Peptidomimetic (r)-2-amino-n-((s)-1-(((s)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide for treating neurodegenerative diseases

Abstract

To provide use of compositions in preparation of medicaments for treating or preventing α-synuclein disease or TDP-43 proteinopathy as well as pharmaceutical compositions.SOLUTION: Disclosed is the use of a composition comprising a therapeutically effective amount of a peptidomimetic, (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide or pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate and / or solvate thereof.SELECTED DRAWING: Figure 1A-1B

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of priority of U.S. Patent Application No. 62 / 878,272, filed Jul. 24, 2019, and U.S. Patent Application No. 63 / 046,292, filed Jun. 30, 2020, the contents of which are hereby incorporated by reference in their entirety.

[0002] The present technology generally relates to compositions and methods for improving or treating amyotrophic lateral sclerosis (ALS). The present technology also generally relates to compositions and methods for improving or treating other neurodegenerative conditions, including frontotemporal lobar degeneration (FTLD), Parkinson's disease (PD), PD with dementia, Lewy body dementia, and multiple system atrophy, such as alpha - synucleinopathies or TDP - 43 proteinopathies. Additionally, the present technology relates to administering to a subject having or at risk of having ALS, an alpha - synucleinopathy, or a TDP - 43 proteinopathy, an effective amount of a mitochondrial - targeting peptide - mimetic compound such as (R)-2 - amino - N - ((S)-1 - (((S)-5 - amino - 1-(3 - benzyl - 1,2,4 - oxadiazol - 5 - yl)pentyl)amino)-3-(4 - hydroxy - 2,6 - dimethylphenyl)-1 - oxopropan - 2 - yl)-5 - guanidinopentanamide, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof.

Background Art

[0003] The following description is provided to assist the reader's understanding. None of the information provided or references cited are admitted to be prior art to the compositions and methods disclosed herein.

[0004] Neurodegenerative diseases and disorders affect physical activities such as balance, movement, speech, breathing, and / or heart function. Neurodegenerative diseases and disorders are generally incurable and debilitating conditions that result in the progressive degeneration and / or death of nerve cells. Some examples of neurodegenerative diseases and disorders include the following. Amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration (FTLD), Parkinson's disease (PD), PD with dementia, Lewy body dementia, and multiple system atrophy (MSA). Some neurodegenerative diseases can be characterized as α-synucleinopathies or TDP-43 proteinopathies.

[0005] Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder that results in the death of motor neurons in the brain and spinal cord. This disorder generally onset in middle age and typically leads to paralysis and death within 3 to 5 years from diagnosis. Up to 10% of ALS is familial and is usually autosomal dominant. Several causative genes are known, among which mutant superoxide dismutase 1 (SOD1) and mutant C9orf72 (i.e., G4C2 hexanucleotide repeat in the C9orf72 gene) are most frequently found. Mutations in the TARDBP gene that result in alterations of TAR DNA-binding protein 43 (TDP-43) are also known to cause familial ALS (see Sreedharan et al., Science (2008), 319(5870):1668-1672). There is no effective treatment for ALS. Therefore, there is a need in the art to develop treatment options for ALS. SUMMARY OF THE INVENTION

[0006] In one aspect, the present disclosure provides a method for treating or preventing amyotrophic lateral sclerosis (ALS) or frontotemporal lobar degeneration (FTLD) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a peptidomimetic such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof.

[0007] In some embodiments, the subject is diagnosed as having ALS or FTLD. In some embodiments, ALS is familial. In some embodiments, familial ALS is caused by a mutation in the superoxide dismutase 1 (SOD1) gene, or a mutation in the TARDBP gene, resulting in a modification of the TAR DNA-binding protein (TDP-43).

[0008] In some embodiments, the peptidomimetic is administered daily for at least two weeks. In some embodiments, the peptidomimetic is administered daily for at least twelve weeks.

[0009] In some embodiments, the treatment or prevention includes the treatment or prevention of one or more signs or symptoms of ALS or FTLD, including one or more of muscle weakness, muscle wasting (atrophy), muscle spasm, muscle contracture, bradykinesia, poor balance, impaired coordination, change in voice quality, dysarthria, dysphagia, incomplete eyelid closure, drooling, emotional regulation disorder, premature death, and increased expression of translocator protein-18 kDa (TSPO) in the brain. In some embodiments, the treatment or prevention includes the treatment or prevention of plasma accumulation of neurofilament light chain (NfL). In some embodiments, the treatment or prevention includes demonstrating an improvement (e.g., an increase) in neurite length in a treated subject as compared to a subject not treated with the peptidomimetic. In some embodiments, the treatment or prevention includes extending the lifespan of a treated subject as compared to a subject not treated with the peptidomimetic. In some embodiments, the treatment or prevention includes protection from axonal injury in the central nervous system (CNS). In some embodiments, the treatment or prevention includes delaying the progression of onset of neurological symptoms in a treated subject as compared to a subject not treated with the peptidomimetic.

[0010] In some embodiments, the subject is a mammal. In some embodiments, the mammalian subject is a human.

[0011] In some embodiments, the peptidomimetic is administered orally. In some embodiments, the peptidomimetic is administered subcutaneously. In some embodiments, the peptidomimetic is administered locally, intranasally, systemically, intravenously, intraperitoneally, intradermally, intravitreally, ophthalmically, intrathecally, intraventricularly, iontophoretically, transmucosally, intravitreally, or intramuscularly.

[0012] In some embodiments, the method further comprises administering additional treatment to the subject, separately, sequentially, or simultaneously. In some embodiments, the additional treatment comprises administration of a therapeutic agent. In some embodiments, the therapeutic agent is selected from the group consisting of riluzole (Rilutek®), edaravone (Radicava®), mecasermin, baclofen (Lioresal®), diazepam (Valium®), dantrolene (Dantrium®), non-steroidal anti-inflammatory agents, anti-spasmodics (e.g., carbamazepine (Tegretol) or phenytoin (Dilantin®)), amitriptyline (Elavil®), nortriptyline (Pamelor™), and lorazepam (Ativan®). In some embodiments, the therapeutic agent is elamipretide (also known as SS-31 or Bendavia). In some embodiments, the combination of the peptidomimetic and the additional therapeutic treatment has a synergistic effect in the prevention or treatment of ALS or FTLD.

[0013] In some embodiments, the pharmaceutically acceptable salts of the peptidomimetic include tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate, or maleate (mono, bis, or tris (tris) salts in each case). In some embodiments, the pharmaceutically acceptable salts include monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride, monotosylate, bistosylate, or tritosylate. In some embodiments, the peptidomimetic is formulated as a tris HCl salt, bis HCl salt, or mono HCl salt.

[0014] In one aspect, the present disclosure provides the use of a composition in the preparation of a medicament for treating or preventing amyotrophic lateral sclerosis (ALS) or frontotemporal lobar degeneration (FTLD) in a subject in need thereof, the composition comprising a therapeutically effective amount of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, such as a peptidomimetic.

[0015] In some embodiments, the subject is diagnosed with having ALS or FTLD. In some embodiments, ALS is familial. In some embodiments, familial ALS is caused by a mutation in the superoxide dismutase 1 (SOD1) gene, or a mutation in the TARDBP gene, resulting in a modification of the TAR DNA-binding protein (TDP-43).

[0016] In some embodiments, the peptidomimetic is intended to be administered daily for 2 weeks or more. In some embodiments, the peptidomimetic is intended to be administered daily for 12 weeks or more.

[0017] In some embodiments, the treatment or prevention includes treating or preventing one or more signs or symptoms of ALS or FTLD, including one or more of muscle weakness, muscle wasting (atrophy), muscle spasm, muscle contracture, bradykinesia, poor balance, impaired coordination, change in voice quality, dysarthria, dysphagia, incomplete eye closure, drooling, emotional regulation disorder, premature death, and increased expression of translocator protein-18 kDa (TSPO) in the brain. In some embodiments, the treatment or prevention includes treating or preventing plasma accumulation of neurofilament light chain (NfL). In some embodiments, the treatment or prevention includes demonstrating improvement (e.g., increase) in neurite outgrowth in a treated subject as compared to a subject not treated with the peptidomimetic. In some embodiments, the treatment or prevention includes extending the lifespan of a treated subject as compared to a subject not treated with the peptidomimetic. In some embodiments, the treatment or prevention includes protecting against axonal injury in the central nervous system (CNS). In some embodiments, the treatment or prevention includes delaying the progression of onset of neurological symptoms in a treated subject as compared to a subject not treated with the peptidomimetic.

[0018] In some embodiments, the subject is a mammal. In some embodiments, the mammalian subject is a human.

[0019] In some embodiments, the peptidomimetic is formulated for oral administration. In some embodiments, the peptidomimetic is formulated for subcutaneous administration. In some embodiments, the peptidomimetic is formulated for topical, intranasal, systemic, intravenous, intraperitoneal, intradermal, intraocular, ophthalmic, intrathecal, intraventricular, iontophoretic, transmucosal, intravitreal, or intramuscular administration.

[0020] In some embodiments, the peptidomimetic is intended to be used separately from, sequentially with, or simultaneously with additional therapy. In some embodiments, the additional therapy includes the use of a therapeutic agent. In some embodiments, the therapeutic agent is selected from the group consisting of riluzole (Rilutek®), edaravone (Radicava®), mecasermin, baclofen (Lioresal®), diazepam (Valium®), dantrolene (Dantrium®), non-steroidal anti-inflammatory agents, antispasmodics (e.g., carbamazepine (Tegretol) or phenytoin (Dilantin®)), amitriptyline (Elavil®), nortriptyline (Pamelor™), and lorazepam (Ativan®). In some embodiments, the therapeutic agent is elamipretide (also known as SS-31 or Bendavia). In some embodiments, the combination of the peptidomimetic and the additional therapy has a synergistic effect in the prevention or treatment of ALS or FTLD.

[0021] In some embodiments, the pharmaceutically acceptable salts include tartrates, fumarates, citrates, benzoates, succinates, suberates, lactates, oxalates, phthalates, methanesulfonates, benzenesulfonates, or maleates (mono, bis, or tris (tris) salts in each case). In some embodiments, the pharmaceutically acceptable salts include monoacetates, bisacetates, triacetates, monotrifluoroacetates, bistrifluoroacetates, tristrifluoroacetates, monohydrochlorides, dihydrochlorides, trihydrochlorides, monotosylates, bistosylates, or tritosylates. In some embodiments, the peptidomimetic is formulated as a tris HCl salt, bis HCl salt, or mono HCl salt.

[0022] In one aspect, the present disclosure provides a peptidomimetic such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof for use in treating or preventing amyotrophic lateral sclerosis (ALS) or frontotemporal lobar degeneration (FTLD) in a subject in need of treatment or prevention of ALS or FTLD.

[0023] In some embodiments, the subject is diagnosed with ALS or FTLD. In some embodiments, ALS is familial. In some embodiments, familial ALS is caused by a mutation in the superoxide dismutase 1 (SOD1) gene, or a mutation in the TARDBP gene, resulting in a modification of the TAR DNA-binding protein (TDP-43).

[0024] In some embodiments, the peptidomimetic is contemplated to be administered daily for 2 weeks or more. In some embodiments, the peptidomimetic is contemplated to be administered daily for 12 weeks or more.

[0025] In some embodiments, the treatment or prevention includes treating or preventing one or more signs or symptoms of ALS or FTLD, including one or more of muscle weakness, muscle wasting (atrophy), muscle spasm, muscle contracture, bradykinesia, poor balance, impaired coordinated movement, change in voice quality, dysarthria, dysphagia, incomplete eyelid closure, drooling, impaired emotional regulation, premature death, and increased expression of translocator protein-18 kDa (TSPO) in the brain. In some embodiments, the treatment or prevention includes treating or preventing plasma accumulation of neurofilament light chain (NfL). In some embodiments, the treatment or prevention includes demonstrating improvement (e.g., an increase) in neurite outgrowth in a treated subject as compared to a subject not treated with a peptidomimetic. In some embodiments, the treatment or prevention includes extending the lifespan of a treated subject as compared to a subject not treated with a peptidomimetic. In some embodiments, the treatment or prevention includes protecting against axonal injury in the central nervous system (CNS). In some embodiments, the treatment or prevention includes delaying the progression of onset of neurological symptoms in a treated subject as compared to a subject not treated with a peptidomimetic.

[0026] In some embodiments, the subject is a mammal. In some embodiments, the mammalian subject is a human.

[0027] In some embodiments, the peptidomimetic is formulated for oral administration. In some embodiments, the peptidomimetic is formulated for subcutaneous administration. In some embodiments, the peptidomimetic is formulated for topical, intranasal, systemic, intravenous, intraperitoneal, intradermal, intraocular, ophthalmic, intrathecal, intraventricular, iontophoretic, transmucosal, intravitreal, or intramuscular administration.

[0028] In some embodiments, the peptidomimetic is intended to be used separately from, sequentially, or simultaneously with additional therapies. In some embodiments, the additional therapy includes the use of a therapeutic agent. In some embodiments, the therapeutic agent is selected from the group consisting of riluzole (Rilutek®), edaravone (Radicava®), mexacermin, baclofen (Lioresal®), diazepam (Valium®), dantrolene (Dantrium®), non-steroidal anti-inflammatory agents, anti-spasmodics (e.g., carbamazepine (Tegretol) or phenytoin (Dilantin®)), amitriptyline (Elavil®), nortriptyline (Pamelor™), and lorazepam (Ativan®). In some embodiments, the therapeutic agent is elamipretide (also known as SS-31 or Bendavia). In some embodiments, the combination of the peptidomimetic and the additional therapy has a synergistic effect in the prevention or treatment of ALS or FTLD.

[0029] In some embodiments, the pharmaceutically acceptable salts include tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate, or maleate (mono-, bis- or tris-(tris) salts in each case). In some embodiments, the pharmaceutically acceptable salts include monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride, monotosylate, bistosylate, or tritosylate. In some embodiments, the peptidomimetic is formulated as a tris HCl salt, bis HCl salt, or mono HCl salt.

[0030] In one aspect, the present disclosure provides a method for treating or preventing α-synucleinopathy or TDP-43 proteinopathy in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a peptidomimetic such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof. In some embodiments, the subject is diagnosed with having α-synucleinopathy or TDP-43 proteinopathy.

[0031] In some embodiments, the α-synucleinopathy is Parkinson's disease (PD), PD with dementia, Lewy body dementia, or multiple system atrophy, and the TDP-43 proteinopathy is amyotrophic lateral sclerosis (ALS) or frontotemporal lobar degeneration (FTLD).

[0032] In some embodiments, the peptidomimetic is administered daily for 2 weeks or more. In some embodiments, the peptidomimetic is administered daily for 12 weeks or more.

[0033] In some embodiments, treatment or prevention of α-synucleinopathy comprises attenuating loss of dopaminergic neurons in the subject as compared to an untreated control. In some embodiments, treatment or prevention of TDP-43 proteinopathy comprises increasing neurite length in the subject as compared to an untreated control.

[0034] In some embodiments, the subject is a mammal. In some embodiments, the mammalian subject is a human.

[0035] In some embodiments, the peptidomimetic is administered orally. In some embodiments, the peptidomimetic is administered subcutaneously. In some embodiments, the peptidomimetic is administered locally, intranasally, systemically, intravenously, intraperitoneally, intradermally, intravitreally, ophthalmically, intrathecally, intraventricularly, iontophoretically, transmucosally, intravitreally, or intramuscularly.

[0036] In some embodiments, the method further comprises administering additional treatment to the subject, separately, sequentially, or simultaneously. In some embodiments, the additional treatment comprises administration of a therapeutic agent. In some embodiments, the therapeutic agent comprises levodopa for the treatment of α-synucleinopathy, and the therapeutic agent comprises a selective serotonin reuptake inhibitor (SSRI) antidepressant for the treatment of TDP-43 proteinopathy. In some embodiments, the combination of the peptidomimetic and the additional therapeutic treatment has a synergistic effect in the prevention or treatment of α-synucleinopathy or TDP-43 proteinopathy.

[0037] In some embodiments, the pharmaceutically acceptable salts include tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate, or maleate (mono, bis or tris (tris) salts in each case). In some embodiments, the pharmaceutically acceptable salts include monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride, monotosylate, bistosylate, or tritosylate. In some embodiments, the peptidomimetic is formulated as a tris HCl salt, bis HCl salt, or mono HCl salt.

[0038] In one aspect, the present disclosure provides the use of a composition in the preparation of a medicament for treating or preventing α-synucleinopathy or TDP-43 proteinopathy in a subject in need thereof, the composition comprising a therapeutically effective amount of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, such as a peptidomimetic.

[0039] In some embodiments, the subject is diagnosed with having α-synucleinopathy or TDP-43 proteinopathy. In some embodiments, α-synucleinopathy is Parkinson's disease (PD), PD with dementia, Lewy body dementia, or multiple system atrophy, and TDP-43 proteinopathy is amyotrophic lateral sclerosis (ALS) or frontotemporal lobar degeneration (FTLD).

[0040] In some embodiments, the peptidomimetic is contemplated to be administered daily for 2 weeks or more. In some embodiments, the peptidomimetic is contemplated to be administered daily for 12 weeks or more.

[0041] In some embodiments, the treatment or prevention of α-synucleinopathy includes attenuating the loss of dopaminergic neurons in the subject as compared to an untreated control. In some embodiments, the treatment or prevention of TDP-43 proteinopathy includes increasing neurite outgrowth in the subject as compared to an untreated control.

[0042] In some embodiments, the subject is a mammal. In some embodiments, the mammalian subject is a human.

[0043] In some embodiments, the peptidomimetic is formulated for oral administration. In some embodiments, the peptidomimetic is formulated for subcutaneous administration. In some embodiments, the peptidomimetic is formulated for topical, intranasal, systemic, intravenous, intraperitoneal, intradermal, intraocular, ophthalmic, intrathecal, intracerebroventricular, iontophoretic, transmucosal, intravitreal, or intramuscular administration.

[0044] In some embodiments, the use further comprises administering an additional treatment to the subject, separately, sequentially, or simultaneously. In some embodiments, the additional treatment comprises administration of a therapeutic agent. In some embodiments, the therapeutic agent comprises levodopa for the treatment of alpha-synucleinopathy, and the therapeutic agent comprises a selective serotonin reuptake inhibitor (SSRI) antidepressant for the treatment of TDP-43 proteinopathy.

[0045] In some embodiments, the combination of the peptidomimetic and the additional therapeutic treatment has a synergistic effect in the prevention or treatment of alpha-synucleinopathy or TDP-43 proteinopathy.

[0046] In some embodiments, the pharmaceutically acceptable salts include tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate, or maleate (mono, bis or tris (tris) salts in each case). In some embodiments, the pharmaceutically acceptable salts include monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride, monotosylate, ditosylate, or tritosylate. In some embodiments, the peptidomimetic is formulated as a tris HCl salt, bis HCl salt, or mono HCl salt.

[0047] In one aspect, the present disclosure provides a peptidomimetic such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide, or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof for use in treating or preventing α-synucleinopathy or TDP-43 proteinopathy in a subject in need of such treatment or prevention of α-synucleinopathy or TDP-43 proteinopathy.

[0048] In some embodiments, the subject is diagnosed with having α-synucleinopathy or TDP-43 proteinopathy. In some embodiments, the α-synucleinopathy is Parkinson's disease (PD), PD with dementia, Lewy body dementia, or multiple system atrophy, and the TDP-43 proteinopathy is amyotrophic lateral sclerosis (ALS) or frontotemporal lobar degeneration (FTLD).

[0049] In some embodiments, the peptidomimetic is intended to be administered daily for 2 weeks or more. In some embodiments, the peptidomimetic is intended to be administered daily for 12 weeks or more.

[0050] In some embodiments, the treatment or prevention of α-synucleinopathy includes attenuating the loss of dopaminergic neurons in the subject as compared to an untreated control. In some embodiments, the treatment or prevention of TDP-43 proteinopathy includes increasing neurite length in the subject as compared to an untreated control.

[0051] In some embodiments, the subject is a mammal. In some embodiments, the mammalian subject is a human.

[0052] In some embodiments, the peptidomimetic is formulated for oral administration. In some embodiments, the peptidomimetic is formulated for subcutaneous administration. In some embodiments, the peptidomimetic is formulated for topical, intranasal, systemic, intravenous, intraperitoneal, intradermal, intraocular, ophthalmic, intrathecal, intraventricular, iontophoretic, transmucosal, intravitreal, or intramuscular administration.

[0053] In some embodiments, the peptidomimetic is intended to be used separately from, sequentially with, or simultaneously with additional therapy. In some embodiments, the additional therapy includes the use of a therapeutic agent. In some embodiments, the therapeutic agent includes levodopa for the treatment of alpha-synucleinopathy, and the therapeutic agent includes a selective serotonin reuptake inhibitor (SSRI) antidepressant for the treatment of TDP-43 proteinopathy. In some embodiments, the combination of the peptidomimetic and the additional therapeutic treatment has a synergistic effect in the prevention or treatment of alpha-synucleinopathy or TDP-43 proteinopathy.

[0054] In some embodiments, pharmaceutically acceptable salts include tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate, or maleate (mono-, bis-, or tris (tris) salts in each case). In some embodiments, pharmaceutically acceptable salts include monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride, monotosylate, ditosylate, or tritosylate. In some embodiments, the peptidomimetic is formulated as a tris HCl salt, bis HCl salt, or mono HCl salt. BRIEF DESCRIPTION OF THE DRAWINGS

[0055]

Fig. 1A-1B

Fig. 1C-1D

Fig. 2A-2B

Fig. 3A-3B

Fig. 4

Fig. 5A

Fig. 5B

Fig. 6A

Fig. 6B

Fig. 6C

Fig. 7

Mode for Carrying Out the Invention

[0056] It should be understood that certain aspects, modes, embodiments, variations and features of the present technology are described below in various levels of detail in order to provide a substantial understanding of the present technology. Definitions of specific terms used herein are provided below. Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this technology belongs.

[0057] In the practice of this technology, many conventional techniques in molecular biology, protein biochemistry, cell biology, immunology, microbiology, and recombinant DNA are used. These techniques are well known and are described, for example, in the following. Respectively, Current Protocols in Molecular Biology, Vols. I-III, Ausubel, Ed. (1997), Sambrook et al., Molecular Cloning: A Laboratory Manual, Second Ed. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989), DNA Cloning: A Practical Approach, Vols. I and II, Glover, Ed. (1985), Oligonucleotide Synthesis, Gait, Ed. (1984), Nucleic Acid Hybridization, Hames & Higgins, Eds. (1985), Transcription and Translation, Hames & Higgins, Eds. (1984), Animal Cell Culture, Freshney, Ed. (1986), Immobilized Cells and Enzymes (IRL Press, 1986), Perbal, A Practical Guide to Molecular Cloning, the series, Meth. Enzymol., (Academic Press, Inc., 1984), Gene Transfer Vectors for Mammalian Cells, Miller & Calos, Eds. (Cold Spring Harbor Laboratory, N Y, 1987), and Meth. Enzymol., Vols. 154 and 155, Wu & Grossman, and Wu, Eds.

[0058] Definitions As used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the content clearly dictates otherwise. For example, reference to "a cell" includes combinations of two or more cells and the like.

[0059] As used herein, "administration" of an agent, drug, therapeutic agent, peptide, or peptidomimetic to a subject includes any route by which a compound is introduced or delivered to the subject to effect its intended function. Administration can be effected by any suitable route, such as oral administration. Administration can be effected subcutaneously. Administration can be effected intravenously. Administration can be effected intravitreally. Administration can be effected systemically. Alternatively, administration can be effected topically, intranasally, intraperitoneally, intradermally, ophthalmically, intrathecally, intracerebroventricularly, iontophoretically, transmucosally, intravitreally, or intramuscularly. Administration includes self-administration and administration by another person.

[0060] As used herein, the term "amino acid" includes both naturally occurring amino acids and non-natural amino acids. The term "amino acid" includes, unless otherwise indicated, both isolated amino acid molecules (i.e., molecules containing both an amino-bonded hydrogen and a carbonyl-carbon-bonded hydroxyl) and residues of amino acids (i.e., molecules in which either or both of the amino-bonded hydrogen or the carbonyl-carbon-bonded hydroxyl have been removed). The amino group can be an alpha amino group, a beta amino group, etc. For example, the term "amino acid alanine" can refer to either the isolated alanine H-Ala-OH, or any one of the alanine residues H-Ala-, -Ala-OH, or -Ala-. Unless otherwise indicated, all amino acids found in the compounds described herein can be in either the D or L configuration. Amino acids in the D configuration can be written such that "D" precedes the abbreviation of the amino acid. For example, "D-Arg" represents arginine in the D configuration. The term "amino acid" includes its salts, including pharmaceutically acceptable salts. Any amino acid can be either protected or unprotected. The protecting group can be attached to the amino group (e.g., the alpha amino group), the backbone carboxyl group, or any functional group on the side chain. By way of example, phenylalanine protected by a benzyloxycarbonyl group (Z) on the alpha amino group would be represented as Z-Phe-OH.

[0061] Except for the N-terminal amino acid, all abbreviations of amino acids (e.g., Phe) in the present disclosure represent the structure of -NH-C(R)(R’)-CO-, where each of R and R’ is independently hydrogen or the side chain of an amino acid (e.g., for Phe, R = benzyl and R’ = H). Thus, phenylalanine is H-Phe-OH. The notation “OH” for these amino acids or peptides (e.g., Lys-Val-Leu-OH) indicates that the C-terminus is a free acid. For example, the notation “NH2” in Phe-D-Arg-Phe-Lys-NH2 indicates that the C-terminus of the protected peptide fragment is amidated. Further, certain R and R’ may contain functional groups that require protection during liquid or solid-phase synthesis, either separately or in combination as a ring structure.

[0062] When an amino acid has an isomeric form, unless specifically indicated as the D-form, e.g., D-Arg, the L-form of the amino acid is represented. In particular, many amino acid residues are commercially available in both D- and L-forms. For example, D-Arg is a commercially available D-amino acid.

[0063] The capital letter “D” used in conjunction with the abbreviation of an amino acid residue refers to the D-form of the amino acid residue.

[0064] The term “DMT” refers to 2,6-di(methyl)tyrosine (e.g., 2,6-dimethyl-L-tyrosine, CAS 123715-02-6).

[0065] As used herein, the phrase “delaying the onset of” refers to delaying, interfering with, or causing one or more symptoms of a disorder, symptom, condition, or sign to occur later than normal relative to an untreated control sample in a statistical sample in a treated sample.

[0066] As used herein, the term "effective amount" refers to an amount sufficient to achieve a desired therapeutic and / or prophylactic effect, e.g., an amount that results in a partial or complete improvement of one or more symptoms of ALS, alpha-synucleinopathy, or TDP-43 proteinopathy. In the context of therapeutic or prophylactic use, in some embodiments, the amount of the composition administered to a subject depends on the type, degree, and severity of the disease, as well as individual characteristics such as general health, age, sex, weight, and drug tolerance. One of ordinary skill in the art will be able to determine appropriate dosages depending on these and other factors. The composition can also be administered in combination with one or more additional therapeutic compounds.In the method described herein, (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (e.g., (Ia) such as tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate, or maleate (mono, bis or tris (tris) salts in each case), monoacetate (i.e., a salt containing one acetic acid moiety), bisacetate (i.e., a salt containing two acetic acid moieties), triacetate (i.e., a salt containing three acetic acid moieties), monotrifluoroacetate (i.e., a salt containing one trifluoroacetic acid moiety), bistrifluoroacetate (i.e., a salt containing two trifluoroacetic acid moieties), tristrifluoroacetate (i.e., a salt containing three trifluoroacetic acid moieties), monohydrochloride (i.e., a salt containing one chloride anion resulting from, or considered to result from, an HCl inclusion, "mono HCl salt"), dihydrochloride (i.e., a salt containing two chloride anions resulting from, or considered to result from, two HCl inclusions, "bis HCl salt"), trihydrochloride (i.e., a salt containing three chloride anions resulting from, or considered to result from, three HCl inclusions, "tri HCl salt"), monotosylate (i.e., a salt containing one tosylate moiety), bistosylate (i.e., a salt containing two tosylate moieties), or tritosylate (i.e., a salt containing three tosylate moieties), etc., etc.) of the mitochondrial targeting peptide mimetic can be administered to a subject having one or more signs, symptoms, or risk factors of ALS or FTLD, including but not limited to muscle weakness, muscle wasting (atrophy), muscle cramps, muscle contractures, slowness of movement, poor balance, impaired coordination, change in voice quality, dysarthria, dysphagia, incomplete eyelid closure, drooling, emotional regulation disorder, premature death, increased expression of brain translocator protein-18 kDa (TSPO), and plasma accumulation of neurofilament light chain (NfL).

[0067] As used herein, the term "hydrate" refers to a compound associated with water. The number of water molecules contained in a hydrate of a compound may (or may not) be in a defined ratio to the number of compound molecules in the hydrate.

[0068] As used herein, the term "peptidomimetic" refers to a small peptidomimetic polymer that contains two or more amino acids and also includes non-peptidic-like modifications. Peptidomimetics can arise by modification of existing peptides or by designing similar molecules that mimic peptide function.

[0069] As used herein, the terms "pharmaceutically acceptable carrier" and "carrier" refer to a diluent, adjuvant, excipient, or vehicle that is administered with a compound or formulated for administration. Non-limiting examples of such pharmaceutically acceptable carriers include liquids such as water, saline, and oils, as well as solids such as acacia gum, gelatin, starch paste, talc, keratin, colloidal silica, urea, and the like. In addition, auxiliary agents, stabilizers, thickeners, lubricants, flavoring agents, and coloring agents may be used. Other examples of suitable pharmaceutical carriers are described in Remington’s Pharmaceutical Sciences by E.W. Martin, which is hereby incorporated by reference in its entirety.

[0070] As used herein, "preventing" or "prevention" of a disorder or condition refers to a compound that, in a statistical sample, reduces the occurrence of a disorder or condition in a treated sample relative to an untreated control sample, or delays the onset of one or more symptoms of a disorder or condition relative to an untreated control sample. As used herein, prevention of ALS, alpha-synucleinopathy, or TDP-43 proteinopathy includes preventing or delaying the onset of symptoms of ALS, alpha-synucleinopathy, or TDP-43 proteinopathy. As used herein, prevention of ALS, alpha-synucleinopathy, or TDP-43 proteinopathy also includes preventing recurrence of one or more signs or symptoms of ALS, alpha-synucleinopathy, or TDP-43 proteinopathy.

[0071] As used herein, the terms "subject" and "patient" are used interchangeably.

[0072] In the context of therapeutic use or administration, the term "separate" or "separately" refers to the administration of at least two active ingredients by different routes, formulations, and / or pharmaceutical compositions.

[0073] The term "simultaneous" therapeutic use refers to the administration of at least two active ingredients simultaneously or at substantially the same time. In some embodiments, simultaneous administration includes administration of a single composition or formulation containing at least two active ingredients, simultaneous administration of at least two separate active ingredients by the same route, and simultaneous administration of at least two separate active ingredients by different routes, but is not limited thereto.

[0074] As used herein, the term "consecutive" therapeutic use refers to the administration of at least two active ingredients at different times, and the administration routes may be the same or different. More specifically, consecutive use refers to the entire administration of one of the active ingredients before the start of the other administration(s). Thus, it is possible to administer one of the active ingredients over minutes, hours, or days before administering the other active ingredient(s). In this case, there is no simultaneous treatment.

[0075] As used herein, the term "subject" refers to a living animal. In various embodiments, the subject is a mammal. In some embodiments, the subject is a non-human mammal including, but not limited to, a mouse, rat, hamster, guinea pig, rabbit, sheep, goat, cat, dog, pig, mini-pig, horse, cow, or non-human primate. In some embodiments, the subject is a human.

[0076] As used herein, the term "solvate" refers to a form of a compound (e.g., a peptide or peptidomimetic) that is usually associated with a solvent by a solvolysis reaction. This physical association may include hydrogen bonding. Conventional solvents include water, methanol, ethanol, isopropanol, acetic acid, ethyl acetate, acetone, hexane, dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), diethyl ether, and the like.

[0077] As used herein, the term "tautomer" refers to an interchangeable form of a particular compound structure, which refers to a compound in which the rearrangement of hydrogen atoms and electrons changes. Thus, the two structures can be in equilibrium via the movement of π electrons and atoms (usually H). For example, enol and ketone are tautomers because they are rapidly interconverted by treatment with either an acid or a base. Tautomeric forms may be relevant to the achievement of the optimal chemical reactivity and biological activity of the compound of interest.

[0078] As used herein, "synergistic therapeutic effect" refers to a therapeutic effect that is produced by the combination of at least two agents and is greater than additive, i.e., greater than the result from the individual administration of the agents. For example, lower doses of one or more agents can be used in the treatment of ALS, α-synucleinopathy, or TDP-43 proteinopathy, resulting in an increase in therapeutic effect and a decrease in side effects.

[0079] As used herein, "treating" or "treatment" or "alleviating" refers to a therapeutic treatment, the purpose of which is to reduce, alleviate or delay the progression or development of a targeted pathological condition or disorder and / or to reverse it. According to the methods described herein, after a subject has received a therapeutically effective amount of a mitochondrial-targeted peptidomimetic such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof such as a tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono-, bis- or tris-(tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride, monotosylate, ditosylate, or tritosylate (e.g., (Ia)), if the subject shows an observable and / or measurable reduction or absence of one or more signs and symptoms of ALS, alpha-synucleinopathy, or TDP-43 proteinopathy, the subject is considered to have been successfully "treated" for ALS, alpha-synucleinopathy, or TDP-43 proteinopathy. For example, in ALS, such signs and symptoms include, but are not limited to, muscle weakness, muscle wasting (atrophy), muscle cramps, muscle fasciculations, bradykinesia, poor balance, impaired coordination, change in voice quality, dysarthria, dysphagia, incomplete eye closure, drooling, emotional regulation disorder, premature death, increased expression of brain translocator protein-18 kDa (TSPO), and plasma accumulation of neurofilament light chain (NfL). In some embodiments, treatment refers to a delay in the onset of the neurological symptoms of ALS as evaluated by the neurological scoring described herein.

[0080] The various modes of treatment or prevention of the medical conditions described herein are intended to mean "substantial", which includes treatment or prevention that is overall but not entirely complete, and it should also be understood that several biologically or medically relevant results are achieved.

[0081] As used herein, the terms "(R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide", "(D-Arg-DMT-NH((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pent-1-yl)", (2R)-2-amino-N-[(1S)-1-{[(1S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl]carbamoyl}-2-(4-hydroxy-2,6-dimethylphenyl)ethyl]-5-carbamimidamidepentanamide, "Compound 7a", and "7a" refer to the same mitochondrial targeting peptide mimetic and are used interchangeably herein to refer to the compound of formula (I) below.

Chemical formula

[0082] "(R)-2-Amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide", (2R)-2-amino-N-[(1S)-1-{[(1S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl]carbamoyl}-2-(4-hydroxy-2,6-dimethylphenyl)ethyl]-5-carbamimidamide pentanamide, "(D-Arg-DMT-NH((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pent-1-yl)", "Compound 7a", and the term "7a (as shown below)" are intended to include pharmaceutically acceptable salt forms such as the tri (or tris) HCl salt of Formula Ia below. [Chemical formula]

[0083] Amyotrophic lateral sclerosis (ALS) Amyotrophic lateral sclerosis (ALS, also known as Lou Gehrig's disease) is a progressive neuromuscular condition characterized by weakness, muscle wasting, contractions, and increased reflexes. Currently, about 30,000 Americans are affected by this disease. The annual incidence rate is 1-2 cases per 100,000 people. This disease is most commonly diagnosed in middle age and affects more men than women. ALS is characterized by the adult-onset, idiopathic, progressive degeneration of the anterior horn cells as well as the upper and lower motor neurons, resulting in progressive muscle weakness, wasting, and contractions. Atrophy of the anterior horn cells and replacement of large motor neurons by fibrous astrocytes (gliosis) cause the anterior and lateral columns of the spinal cord to become stiff, thus giving rise to the term "lateral sclerosis". Typical signs and symptoms of ALS include muscle weakness, muscle wasting (atrophy), muscle cramps, muscle spasms, slow movement, poor balance, impaired coordination, changes in voice quality, dysarthria, dysphagia, incomplete eye closure, drooling, emotional regulation disorders, and premature death.

[0084] The top 10% of ALS cases are familial and are usually autosomal dominant. Several causative genes are known, among which mutant superoxide dismutase 1 (SOD1) and mutant C9orf72 (i.e., G4C2 hexanucleotide repeat in the C9orf72 gene) are the most frequently found in familial ALS (fALS) and sporadic ALS (sALS). Several other genes are known to be causes of classical ALS, but these are less frequent in cases than mutant SOD1. These genes include mutant FUS (fused in sarcoma), the mutant TARDBP gene that results in modifications of TAR DNA-binding protein 43 (TDP-43), and optineurin.

[0085] Clinical findings vary depending on the area of the nervous system affected and the progression of the pathological changes. The classical findings in ALS are insidious, progressive, asymmetric muscle weakness and atrophy, along with neurological signs, particularly spasticity and hyperreflexia. Usually, there are problems with dexterity or walking due to muscle weakness. Difficulty speaking or swallowing is an early symptom of the bulbar form of the disease. Over a period of months or years, patients with ALS develop severe progressive muscle weakness and other symptoms caused by loss of function in both upper and lower motor neurons. Sphincter control, sensory function, intellectual ability, and skin integrity are maintained. Patients become completely disabled and often require ventilatory support and gastrostomy. Death usually occurs within 5 years of diagnosis and is due to respiratory failure or cachexia. The diagnosis of ALS is clinical, based on the characteristic signs of progressive weakness, atrophy, spasticity, and hyperreflexia affecting several areas of the body. Initial differential diagnoses can include musculoskeletal, neurological, or systemic conditions. The cause of the disease is unknown. Current management includes active and individualized palliative care of symptoms and complications. There is no cure for ALS.

[0086] Currently, the only drugs indicated for ALS treatment are riluzole (Rilutek®) and edaravone (Radicava®). At least one other drug (meclofenamine) is being investigated by the US Food and Drug Administration. Various symptomatic treatments may be useful, including baclofen (Lioresal®), diazepam (Valium®), dantrolene (Dantrium®), non-steroidal anti-inflammatory agents, antispasmodics such as carbamazepine (Tegretol) or phenytoin (Dilantin®), amitriptyline (Elavil®), nortriptyline (Pamelor®), and lorazepam (Ativan®).

[0087] Neurofilament light chain (NfL) Biomarkers reflecting the characteristics of ALS may not only assist in disease diagnostic algorithms but may also be valuable in defining homogeneous subgroups of patients. They can also help in tracking disease progression and treatment response. Neurofilaments (NFs) have been extensively studied in various neurological conditions and are considered useful as markers of acute and chronic nerve injury (Bacioglu et al., Neuron 91:56 - 66 (2016)). Neurofilaments are 10 nm intermediate filaments in neurons, composed of heteropolymers of different subunits, neurofilament light chain (NfL), neurofilament medium chain (NfM), and neurofilament heavy chain (NfH) (Lee, Ann. Rev. Neurosci. 19:187 - 217 (1996)). Neurofilament light chain (NfL) is specific to nerve cells, leaks into the cerebrospinal fluid (CSF), and can be detected at low concentrations in peripheral blood. CSF, serum, and plasma NfL levels have been shown to distinguish patients with ALS from healthy controls with high sensitivity and specificity and to correlate with disease progression or survival in patients with ALS (Lu et al., Neurology 2015 Jun 2;84(22):2247 - 57). In an SOD1 mouse model of ALS, motor neuron degeneration has been shown to be accompanied by a progressive increase in blood NF levels, and these levels have been shown to be able to capture treatment response (Lu et al., PLoS ONE 7:e40998 (2012), Boylan et al., J. Neurochem. 111:1182 - 1191 (2009)).

[0088] α - synucleinopathy Synucleinopathy or α-synucleinopathy is a neurodegenerative disease characterized by abnormal accumulation of aggregates of α-synuclein protein in neurons, nerve fibers, or glial cells. These conditions are also associated with loss of substantia nigra dopaminergic neurons. These diseases include Parkinson's disease (PD), PD with dementia, dementia with Lewy bodies, and multiple system atrophy. The neuropathological diagnosis of α-synucleinopathy is based on the detection of altered α-synuclein in tissue and the registration of the neuroanatomical distribution of this change in the brain.

[0089] TDP-43 proteinopathy TAR-DNA-binding protein 43 (TDP-43) proteinopathy includes amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). FTLD refers to a clinically, genetically, and neuropathologically heterogeneous group of neurodegenerative disorders and is the third most common form of dementia after Alzheimer's disease (AD) and dementia with Lewy bodies. Current research criteria divide FTLD into the following three clinical syndromes: frontotemporal dementia, primary progressive non-fluent aphasia, and semantic dementia. The most common clinical form, frontotemporal dementia, presents mainly as changes in personality and behavior, while primary progressive non-fluent aphasia and semantic dementia present mainly as language dysfunction. In addition, patients may develop movement abnormalities such as parkinsonism and motor neuron diseases.

[0090] The term frontotemporal lobar degeneration reflects the prominent frontotemporal atrophy seen in these patients on neuropathological examination. A characteristic feature in most FTLD brains is the formation of abnormal proteinaceous inclusions in neurons and glial cells. TAR-DNA-binding protein 43 (TDP-43) has been identified as the disease protein in FTLD. Mutant TDP-43 has been found to inhibit neurite outgrowth, and overexpression of wild-type (WT) and mutant TDP43 causes toxicity in motor neurons.

[0091] Mitochondrial targeting peptide mimetic In some embodiments, the present disclosure provides a compound of formula (II), or a pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof.

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0092] In some embodiments, AA1 is,

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0093] In some embodiments, AA2 is,

Chemical formula

Chemical formula

[0094] In some embodiments, R 1 is, [Chemical formula] It is. In some embodiments, R 1 is [Chemical formula] It is. In some embodiments, R 1 is [Chemical formula] It is. In some embodiments, R 1 is [Chemical formula] It is. In some embodiments, R 1 is [Chemical formula] It is. In some embodiments, R 1 is [Chemical formula] It is. In some embodiments, R 1 is [Chemical formula] It is. In some embodiments, R 1 is [Chemical formula] It is. In some embodiments, R 1 is [Chemical formula] It is.

[0095] In some embodiments, R 2a is [Chemical formula] It is. In some embodiments, R 2a is

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0096] In some embodiments, R 2b is H. In some embodiments, R 2b is methyl.

[0097] In some embodiments, R 3 is H. In some embodiments, R 3 is (C1-C6) alkyl. In some embodiments, R 3is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, or t-butyl. In some embodiments, R 3 is methyl. In some embodiments, R 3 is ethyl.

[0098] In some embodiments, R 4 is H. In some embodiments, R 4 is (C1-C6) alkyl. In some embodiments, R 4 is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, or t-butyl. In some embodiments, R 4 is methyl. In some embodiments, R 4 is ethyl.

[0099] In some embodiments, R 3 and R 4 are the same. In some embodiments, R 3 and R 4 are different.

[0100] In some embodiments, R 5 is H. In some embodiments, R 5 is methyl.

[0101] In some embodiments, R 6 is H. In some embodiments, R 6 is methyl.

[0102] In some embodiments, R 5 and R 6 are the same. In some embodiments, R 5 and R 6 are different.

[0103] In some embodiments, R 5 and R 6Together with the N atoms to which they are attached, they form a 4- to 6-membered heterocyclyl. In some embodiments, the heterocyclyl is a 4- to 6-membered ring. In some embodiments, the heterocyclyl is azetidinyl, pyrrolidinyl, or piperidinyl.

[0104] In some embodiments, R 7 is H. In some embodiments, R 7 is (C1-C6)alkyl. In some embodiments, R 7 is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, or t-butyl. In some embodiments, R 7 is methyl.

[0105] In some embodiments, R 7 is cycloalkyl. In some embodiments, R 7 is cyclopropyl, cyclobutyl, cyclopropyl, or cyclohexyl. In some embodiments, R 7 is aryl. In some embodiments, R 7 is phenyl.

[0106] In some embodiments, R 8 is H. In some embodiments, R 8 is (C1-C6)alkyl. In some embodiments, R 8 is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, or t-butyl. In some embodiments, R 8 is methyl.

[0107] In some embodiments, R 8 is cycloalkyl. In some embodiments, R 8 is cyclopropyl, cyclobutyl, cyclopropyl, or cyclohexyl. In some embodiments, R 8is aryl. In some embodiments, R 8 is phenyl.

[0108] In some embodiments, R 9 is H. In some embodiments, R 9 is (C1-C6) alkyl. In some embodiments, R 9 is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, or t-butyl. In some embodiments, R 9 is methyl.

[0109] In some embodiments, R 9 is cycloalkyl. In some embodiments, R 9 is cyclopropyl, cyclobutyl, cyclopropyl, or cyclohexyl. In some embodiments, R 9 is aryl. In some embodiments, R 9 is phenyl.

[0110] In some embodiments, R 8 and R 9 are the same. In some embodiments, R 8 and R 9 are different.

[0111] In some embodiments, R 8 and R 9 together with the N atom to which they are attached form a 4- to 6-membered heterocyclyl. In some embodiments, the heterocyclyl is a 4- to 6-membered ring. In some embodiments, the heterocyclyl is azetidinyl, pyrrolidinyl, or piperidinyl.

[0112] In some embodiments, X is

Chemical formula

[0113] In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0114] The chiral centers of the peptidomimetics disclosed herein may be in either the R or S configuration, as will be discussed in more detail below.

[0115] Chiral / Stereochemical Considerations The peptidomimetics described herein may contain one or more chiral centers and, thus, may exist in various isomeric forms, such as enantiomers and / or diastereomers. For example, the compounds described herein may be in the form of individual enantiomers, diastereomers, or geometric isomers, or in the form of mixtures of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers can be isolated from the mixture by methods known to those skilled in the art, including chiral high performance liquid chromatography (HPLC) and formation and crystallization of chiral salts, or the preferred isomers can be prepared by asymmetric synthesis. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley lnterscience, New York, 1981), Wilen et al., Tetrahedron 33:2725 (1977), Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962), and Wilen, Tables of Resolving Agents and Optical Resolutions p.268 (E.L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972). Additionally, peptidomimetics include the compounds described herein as individual isomers that are substantially free of other isomers, alternatively as mixtures of various isomers.

[0116] R (D in the case of amino acids) or S (L in the case of amino acids) As used herein, a pure enantiomeric peptide mimic is substantially free of other enantiomers or stereoisomers of the compound (i.e., is enantiomerically pure). In other words, the "S" form of the compound is substantially free of the "R" form of the compound and is thus enantiomerically pure with respect to the "R" form. With respect to amino acids (more generally described with respect to the "D" and "L" enantiomers), it should be understood that for "D" amino acids the configuration is "R" and for "L" amino acids the configuration is "S". In some embodiments, "substantially free of" means the following. (i) An aliquot of the "R" form compound containing less than 2% of the "S" form, or (ii) an aliquot of the "S" form compound containing less than 2% of the "R" form. The terms "enantiomerically pure" or "pure enantiomer" mean that the compound contains more than 90 wt%, more than 91 wt%, more than 92 wt%, more than 93 wt%, more than 94 wt%, more than 95 wt%, more than 96 wt%, more than 97 wt%, more than 98 wt%, more than 99 wt%, more than 99.5 wt%, or more than 99.9 wt% of the enantiomer. In certain embodiments, the weight is based on the total weight of all enantiomers or stereoisomers of the compound.

[0117] In the compositions provided herein, the enantiomerically pure compounds can be present with other active or inactive ingredients. For example, a pharmaceutical composition containing an enantiomerically pure "R" form compound can contain, for example, about 90% excipient and about 10% enantiomerically pure "R" form compound. In certain embodiments, the enantiomerically pure "R" form compound in such a composition can contain, for example, at least about 95% by weight of the "R" form compound and up to about 5% by weight of the "S" form compound, based on the total weight of the compound. For example, a pharmaceutical composition containing an enantiomerically pure "S" form compound can contain, for example, about 90% excipient and about 10% enantiomerically pure "S" form compound. In certain embodiments, the enantiomerically pure "S" form compound in such a composition can contain, for example, at least about 95% by weight of the "S" form compound and up to about 5% by weight of the "R" form compound, based on the total weight of the compound. In certain embodiments, the active ingredient can be formulated with little or no excipient or carrier.

[0118] The nomenclature used to define the peptide compounds described herein is typically that used in the art where the N-terminal amino group appears on the left and the C-terminal carboxyl group appears on the right, provided, however, that the peptidomimetics disclosed herein are provided to not contain a carboxylic acid or amide moiety at the C-terminus.

[0119] The capital letter "D" used in conjunction with the abbreviation for an amino acid residue refers to the D-form of the amino acid residue. For example, D-Arg is a commercially available D-amino acid.

[0120] The peptidomimetics disclosed herein may exist in an unsolvated form, as well as in solvated forms including a hydrated form. The solvated forms may exist, for example, because it is difficult or impossible to remove all of the solvent from the peptidomimetic after synthesis. Generally, the solvated forms are equivalent to the unsolvated form and are encompassed within the scope of this application. Certain peptidomimetics of this application may exist in multiple crystalline or amorphous forms. Certain peptidomimetics of this application may exist in various tautomeric forms. Certain peptidomimetics of this application may exist in various salt forms. Generally, all physical forms are equivalent for the uses contemplated by this application and are intended to be within the scope of this application.

[0121] In some embodiments, the mitochondrial targeting peptide mimetic disclosed herein, such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono, bis or tris (tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), is for use in treating or preventing ALS, alpha-synucleinopathy, or TDP-43 proteinopathy in a subject in need thereof. In some embodiments, the mitochondrial targeting peptide mimetic is (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (e.g., (Ia)). In some embodiments, the subject has been diagnosed with ALS, alpha-synucleinopathy, or TDP-43 proteinopathy.

[0122] In other embodiments, the mitochondrial targeting peptidomimetics disclosed herein, such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono-, bis- or tris(tri)salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), are for use in improving muscle weakness, muscle wasting (atrophy), muscle cramps, muscle contractures, slowness of movement, poor balance, impaired coordination, change in voice quality, dysarthria, dysphagia, incomplete eye closure, drooling, emotional regulation disorder, premature death, increased expression of brain translocator protein-18 kDa (TSPO), and plasma accumulation of neurofilament light chain (NfL) in a subject having ALS. In some embodiments, the mitochondrial targeting peptidomimetic is (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (e.g., (Ia)). In some embodiments, the subject has been diagnosed as having ALS.

[0123] In some embodiments of the mitochondrial-targeting peptide mimics of the present technology, treatment or prevention includes treatment or prevention of one or more signs or symptoms of ALS, including one or more of muscle weakness, muscle wasting (atrophy), muscle spasms, muscle cramps, slowness of movement, poor balance, impaired coordination, changes in voice quality, dysarthria, dysphagia, incomplete eye closure, drooling, emotional regulation disorders, premature death, increased expression of brain translocator protein-18 kDa (TSPO), and plasma accumulation of NfL. In some embodiments, treatment or prevention refers to a delay in the onset of neurological symptoms of ALS as evaluated by the neurological scoring described herein.

[0124] In some embodiments of the peptide mimetics of the present technology, the mitochondrial-targeted peptide mimetics are intended to be administered to a subject separately, sequentially, or simultaneously with, or formulated to be so administered with, an additional therapeutic agent or additional therapeutic treatment. In some embodiments, the additional therapeutic agent is selected from the group consisting of riluzole (Rilutek®), edaravone (Radicava®), mecaserimin, baclofen (Lioresal®), diazepam (Valium®), dantrolene (Dantrium®), non-steroidal anti-inflammatory agents, anti-spasmodics (e.g., carbamazepine (Tegretol) or phenytoin (Dilantin®)), amitriptyline (Elavil®), nortriptyline (Pamelor™), and lorazepam (Ativan®). In some embodiments, the therapeutic agent is elamipretide (also known as SS-31 or Bendavia). In some embodiments, the peptide mimetic is for use in which the peptide mimetic and an additional therapeutic agent or combination of treatments have a synergistic effect in the prevention or treatment of ALS. In some embodiments, the additional therapeutic agent is levodopa. In some embodiments, the peptide mimetic is for use in which the peptide mimetic and an additional therapeutic agent or combination of treatments have a synergistic effect in the prevention or treatment of α-synucleinopathy. In some embodiments, the additional therapeutic agent is an antidepressant such as a selective serotonin reuptake inhibitor (SSRI) including trazodone. In some embodiments, the peptide mimetic is for use in which the peptide mimetic, an additional therapeutic agent or combination of treatments have a synergistic effect in the prevention or treatment of TDP-43 proteinopathy.

[0125] Synthesis of Mitochondrial-Targeted Peptide Mimetics The peptide mimetic compounds of the present technology can be prepared in whole or in part using peptide synthesis methods such as conventional liquid phase (also known as solution phase) peptide synthesis or solid phase peptide synthesis, or by peptide synthesis using an automated peptide synthesizer (Kelley et al., Genetics Engineering Principles and Methods, Setlow, J.K. eds., Plenum Press NY. (1990) Vol. 12, pp. 1 to 19, Stewart et al., Solid-Phase Peptide Synthesis (1989) W.H., Houghten, Proc. Natl. Acad. Sci. USA (1985) 82: p. 5132). The peptide mimetics thus produced can be collected or purified by conventional methods such as chromatography, for example, gel filtration chromatography, ion exchange column chromatography, affinity chromatography, reverse phase column chromatography, and HPLC, ammonium sulfate fractionation, ultrafiltration, and immunoabsorption.

[0126] In solid phase peptide synthesis, peptides are typically synthesized from the carbonyl group side (C-terminus) to the amino group side (N-terminus) of the amino acid chain. In certain embodiments, an amino-protected amino acid is covalently attached to a solid support material via the carboxyl group of the amino acid, typically via an ester or amide bond, and optionally via a linking group. The amino group is deprotected and reacted (i.e., "coupled") with the carbonyl group of a second amino-protected amino acid using a coupling reagent to produce a dipeptide attached to the solid support. After coupling, the resin is optionally treated with a capping reagent to cap any unreacted amine groups (rendering them inert towards subsequent coupling steps). These steps (i.e., deprotection, coupling, and optionally capping) can be repeated to form the desired peptide chain. Once the desired peptide chain is complete, the peptide can be cleaved from the solid support.

[0127] In certain embodiments, protecting groups used on the amino groups of amino acid residues (peptides and / or peptidomimetics) include the 9-fluorenylmethyloxycarbonyl group (Fmoc) and the t-butyloxycarbonyl (Boc). The Fmoc group is removed from the amino terminus with base, and the Boc group is removed with acid. In alternative embodiments, the amino protecting group can be an aralkyloxycarbonyl type of substituted or unsubstituted group such as formyl, acrylyl (Acr), benzoyl (Bz), acetyl (Ac), trifluoroacetyl, benzyloxycarbonyl (Z), p-chlorobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, benzhydryloxycarbonyl, 2(p-biphenylyl)isopropyloxycarbonyl, 2-(3,5-dimethoxyphenyl)isopropyloxycarbonyl, p-phenylazobenzyloxycarbonyl, triphenylphosphonoethyloxycarbonyl or 9-fluorenylmethyloxycarbonyl group (Fmoc), an alkyloxycarbonyl type of substituted or unsubstituted group such as tert-butyloxycarbonyl (BOC), tert-amyloxycarbonyl, diisopropylmethyloxycarbonyl, isopropyloxycarbonyl, ethyloxycarbonyl, allyloxycarbonyl, 2methylsulfonylethyloxycarbonyl or 2,2,2-trichloroethyloxycarbonyl group, a cycloalkyloxycarbonyl type of group such as cyclopentyloxycarbonyl, cyclohexyloxycarbonyl, adamantyloxycarbonyl or isobornyloxycarbonyl group, and a group containing a heteroatom such as benzenesulfonyl, p-toluenesulfonyl, mesitylenesulfonyl, methoxymethylphenylsulfonyl, 2-nitrobenzenesulfonyl, 2-nitrobenzenesulfenyl, 4-nitrobenzenesulfonyl or 4-nitrobenzenesulfenyl group.

[0128] Many amino acids have reactive functional groups on their side chains. In certain embodiments, such functional groups are protected to prevent the incoming amino acid from reacting with the functional group. Protecting groups used with these functional groups must be stable to the conditions of peptide and / or peptidomimetic synthesis, but can be removed before, after, or concomitant with cleavage of the peptide from the solid support (if the support is attached), or upon final deprotection in solution-phase synthesis. Further reference is also made to the following. As a comprehensive review of protecting groups commonly used in peptide synthesis, see Isidro-Llobet, A., Alvarez, M., Albericio, F., “Amino Acid-Protecting Groups”; Chem. Rev., 109:2455-2504 (2009) (this protecting group can also be used in the synthesis of peptidomimetics that contain functional groups found in peptides).

[0129] In certain embodiments, the solid support material used in solid-phase peptide synthesis methods is a gel-type support such as polystyrene, polyacrylamide, or polyethylene glycol. Alternatively, materials such as controlled pore glass, cellulose fibers, or polystyrene can be functionalized on their surfaces to provide a solid support for peptide synthesis.

[0130] Coupling reagents that can be used in the solid-phase (or solution-phase) peptide synthesis described herein are typically carbodiimide reagents. Examples of carbodiimide reagents include N,N'-dicyclohexylcarbodiimide (DCC), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and its HCl salt (EDC·HCl), N-cyclohexyl-N'-isopropylcarbodiimide (CIC), N,N'-diisopropylcarbodiimide (DIC), N-tert-butyl-N'-methylcarbodiimide (BMC), N-tert-butyl-N'-ethylcarbodiimide (BEC), bis[[4-(2,2-dimethyl-1,3-dioxolyl)]-methyl]carbodiimide (BDDC), and N,N-dicyclopentylcarbodiimide, but are not limited thereto. DCC is a preferred coupling reagent. Other coupling agents include HATU and HBTU, which are generally used in combination with organic bases such as DIEA and hindered pyridine-type bases such as lutidine or collidine.

[0131] In some embodiments, as described in Fuller et al., Urethane-Protected α-Amino Acid N-Carboxyanhydrides and Peptide Synthesis, Biopolymers (Peptide Science), Vol. 40, 183-205 (1996) and WO2018 / 034901, amino acids can be activated for coupling to peptides or peptidomimetics by forming N-carboxyanhydrides.

[0132] In certain exemplary embodiments, compounds useful in the therapeutic methods described herein can be synthesized by a convergent method according to the solid-phase synthesis shown in Scheme 1. For reference in the following schemes,

Chemical formula

Chemical formula

Chem.

Chem.

[0133] For example, the compounds shown below can be synthesized in such a manner as shown in Scheme 2.

Chem.

[0134] For reference in the following scheme,

Chem.

Chem.

Chem.

Chem.

[0135] The compounds of the present technology can also be synthesized according to conventional liquid-phase peptide synthesis routes, for example, according to Scheme 3.

Chem.

[0136] For example, the compounds shown below can be synthesized in such a manner as shown in Scheme 4.

Chem.

Chem.

[0137] In some embodiments, compound 7a can be synthesized as shown in Scheme 5 below (see also WO2019 / 118878, which is incorporated herein by reference). Compound 12a can be prepared as shown in Scheme 6 below.

Chemical formula

Chemical formula

[0138] Step a: Synthesis of (S)-2-((R)-2-((tert-Butoxycarbonyl)amino)-5-guanidinopentanamido)-3-(4-hydroxy-2,6-dimethylphenyl)propanoate (3a). To a suspension (800 mL) of 2,6-Dmt-OBn·HCl (2a, 45.0 g, 134 mmol) in ACN, NMM (32.7 mL, 298 mmol) was added at 0 °C. The reaction mixture was stirred until it became clear. Next, Boc-D-Arg-OH·HCl (1a, 46.3 g, 149 mmol) and HOBt·H2O (9.11 g, 59.5 mmol) were added to the reaction mixture and stirred for 15 minutes. Finally, EDC·HCl (38.5 g, 201 mmol) was added and the mixture was stirred at 0 °C for 4 hours. Then, EtOAc (450 mL) and 1 N HCl in brine (300 mL) were added. The combined organic extracts were washed with 1 N HCl in brine (7 × 150 mL), NaHCO3 / brine (300 mL, and until the pH of the aqueous layer reached about pH = 6 - 7), dried over Na2SO4, filtered, and concentrated to give 86.0 g (97%) of Boc-D-Arg-DMT-OBn (3a), which was used without further purification. 1 H-NMR (400 MHz, methanol-d4) δ 7.33 - 7.18 (m, 5H), 6.43 (s, 2H), 5.06 (s, 2H) 4.71 (t, J = 7.8 Hz, 1H), 4.07 (t, J = 6.7 Hz, 1H), 3.19 - 3.09 (m, 3H), 3.03 - 2.97 (m, 1H), 2.23 (s, 6H), 1.72 - 1.65 (m, 1H), 1.54 - 1.43 (m, 3H), 1.45 (s, 9H).

[0139] Step b: Synthesis of (S)-2-((R)-2-((tert-Butoxycarbonyl)amino)-5-guanidinopentanamido)-3-(4-hydroxy-2,6-dimethylphenyl)propanoic acid (4a). Pd / C (10% w / w, 14.0 g) was added to a solution (1000 mL) of Boc-D-Arg-DM-Tyr-OBn (3a, 84.0 g, 142 mmol) in MeOH. Hydrogen was purged into the reaction mixture at room temperature for 4 hours. The reaction mixture was then filtered through filter paper and washed with MeOH (150 mL). The solvent was removed by evaporation. The white foamy product 4a was obtained (74.0 g, 93%) and used without further purification. 1 H-NMR (400 MHz, methanol-d4) δ 6.44 (s, 2H), 4.68 (t, J = 7.2 Hz, 1H), 4.04 (t, J = 6.8 Hz, 1H), 3.15 - 3.09 (m, 3H), 3.02 - 2.94 (m, 1H), 2.29 (s, 6H), 1.74 - 1.59 (m, 1H), 1.54 - 1.43 (m, 1H), 1.45 (s, 9H).

[0140] Step c: Synthesis of tert-butyl ((6R,9S,12S)-1-amino-12-(3-benzyl-1,2,4-oxadiazol-5-yl)-9-(4-hydroxy-2,6-dimethylbenzyl)-1-imino-20,20-dimethyl-7,10,18-trioxo-19-oxa-2,8,11,17-tetraazhenicosan-6-yl)carbamate (6a). DMF (200 mL) was added to 4a (11.17 g, 24 mmol), and the mixture was stirred at room temperature for 15 minutes. 12a (10.65 g, 20 mmol) was added to the resulting suspension, and the mixture was stirred at room temperature for 20 minutes. After the addition of HOBt (612 mg, 4.00 mmol), the suspension was cooled in an ice bath. EDC·HCl (5.38 g, 28 mmol) was added all at once, and the reaction mixture was stirred for 2.5 hours in an ice bath and then for 4.5 hours at room temperature. The almost homogeneous reaction mixture was quenched with EtOAc (1500 mL), and the resulting solution was washed 10 times with brine / 0.5 M aqueous HCl solution (1:1, 400 mL). During the 6th and 9th washings, a gel formed in the aqueous phase. iPrOH (40 mL in each case) was added, and after repeated shaking, the layers became clear again. Thereafter, the organic phase was washed 6 times with brine / saturated aqueous NaHCO3 solution (9:1, 400 mL). During the 4th washing, a gel formed in the aqueous phase. iPrOH (40 mL) was added, and after repeated shaking, the layers separated easily. The organic phase was washed with brine (200 mL) and water (100 mL), and the solvent was removed under reduced pressure. To avoid difficulties in phase separation, vigorous shaking was not carried out during washing with water. As a result, 16.8 g of the crude product was obtained (6a, purity 97.0% by HPLC, white amorphous solid). 1 1H-NMR (300 MHz, methanol-d4) ppm: δ = 7.33 - 7.16 (m, 5H), 6.38 (s, 2H), 5.18 - 5.07 (m, 1H), 4.64 - 4.55 (m, 1H), 4.10 - 3.92 (m, 3H), 3.18 - 2.77 (m, 6H), 2.20 (s, 6H), 1.97 - 1.76 (m, 2H), 1.75 - 1.14 (m, 8H), 1.43 (s, 9H), 1.41 (s, 9H).

[0141] Step d: Synthesis of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (7a, also referred to herein as the trihydrochloride salt of (Ia-Compound I)). 6a (16.8 g) was dissolved in DCM (100 mL), cooled to 0 °C, and then TFA (20 mL) was added dropwise. The solution was stirred at 0 °C for 10 minutes and then at room temperature for 3 hours (LC / MS showed no starting material). The reaction mixture was then evaporated (0 - 5 °C) and additionally re-evaporated from DCM (100 mL, 0 - 5 °C). Purification by reverse-phase flash chromatography (cartridge C-18, 120G) was performed on the crude material divided into four portions. Then, all solvents were evaporated under reduced pressure at <40 °C. The white foam was dissolved in isopropanol (100 mL), and HCl (5 - 6 M) in 5 mL of isopropanol was added at 0 °C and evaporated under reduced pressure. This step was repeated three times. Additionally, 100 mL of ACN was added and the suspension was evaporated once more. As a result, a white powder of 7a was obtained as the trihydrochloride salt. 1 H-NMR (300 MHz, methanol-d4) δ 7.36 - 7.14 (m, 5H), 6.40 (s, 2H), 5.15 (dd, J = 8.5, 6.3 Hz, 1H), 4.68 (dd, J = 8.7, 7.5 Hz, 1H), 4.07 (s, 2H), 3.97 (t, J = 6.3 Hz, 1H), 3.18 (t, J = 6.9 Hz, 2H), 3.11 (dd, J = 14.2, 8.8 Hz, 1H), 2.95 - 2.84 (m, 3H), 2.22 (s, 6H), 2.02 - 1.59 (m, 6H), 1.57 - 1.28 (m, 4H). MS: EI-MS: m / z 608.4 [M + 1].

[0142] Synthesis of (S)-1-(3-benzyl-1,2,4-oxadiazol-5-yl)-5-((tert-butoxycarbonyl)amino)pentan-1-aminium 4-methylbenzenesulfonate (12a)

Chemical Structure

[0143] Step b: Synthesis of (9H-fluoren-9-yl)methyl tert-butyl (1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentane-1,5-diyl) (S)-dicarbamate (11a). To a solution of protected enantiomerically pure N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N6-(tert-butoxycarbonyl)-L-lysine (10a, 4.31 kg, 9.2 mol) and hydroxyimidamide 9a (1.1 equiv. “equivalent (equiv.)” or “equivalent (eq.)”) in ethyl acetate, NaHCO3 (3.0 equiv.) was added. The mixture was stirred at 25 °C for 20 min (min.). Then, propanephosphonic anhydride (T3P, 50% solution in ethyl acetate, 3.0 equiv. (equiv.)) was added, and the reaction mixture was heated to 80 °C and stirred for 4 h (conversion of compound 10a based on HPLC was about 60%). Then, compound 9a (1.1 equiv.) was added, and the reaction mixture was stirred at 80 °C for an additional 20 h. (About 10% of compound 10a remained). The reaction mixture was cooled to room temperature, saturated aqueous NaHCO3 solution (2.0 L) was added, and the mixture was extracted with ethyl acetate (3 × 1.0 L). Then, the combined organic layers were washed with brine (1 L), dried over anhydrous Na2SO4, filtered, and concentrated to obtain a crude residue, which was generally purified by silica gel column chromatography (petroleum ether (PE):EtOAc = 5:1) to give a crude product in ACN, (9H-fluoren-9-yl)methyl tert-butyl (1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentane-1,5-diyl) (S)-dicarbamate (11a), solution (19.7 kg, assay = 20%, chiral HPLC purity = 99.12A%, yield = 73%). 1 H-NMR (300 MHz, CDCl3): δ 7.78 (d, J = 7.5 Hz, 2H), 7.61 (d, J = 6.3 Hz, 2H), 7.42 (t, J = 7.5 Hz, 2H), 7.35 - 7.30 (m, 7H), 5.52 (br, 1H), 5.09 - 5.05 (m, 1H), 4.56 - 4.37 (m, 3H), 4.22 (t, J = 6.6 Hz, 1H), 4.08 (s, 2H), 1.95 - 1.86 (m, 2H), 1.48 - 1.42 (m, 11H) ppm. MS: (M - 100 + H)+ : m / z = 483.2

[0144] Step c: Synthesis of tert-butyl (S)-(5-amino-5-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)-carbamate (5a). TEA (2.5 eq) was added to a solution of the compound (9H-fluoren-9-yl)methyl tert-butyl (1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentane-1,5-diyl)(S)-dicarbamate (11a). The mixture was stirred at 20 - 25 °C for 15 h with a mechanical stirrer. The reaction mixture was diluted with tap water and MTBE. The separated aqueous layer was extracted once with MTBE. Both MTBE layers were combined and then washed with NH4Cl. Then, anhydrous Na2SO4 was added and the solution was stirred for at least 2 h, then filtered and washed with MTBE to obtain a solution of tert-butyl (S)-(5-amino-5-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)-carbamate (5a) in MTBE (32.9 kg, assay = 6.5%, yield = 88%). 1 1H-NMR (300 MHz, DMSO-d6): δ 7.33 - 7.25 (m, 5H), 6.78 (br, 1H), 5.09 - 5.05 (m, 1H), 4.56 - 4.37 (m, 3H), 4.06 (s, 2H), 3.98 (t, J = 6.6 Hz, 1H), 2.87 - 2.84 (m, 2H), 2.10 (s, 2H), 1.38 - 1.34 (m, 2H), 1.24 (s, 9H), 1.20 - 1.15 (m, 2H) ppm. MS: (M + H) + : m / z = 361.1

[0145] Step d: Synthesis of (S)-1-(3-benzyl-1,2,4-oxadiazol-5-yl)-5-((tert-butoxycarbonyl)amino)pentan-1-aminium 4-methylbenzenesulfonate (12a). p-Toluenesulfonic acid (PTSA) was added to a solution of crude tert-butyl (S)-(5-amino-5-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)-carbamate (5a) in MTBE to give (S)-1-(3-benzyl-1,2,4-oxadiazol-5-yl)-5-((tert-butoxycarbonyl)amino)pentan-1-aminium 4-methylbenzenesulfonate (12a) as a white solid (2.7 kg, yield = 85%, HPLC purity > 99%, ee > 99%). 1 1H-NMR (400 MHz, DMSO-d6): δ 8.74 (br, 3H), 7.48 (d, J = 8.0 Hz, 2H), 7.37 - 7.26 (m, 5H), 7.11 (d, J = 8.0 Hz, 2H), 6.77 (t, J = 5.2 Hz, 1H), 4.82 (t, J = 6.8 Hz, 1H), 4.17 (s, 2H), 2.90 - 2.86 (m, 2H), 2.29 (s, 3H), 1.39 - 1.36 (m, 11H), 1.35 - 1.28 (m, 2H) ppm. MS: (M - 172 + H) + : m / z = 361.1.

[0146] Treatment method The following discussion is presented by way of example only and is not intended to be limiting.

[0147] One aspect of the present technology includes a method for treating ALS, alpha-synucleinopathy, or TDP-43 proteinopathy in a subject diagnosed with, suspected of having, or at risk of having ALS, alpha-synucleinopathy, or TDP-43 proteinopathy. In therapeutic use, a composition or agent comprising a mitochondrial-targeted peptidomimetic such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (in each case mono-, bis- or tri(tris) salt), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.) is administered in an amount sufficient to cure or at least partially prevent the symptoms of the disease, including its complications and intermediate pathological phenotypes in the development of the disease, in a subject suspected of having or already suffering from ALS, alpha-synucleinopathy, or TDP-43 proteinopathy. In some embodiments of the methods of the present technology, the mitochondrial-targeted peptidomimetic is (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (e.g., (Ia)).

[0148] Other aspects of the technology include the use of a composition in the preparation of a medicament for treating or preventing ALS, alpha-synucleinopathy, or TDP-43 proteinopathy in a subject in need thereof. A composition or medicament comprising a mitochondrial-targeted peptidomimetic such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono-, bis- or tris-(tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.) is suitable for administration to a subject suspected of having or already suffering from ALS, alpha-synucleinopathy, or TDP-43 proteinopathy in an amount sufficient to alleviate one or more signs or symptoms of ALS, alpha-synucleinopathy, or TDP-43 proteinopathy in the subject. In some embodiments of the methods of the technology, the mitochondrial-targeted peptidomimetic is (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (e.g., (Ia)).

[0149] Subjects suffering from ALS, alpha-synucleinopathy, or TDP-43 proteinopathy can be identified by any or a combination of diagnostic or prognostic assays known in the art.

[0150] For therapeutic use, the mitochondrial-targeting peptidomimetic disclosed herein, such as 2(R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono-, bis- or tris(tri)salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), is administered to a subject. In some embodiments, the peptidomimetic composition is administered 1, 2, 3, 4, or 5 times per day. In some embodiments, the peptidomimetic composition is administered more than 5 times per day. Additionally or alternatively, in some embodiments, the peptidomimetic composition is administered daily, every other day, every 3 days, every 4 days, every 5 days, or every 6 days. In some embodiments, the peptidomimetic composition is administered weekly, every 2 weeks, every 3 weeks, or monthly. In some embodiments, the peptidomimetic composition is administered for a period of 1, 2, 3, 4, or 5 weeks. In some embodiments, the peptidomimetic is administered for 6 weeks or more. In some embodiments, the peptidomimetic is administered for 12 weeks or more. In some embodiments, the peptidomimetic is administered for a period of less than 1 year. In some embodiments, the peptidomimetic is administered for a period exceeding 1 year, or until the signs or symptoms of ALS, alpha-synucleinopathy, or TDP-43 proteinopathy are alleviated in the subject. In some embodiments, the peptidomimetic is administered according to a protocol recommended by a physician from the time of diagnosis of a subject having, suspected of having, or at risk of having ALS, alpha-synucleinopathy, or TDP-43 proteinopathy until the end of life.

[0151] Subjects to be treated according to the present treatment method can be any mammal, including, for example, farm animals such as sheep, pigs, cows, and horses; pet animals such as dogs and cats; and laboratory animals such as rats, mice, and rabbits. In some embodiments, the mammal is a human.

[0152] In some embodiments, treatment of a subject diagnosed with or suspected of having ALS with one or more mitochondrial targeting peptide mimetics improves or eliminates one or more of the following symptoms of ALS. Muscle weakness, muscle wasting (atrophy), muscle spasms, muscle cramps, slowness of movement, poor balance, impaired coordination, change in voice quality, dysarthria, dysphagia, incomplete eye closure, drooling, emotional regulation disorder, and premature death. In some embodiments, treatment of a subject diagnosed with or suspected of having ALS with one or more mitochondrial targeting peptide mimetics improves or reduces the increase in brain translocator protein-18 kDa (TSPO) expression. In some embodiments, treatment of a subject diagnosed with or suspected of having ALS with one or more mitochondrial targeting peptide mimetics improves or eliminates the plasma accumulation of NfL. In some embodiments, treatment of a subject diagnosed with or suspected of having ALS with one or more mitochondrial targeting peptide mimetics increases the survival / lifespan of the subject. In some embodiments, the success of treatment with mitochondrial targeting peptide mimetics is determined by detecting improvement in the subject's symptoms compared to one or more of the following. (1) Baseline measurements or symptom levels detected before or at the start of treatment, (2) measurements or symptom levels from control subjects or a population of control subjects, where the control subjects exhibit one or more symptoms of ALS and (i) have not been administered a mitochondrial targeting peptide mimetic or (ii) have been administered a control peptide or peptide mimetic, or (3) a standard substance.

[0153] In some embodiments, treatment of a subject diagnosed with or suspected of having alpha-synucleinopathy with one or more mitochondrial targeting peptide mimetics ameliorates or eliminates one or more of the symptoms of alpha-synucleinopathy, including but not limited to loss of dopaminergic neurons in the subject.

[0154] In some embodiments, treatment of a subject diagnosed with or suspected of having TDP-43 proteinopathy with one or more mitochondrial targeting peptide mimetics ameliorates or eliminates one or more of the symptoms of TDP-43 proteinopathy, including but not limited to reduction of neurite length in the subject.

[0155] Preventive method In one aspect, the technology of the present invention provides a method for preventing or delaying the onset of ALS, alpha-synucleinopathy, or TDP-43 proteinopathy, or one or more symptoms of ALS, alpha-synucleinopathy, or TDP-43 proteinopathy, in a subject having or at risk of developing ALS, alpha-synucleinopathy, or TDP-43 proteinopathy. In prophylactic use, a pharmaceutical composition or agent of a mitochondrial-targeted peptide mimetic such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono, bis or tris (tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.) is administered in an amount sufficient to eliminate or reduce the risk of the disease, including the biochemical, histological and / or behavioral symptoms of the disease, its complications, and the intermediate pathological phenotypes that appear during the onset of the disease, in a subject who is highly susceptible to or otherwise at risk of ALS, alpha-synucleinopathy, or TDP-43 proteinopathy, or delay the onset of the disease. In some embodiments of the methods of the present technology, the mitochondrial-targeted peptide mimetic is (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (e.g., (Ia)).

[0156] Administration of a prophylactic mitochondrially targeted peptide mimetic can be carried out prior to the manifestation of characteristic symptoms of a disease or disorder such that the disease or disorder is prevented or alternatively its progression is delayed.

[0157] For prophylactic use, (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (in each case mono-, bis- or tri-(tris) salts), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.) and other mitochondrial targeting peptide mimetics are administered to a subject. In some embodiments, the peptide mimetic composition is administered once, twice, three times, four times, or five times a day. In some embodiments, the peptide mimetic composition is administered more than five times a day. Additionally or alternatively, in some embodiments, the peptide mimetic composition is administered daily, every other day, every three days, every four days, every five days, or every six days. In some embodiments, the peptide mimetic composition is administered weekly, every two weeks, every three weeks, or monthly. In some embodiments, the peptide mimetic composition is administered for a period of 1, 2, 3, 4, or 5 weeks. In some embodiments, the peptide mimetic is administered for a period of 6 weeks or more. In some embodiments, the peptide mimetic is administered for a period of 12 weeks or more. In some embodiments, the peptide mimetic is administered for a period of less than 1 year. In some embodiments, the peptide mimetic is administered for a period of more than 1 year. In some embodiments, the peptide mimetic is administered according to a protocol recommended by a physician from the time of diagnosis of a subject having, suspected of having, or at risk of having ALS, alpha-synucleinopathy, or TDP-43 proteinopathy until the end of life.

[0158] In some embodiments, treatment with a mitochondrial-targeting peptide mimetic will prevent or delay the onset of one or more of the following conditions. Muscle weakness, muscle wasting (atrophy), muscle cramps, muscle contractures, slowness of movement, poor balance, impaired coordination, change in voice quality, dysarthria, dysphagia, incomplete eyelid closure, drooling, emotional regulation disorder, and / or premature death. In some embodiments, treatment with a mitochondrial-targeting peptide mimetic will prevent or delay the onset of increased brain translocator protein-18 kDa (TSPO) expression. In some embodiments, treatment with a mitochondrial-targeting peptide mimetic will prevent or delay the onset of plasma accumulation of neurofilament light chain (NfL). In some embodiments, treatment with a mitochondrial-targeting peptide mimetic will prevent or delay premature death. In some embodiments, treatment refers to delaying the onset of neurological symptoms of ALS as evaluated by the neurological scoring described herein.

[0159] In some embodiments, treatment with a mitochondrial-targeting peptide mimetic will prevent, delay, or attenuate the loss of dopaminergic neurons in a subject.

[0160] In some embodiments, treatment with a mitochondrial-targeting peptide mimetic will prevent or delay the reduction of neurite length in a subject.

[0161] The mammal treated according to this prevention method can be any mammal, including, for example, farm animals such as sheep, pigs, cows, and horses; pet animals such as dogs and cats; and laboratory animals such as rats, mice, and rabbits. In some embodiments, the mammal is a human.

[0162] Determination of the biological effects of mitochondrial-targeting peptide mimetic-based therapeutics In various embodiments, appropriate in vitro or in vivo assays are performed to determine the effect of a particular mitochondria-targeting peptide mimetic-based therapeutic agent and whether its administration is indicated for treatment. In various embodiments, in vitro assays are performed in representative animal models to determine whether a given mitochondria-targeting peptide mimetic-based therapeutic agent exerts the desired effect of reducing or eliminating the signs and / or symptoms of ALS, alpha-synucleinopathy, or TDP-43 proteinopathy.

[0163] Animal model The compounds for use in therapy can be tested in suitable animal model systems including, but not limited to, rats, mice, chickens, cows, monkeys, rabbits, etc. prior to testing in human subjects. Similarly, any of the animal model systems known in the art can be used prior to administration to human subjects for in vivo testing. In some embodiments, the in vitro or in vivo testing is directed to the biological function of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono, bis or tris (tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.). In some embodiments, the animal model is the SOD1 G93A mouse model of ALS. In some embodiments, the animal model is a Sprague Dawley rat. In some embodiments, the animal model is a mutant α-synuclein transgenic mouse. In some embodiments, the animal model is a prp-TDP-43 A315T -UeGFP mouse model (Gautam, et al. Acta Neuropathol. 2019 Jan;137(1):47-69).

[0164] Mode of administration and effective dosage A cell, organ, or tissue can be contacted with a mitochondrial-targeting peptidomimetic of the present technology, such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono, bis or tris (tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, acetate, tartrate, trifluoroacetate, chloride salt, tris HCl salt, di HCl salt, mono HCl salt or tosylate, etc.) by any method known to those skilled in the art. In some embodiments of the methods of the present technology, the mitochondrial-targeting peptidomimetic is (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (e.g., (Ia)). Suitable methods include in vitro, ex vivo, or in vivo methods. In vivo methods typically involve administering the mitochondrial-targeting peptidomimetic to a mammal, suitably a human.When used in vivo for therapy, (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono, bis or tris (tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), such as mitochondrial targeting peptide mimetics, are administered to a subject in an effective amount (i.e., an amount having the desired therapeutic effect). The dosage and administration regimen will depend on the degree of the disease, disorder or condition in the subject, the characteristics of the particular mitochondrial targeting peptide mimetic used, such as its therapeutic index, the subject, and the medical history of the subject.

[0165] The effective amount can be determined by methods well known to physicians and clinicians during preclinical and clinical trials. The effective amount of the peptide mimetic useful in this method can be administered to a mammal in need thereof by any of several well-known methods for administering pharmaceutical compounds. The peptide mimetic can be administered systemically or locally.

[0166] The peptidomimetics can be formulated as pharmaceutically acceptable salts. The term "pharmaceutically acceptable salts" means salts prepared from bases or acids acceptable for administration to patients such as mammals (e.g., salts having mammalian safety acceptable for a given dosing regimen). However, it should be understood that salts such as salts of intermediate compounds not intended for administration to patients need not be pharmaceutically acceptable salts. Pharmaceutically acceptable salts can be derived from pharmaceutically acceptable inorganic or organic bases, and pharmaceutically acceptable inorganic or organic acids. In addition, when a peptide or peptidomimetic contains both a basic moiety such as an amine, pyridine or imidazole, and an acidic moiety such as a carboxylic acid or tetrazole, zwitterions can be formed and are included within the term "salt" as used herein. Salts derived from pharmaceutically acceptable inorganic bases include ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, and zinc salts, etc. Salts derived from pharmaceutically acceptable organic bases include arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-methylmorpholine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperadine, polyamine resin, procaine, purine, theobromine, trimethylamine (NEt3), trimethylamine, tripropylamine, tromethamine, etc., organic bases in protonated form (e.g., [HNEt3] +When ) contains a salt, salts of primary, secondary, and tertiary amines including substituted amines, cyclic amines, natural amines, etc. are included. Salts derived from pharmaceutically acceptable inorganic acids include salts of boric acid, carbonic acid, hydrohalic acids (hydrobromic acid, hydrochloric acid, hydrofluoric acid or hydroiodic acid), nitric acid, phosphoric acid, sulfamic acid and sulfuric acid. Salts derived from pharmaceutically acceptable organic acids include aliphatic hydroxy acids (e.g., citric acid, gluconic acid, glycolic acid, lactic acid, lactobionic acid, malic acid, and tartaric acid), aliphatic monocarboxylic acids (e.g., acetic acid, butyric acid, formic acid, propionic acid, and trifluoroacetic acid), amino acids (e.g., aspartic acid and glutamic acid), aromatic carboxylic acids (e.g., benzoic acid, p-chlorobenzoic acid, diphenylacetic acid, gentisic acid, hippuric acid, and triphenylacetic acid), aromatic hydroxy acids (e.g., o-hydroxybenzoic acid, p-hydroxybenzoic acid, 1-hydroxynaphthalene-2-carboxylic acid and 3-hydroxynaphthalene-2-carboxylic acid), ascorbic acid, dicarboxylic acids (e.g., fumaric acid, maleic acid, oxalic acid, and succinic acid), glucuronic acid, mandelic acid, mucic acid, nicotinic acid, orotic acid, pamoic acid, pantothenic acid, sulfonic acids (e.g., benzenesulfonic acid, camphorsulfonic acid, edetic acid, ethanesulfonic acid, isethionic acid, methanesulfonic acid, naphthalenesulfonic acid, naphthalene-1,5-disulfonic acid, naphthalene-2,6-disulfonic acid and p-toluenesulfonic acid (PTSA)), and xinafoic acid.In some embodiments, the pharmaceutically acceptable counterion is selected from the group consisting of acetate, benzoate, besylate, bromide, camphorsulfonate, chloride, chlorotheophyllinate, citrate, ethanedisulfonate, fumarate, gluceptate, gluconate, glucuronate, hippurate, iodide, isethionate, lactate, lactobionate, lauryl sulfate, malate, maleate, mesylate, methyl sulfate, naphthoate, sapsylate, nitrate, octadecanoate, oleate, oxalate, pamoate, phosphate, polygalacturonate, succinate, sulfate, sulfosalicylate, tartrate, tosylate, and trifluoroacetate. In some embodiments, the salt is tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (in each case mono, bis or tri(tris) salt), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, bistosylate, or tritosylate. In some embodiments, the peptidomimetic is formulated as mono HCl, bis HCl salt, or tri(or tris) HCl salt (e.g., (Ia)).

[0167] (R)-2-Amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono-, bis- or tris(tri)salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.) such as the mitochondrial-targeted peptidomimetics described herein can be incorporated into a pharmaceutical composition for administration to a subject, alone or in combination, for the treatment or prevention of the diseases, disorders or conditions described herein. The peptidomimetics can be formulated with other compounds such as therapeutic agents, peptides, other peptidomimetics, or mixtures thereof. In some embodiments of the methods of the present technology, the mitochondrial-targeted peptidomimetics are (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (e.g., (Ia)). Such compositions typically comprise an active agent and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition can be used as an agent for administration to a subject suffering from ALS, alpha-synucleinopathy, or TDP-43 proteinopathy, or in the preparation of an agent. Pharmaceutically acceptable carriers include, for example, saline, solvents, dispersion media, coating agents, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc., which are compatible with pharmaceutical administration. Additionally, auxiliary active compounds can also be incorporated into the composition.

[0168] Pharmaceutical compositions are typically formulated to be compatible with their intended route of administration. Examples of routes of administration include parenteral (e.g., intravenous, intradermal, intraperitoneal, or subcutaneous), oral, intravitreal, inhalation, transdermal (topical), intraocular, ophthalmic, intrathecal, intracerebroventricular, iontophoresis, and transmucosal administration. In some embodiments, the route of administration is oral. In some embodiments, the route of administration is subcutaneous. As solutions or suspensions for parenteral, intradermal, or subcutaneous application, they can contain the following components. Sterile diluents such as water for injection, aqueous saline solution, fixed oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antibacterial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetate, citrate, phosphate, and agents for adjusting tonicity such as sodium chloride or dextrose. The pH can be adjusted with an acid or base such as hydrochloric acid or sodium hydroxide. Parenteral preparations can be enclosed in ampoules, disposable syringes, or multiple-dose vials made of glass or plastic. For the convenience of the patient or treating physician, the dosage formulation can be provided alone or in a kit containing all the equipment necessary for the progress of the treatment (e.g., vials of the drug, vials of diluent, syringes, and needles) (e.g., for 7 days of treatment).

[0169] Pharmaceutical compositions suitable for injectable use can contain sterile aqueous solutions (when water-soluble), or dispersions and sterile powders for the immediate preparation of sterile injectable solutions or dispersions. In the case of intravenous administration, suitable carriers include saline, bacteriostatic water, CREMOPHOR EL (trademark) (BASF, Parsippany, N.J.), or phosphate-buffered saline (PBS). In any case, the compositions for parenteral administration must be sterile and fluid to the extent that they are easily injectable. The compositions must be stable under the conditions of manufacture and storage and must be protected against the contaminating action of microorganisms such as bacteria and fungi.

[0170] Compositions containing mitochondrially targeted peptide mimetics can include a carrier, which can be, for example, a solvent or dispersion medium including water, ethanol, polyols (such as glycerol, propylene glycol, and liquid polyethylene glycol, etc.), and suitable mixtures thereof. Suitable fluidity can be maintained, for example, by the use of coatings such as lecithin, by maintaining the required particle size in the case of dispersion, and by the use of surfactants. Prevention of microbial action can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc. Oxidation can be prevented by including glutathione and other antioxidants. In many cases, it is advantageous to include in the composition isotonic agents, such as sugars, polyhydric alcohols such as mannitol, sorbitol, etc., or sodium chloride. Sustained absorption of injectable compositions can be brought about by including in the composition an agent that delays absorption, such as aluminum monostearate or gelatin.

[0171] Sterile injectable solutions can be prepared by incorporating the required amount of the active compound into a suitable solvent containing, as required, one or a combination of the ingredients listed above, followed by filter sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and the other ingredients required from those listed above. In the case of sterile powders for preparing sterile injectable solutions, typical methods of preparation include vacuum drying and lyophilization from which powders of the active ingredient plus any additional desired ingredients can be obtained from its solution that has been pre-sterilized by filtration.

[0172] Oral compositions generally contain an inert diluent or an edible carrier. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules, such as gelatin capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash. Pharmaceutically compatible binders and / or adjuvant materials can be included as part of the composition. Tablets, pills, capsules, troches, etc. can contain any of the following ingredients, or compounds of similar nature. Binders such as microcrystalline cellulose, tragacanth gum or gelatin; Excipients such as starch or lactose, Disintegrants such as alginic acid, primogel, or corn starch; Lubricants such as magnesium stearate or stearates; Glidants such as colloidal silicon dioxide; Sweetening agents such as sucrose or saccharin; or Flavoring agents such as peppermint, methyl salicylate, or orange flavor.

[0173] One is that a compound, therapeutic agent, peptide, peptidomimetic, or a mixture thereof can be diluted with an inert material or the volume can be increased. These diluents can include carbohydrates, especially mannitol, lactose, lactose anhydrous, cellulose, sucrose, modified dextran, and starch. Certain inorganic salts including tricalcium phosphate, magnesium carbonate and sodium chloride can also be used as fillers. Some commercially available diluents are Fast-Flo, Emdex, STA-Rx1500, Emcompress and Avicel.

[0174] Disintegrants can be included in the formulation of a solid dosage form of a compound, therapeutic agent, peptide, peptidomimetic, or a mixture thereof with an inert substance. Materials used as disintegrants include, but are not limited to, starches including the commercially available disintegrant Explotab based on starch. Sodium starch glycolate, Amberlite, sodium carboxymethyl cellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acidic carboxymethyl cellulose, natural sponges, and bentonite can all be used. Another form of disintegrant is an insoluble cation exchange resin. Powder gums can be used as disintegrants and binders and can include powder gums such as agar, karaya, or tragacanth. Alginate and its sodium salts are also useful as disintegrants.

[0175] Binders are used to hold a compound, therapeutic agent, peptide, peptidomimetic, or a mixture thereof together with an inert material to form hard tablets and can include materials derived from natural products such as acacia, tragacanth, starch, and gelatin. Others include methylcellulose (MC), ethylcellulose (EC), and carboxymethylcellulose (CMC). Polyvinylpyrrolidone (PVP) and hydroxypropylmethylcellulose (HPMC) can both be used in an alcohol solution to granulate a therapeutic agent.

[0176] Antifriction agents may be included in the formulation of a compound, therapeutic agent, peptide, peptidomimetic, or a mixture thereof to prevent adhesion during the formulation process. Lubricants can be used as a layer between the therapeutic agent and the wall of the die and can include, but are not limited to, stearic acid including magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils, and waxes. Soluble lubricants such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycols of various molecular weights, and Carbowax 4000 and 6000 can also be used.

[0177] During formulation, a flow promoter that can improve the flow properties of the drug and assist in rearrangement during compression can be added. The flow promoter may include starch, talc, fumed silica, calcined silica, and hydrated silicoaluminate.

[0178] Surfactants may be added as wetting agents to assist in dissolving the compound, therapeutic agent, peptide, peptidomimetic, or mixtures thereof in an aqueous environment. The surfactants may include anionic detergents such as sodium lauryl sulfate, sodium dioctyl sulfosuccinate, and sodium dioctyl sulfonate. Cationic detergents that can be used and can include benzalkonium chloride and benzethonium chloride. Potential nonionic detergents that can be included in the formulation as surfactants include lauromacrogol 400, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50, and 60, glyceryl monostearate, polysorbate 40, 60, 65, and 80, sucrose fatty acid esters, methylcellulose, and carboxymethylcellulose. These surfactants may be present in the formulation of the compounds, therapeutic agents, peptides, peptidomimetics, or mixtures or derivatives thereof of this technology either alone or as mixtures in different ratios.

[0179] Pharmaceuticals that can be used orally include push-fit capsules made of gelatin, as well as soft sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. The push-fit capsules can contain the active ingredient in the active ingredient mixed with a filler such as lactose, a binder such as starch, and / or a lubricant such as talc or magnesium stearate, and optionally a stabilizer. In soft capsules, the active compound can be dissolved or suspended in a suitable liquid such as fatty oil, liquid paraffin, or liquid polyethylene glycol. In addition, a stabilizer may be added. Microspheres formulated for oral administration can also be used. Such microparticles are well defined in the art. All formulations for oral administration need to be in a dosage appropriate for such administration.

[0180] In the case of administration by inhalation, the compounds, therapeutic agents, peptides, peptidomimetics or mixtures thereof for use according to the present application can be conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or nebulizer using a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In some embodiments, the compound can be delivered in the form of an aerosol spray from a pressurized container or dispenser or nebulizer containing a suitable propellant, such as a gas like carbon dioxide. Such methods include those described in U.S. Patent No. 6,468,798. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve for delivering a metered amount. Capsules and cartridges, for example of gelatin, for use in an inhaler or injector can be formulated to contain a powder mixture of the compound and a suitable powder base, such as lactose or starch.

[0181] Compounds, therapeutic agents, peptides, peptidomimetics, or mixtures thereof can be delivered to the mammalian lung during inhalation and cross the pulmonary epithelium into the bloodstream. Other reports of inhaled molecules include Adjei et al., Pharm Res 7:565-569 (1990), Adjei et al., Int J Pharmaceutics 63:135-144 (1990) (leuprolide acetate), Braquet et al., J Cardiovasc Pharmacol 13(suppl.5):143-146 (1989) (endothelin-1), Hubbard et al., Annal Int Med 3:206-212 (1989) (antitrypsin), Smith et al., 1989, J Clin Invest 84:1145-1146 (a-1-proteinase), Oswein et al., 1990, “Aerosolization of Proteins”, Proceedings of Symposium on Respiratory Drug Delivery II, Keystone, Colorado, March, (recombinant human growth hormone), Debs et al., 1988, J Immunol 140:3482-3488 (interferon gamma and tumor necrosis factor alpha) and Platz et al U.S. Patent No. 5,284,656 (granulocyte colony stimulating factor, incorporated by reference). Methods and compositions for pulmonary delivery of drugs for systemic effects are described in U.S. Patent No. 5,451,569, issued September 19, 1995 to Wong et al. (incorporated by reference).

[0182] Contemplated for use in the practice of this technology are a wide range of mechanical devices designed for the pulmonary delivery of therapeutic products, including but not limited to nebulizers, metered dose inhalers, and powder inhalers, all of which are familiar to those skilled in the art.

[0183] Some specific examples of commercially available devices suitable for practicing the present technology are the Ultravent nebulizer manufactured by Mallinckrodt, Inc., St. Louis, Mo., the Acorn II nebulizer manufactured by Marquest Medical Products, Englewood, Colo., the Ventolin metered dose inhaler manufactured by Glaxo, Inc., Research Triangle Park, North Carolina, and the Spinhaler powder inhaler manufactured by Fisons Corp., Bedford, Mass.

[0184] In the case of ophthalmic or intraocular preparations, any suitable mode for delivering to the eye or the region near the eye the mitochondrial targeting peptide mimetic described herein (with or without a therapeutic agent, peptide or other peptide mimetic), (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), its pharmaceutically acceptable salts (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono-, bis- or tris-(tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.) or pharmaceutical compositions thereof may be used. For ophthalmic preparations, see generally Mitra (ed.), Ophthalmic Drug Delivery Systems, Marcel Dekker, Inc., New York, N.Y. (1993) and / or Havener, W.H., Ocular Pharmacology, C.V. Mosby Co., St. Louis (1983). Non-limiting examples of suitable formulations for administration into or near the eye include, but are not limited to, ophthalmic inserts, minitablets, and topical formulations such as eye drops, ointments, and in situ gels.In one embodiment, the contact lens is coated with a mitochondrial targeting peptide mimetic as described herein, such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono-, bis- or tris(tri)salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.). In some embodiments, a single dose comprises from 0.1 ng to 5000 μg, from 1 ng to 500 μg, or from 10 ng to 100 μg of the mitochondrial targeting peptide mimetic administered to the eye.

[0185] The eye drops contain a sterile liquid preparation that can be administered directly to the eye. In some embodiments, the eye drops containing one or more of the mitochondrial targeting peptide mimetics described herein, such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono-, bis- or tris-(tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), further contain one or more preservatives. In some embodiments, the optimal pH of the eye drops is equal to the pH of the tear fluid and is about 7.4.

[0186] In situ gel is a viscous liquid that exhibits the ability to transition from a sol to a gel when affected by external factors such as appropriate pH, temperature, and the presence of electrolytes. This property delays drug excretion from the ocular surface and increases the bioavailability of the active ingredient. Polymers commonly used in in situ gel formulations include, but are not limited to, gellan gum, poloxamer, and cellulose acetate phthalate.

[0187] In the case of topical administration, the compound, therapeutic agent, peptide, peptidomimetic, or mixtures thereof can be formulated as a solution, gel, ointment, cream, suspension, etc., as is well known in the art. An ointment is a semi-solid dosage form for external use such as for topical use on the eye or skin. In some embodiments, the ointment comprises a solid or semi-solid hydrocarbon base material having a melting point or softening point close to the core temperature of a human. In some embodiments, an ointment applied to the eye breaks down into small droplets, which remain in the conjunctival sac for a longer period of time and thus increase bioavailability.

[0188] An ocular insert is a solid or semi-solid dosage form that does not have the drawbacks of conventional ophthalmic drug forms. They are less affected by defense mechanisms such as outflow through the nasolacrimal duct, show the ability to remain in the conjunctival sac for a long time, and are more stable than conventional dosage forms. They also offer advantages such as accurate dosing of one or more mitochondrial targeting peptide mimics, slow release of one or more mitochondrial targeting peptide mimics at a constant rate, and limitation of systemic absorption of one or more mitochondrial targeting peptide mimics. In some embodiments, the ocular insert comprises one or more mitochondrial targeting peptide mimics described herein, such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate salt, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono, bis or tris (tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.) and one or more polymer materials. Polymer materials include, but are not limited to, methylcellulose and its derivatives (e.g., hydroxypropylmethylcellulose (HPMC)), ethylcellulose, polyvinylpyrrolidone (PVP K-90), polyvinyl alcohol, chitosan, carboxymethylchitosan, gelatin, and various mixtures of the aforementioned polymers.

[0189] The mini-tablet is a biodegradable solid dosage form that migrates into a gel after application to the conjunctival sac, thereby extending the contact period between the active ingredient and the ocular surface and then increasing the bioavailability of the active ingredient. The advantages of the mini-tablet include easy application to the conjunctival sac, resistance to defense mechanisms such as tears or outflow through the nasolacrimal duct, longer contact with the cornea caused by the presence of a mucoadhesive polymer, and gradual release of the active ingredient from the formulation at the application site due to the swelling of the outer carrier layer. The mini-tablet contains one or more mitochondrial targeting peptidomimetics described herein, such as (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono, bis or tri(tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.) and one or more polymers. Non-limiting examples of polymers suitable for use in the mini-tablet formulation include, for example, cellulose derivatives such as hydroxypropylmethylcellulose (HPMC), hydroxyethylcellulose (HEC), sodium carboxymethylcellulose, ethylcellulose, acrylates (e.g., polyacrylic acid and its cross-linked forms), carbopol or carbomer, chitosan, and starch (e.g., drum-dried waxy corn starch). In some embodiments, the mini-tablet further comprises one or more excipients. Non-limiting examples of excipients include mannitol and magnesium stearate.

[0190] Ophthalmic or intraocular preparations may contain antibacterial components that are non - injurious during use, such as non - toxic auxiliary substances like thimerosal, benzalkonium chloride, methyl and propyl parabens, benzalkonium decynium bromide, benzyl alcohol, or phenylethanol; buffer components such as sodium chloride, sodium borate, sodium acetate, sodium citrate, or gluconic acid buffer; and other conventional components such as sorbitan monolaurate, triethanolamine, polyoxyethylene sorbitan monopalmitate, ethylenediaminetetraacetic acid, etc.

[0191] In some embodiments, the viscosity of an ophthalmic preparation containing one or more of the mitochondrial - targeting peptide mimetics described herein, such as (R)-2 - amino - N - ((S)-1 - (((S)-5 - amino - 1-(3 - benzyl - 1,2,4 - oxadiazol - 5 - yl)pentyl)amino)-3-(4 - hydroxy - 2,6 - dimethylphenyl)-1 - oxopropan - 2 - yl)-5 - guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (in each case mono, bis or tri(tris) salt), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, bistosylate, or tritosylate, etc.), is increased to improve contact with the cornea and bioavailability in the eye. The viscosity can be increased by adding high - molecular - weight hydrophilic polymers that do not diffuse through biological membranes and form a three - dimensional network in water. Non - limiting examples of such polymers include polyvinyl alcohol, poloxamer, hyaluronic acid, carbomer, and polysaccharides, cellulose derivatives, gellan gum, and xanthan gum.

[0192] Systemic administration of the compounds, therapeutic agents, peptides, peptidomimetics, or mixtures thereof described herein can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, permeation enhancers suitable for the permeated barrier are used in the formulation. Such permeation enhancers are generally known in the art and include, for example, detergents, bile salts, and fusidic acid derivatives for transmucosal administration. Transmucosal administration can be achieved via the use of nasal sprays. In the case of transdermal administration, the active compound is generally formulated in ointments, salves, gels, or creams known in the art. In one embodiment, transdermal administration may be carried out by iontophoresis.

[0193] A compound, therapeutic agent, peptide, peptidomimetic, or mixture thereof can be formulated in a carrier system. The carrier can be a colloidal system. The colloidal system can be a liposome, a phospholipid bilayer vehicle. In one embodiment, the compound, therapeutic agent, peptide, peptidomimetic, or mixture thereof is encapsulated within a liposome while maintaining the integrity of the compound, therapeutic agent, peptide, peptidomimetic, or mixture thereof. Those skilled in the art will understand that there are various methods for preparing liposomes. (See Lichtenberg, et al., Methods Biochem. Anal., 33:337-462 (1988), Anselem, et al., Liposome Technology, CRC Press (1993)). Liposome formulations can delay clearance and increase cellular uptake. (See Reddy, Ann. Pharmacother., 34(7-8):915-923 (2000)). The active agent can also be carried within particles prepared from pharmaceutically acceptable components including, but not limited to, soluble, insoluble, permeable, impermeable, biodegradable, or digestibility enhancing polymers or liposomes. Such particles include, but are not limited to, nanoparticles, biodegradable nanoparticles, microparticles, biodegradable microparticles, nanospheres, biodegradable nanospheres, microspheres, biodegradable microspheres, capsules, emulsions, liposomes, micelles, and virus vector systems.

[0194] The carrier can also be a polymer, such as a biodegradable biocompatible polymer matrix. In one embodiment, the compound, therapeutic agent, peptide, peptidomimetic, or a mixture thereof can be embedded in the polymer matrix while maintaining the integrity of the composition. The polymer can be a natural product such as a polypeptide, protein or polysaccharide, or a synthetic product such as a polyα-hydroxy acid. Examples include carriers made of, for example, collagen, fibronectin, elastin, cellulose acetate, cellulose nitrate, polysaccharides, fibrin, gelatin, and combinations thereof. In one embodiment, the polymer is polylactic acid (PLA) or copolylactic / glycolic acid (PLGA). The polymer matrix can be prepared and isolated in various forms and sizes including microspheres and nanospheres. Polymer formulations can result in an extended duration of therapeutic effect. (See Reddy, Ann. Pharmacother., 34(7-8):915-923(2000)). Polymer formulations for human growth hormone (hGH) have been used in clinical trials. (See Kozarich and Rich, Chemical Biology, 2:548-552(1998)).

[0195] Examples of polymer microsphere sustained release formulations are described in PCT Publication WO99 / 15154 (Tracy et al.), U.S. Pat. Nos. 5,674,534 and 5,716,644 (both Zale et al.), PCT Publication WO96 / 40073 (Zale et al.), and PCT Publication WO00 / 38651 (Shah et al.). U.S. Pat. Nos. 5,674,534 and 5,716,644, and PCT Publication WO96 / 40073 describe polymer matrices containing particles of erythropoietin stabilized against aggregation with salts.

[0196] In some embodiments, the therapeutic compound is prepared with a carrier that protects the therapeutic compound from rapid elimination from the body, such as a controlled release formulation that includes an implant and a microencapsulation delivery system. Biodegradable biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Such formulations can be prepared using known techniques. The materials can also be obtained commercially, for example, from Alza Corporation and Nova Pharmaceuticals, Inc. Liposome suspensions, including liposomes targeted to specific cells having monoclonal antibodies to cell-specific antigens, can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Patent No. 4,522,811.

[0197] The therapeutic compound can also be formulated to enhance intracellular delivery. For example, liposome delivery systems are known in the art, see, for example, Chonn and Cullis, “Recent Advances in Liposome Drug Delivery Systems,” Current Opinion in Biotechnology 6:698-708 (1995), Weiner, “Liposomes for Protein Delivery: Selecting Manufacture and Development Processes,” Immunomethods, 4(3):201-9 (1994), and Gregoriadis, “Engineering Liposomes for Drug Delivery: Progress and Problems,” Trends Biotechnol., 13(12):527-37 (1995). Mizguchi, et al., Cancer Lett., 100:63-69 (1996) describes the use of fusogenic liposomes for delivering proteins to cells both in vivo and in vitro.

[0198] In addition to the above formulations, compounds, therapeutic agents, peptides, peptidomimetics, or mixtures thereof may also be formulated as depot preparations. Such long-acting formulations can be formulated with suitable polymers or hydrophobic materials (e.g., as emulsions in acceptable oils) or ion exchange resins, or as poorly soluble derivatives, e.g., as poorly soluble salts.

[0199] Compounds, therapeutic agents, peptides, peptidomimetics, or mixtures thereof can be provided in particles or polymeric microspheres. Examples of polymeric microsphere sustained release formulations are described in PCT Publication WO99 / 15154 (Tracy et al.), U.S. Patent Nos. 5,674,534 and 5,716,644 (both Zale et al.), PCT Publication WO96 / 40073 (Zale et al.), and PCT Publication WO00 / 38651 (Shah et al.). U.S. Patent Nos. 5,674,534 and 5,716,644, and PCT Publication WO96 / 40073 describe polymeric matrices containing particles of erythropoietin stabilized against aggregation with salts. The particles can contain a therapeutic agent within a central portion surrounded by a coating including, but not limited to, enteric coatings. Compounds, therapeutic agents, peptides, peptidomimetics, or mixtures thereof can also be dispersed throughout the particles. Compounds, therapeutic agents, peptides, peptidomimetics, or mixtures thereof can also be adsorbed within the particles. The particles can have any order of release kinetics including, but not limited to, zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, and any combination thereof. The particles can contain, in addition to the compound, any of those materials customarily used in the fields of pharmacy and medicine, including therapeutic agents, peptides, peptidomimetics, or mixtures thereof, and these include, but are not limited to, erosive, non-erosive, biodegradable, or non-biodegradable materials, or combinations thereof. The particles can be microcapsules containing the compounds of the present technology in solution or in a semi-solid state. The particles can be substantially any shape.

[0200] Both non-biodegradable and biodegradable polymer materials can be used in the manufacture of particles for delivering compounds, therapeutic agents, peptides, peptidomimetics or mixtures thereof. Such polymers can be natural or synthetic polymers. The polymer may be a natural product such as a polypeptide, protein or polysaccharide, or a synthetic product such as a polyα-hydroxy acid. Examples include carriers made of, for example, collagen, fibronectin, elastin, cellulose acetate, cellulose nitrate, polysaccharides, fibrin, gelatin, and combinations thereof. Specific purpose bioadhesive polymers include the bioerodible hydrogels described in Sawhney H S et al. (1993) Macromolecules 26:581-7, the teachings of which are incorporated herein. These include polyhyaluronic acid, casein, gelatin, gluten, polyanhydrides, polyacrylic acid, alginates, chitosan, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate) and polycaprolactone.

[0201] A compound, therapeutic agent, peptide, peptidomimetic, or mixture thereof can be included in a controlled release system. The term "controlled release" is intended to refer to a formulation containing any drug in which the mode and profile of drug release from the formulation are controlled. This refers to both immediate and non-immediate release formulations, and non-immediate release formulations include, but are not limited to, sustained release and delayed release formulations. The term "sustained release" (also referred to as "extended release") is used in its conventional meaning and refers to a drug formulation that provides a gradual release of the drug over an extended period of time, preferably, but not necessarily, resulting in a substantially constant blood level of the drug over an extended period of time. The term "delayed release" is used in its conventional meaning and refers to a drug formulation in which there is a time delay between administration of the formulation and release of the drug therefrom. "Delayed release" may or may not be accompanied by a gradual release of the drug over an extended period of time and thus may or may not be "sustained release".

[0202] The use of long-term sustained release implants may be particularly suitable for the treatment of chronic diseases. "Long-term" release, as used herein, means that the implant (depot) is constructed and arranged to deliver a therapeutically effective level of the active ingredient (i.e., a compound, therapeutic agent, peptide, peptidomimetic or mixture thereof) for at least 7 days, preferably 30 to 60 days. Long-term sustained release implants are well known to those skilled in the art and include some of the release systems described above.

[0203] The dosage, toxicity and therapeutic efficacy of any compound, therapeutic agent, peptide, peptidomimetic or mixtures thereof can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dosage ratio between toxic and therapeutic effects is the therapeutic index and can be expressed as the ratio of LD50 / ED50. Compounds showing a high therapeutic index are advantageous. Compounds that exhibit toxic side effects may be used, but care should be taken to design a delivery system that targets such compounds to the site of the affected tissue in order to minimize potential damage to uninfected cells and thereby reduce side effects.

[0204] Data obtained from cell culture assays and animal studies can be used in formulating a range of dosages for use in humans. The dosage of such compounds can be within the range of circulating concentrations that include an ED50 with little or no toxicity. The dosage can vary within this range depending on the dosage form employed and the route of administration utilized. For any compound used in the methods described herein, the therapeutically effective dosage can be estimated initially from cell culture assays. The dosage can be formulated in animal models to achieve a range of circulating plasma concentrations that include the IC50 (i.e., the concentration of the test compound that achieves half-maximal inhibition of the symptoms) determined in cell culture. Such information can be used to accurately determine useful dosages in humans. Levels in the plasma can be measured, for example, by high performance liquid chromatography.

[0205] Typically, an effective amount of the mitochondrial targeting peptide mimetic sufficient to achieve a therapeutic or prophylactic effect ranges from about 0.000001 mg per kilogram of body weight per day to about 10,000 mg per kilogram of body weight per day. Suitably, the dosage range is from about 0.0001 mg per kilogram of body weight per day to about 100 mg per kilogram of body weight per day. For example, the dosage can be within the range of 1 mg / kg body weight or 10 mg / kg body weight daily, every two days or every three days, or within the range of 1 - 10 mg / kg weekly, every two weeks or every three weeks. In one embodiment, the single dosage of the peptide or peptide mimetic ranges from 0.001 to 10,000 micrograms per kilogram of body weight. In one embodiment, the concentration of the mitochondrial targeting peptide mimetic in the carrier ranges from 0.2 to 2000 micrograms per milliliter delivered. Exemplary treatment regimens involve administration once a day or once a week. In therapeutic use, relatively high dosages at relatively short intervals may be required until the progression of the disease is reduced or halted, or until the subject shows a partial or complete improvement of the symptoms of the disease. Thereafter, the patient may be placed on a prophylactic regimen.

[0206] In some embodiments, a therapeutically effective amount of the mitochondrial targeting peptide mimetic can be defined as a concentration of the peptide mimetic in the target tissue of 10 -12 ~10 -6 moles, for example, approximately 10 -7 moles. This concentration can be delivered by a systemic dose of 0.001 - 100 mg / kg or an equivalent dose based on body surface area. The dosing schedule will be optimized to maintain a therapeutic concentration in the target tissue, such as by single daily or weekly single administrations, but continuous administration (e.g., intravenous infusion or transdermal application) is also included.

[0207] One of ordinary skill in the art will understand that certain factors can affect the dosage amount and timing required to effectively treat a subject. This includes, but is not limited to, the severity of the disease or disorder, previous treatments, the general health and / or age of the subject, and other diseases present. Further, treatment of a subject with a therapeutically effective amount of a compound, therapeutic agent, peptide, peptidomimetic, or mixture thereof described herein can include a single treatment or a series of treatments.

[0208] Combination therapy In some embodiments, (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (in each case mono-, bis- or tri-(tris) salts), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), such as mitochondrial targeting peptide mimics, can be combined with one or more additional therapies for the prevention or treatment of ALS. In some embodiments of the methods of the present technology, the mitochondrial targeting peptide mimic is (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (e.g., (Ia)). In some embodiments, the additional therapies include, but are not limited to, administration of riluzole (Rilutek®), edaravone (Radicava®), mecaserimin, baclofen (Lioresal®), diazepam (Valium®), dantrolene (Dantrium®), non-steroidal anti-inflammatory agents, anticonvulsants (e.g., carbamazepine (Tegretol) or phenytoin (Dilantin®)), amitriptyline (Elavil®), nortriptyline (Pamelor™), and lorazepam (Ativan®). In some embodiments, the additional therapy includes co-administration of elamipretide (also known as SS-31 or Bendavia).

[0209] In some embodiments, riluzole is administered separately from, simultaneously with, or sequentially with a mitochondrial targeting peptide mimetic. In some embodiments, the dosage of riluzole is from about 0.5 mg / kg to about 2 mg / kg, from about 1 mg / kg to about 2 mg / kg, from about 0.5 mg / kg to about 5 mg / kg, from about 5 mg / kg to about 100 mg / kg, from about 10 mg / kg to about 75 mg / kg, or from about 25 mg / kg to about 50 mg / kg. In some embodiments, the dosage of resveratrol is 0.8 mg / kg, about 5 mg / kg, about 10 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 75 mg / kg, about 80 mg / kg, about 90 mg / kg, about 100 mg / kg, about 110 mg / kg, about 120 mg / kg, about 125 mg / kg, about 130 mg / kg, about 140 mg / kg, about 150 mg / kg, about 160 mg / kg, about 175 mg / kg, about 180 mg / kg, about 190 mg / kg, about 200 mg / kg, or more. In some embodiments, riluzole is administered twice daily, daily, every 48 hours, every 72 hours, twice a week, once a week, once every two weeks, once a month, once every two months, once every three months, or once every six months. In some embodiments, the dosage of riluzole depends on the weight and / or age of the subject.

[0210] In some embodiments, Mecasermin is administered separately from, simultaneously with, or sequentially to the mitochondrial targeting peptide mimetic. In some embodiments, the dosage of Mecasermin is from about 0.5 mg / kg to about 2 mg / kg, from about 1 mg / kg to about 2 mg / kg, from about 0.5 mg / kg to about 5 mg / kg, from about 5 mg / kg to about 100 mg / kg, from about 10 mg / kg to about 75 mg / kg, or from about 25 mg / kg to about 50 mg / kg. In some embodiments, the dosage of Resveratrol is 0.8 mg / kg, about 5 mg / kg, about 10 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 75 mg / kg, about 80 mg / kg, about 90 mg / kg, about 100 mg / kg, about 110 mg / kg, about 120 mg / kg, about 125 mg / kg, about 130 mg / kg, about 140 mg / kg, about 150 mg / kg, about 160 mg / kg, about 175 mg / kg, about 180 mg / kg, about 190 mg / kg, about 200 mg / kg, or more. In some embodiments, Mecasermin is administered twice a day, daily, every 48 hours, every 72 hours, twice a week, once a week, once every two weeks, once a month, once every two months, once every three months, or once every six months. In some embodiments, the dosage of Mecasermin depends on the weight and / or age of the subject.

[0211] In one embodiment, the additional therapeutic agent is administered to a subject in combination with at least one mitochondrial targeting peptide mimetic such that a synergistic therapeutic effect is produced. For example, administering at least one mitochondrial targeting peptide mimetic together with one or more additional therapeutic agents for the prevention or treatment of ALS will likely be greater than the additive effect in the prevention or treatment of the disease. Thus, one or more of any of the individual therapeutic agents at a lower dose can be used in the treatment or prevention of ALS, resulting in an increase in therapeutic efficacy and a decrease in side effects. In some embodiments, the at least one mitochondrial targeting peptide mimetic is administered in combination with one or more of riluzole (Rilutek®), edaravone (Radicava®), mecobalamin, baclofen (Lioresal®), diazepam (Valium®), dantrolene (Dantrium®), non-steroidal anti-inflammatory agents, antispasmodics (e.g., carbamazepine (Tegretol) or phenytoin (Dilantin®)), amitriptyline (Elavil®), nortriptyline (Pamelor™), or lorazepam (Ativan®) such that a synergistic effect in the prevention or treatment of ALS is provided. In some embodiments, the additional therapeutic agent is elamipretide (also known as SS-31 or Bendavia).

[0212] In some embodiments, (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono, bis or tris (tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), such as mitochondrial targeting peptide mimics, can be combined with one or more additional therapies for the prevention or treatment of alpha-synucleinopathy. In some embodiments of the methods of the present technology, the mitochondrial targeting peptide mimic is (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (e.g., (Ia)). In some embodiments, the additional therapy includes, but is not limited to, the administration of levodopa. In one embodiment, the additional therapeutic agent is administered to a subject in combination with at least one mitochondrial targeting peptide mimic such that a synergistic therapeutic effect is produced. For example, administering at least one mitochondrial targeting peptide mimic together with one or more additional therapeutic agents for the prevention or treatment of alpha-synucleinopathy will be greater than the additive effect in the prevention or treatment of the disease. Thus, one or more of any of the individual therapeutic agents at a lower dose can be used in the treatment or prevention of alpha-synucleinopathy, resulting in an increase in therapeutic efficacy and a decrease in side effects.

[0213] In some embodiments, (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono, bis or tris (tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.) such as mitochondrial targeting peptide mimics can be combined with one or more additional therapies for the prevention or treatment of TDP-43 proteinopathy. In some embodiments of the methods of the present technology, the mitochondrial targeting peptide mimics are (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (e.g., (Ia)). In some embodiments, the additional therapy includes, but is not limited to, administration of antidepressants such as SSRI antidepressants including trazodone. In one embodiment, the additional therapeutic agent is administered to a subject in combination with at least one mitochondrial targeting peptide mimic such that a synergistic therapeutic effect is produced. For example, administering at least one mitochondrial targeting peptide mimic together with one or more additional therapeutic agents for the prevention or treatment of TDP-43 proteinopathy will be greater than the additive effect in the prevention or treatment of the disease. Thus, one or more of any of the individual therapeutic agents at a lower dose can be used in the treatment or prevention of TDP-43 proteinopathy, resulting in increased therapeutic efficacy and decreased side effects.

[0214] In some embodiments, the plurality of therapeutic agents can be administered in any order or even simultaneously. In the case of simultaneous administration, the plurality of therapeutic agents can be provided in a single, unified form or in multiple forms (by way of example only, either as a single pill or as two separate pills). One of the therapeutic agents can be given in multiple doses or both can be given as multiple doses. When not simultaneous, the timing between multiple administrations can vary from greater than 0 weeks to less than 4 weeks. Additionally, the methods, compositions, and formulations of the combination are not limited to the use of only two agents.

Example

[0215] The present technology is further illustrated by the following examples, which should not be construed as limiting in any way.

[0216] Example 1 - Use of a Mitochondria-Targeted Peptide Mimetic Compound in the Treatment of ALS in an Animal Model This example demonstrates the use of a mitochondria-targeted peptide mimetic compound, (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono, bis or tris (tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), stereoisomers, tautomers, hydrates, and / or solvates, etc., in the treatment of ALS in an animal model of the disease.

[0217] Method Study design. Three experimental groups of n = 20 (10 males, 10 females) SOD1 G93A high-copy transgenic mice were dosed daily via intraperitoneal administration with vehicle control, 0.5 mg / kg of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (as its tris HCl salt (Ia)), or 5.0 mg / kg of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (as its tris HCl salt (Ia)). Animals were dosed from 8 weeks of age until the humane end-of-life stage (defined by the animal being unable to right itself from either side within 30 seconds when placed on its side). The experimental endpoints and in-life collections and observations were as follows. 1. Body weight measurement. Daily recording of body weight. 2. Neurological scoring. Weekly NeuroScore is a qualitative assessment of the progression of neurological disease based on a 5-point grading system. Scoring is defined as follows. Score 0: When the mouse is suspended by its tail, the hind legs are fully extended from the midline and the mouse can hold this extension for 2 seconds and be suspended three times in succession. Score 1: During suspension by the tail, the extended part of the leg falls or partially falls towards the midline or the hind legs tremble. Score 2: During 12-inch walking, the toes bend down at least twice or any part of the foot is dragged along the bottom of the cage or the table. Score 3: Rigid paralysis or minimal joint movement, the feet are not used for forward movement. Score 4: The mouse is unable to right itself from either side within 30 seconds. 3. Grip strength test. Weekly forelimb grip strength was performed as follows. a. Measure the weight of the subject and allow it to adapt to the laboratory for at least 60 minutes. b. Apparatus: Bioseb grip strength meter equipped with a grip grid suitable for mice. c. The mouse can be visually positioned and is lowered by its tail towards the grid so that the mouse grasps the grid with its front paws. d. The subject is firmly pulled horizontally away from the grid (parallel to the bench) during three consecutive trials, with a short rest period (about 30 seconds) on the bench between trials. e. Analyze the average force in grams of three front paw and three whole paw trials with and without normalization to body weight. 4. Retro-orbital blood collection. Blood is collected from the retro-orbital sinus every two weeks to determine the levels of drug exposure and neurofilament light chain, a biomarker of axonal injury that has been shown to correlate with the progression of ALS disease in human patients. Briefly, mice are sedated with isoflurane (5% induction, 2% maintenance) in O2. When the plane of anesthesia is reached, the mice are disconnected from isoflurane and blood is collected from the retro-orbital sinus with a 25 μL glass capillary tube, bleeding the sides alternately and continuously. 200 μL of whole blood is collected into a BD K2EDTA microtainer collection tube containing 3.5 μL of 25X HALT protease inhibitor cocktail and kept on ice for processing. The mice are allowed to recover from anesthesia and then returned to their home cages.

[0218] Results (R)-2-Amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide administered systemically delays the progression of neurological symptoms in ALS. As described above, mice were treated daily from 8 weeks of age until the end of their humane lifespan by intraperitoneal injection of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (Ia), or vehicle control. Progression of the neurological disease was measured weekly using a 5-point neurological scoring scale (see the study design for the scoring criteria shown above). (R)-2-Amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (as its tris HCl salt (Ia)) at a dose of 5.0 mg / kg delayed the progression of onset of neurological symptoms in male animals relative to vehicle-treated animals as determined by two-way ANOVA (Figure 1A). In this transgenic model, there was no effect in female mice that exhibit a milder phenotype (Figure 1B).

[0219] (R)-2-Amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide administration systemically extends lifespan in an ALS mouse model. The lifespan of male animals dosed with (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide at 5.0 mg / kg was significantly increased compared to vehicle control (Figure 1C). In this transgenic model, there was no effect in female mice that showed a milder disease phenotype (Figure 1D).

[0220] (R)-2-Amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide administered systemically attenuates the decline in muscle strength in an ALS mouse model. As shown in FIGS. 2A and 1A, systemic administration of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (as its tris HCl salt (Ia)) attenuates the loss of grip strength in male SOD1 G93A transgenic mice. Grip strength values were determined through the baseline (week 8) and end-of-life of each animal. The average of the decrease for individual animals is shown for animals 10 weeks after drug. Male animals treated with high-dose (5.0 mg / kg) (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (as its tris HCl salt (Ia)) showed a trend towards protection from loss of grip strength, although the magnitude of the effect was slightly below statistical significance (p = 0.08).

Table 1

Table 2

[0221] (R)-2-Amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide administration systemically reduces the plasma accumulation of neurofilament light chain (NfL) in ALS mouse models. Systemic administration of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide reduces the accumulation of neurofilament light chain (NfL) in the plasma of male SOD1 G93A transgenic mice. Plasma levels of NfL, a marker of axonal injury 10 weeks after administration of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide or vehicle control, are shown in FIGS. 3A and 3B (and Tables 2A-2B). (R)-2-Amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (as its tris HCl salt (Ia)) at 5.0 mg / kg had a statistically significant effect on the accumulation of NfL in male transgenic mice, suggesting protection from axonal injury in the CNS (FIG. 3A). It should be noted that female mice had lower overall levels of NfL, consistent with the milder disease phenotype associated with female mice in this model (FIG. 3B).

Table 3

Table 4

[0222] As shown in FIG. 4 (and Table 3), in male SOD1 G93A transgenic mice, there is a significant correlation between plasma neurofilament levels and animal survival rate. Depicted are the plasma NfL levels for all male mice in this study plotted as a function of their age at the humane end-stage of life. The correlation between the accumulation of axonal injury biomarkers and the lifespan of the animals is highly significant, and (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (Ia) is shown to at least partially extend the lifespan of male SOD1 G93A mice through protection against axonal loss or damage in the CNS.

Table 5

[0223] These results indicate that the mitochondrial-targeting peptidomimetic compound of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono, bis or tris (tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), stereoisomers, tautomers, hydrates, and / or solvates, etc., is useful for improving one or more of the following symptoms, and thus useful in the treatment of ALS. Delayed onset of neurological symptoms of ALS, increased survival rate, attenuation of muscle strength decline, and / or decrease in plasma neurofilament light chain (NfL) levels. Accordingly, (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide is useful in a method for the treatment of ALS in a subject in need thereof.

[0224] Example 2 - Use of Mitochondrial-Targeting Peptidomimetic Compounds in the Treatment of ALS In the treatment of ALS in a subject in need of treatment of ALS, the use of the mitochondrial targeting peptide mimetic compound (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono-, bis- or tris(tri)salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), stereoisomers, tautomers, hydrates, and / or solvates, etc. is predictively shown.

[0225] Method Subjects suspected of having or diagnosed with ALS receive daily administration of 1 mg / kg body weight of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (in each case mono-, bis- or tri-(tris) salt), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tritrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), stereoisomers, tautomers, hydrates, and / or solvates, alone or in combination with one or more additional therapeutic agents for the treatment or prevention of ALS. The peptidomimetics and / or additional therapeutic agents are administered orally, topically, systemically, intravenously, subcutaneously, intravitreally, intraperitoneally, or intramuscularly according to methods known in the art. The subject will be evaluated weekly for the presence and / or severity of signs and symptoms associated with ALS, including but not limited to muscle weakness, muscle wasting (atrophy), muscle cramps, muscle spasticity, slowness of movement, poor balance, impaired coordination, change in voice quality, dysarthria, dysphagia, incomplete eyelid closure, drooling, emotional regulation disorder, premature death, increased expression of brain translocator protein-18 kDa (TSPO), and plasma accumulation of neurofilament light chain (NfL). Treatment is maintained until one or more signs or symptoms of ALS are improved or eliminated.

[0226] Results Subjects suspected of having ALS or diagnosed with ALS and receiving a therapeutically effective amount of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono, bis or tris (tris) salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), stereoisomers, tautomers, hydrates, and / or solvates are predicted to show a reduction or disappearance in the severity of one or more symptoms associated with ALS. Administration of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I) in combination with one or more additional therapeutic agents is further expected to have a synergistic effect in this regard as compared to that observed in subjects treated with a mitochondrial targeting peptide mimetic compound or the additional therapeutic agent alone.

[0227] These results would indicate that (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono-, bis- or tris(tri)salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), stereoisomers, tautomers, hydrates, and / or solvates are useful in the treatment of ALS. These results would indicate that (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (I), or a pharmaceutically acceptable salt thereof (tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate or maleate (mono-, bis- or tris(tri)salts in each case), monoacetate, bisacetate, triacetate, monotrifluoroacetate, bistrifluoroacetate, tristrifluoroacetate, monohydrochloride, dihydrochloride, trihydrochloride (e.g., (Ia)), monotosylate, ditosylate, or tritosylate, etc.), stereoisomers, tautomers, hydrates, and / or solvates are useful in improving one or more of the following symptoms.Muscle weakness, muscle wasting (atrophy), muscle spasm, muscle contracture, slow movement, poor balance, impaired coordination, change in voice quality, dysarthria, dysphagia, incomplete eye closure, drooling, emotional regulation disorder, early death, increased expression of translocator protein-18 kDa (TSPO) in the brain, and plasma accumulation of neurofilament light chain (NfL). Accordingly, the peptidomimetic is useful in a method of treating a subject in need thereof for the treatment of ALS.

[0228] Example 3 - The mitochondrially targeted peptidomimetic compound of the present technology shows high brain exposure and a mitochondrial protective effect compared to elamipretide. This example concerns the pharmacological effect of compound Ia, (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (Ia), compared to elamipretide, with respect to reducing brain uptake and mitochondrial reactive oxygen species (ROS) and maintaining ATP production under conditions of oxidative stress in a rat model.

[0229] Method Sprague Dawley rats were subcutaneously injected with 5 mg / kg of compound Ia or elamipretide (n = 4 per time point). Animals were sacrificed at the indicated time points and subjected to cardiac perfusion. Drug levels in whole brain homogenates were determined by LC-MS / MS. The results are shown in Figure 5A.

[0230] Ischemic stroke was induced in Sprague Dawley rats via middle cerebral artery occlusion with the vasoconstrictor peptide endothelin-1 (ET-1, 240 pmol per injection). Compounds Ia were administered to each rat via subcutaneous injection at 5 mg / kg, 24 hours and 4 hours before the onset of ischemia. High-resolution respirometry (OxyGraph O2K) was used to measure mitochondrial respiration in brain homogenates excised from the infarct area 24 hours after the onset of ischemia. The respiratory control ratio was calculated as the ratio of oxidative phosphorylation supported by complex I to leak respiration coupled to complex I. The results are shown in Figure 5B, **p < 0.01, one-way ANOVA.

[0231] Results Exposure of compound Ia to the brain was higher than that of elamipretide (Figure 5A), and compound Ia restored mitochondrial respiration in the brain under conditions of oxidative stress (Figure 5B). These findings indicate that compound Ia is suitable for the treatment of neurodegenerative diseases in which mitochondrial damage in the central nervous system contributes to the pathological mechanism.

[0232] Example 4 - Mitochondria-targeted peptide mimetic compounds attenuate dopaminergic neuron loss in the substantia nigra of mutant α-synuclein transgenic mice This example demonstrates the use of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (Ia) in a mouse model of mutant alpha-synuclein-induced dopaminergic neuron loss as a means to examine possible drug effects in a model of neurodegenerative disease.

[0233] Methods Using AAV9-mediated viral delivery of human alpha-synuclein carrying the pathogenic A53T mutation, dopaminergic neuron loss in the substantia nigra pars compacta (SNc) was induced in wild-type C57BL / 6 mice. 4 × 10 10Individual virus particles were injected bilaterally via stereotactic surgery. The animals were 9 weeks old at the time of viral transduction. The animals were treated daily with intraperitoneal administration of (Ia) starting 24 hours before viral transduction and continuing for 5 weeks. The experimental groups were as follows. 1. Group A - AAV A53T injection, vehicle treatment (n = 7) 2. Group B1 - AAV A53T injection, (Ia) treatment at 0.5 mg / kg (n = 8) 3. Group B2 - AAV A53T injection, (Ia) treatment at 5.0 mg / kg (n = 8) 4. Group C1 - sham AAV9 injection, (Ia) treatment at 0.5 mg / kg (n = 6) 5. Group C2 - sham AAV9 injection, (Ia) treatment at 5.0 mg / kg (n = 7)

[0234] Five weeks after viral transduction, the animals were sacrificed, the brains were removed, processed for immunohistochemistry, and cell counting in the substantia nigra pars compacta (i.e., the number of TH - positive neurons) was performed via automated stereological analysis. Serial sections were cut and stained with tyrosine hydroxylase (TH) to count the number of dopaminergic neurons. The results are shown in Figures 6A and 6B, **p < 0.01 vs. group, ***p < 0.001 vs. group A; OO p < 0.001 vs. groups C1 and C2.

[0235] Plasma neurofilament light chain levels were determined by SIMOA assay from blood collected at sacrifice, and the results are shown in Figure 6C. No statistical significance by one - way ANOVA between groups was found.

[0236] Results The mutant alpha-synuclein-induced loss of dopaminergic neurons in the substantia nigra was significantly attenuated by compound Ia, Figures 6A and 6B. Neurofilament levels were not increased by this induction protocol (Figure 6C). Thus, treatment with compound Ia prevented the loss of dopaminergic neurons in the substantia nigra following mutant alpha-synuclein toxicity. Thus, these data indicate that compound Ia may be useful in methods for the treatment or prevention of neurodegenerative diseases caused by alpha-synucleinopathies such as Parkinson's disease (PD), PD with dementia, Lewy body dementia, and multiple system atrophy.

[0237] Example 5 - Mitochondria-targeted peptide mimetic compounds are neuroprotective in primary mutant TDP43 expressing upper motor neurons This example is prp-TDP-43 A315T -Examine the neuroprotective effect of (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide (Ia) in primary cells derived from the prp-TDP-43

[0238] Method prp-TDP-43 A315T-Research was conducted on primary cells derived from the UeGFP mouse model (Gautam, et al. Acta Neuropathol. 2019 Jan;137(1):47-69). Briefly, these mice express a pathogenic human TDP43 transgene, which, in the background of eGFP reporter mice, causes severe structural changes in mitochondria, nuclei, and the endoplasmic reticulum. Both the transgene (prp) and the reporter (UCHL1) are driven by tissue-specific promoter elements, resulting in the generation of fluorescent green corticospinal motor neurons carrying the TDP43 mutation. Using the endogenous fluorescence of the EGFP reporter, corticospinal motor neurons can be directly visualized in situ. Mixed cortical cultures were prp-TDP-43 A315T -Derived from the brains of UeGFP mice and split in serum-free minimal medium for three cell divisions. Medium + / − drugs was removed and replenished daily. The doses of compound Ia used in the experiment were 10 nM, 100 nM, and 1000 nM. The drugs were prepared in DMSO vehicle, and DMSO was used as a vehicle control (<1% volume / volume). Cells were imaged using standard fluorescence microscopy. Neurite length was calculated using automated imaging software (NIH Image J) for GFP-expressing cells. Three independent biological replicates were performed, and at least 10 motor neurons were imaged for each replicate. The average neurite length per cell is shown in Figure 7. Statistical analysis was performed by one-way ANOVA using Dunnett's multiple comparison test. ***p < 0.001 vs. vehicle, ****p < 0.0001 vs. vehicle.

[0239] Results Treatment with compound Ia improved neurite length at all doses evaluated in primary upper motor neuron cultures derived from A315T mutant TDP43 transgenic mice. Thus, these data indicate that compound Ia may be effective in methods for treating or preventing diseases of TDP-43 proteinopathies, including ALS and frontotemporal lobar degeneration (FTLD).

[0240] Equivalents This technology is not limited to the specific embodiments described in this application, and is contemplated as a single illustration of individual aspects of this technology. As will be apparent to those skilled in the art, many modifications and variations of this technology can be made without departing from its spirit and scope. In addition to those listed herein, functionally equivalent methods and apparatuses within the scope of this technology will be apparent to those skilled in the art from the foregoing description. Such modifications and variations are intended to fall within the scope of the appended claims. This technology is limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled. This technology is not limited to a particular method, reagent, compound composition, or biological system, and of course, it should be understood that it can vary. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0241] In addition, when a feature or aspect of the present disclosure is described in terms of a Markush group, those skilled in the art will recognize thereby that the present disclosure is also described in terms of any individual member of the Markush group or a subgroup of members.

[0242] As will be understood by those skilled in the art, for all purposes, particularly in terms of providing a written description, all ranges disclosed herein also include any and all possible sub-ranges and combinations of sub-ranges thereof. Any recited range can be readily recognized as being such that the same range can be adequately described and made possible to be decomposed into at least equal halves, thirds, quarters, fifths, tenths, etc. of the range. By way of non-limiting example, each range discussed herein can be readily decomposed into lower thirds, middle thirds, upper thirds, etc. Also, as will be understood by those skilled in the art, all language such as "maximum," "at least," "greater than," "less than," etc. includes the recited number and refers to a range that can then be decomposed into the sub-ranges described above. Finally, as will be understood by those skilled in the art, a range includes each individual member. Thus, for example, a group having 1 to 3 cells refers to a group having 1, 2, or 3 cells. Similarly, a group having 1 to 5 cells refers to a group having 1, 2, 3, 4, or 5 cells.

[0243] All patents, patent applications, provisional applications, and publications referenced or cited herein are hereby incorporated by reference in their entirety, including all figures and tables, to the extent that they are not inconsistent with the explicit teachings herein.

[0244] Other embodiments are within the scope of the following claims.

Claims

1. 1. A method for treating or preventing amyotrophic lateral sclerosis (ALS) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a peptidomimetic, (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof.

2. 10. The method of claim 1, wherein the subject has been diagnosed with ALS.

3. 2. The method of any one of the preceding claims, wherein the ALS is familial.

4. 4. The method of claim 3, wherein the familial ALS is caused by a mutation in the superoxide dismutase 1 (SOD1) gene or the TARDBP gene.

5. 13. The method of any one of the preceding claims, wherein the peptidomimetic is administered daily for two weeks or more.

6. 2. The method of any one of the preceding claims, wherein the peptidomimetic is administered daily for 12 weeks or more.

7. 10. The method of any one of the preceding claims, wherein said treatment or prevention comprises said treatment or prevention of one or more signs or symptoms of ALS comprising one or more of the following: muscle weakness, muscle wasting (atrophy), muscle cramps, muscle spasms, slow movement, poor balance, loss of coordination, changes in voice quality, dysarthria, dysphagia, incomplete eye closure, salivation, emotional dysregulation, premature death, and increased brain translocator protein-18 kDa (TSPO) expression.

8. 10. The method of any one of the preceding claims, wherein the treatment or prevention comprises the treatment or prevention of plasma accumulation of neurofilament light chain (NfL).

9. 10. The method of any one of the preceding claims, wherein the subject is a mammal.

10. 10. The method of claim 9, wherein the mammalian subject is a human.

11. 2. The method of any one of the preceding claims, wherein the peptidomimetic is administered orally or subcutaneously.

12. 12. The method of any one of claims 1 to 11, wherein the peptidomimetic is administered topically, intranasally, systemically, intravenously, intraperitoneally, intradermally, intraocularly, ophthalmologically, intrathecally, intracerebroventricularly, iontophoretically, transmucosally, intravitreally, or intramuscularly.

13. 13. The method of any one of the preceding claims, further comprising administering to the subject an additional treatment, separately, sequentially or simultaneously.

14. The method of claim 13 , wherein the additional treatment comprises administration of a therapeutic agent.

15. 15. The method of claim 14, wherein the therapeutic agent is selected from the group consisting of riluzole (Rilutek®), edaravone (Radicava®), mecasermin, baclofen (Lioresal®), diazepam (Valium®), dantrolene (Dantrium®), nonsteroidal anti-inflammatory agents, anticonvulsants (e.g., carbamazepine (Tegretol®) or phenytoin (Dilantin®)), amitriptyline (Elavil®), nortriptyline (Pamelor™), and lorazepam (Ativan®).

16. 16. The method of claim 15, wherein the combination of the peptidomimetic and the additional therapeutic treatment has a synergistic effect in the prevention or treatment of the ALS.

17. 2. The method of any one of the preceding claims, wherein the pharma- ceutically acceptable salt comprises a tartrate, a fumarate, a monoacetate, a bisacetate, a triacetate, a monotrifluoroacetate, a bistrifluoroacetate, a trifluoroacetate, a monohydrochloride, a bishydrochloride, a trihydrochloride, a monotosylate, a bistosylate, or a tritosylate.

18. The method of any one of claims 1 to 16, wherein the peptidomimetic is formulated as a Tris-HCl salt, a Bis-HCl salt, or a Mono-HCl salt.

19. 1. Use of a composition in the preparation of a medicament for treating or preventing amyotrophic lateral sclerosis (ALS) in a subject in need thereof, said composition comprising a therapeutically effective amount of a peptidomimetic, (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof.

20. 20. The use of claim 19, wherein the subject has been diagnosed with ALS.

21. 21. The use according to claim 19 or 20, wherein the ALS is familial.

22. 22. The use according to claim 21, wherein the familial ALS is caused by a mutation in the superoxide dismutase 1 (SOD1) gene or the TARDBP gene.

23. The use according to any one of claims 19 to 22, wherein the peptidomimetic is intended to be administered daily for more than two weeks.

24. The use according to any one of claims 19 to 22, wherein the peptidomimetic is intended to be administered daily for 12 weeks or more.

25. 25. The use according to any one of claims 19 to 24, wherein said treatment or prevention comprises said treatment or prevention of one or more signs or symptoms of ALS comprising one or more of the following: muscle weakness, muscle wasting (atrophy), muscle cramps, muscle spasms, slow movement, poor balance, loss of coordination, change in voice quality, dysarthria, dysphagia, incomplete eye closure, salivation, emotional dysregulation, premature death, and increased brain translocator protein-18 kDa (TSPO) expression.

26. The use according to any one of claims 19 to 24, wherein the treatment or prevention comprises the treatment or prevention of plasma accumulation of neurofilament light chain (NfL).

27. The use according to any one of claims 19 to 26, wherein the subject is a mammal.

28. 28. The use according to claim 27, wherein the mammalian subject is a human.

29. The use according to any one of claims 19 to 28, wherein the peptidomimetic is formulated for oral or subcutaneous administration.

30. 29. The use according to any one of claims 19 to 28, wherein the peptidomimetic is formulated for topical, intranasal, systemic, intravenous, intraperitoneal, intradermal, intraocular, ophthalmic, intrathecal, intraventricular, iontophoretic, transmucosal, intravitreal, or intramuscular administration.

31. The use according to any one of claims 19 to 30, wherein it is contemplated that the peptidomimetic is used separately, sequentially or simultaneously with an additional treatment.

32. 32. The use of claim 31 , wherein the additional treatment comprises the use of a therapeutic agent.

33. 33. The use of claim 32, wherein the therapeutic agent is selected from the group consisting of riluzole (Rilutek®), edaravone (Radicava®), mecasermin, baclofen (Lioresal®), diazepam (Valium®), dantrolene (Dantrium®), nonsteroidal anti-inflammatory agents, anticonvulsants (e.g., carbamazepine (Tegretol) or phenytoin (Dilantin®)), amitriptyline (Elavil®), nortriptyline (Pamelor™), and lorazepam (Ativan®).

34. The use according to any one of claims 31 to 33, wherein the combination of the peptidomimetic and the additional treatment has a synergistic effect in the prevention or treatment of ALS.

35. 35. The use of any one of claims 19 to 34, wherein the pharma- ceutically acceptable salt comprises a tartrate, a fumarate, a monoacetate, a bisacetate, a triacetate, a monotrifluoroacetate, a bistrifluoroacetate, a trifluoroacetate, a monohydrochloride, a bishydrochloride, a trihydrochloride, a monotosylate, a bistosylate, or a tritosylate.

36. The use of any one of claims 19 to 34, wherein the peptidomimetic is formulated as a Tris-HCl salt, a Bis-HCl salt, or a Mono-HCl salt.

37. 2. The peptidomimetic (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, for use in treating or preventing amyotrophic lateral sclerosis (ALS) in a subject in need thereof.

38. 38. The peptidomimetic of claim 37, for use wherein the subject has been diagnosed with ALS.

39. 39. The peptidomimetic of claim 37 or 38, wherein the ALS is familial.

40. 40. The peptidomimetic of claim 39, wherein the familial ALS is caused by a mutation in the superoxide dismutase 1 (SOD1) gene or the TARDBP gene.

41. The peptidomimetic of any one of claims 37 to 40 for use, wherein the peptidomimetic is intended to be administered daily for two weeks or more.

42. The peptidomimetic of any one of claims 37 to 40 for use, wherein the peptidomimetic is intended to be administered daily for 12 weeks or more.

43. 43. The peptidomimetic of any one of claims 37 to 42 for use comprising said treatment or prevention of one or more signs or symptoms of ALS comprising one or more of muscle weakness, muscle wasting (atrophy), muscle cramps, muscle spasms, slow movement, poor balance, impaired coordination, change in voice quality, dysarthria, dysphagia, incomplete eye closure, salivation, emotional dysregulation, premature death, and increased brain translocator protein-18 kDa (TSPO) expression.

44. 43. The use according to any one of claims 37 to 42, wherein said treatment or prevention comprises said treatment or prevention of plasma accumulation of neurofilament light chain (NfL).

45. The peptidomimetic of any one of claims 37 to 44 for use, wherein the subject is a mammal.

46. 46. ​​The peptidomimetic of claim 45 for use wherein the mammalian subject is a human.

47. The peptidomimetic of any one of claims 37 to 46 for use, wherein the peptidomimetic is formulated for oral or subcutaneous administration.

48. 47. The peptidomimetic of any one of claims 37 to 46, for use, wherein the peptidomimetic is formulated for topical, intranasal, systemic, intravenous, intraperitoneal, intradermal, intraocular, ophthalmic, intrathecal, intraventricular, iontophoretic, transmucosal, intravitreal, or intramuscular administration.

49. The peptidomimetic of any one of claims 37 to 48 for use, wherein said peptidomimetic is intended to be used separately, sequentially or simultaneously with an additional treatment.

50. 50. The peptidomimetic of claim 49 for use, wherein the additional treatment comprises the use of a therapeutic agent.

51. 51. The peptidomimetic of claim 50, for use wherein the therapeutic agent is selected from the group consisting of riluzole (Rilutek®), edaravone (Radicava®), mecasermin, baclofen (Lioresal®), diazepam (Valium®), dantrolene (Dantrium®), nonsteroidal anti-inflammatory agents, anticonvulsants (e.g., carbamazepine (Tegretol) or phenytoin (Dilantin®)), amitriptyline (Elavil®), nortriptyline (Pamelor™), and lorazepam (Ativan®).

52. 52. The peptidomimetic of any one of claims 49 to 51 for use, wherein the combination of the peptidomimetic and an additional treatment has a synergistic effect in the prevention or treatment of ALS.

53. 53. The peptidomimetic of any one of claims 37 to 52 for use, wherein the pharma- ceutically acceptable salt comprises a tartrate, a fumarate, a monoacetate, a bisacetate, a triacetate, a monotrifluoroacetate, a bistrifluoroacetate, a trifluoroacetate, a monohydrochloride, a bishydrochloride, a trihydrochloride, a monotosylate, a bistosylate, or a tritosylate.

54. 54. The peptidomimetic of any one of claims 37 to 53 for use, wherein the peptidomimetic is formulated as a Tris-HCl salt, a Bis-HCl salt, or a Mono-HCl salt.

55. 1. A method for treating or preventing an alpha-synucleinopathy or a TDP-43 proteinopathy in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a peptidomimetic (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof.

56. 56. The method of claim 55, wherein the subject has been diagnosed with an alpha-synucleinopathy or a TDP-43 proteinopathy.

57. 57. The method of claim 55 or 56, wherein the alpha-synucleinopathy is Parkinson's disease (PD), PD with dementia, dementia with Lewy bodies, or multiple system atrophy (MSA), and the TDP-43 proteinopathy is amyotrophic lateral sclerosis (ALS) or frontotemporal lobar degeneration (FTLD).

58. 58. The method of any one of claims 55 to 57, wherein the peptidomimetic is administered daily for two weeks or more.

59. 59. The method of any one of claims 55 to 58, wherein the peptidomimetic is administered daily for 12 weeks or more.

60. 60. The method of any one of claims 55 to 59, wherein the treating or preventing an alpha-synucleinopathy comprises attenuating loss of dopaminergic neurons in the subject compared to an untreated control.

61. 60. The method of any one of claims 55 to 59, wherein the treatment or prevention of the TDP-43 proteinopathy comprises increasing neurite length in the subject compared to an untreated control.

62. The method of any one of claims 55 to 61, wherein the subject is a mammal.

63. 63. The method of claim 62, wherein the mammalian subject is a human.

64. 64. The method of any one of claims 55 to 63, wherein the peptidomimetic is administered orally or subcutaneously.

65. 64. The method of any one of claims 55-63, wherein the peptidomimetic is administered topically, intranasally, systemically, intravenously, intraperitoneally, intradermally, intraocularly, ophthalmologically, intrathecally, intraventricularly, iontophoretically, transmucosally, intravitreally, or intramuscularly.

66. 66. The method of any one of claims 55 to 65, further comprising administering to the subject an additional treatment, either separately, sequentially or simultaneously.

67. 67. The method of claim 66, wherein the additional treatment comprises administration of a therapeutic agent.

68. 68. The method of claim 67, wherein the therapeutic agent comprises levodopa for the treatment of the alpha-synucleinopathy and the therapeutic agent comprises a selective serotonin reuptake inhibitor (SSRI) antidepressant for the treatment of the TDP-43 proteinopathy.

69. The method of any one of claims 66 to 68, wherein the combination of the peptidomimetic and the additional therapeutic treatment has a synergistic effect in the prevention or treatment of an alpha-synucleinopathies or TDP-43 proteinopathy.

70. 70. The method of any one of claims 55-69, wherein the pharma- ceutically acceptable salt comprises a tartrate, a fumarate, a monoacetate, a bisacetate, a triacetate, a monotrifluoroacetate, a bistrifluoroacetate, a trifluoroacetate, a monohydrochloride, a bishydrochloride, a trihydrochloride, a monotosylate, a bistosylate, or a tritosylate.

71. 71. The method of any one of claims 55-70, wherein the peptidomimetic is formulated as a Tris-HCl salt, a Bis-HCl salt, or a Mono-HCl salt.

72. 1. Use of a composition in the preparation of a medicament for treating or preventing an alpha-synucleinopathy or a TDP-43 proteinopathy in a subject in need thereof, said composition comprising a therapeutically effective amount of the peptidomimetic (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof.

73. 73. The use of claim 72, wherein the subject has been diagnosed with an alpha-synucleinopathy or a TDP-43 proteinopathy.

74. 74. The use of claim 72 or 73, wherein the alpha-synucleinopathy is Parkinson's disease (PD), PD with dementia, dementia with Lewy bodies, or multiple system atrophy (MSA) and the TDP-43 proteinopathy is amyotrophic lateral sclerosis (ALS) or frontotemporal lobar degeneration (FTLD).

75. The use according to any one of claims 72 to 74, wherein the peptidomimetic is intended to be administered daily for two weeks or more.

76. The use according to any one of claims 72 to 75, wherein the peptidomimetic is intended to be administered daily for 12 weeks or more.

77. 77. The use according to any one of claims 72 to 76, wherein the treatment or prevention of an alpha-synucleinopathic disease comprises attenuating loss of dopaminergic neurons in the subject compared to an untreated control.

78. 77. The use according to any one of claims 72 to 76, wherein the treatment or prevention of the TDP-43 proteinopathy comprises an increase in neurite length in the subject compared to an untreated control.

79. The use according to any one of claims 72 to 78, wherein the subject is a mammal.

80. 80. The use of claim 79, wherein the mammalian subject is a human.

81. The use according to any one of claims 72 to 80, wherein the peptidomimetic is formulated for oral or subcutaneous administration.

82. 81. The use of any one of claims 72 to 80, wherein the peptidomimetic is formulated for topical, intranasal, systemic, intravenous, intraperitoneal, intradermal, intraocular, ophthalmic, intrathecal, intraventricular, iontophoretic, transmucosal, intravitreal, or intramuscular administration.

83. The use according to any one of claims 72 to 82, further comprising administering to the subject an additional treatment, either separately, sequentially or simultaneously.

84. 84. The use of claim 83, wherein the additional treatment comprises administration of a therapeutic agent.

85. 85. The use of claim 84, wherein the therapeutic agent comprises levodopa for the treatment of the alpha-synucleinopathy and the therapeutic agent comprises a selective serotonin reuptake inhibitor (SSRI) antidepressant for the treatment of the TDP-43 proteinopathy.

86. 86. The use according to any one of claims 83 to 85, wherein the combination of the peptidomimetic and the additional therapeutic treatment has a synergistic effect in the prevention or treatment of an alpha-synucleinopathies or TDP-43 proteinopathies.

87. 87. The use of any one of claims 72 to 86, wherein the pharma- ceutically acceptable salt comprises a tartrate, a fumarate, a monoacetate, a bisacetate, a triacetate, a monotrifluoroacetate, a bistrifluoroacetate, a trifluoroacetate, a monohydrochloride, a bishydrochloride, a trihydrochloride, a monotosylate, a bistosylate, or a tritosylate.

88. 88. The use of any one of claims 72 to 87, wherein the peptidomimetic is formulated as a Tris-HCl salt, a Bis-HCl salt, or a Mono-HCl salt.

89. 2. The peptidomimetic (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide, or a pharma- ceutically acceptable salt, stereoisomer, tautomer, hydrate, and / or solvate thereof, for use in treating or preventing an alpha-synucleinopathy or a TDP-43 proteinopathy in a subject in need thereof.

90. 90. The peptidomimetic of claim 89, for use wherein the subject has been diagnosed with an alpha-synucleinopathy or a TDP-43 proteinopathy.

91. 91. The peptidomimetic of claim 89 or 90, wherein the alpha-synucleinopathic disease is Parkinson's disease (PD), PD with dementia, dementia with Lewy bodies, or multiple system atrophy (MSA) and the TDP-43 proteinopathy is amyotrophic lateral sclerosis (ALS) or frontotemporal lobar degeneration (FTLD).

92. The peptidomimetic of any one of claims 89 to 91 for use, wherein the peptidomimetic is intended to be administered daily for two weeks or more.

93. The peptidomimetic of any one of claims 89 to 92 for use, wherein the peptidomimetic is intended to be administered daily for 12 weeks or more.

94. 94. The peptide mimetic of any one of claims 89 to 93 for use, wherein the treatment or prevention of an alpha-synucleinopathic disease comprises attenuating loss of dopaminergic neurons in the subject compared to an untreated control.

95. 94. The peptidomimetic of any one of claims 89 to 93 for use, wherein the treatment or prevention of the TDP-43 proteinopathy comprises an increase in neurite length in the subject compared to an untreated control.

96. 96. The peptidomimetic of any one of claims 89 to 95 for use, wherein the subject is a mammal.

97. 97. The peptidomimetic of claim 96, for use wherein the mammalian subject is a human.

98. 98. The peptidomimetic of any one of claims 89 to 97, for use, wherein the peptidomimetic is formulated for oral or subcutaneous administration.

99. 99. The peptidomimetic of any one of claims 89 to 98, for use, wherein the peptidomimetic is formulated for topical, intranasal, systemic, intravenous, intraperitoneal, intradermal, intraocular, ophthalmic, intrathecal, intraventricular, iontophoretic, transmucosal, intravitreal, or intramuscular administration.

100. The peptidomimetic of any one of claims 89 to 99 for use, wherein said peptidomimetic is intended to be used separately, sequentially or simultaneously with an additional treatment.

101. The peptide mimetic of claim 100 for use, wherein the additional treatment comprises the use of a therapeutic agent.

102. 102. The peptidomimetic of claim 101, for use wherein said therapeutic agent comprises levodopa for the treatment of said alpha-synucleinopathies and said therapeutic agent comprises a selective serotonin reuptake inhibitor (SSRI) antidepressant for the treatment of said TDP-43 proteinopathy.

103. 103. The peptidomimetic of any one of claims 100 to 102 for use, wherein the combination of the peptidomimetic and the additional therapeutic treatment has a synergistic effect in the prevention or treatment of an alpha-synucleinopathies or TDP-43 proteinopathies.

104. 104. The peptidomimetic of any one of claims 89 to 103 for use, wherein the pharma- ceutically acceptable salt comprises a tartrate, a fumarate, a monoacetate, a bisacetate, a triacetate, a monotrifluoroacetate, a bistrifluoroacetate, a trifluoroacetate, a monohydrochloride, a bishydrochloride, a trihydrochloride, a monotosylate, a bistosylate, or a tritosylate.

105. 105. The peptidomimetic of any one of claims 89 to 104 for use, wherein the peptidomimetic is formulated as a Tris-HCl salt, a Bis-HCl salt, or a Mono-HCl salt.

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