 mg per day. In some embodiments, the effective amount of the composition is greater than or equal to about 1 mg, greater than or equal to about 20 mg, greater than or equal to about 40 mg, greater than or equal to about 60 mg, greater than or equal to about 80 mg, or greater than or equal to about 100 mg. In certain embodiments, the effective amount is Img to 500 mg, 5mg to 300 mg, 10 mg to 250 mg, 20 mg to 200 mg, 50 mg to 150 mg, or 60 mg to 120 mg.

[0068] In some embodiments, the composition is administered orally. In some embodiments, the composition is administered parenterally, e.g., intravenously, rectally, or by injection. In some embodiments, the composition is administered locally, e.g., topically or intramuscularly.1331222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013In some embodiments, the composition is administered to target tissues. The skilled artisan can determine an appropriate site and route of administration based on factors including, but not limited to, the disease or condition being treated.

[0069] In some embodiments, the composition is administered once or twice per day. The skilled artisan can determine an appropriate dosage regimen based on factors including, but not limited to, the nature of the disease or condition, the characteristics of the subject, and the subject's history.

[0070] In some embodiments, the composition is administered for at least about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 2 months, at least about 3 months, at least about 4 months, at least about 6 months, at least about 9 months, at least about 12 months, at least about 15 months, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years. The skilled artisan can determine an appropriate duration of treatment based on factors including, but not limited to, the nature of the disease or condition, the characteristics of the subject, and the subject's history.

[0071] The compositions described herein may be administered chronically (“chronic administration’7). Chronic administration refers to administration of a compound or pharmaceutical composition thereof over an extended period of time, e.g., for example, over 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc., or may be continued indefinitely, for example, for the rest of the subject’s life. In certain embodiments, the chronic administration is intended to provide a constant level of the compound in the blood, e.g., within the therapeutic window over the extended period of time.

[0072] The pharmaceutical compositions described herein may be presented in unit dosage forms to facilitate accurate dosing. The term “unit dosage forms” refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, premeasured ampules or syringes of the liquid compositions or pills, tablets, capsules or the like in the case of solid compositions.

[0073] In certain embodiments, the pharmaceutical compositions provided herein are administered to the patient as a solid dosage form. In certain embodiments, the solid dosage form is a capsule.

[0074] In some aspects, the present disclosure provides a composition for use in treating or 1431222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013preventing a disease or disorder disclosed herein. In some aspects, the present disclosure provides use of a composition of the present disclosure in the manufacture of a medicament for treating or preventing a disease or disorder disclosed herein.

[0075] Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and / or perform such modification with ordinary experimentation. General considerations in the formulation and / or manufacture of pharmaceutical compositions can be found, for example, in Remington: The Science and Practice of Pharmacy 21sted., Lippincott Williams & Wilkins, 2005.

[0076] Examples of carriers, diluents and excipients include, but are not limited to. water, saline, Ringer's solutions, dextrose solution, and 5% human serum albumin. Solutions or suspensions used for the formulations can include a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial compounds such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating compounds such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates; detergents such as Tween 20 to prevent aggregation; and compounds for the adjustment of tonicity such as sodium chloride or dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. In particular embodiments, the pharmaceutical compositions are sterile.

[0077] Pharmaceutical compositions may further include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, or phosphate buffered saline (PBS). In some embodiments, the composition is sterile and may be fluid such that it can be drawn into a syringe or delivered to a subject from a syringe. The carrier can be, e g., a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the 1531222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013required particle size in the case of dispersion and by the use of surfactants. In certain embodiments, the pharmaceutical composition is stable under the conditions of manufacture and storage and is preserved against the contaminating action of microorganisms such as bacteria and fungi. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the internal compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

[0078] In some embodiments, the pharmaceutical composition further comprises an additional therapeutic agent disclosed herein.

[0079] In some embodiments, the method results in decreased PMP22 expression (e.g., decreased PMP22 transcript and / or protein levels) in the subject compared to the expression of PMP22 in the subject prior to administration. In some embodiments, expression of PMP22 is decreased 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 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% compared to the expression of PMP22 in the subject prior to administration.

[0080] In some embodiments, PMP22 expression in the subject is decreased to a level comparable to that of a subject does not comprise a mutation in either allele of the PMP22 gene.

[0081] In some embodiments, the method results in plasma levels of omaveloxolone of at least 10 nM, at least 20 nM, at least 30 nM, at least 40 nM, at least 50 nM, at least 60 nM, at least 70 nM, at least 80 nM, at least 90 nM, at least 100 nM, at least 150 nM, at least 200 nM, at least 250 nM, at least 300 nM, at least 350 nM, at least 400 nM, at least 450 nM, or at least 500 nM. In some embodiments, the method results in plasma levels of omaveloxolone of at least 100 nM, at least 150 nM, at least 200 nM, at least 250 nM, at least 300 nM, at least 350 nM, at least 400 nM, at least 450 nM, or at least 500 nM. In some embodiments, the method results in plasma levels of omaveloxolone of at least 100 nM, In some embodiments, the method results in plasma levels of omaveloxolone of at least 200 nM, In some embodiments, the method results in plasma levels of omaveloxolone of at least 300 nM. In some embodiments, the1631222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013method results in plasma levels of omaveloxolone of at least 400 nM, In some embodiments, the method results in plasma levels of omaveloxolone of at least 500 nM.

[0082] In some embodiments, the method results in reduction or improvement of at least one symptom of the disease or condition. In some embodiments, the method results in prevention of at least one symptom of the disease or condition. Non-limiting examples of symptoms of the disease or condition include absent deep tendon reflexes, postural tremor, slowed nerve conduction velocity, reduced amplitude of sensory and / or motor responses, weakness of muscles in the foot and / or lower leg, paralysis of muscles in the foot and / or lower leg, abnormal gait (e.g., high-stepped gait), tripping and / or falling, impaired balance, abnormal foot formation (e.g., high arches and / or curled toes), decreased muscle bulk in legs, decreased sensation in legs and / or feet, weakness and / or atrophy in hands, impaired fine motor skills, decreased sense of vibration and position (proprioception), curvature of the spine (scoliosis), hip displacement, contractures (chronic shortening of muscles or tendons around joints, muscle cramping, nen e pain, and / or decreased mobility,.

[0083] In some embodiments, the subject is a mammal. In some embodiments, the subject is a human. In some embodiments, the human is an adult human.

[0084] In some embodiments, the composition additionally comprises a pharmaceutically acceptable carrier. Non-limiting examples of pharmaceutically acceptable carriers include a pharmaceutically acceptable excipient, binder, and / or diluent.

[0085] In one aspect, the present disclosure provides methods of decreasing the level of PMP22 transcript and / or protein in a cell, wherein the method comprises contacting a cell with an effective amount of a composition comprising omaveloxolone, or a pharmaceutically acceptable salt thereof. In some embodiments, the method comprises decreasing the level of PMP22 transcript in the cell. In some embodiments, the method comprises decreasing the level of PMP22 protein in the cell.

[0086] In some embodiments, the cell comprises one or more mutations in one allele of the PMP22 gene described herein. In some embodiments, the one or more mutations comprise a duplication of the PMP22 gene, a duplication upstream of the PMP22 gene, or a point mutation.

[0087] In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by about 10% to about 90%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 1731222371Attorney Docket No.: RRBS-007 / 01WO 353016-201310%, 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 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% compared to a cell treated with vehicle / control.

[0088] In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 10%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 15%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 20%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 25%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 30%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcnpt and / or protein in the cell is decreased by at least 35%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 40%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 45%. compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 50%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 55%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 60%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 65%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 70%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 75%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 80%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 85%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 90%, compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by at least 95%, compared to a cell 1831222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013treated with vehicle / control.

[0089] In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 10% to 95%, 15% to 95%, 20% to 95%, 25% to 95%, 30% to 95%, 35% to 95%, 40% to 95%. 45% to 95%, 50% to 95%, 55%, to 95%. 60% to 95%. 65% to 95%. 70% to 95%, 75% to 95%, 80% to 95%, 85% to 95%, or 90% to 95%, compared to a cell treated with vehicle / control.

[0090] In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 10% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 15% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 20% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 25% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 30% to 95% compared a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 35% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 40% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 45% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 50% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 55% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 60% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 65% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 70% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 75% to 95% compared a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 80% to 95% compared to a cell treated with vehicle / control. In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 85% to 95% compared to a cell treated with vehicle / control. In some 1931222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013embodiments, the level of PMP22 transcript and / or protein in the cell is decreased by 90% to 95% compared to a cell treated with vehicle / control.

[0091] In some embodiments, the level of PMP22 transcript and / or protein in the cell is decreased to a level comparable to that of a cell that does not comprise a mutation in either allele of the PMP22 gene.

[0092] In some embodiments, the cell is a glia cell. In some embodiments, the cell is a neuron. In some embodiments, the cell is a Schwann cell. In some embodiments, the contact occurs in vivo or ex vivo.

[0093] In another aspect, the present disclosure provides a composition comprising omaveloxolone, or a pharmaceutically acceptable salt thereof, for use in decreasing the level of PMP22 transcript and / or protein in a cell.

[0094] In another aspect, the present disclosure provides a composition comprising omaveloxolone, or a pharmaceutically acceptable salt thereof, for use in the treatment of a disease or condition. In some embodiments, the disease or condition is Charcot Marie Tooth disease.EXAMPLESExample 1. High-Throughput Screening of Small Molecule Modulators of PMP22

[0095] Small molecules were screened using human neurons derived from the induced pluripotent stem cells (iPSCs) of a presumably healthy donor. iPSCs were directly differentiated into i-Ngn2 neurons (which are primarily glutamatergic) by overexpression of the transcription factor NGN2 as described in Wang et al. Stem Cell Reports. 2017 Oct 10; 9(4), 1221-1233. Briefly, iPHN healthy control iPSCs were directly differentiated into glutamatergic neurons by overexpression of NGN2 via doxycycline treatment on Days 1-3 (neuron induction). On day 4, the cells were replated into 384-well plates and cultured in neuron maturation media until day 17.

[0096] A library of 389 FDA-approved and 45 investigational small molecules was screened in the iPSC-derived motor neurons. Compounds were added at a 10-point half-logarithmic dilution series (lOpM - 0.0005pM) in quadruplicate on day 17 of neuronal differentiation, and treatment was carried out for 24 hours.Gene expression was measured by 3’RNA-seq in cells from each treatment condition. Following data analysis, candidate compound Omaveloxolone was identified for further 2031222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013validation. This molecule decreased the expression levels of PMP22 relative to vehicle-treated cells in a dose-dependent manner. This was determined using a non-linear least squares fit to a 4-parameter Hill Equation for the binding of a ligand to a macromolecule (FIG. 1). Data are show n as a ten-point concentration-response curve (CRC) demonstrating Log2 fold changes in expression in PMP22 in Omaveloxolone-treated control motor neurons (treatment doses 0.0005pM - lOpM).Example 2. Validation with qPCR and ELISA

[0097] To confirm the decrease in PMP22 expression following Omaveloxolone treatment at both the RNA and protein levels, multiple concentrations of Omaveloxolone were tested on human fibroblasts with a confirmed PMP22 duplication (hereafter referred to as “disease model fibroblasts”) obtained from the Coriell Institute for Medical Research.

[0098] Disease model fibroblasts were treated once with Omaveloxolone at 2 concentrations: 125 nM and 250 nM. On day 2, cells were lysed, and RNA was extracted. Gene expression of PMP22 was measured by qPCR and normalized to that of a housekeeping gene (ACTB). Omaveloxolone treatment significantly decreased PMP22 expression in fibroblasts harboring a PMP22 duplication at 250 nM when measured by qPCR (****p<0.0001) (FIG. 2A). Data are shown as fold change in the omaveloxolone treated cells relative to cells treated with vehicle control (DMSO); *** indicates p<0.001 by one-way ANOVA with Dunnett’s multiple comparisons test (n=8).

[0099] PMP22 protein expression levels were tested after daily treatment of disease model fibroblasts with vehicle (0.01% DMSO) or omaveloxolone at two concentrations: 125 nM and 250 nM, for three days. On day 4, the cells were lysed, and protein was extracted. PMP22 protein expression levels were measured using a by ELISA (LS Bio. LS-F34822) according to manufacturer’s instructions. After overnight incubation, a biotinylated detection antibody w?as added to each well, followed by a horseradish peroxidase streptavi din-conjugated secondary antibody. TMB substrate w?as subsequently added and allowed to develop at 37°C. A stop solution was used to terminate color development, and the optical density was measured using a Flexstation 3 plate reader (Molecular Devices, Flex3). Omaveloxolone treatment at both 125 nM and 250 nM significantly decreased PMP22 protein expression in disease model fibroblasts harboring a PMP22 duplication, as measured by ELISA (FIG. 2B). Data are shown as fold change in the omaveloxolone treated cells relative to cells treated with vehicle control (DMSO); ** indicates P<0.01 by one-way ANOVA with Dunnett’s multiple comparisons test 2131222371Attorney Docket No.: RRBS-007 / 01WO 353016-2013(n=3 to 6).Example 3. Candidate testing in human iPSC-derived Schwann cells

[0100] PMP22 protein expression levels were also tested after daily treatment of wildtype Schwann Cells with vehicle (0.01 % DMSO) or omaveloxolone at two concentrations: 250 nM and 500 nM, for three days. On day 4, the cells were lysed, and protein was extracted. Treatment with 250 nM omaveloxolone significantly decreased PMP22 protein expression in wild-type Schwann cells, as measured by ELISA (FIG. 3). Data are shown as fold change in the omaveloxolone treated cells relative to cells treated with vehicle control (DMSO): * indicates P<0.05 by one-way ANOVA with Dunnett’s multiple comparisons test (n=4 to 5).2231222371

Claims

Attorney Docket No.: RRBS-007 / 01WO 353016-2013CLAIMS1. A method of treating or preventing a disease or condition characterized by increased expression of PMP22. in a subject in need thereof, comprising administering an effective amount of a composition comprising omaveloxolone, or a pharmaceutically acceptable salt thereof.

2. The method of claim 1, wherein the subject comprises one or more mutations in at least one allele of the PMP22 gene.

3. The method of claim 2, wherein the one or more mutations comprise a duplication of the PMP22 gene, a duplication upstream of the PMP22 gene, or a point mutation.

4. The method of any one of claims 1-3, wherein the subject is determined to have the one or more mutations by analysis of the subject’s PMP22 genome sequence.

5. The method of claim 4, wherein the analysis of the subject's PMP22 genome sequence is performed using a method selected from the group consisting of PMP22 gene dosage analysis, sequence analysis of the entire PMP22 coding region, sequence analysis of select exons, mutation scanning of select exons, mutation scanning of the entire PMP22 coding region, or PMP22 RNA analysis.

6. The method of any one of claims 1-5, wherein the disease or condition is Charcot Marie Tooth disease.

7. The method of any one of claims 1-6, wherein the effective amount of the composition is about 0.5 mg / kg to about 20 mg / kg per day.8 The method of claim 7, wherein the effective amount of the composition is about 1 mg / kg to about 5 mg / kg per day.

9. The method of claim 7, wherein the effective amount of the composition is about 1 mg / kg, about 2 mg / kg. about 3 mg / kg, about 4 mg / kg, about 5 mg / kg per day, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, or about 10 mg / kg per day.

10. The method of any one of claims 1-9, wherein the composition is administered orally.2331222371Attorney Docket No.: RRBS-007 / 01WO 353016-201311. The method of any one of claims 1-10, wherein the composition is administered once or twice per day.

12. The method of any of claims 1-11, wherein the composition is administered for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 6 months, at least about 9 months, at least about 12 months, at least about 15 months, at least about 18 months, at least about 2 years, at least about 3 years, at least about 4 years, or at least about 5 years.

13. The method of any one of claims 1-12, wherein the method results in reduction or improvement of at least one symptom of the disease or condition.

14. The method of claim 13, wherein the method results in prevention of at least one symptom of the disease or condition.

15. The method of any one of claims 1-14, wherein the subject is a mammal.

16. The method of claim 15, wherein the subject is a human.

17. The method of any one of claims 1-16, wherein the composition additionally comprises a pharmaceutically acceptable carrier.

18. A method of decreasing the level of PMP22 transcript and / or protein in a cell, wherein the method comprises contacting a cell with an effective amount of a composition comprising omaveloxolone, or a pharmaceutically acceptable salt thereof.

19. The method of claim 18, wherein the cell comprises one or more mutations in one allele of the PMP22 gene.

20. The method of claim 19, wherein the one or more mutations comprise a duplication of the PMP22 gene, a duplication upstream of the PMP22 gene, or a point mutation.

21. The method of any one of claims 18-20, wherein the level of PMP22 transcript and / or protein in the cell is decreased by about 10% to about 90%, compared to a cell treated with vehicle / control.2431222371Attorney Docket No.: RRBS-007 / 01WO 353016-201322. The method of any one of claims 18-21, wherein the level of PMP22 transcript and / or protein in the cell is decreased to a level comparable to that of a cell that does not comprise a mutation in either allele of the PMP22 gene.

23. The method of any one of claims 18-22, wherein the cell is a glia cell or a neuron.

24. The method of any one of claims 18-23, wherein the contact occurs in vivo or ex vivo.

25. A composition comprising omav el oxoIone, or a pharmaceutically acceptable salt thereof, for use in the treatment of a disease or condition.

26. The composition for use of claim 25, wherein the disease or condition is Charcot Marie Tooth disease.2531222371

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