Use of small molecules that target karyopherin beta 1 (KPNβ1) for treatment of neurodegenerative diseases

Abstract

Disclosed herein methods of treating proteinopathies involving proteins that are targeted by karyopherin beta 1's (Kpnβ1) chaperoning function. Therefore the methods of the present application are for increasing Kpnβ1's chaperoning function and comprise administering to a subject in need thereof, an effective amount of one ormore compounds of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, and XVI. The methods can be used to, for example, to treat proteinopathies of TDP- 43, SOD1 or a-synuclein, or fragments and / or isoforms thereof, and / or proteinopathies other proteins that arc targeted by Kpnβ1's chaperoning function, and therefore to treat subjects with neurodegenerative disorders. Formula (I)

Description

USE OF SMALL MOLECULES THAT TARGET KARYOPHERIN BETA 1 (KPN 1 ) FOR TREATMENT OF NEURODEGENERATIVE DISEASESCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the benefit of priority from co-pending U.S. provisional patent application no. 63 / 542,593 filed on October 5, 2023, the contents of which are incorporated by reference in their entirety.FIELD

[0002] The present application relates to the use of compounds for the treatment of proteinopathies, in particular proteinopathies involving proteins that are targeted by karyopherin beta 1’s (Kpn|31 ; also known as Importin |31) chaperoning function.BACKGROUND

[0003] TAR-DNA-binding protein 43 kDa (TDP-43) mislocalization from the nucleus to the cytoplasm and formation of ubiquitinated and phosphorylated aggregates is the pathological hallmark in most amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) cases and is also present in other neurodegenerative diseases (de Boer et al., 2021). The biochemical signature of TDP-43 pathology is the presence of phosphorylated TDP-43 and lower molecular weight fragments of TDP-43 (e.g. TDP-25 and TDP-35 C-terminal fragments, and TDP-C-spl-272 an N-terminal fragment) (Arai et al., 2006; Neumann et al., 2006; Shenouda, Xiao, MacNair, Lau, & Robertson, 2022). In ALS / FTLD tissue, TDP-43 aggregates were shown to contain nucleocytoplasmic (NCT) proteins, and some of these, such as Kpn|31 , were shown to regulate TDP-43 aberrant phase transitions in vitro (Chou et al., 2018; Khalil et al., 2022). Prevention and / or removal of TDP-43 aggregates is a key therapeutic approach, and numerous methodologies have been investigated (Francois-Moutal et al. 2021 ; Buratti, 2021). This includes antibodies targeting misfolded TDP- 43, antisense oligonucleotides to downregulate expression of the TARDBP gene encoding TDP-43, and small molecules that interfere with the protein aggregation process (Francois- Moutal, Scott, & Khanna, 2021). Another approach is to harness the intrinsic cell machinery to upregulate molecular chaperones, which prevent / reverse protein misfolding and targets TDP-43 for degradation (Sinnige, Yu, & Morimoto, 2020). Upregulating or eliciting a heat shock response is one such strategy, with several heat shock proteins shown to ameliorate protein aggregation in vitro and in vivo (Gomez-Pastor, Burchfiel, & Thiele, 2018). However, to date, translation of using heat shock proteins to therapeutically target protein aggregates in neurodegenerative diseases has thus far failed to deliver in the clinic (Benatar et al., 2018; Crippa et al., 2016; Ganassi et al., 2016; Mandrioli et al., 2019).

[0004] It has become evident that other proteins also have chaperone properties. One such protein is karyopherin betal (Kpn|31) which is canonically associated with transferringcargoes from the cytoplasm to the nucleus, a process called nucleocytoplasmic transport (NCT) (Kimura & Imamoto, 2014). During classical NCT, karyopherin alpha (Kpnal) also known as Importin a binds to the nuclear location sequence (NLS) of cargo proteins, with Kpnpi binding to Kpna facilitating nuclear import through the nuclear pores (Kimura & Imamoto, 2014). In the nucleus, interaction with RanGTP releases Kpn|31 , and RanGTP complexed with Kpn|31 is returned to the cytoplasm, where a nucleotide exchange converting RanGTP to RanGDP releases Kpn|31 for another round of NCT (Kimura & Imamoto, 2014). In addition to this role in NCT, ‘free’ Kpn|31 and other importins can act as a chaperones, binding to cytoplasmic proteins with low complexity domains, such as TDP-43, FUS, hnRNPAI , and hnRNPA2, preventing inappropriate associations and counteracting protein aggregation (Guo, Fare, & Shorter, 2019; Jakel, Mingot, Schwarzmaier, Hartmann, & Gbrlich, 2002; Khalil et al., 2022; Springhower, Rosen, & Chook, 2020).

[0005] U.S. Patent No. 9,931 ,339 discloses certain compounds that bind to and inhibit the nuclear transport activity of Kpn|31 for use in the treatment of cancer. This activity is based on the knowledge that Kpn|31 mRNA and protein is expressed in elevated levels in certain cancer cells. The elevated levels of Kpn|31 may suggest that the Kpn[31 protein is associated with cellular transformation and cancer, and it has been shown that inhibition of Kpn|31 protein expression in cancer cells leads to apoptosis, emphasizing the role of this protein in cancer development. Inhibition of Kpn|31 protein expression in non-cancer cells has only a minor effect on cell viability. The compounds in U.S. Patent No. 9,831 ,339 were designed to bind amino acid residues 331-364 of Kpn|31 as this region was identified as a common region for the binding of Kpn|31 to RanGTP, SREBP-2, Kpna and Kpn|32 for the transport of classical NLS-containing cargos.

[0006] The background herein is included solely to explain the context of the disclosure. This is not to be taken as an admission that any of the material referred to was published, known, or part of the common general knowledge as of the priority date.SUMMARY

[0007] Through investigating the mechanisms of nuclear import of TDP-43, compounds have been identified as a small molecules that abrogate aggregation of TDP-43 fragments and / or isoforms, and it has been established that the compounds act by increasing cytoplasmic levels of Kpn|31 , promoting its chaperone activity. Kpn|31 levels also modulate aggregation of TDP-43 fragments and / or isoforms and nuclear localization, and the compounds, through ‘freeing up’ cytoplasmic Kpn[31 , have been shown to prevent and reverse aggregates of TDP-43 fragments. The compounds reduce aggregation of TDP-43 fragments in a dose-dependent manner in cells, primary cortical neurons, human-derived cortical neurons and in a TDP-43 proteinopathy mouse model. As this therapeutic mechanism is beingincreasingly shown to alleviate aggregation of different proteins, compounds of the application have the potential to possess therapeutic potential beyond TDP-43-centric proteinopathies. It was also shown that the identified compounds can harnesses Kpn|31 intrinsic chaperoning function to prevent / reverse aggregation of the SOD1 mutant, SOD1 G93A, and alpha- synuclein.

[0008] Therefore, the present application includes a method of increasing Kpnpi ’s chaperoning function in a subject comprising administering to a subject in need thereof, an effective amount of a compound of the application as defined herein.

[0009] The present application also includes a method of treating or preventing a proteinopathy comprising administering to a subject in need thereof an effective amount of a compound of the application as defined herein, wherein the proteinopathy comprises an abnormal configuration of one or more proteins that are chaperoned by Kpn[31 .

[0010] In some embodiments, the protein that is chaperoned by Kpn|31 is TDP-43, a fragment and / or isoform thereof, and the chaperoning prevents or reverses aggregation of the protein. Accordingly, the present application also includes a method of preventing or reducing the accumulation of pathological aggregates of TDP-43, a fragment and / or isoform thereof, or preventing or reducing pathological distribution of TDP-43, a fragment and / or isoform thereof, in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of the application as defined herein.

[0011] In some embodiments, the protein that is chaperoned by Kpn|31 is SOD1 , a fragment and / or isoform thereof, and the chaperoning prevents or reverses aggregation of the protein. Accordingly, the present application also includes a method of preventing or reducing the accumulation of pathological aggregates of SOD1 , a fragment and / or isoform thereof, or preventing or reducing pathological distribution of SOD1 , a fragment and / or isoform thereof, in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of the application as defined herein.

[0012] In some embodiments, the protein that is chaperoned by Kpn|31 is alpha- synuclein, a fragment and / or isoform thereof, and the chaperoning prevents or reverses aggregation of the protein. Accordingly, the present application also includes a method of preventing or reducing the accumulation of pathological aggregates of alpha-synuclein, a fragment and / or isoform thereof, or preventing or reducing pathological distribution of alpha- synuclein, a fragment and / or isoform thereof, in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of the application as defined herein.

[0013] In some embodiments, the subject is suffering from a neurodegenerative disorder. In some embodiments, the neurodegenerative disorder is argyrophilic grain disease, Alzheimer’s disease, amyotrophic lateral sclerosis (ALS), ALS-Parkinsonism dementia complex of Guam, corticobasal degeneration, Dementia with Lewy bodies, Huntington’s disease, Lewy body disease, motor neuron disease, frontotemporal lobar degeneration (FTLD), frontotemporal dementia, frontotemporal lobar degeneration with ubiquitin-positive inclusions, hippocampal sclerosis, inclusion body myopathy, inclusion body myositis, Parkinson’s disease, Parkinson’s disease dementia, Parkinson-dementia complex in Kii peninsula, chronic traumatic encephalopathy, Limbic-predominant age-related TDP- 43 encephalopathy (LATE), Pick's disease, or Creutzfeldt-Jakob disease, or a combination thereof. In some embodiments, the neurodegenerative disorder is ALS, frontotemporal dementia, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, chronic traumatic encephalopathy or LATE. In some embodiments, neurodegenerative disorder is REM sleep behavior disorder (RBD), multiple systems atrophy (MSA) or pure autonomic failure (PAF), or a combination thereof, or a combination with any of the above listed disorders).

[0014] In some embodiments, the methods and uses of the application reduce the number or frequency of neurons in the brain or spinal cord in the subject in need thereof that include mislocalized and / or misfolded proteins that are chaperoned by Kpn|31 , or fragments and / or isoforms thereof. In some embodiments, the methods and uses of the application reduce the amount or concentration of misfolded neuronal proteins that are chaperoned by Kpn p 1 , or fragments and / or isoforms thereof, in the brain or spinal cord of the subject in need thereof. In some embodiments, the methods and uses of the application reduce the amount or concentration of mislocalized neuronal proteins that are chaperoned by Kpn|31 , or fragments and / or isoforms thereof, in the brain or spinal cord of the subject in need thereof.

[0015] Other features and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating embodiments of the disclosure, are given by way of illustration only and the scope of the claims should not be limited by these embodiments, but should be given the broadest interpretation consistent with the description as a whole.DRAWINGS

[0016] Certain embodiments of the disclosure will now be described in greater detail with reference to the attached drawings in which:

[0017] Figure 1 shows exemplary compound JRMS prevents / reverses aggregation of TDP-43 fragments and / or isoforms allowing their nuclear localization in cells. (A) Immunofluorescence image of HeLa cells expressing GFP-TDP-25, GFP-TDP-35, GFP-TDP- 43ANLS and GFP-TDP-C-spl-272 treated with 15pM JRMS showing decreased aggregates (arrow) compared to DMSO control. Scale bar = 4pm. The percentage of cells with TDP-25 aggregates (B), TDP-35 aggregates (C), TDP-43 ANLS aggregates, and TDP-C-spl-272 (E) is significantly decreased. JRMS treatment was also shown to enhance TDP-43 fragments nuclear localization in (A), the Nuclear:Cytoplasmic ratio of fragments is quantified in (F-l). JRMS leads to a significant increase in nuclear localization of TDP-35 (G) and TDP-C-spl-272 (I), and an upward trend for TDP-25 (F) and TDP-43 ANLS (H). N=3 biological replicates p<0.05"p<0.01mp<0.001

[0018] Figure 2 shows exemplary compound JRMS prevents / reverses aggregation of TDP-25 in a dose-dependent manner in cells. (A) Immunofluorescence image of HeLa cells expressing GFP-TDP-25 treated with 15pM JRMS showing decreased aggregates (arrow) compared to DMSO control. Scale bar = 15pm. (B) GFP-TDP-25 aggregates volume are decreased upon JRMS treatment compared to DMSO control. (C) Immunoblot of solubility fractionation of cell lysates expressing TDP-25 treated with DMSO control or 15pM JRMS showing decreased insoluble fraction, quantified in (D). (E) Acute dose response curve showing the TDP-25 aggregates volume decreasing with increasing concentration of JRMS. (F) Chronic dose response curve showing the percentage of cells with TDP-25 aggregates decreasing with increasing concentration of JRMS compared to DMSO control. (G) Chronic dose response curve showing TDP-25 aggregates volume decreasing with increasing concentration of JRMS compared to DMSO control. N=3 biological replicates 'p<0.05 "p<0.01mp<0.001

[0019] Figure 3 shows exemplary compound JRMS prevents / reverses co-aggregation of TDP-25 and TDP-43ANLS in cells. (A) Immunofluorescence image of HeLa cells expressing GFP-TDP-43ANLS and RFP-TDP-25 treated with 15pM JRMS show a decrease in TDP- 43ANLS & TDP-25 aggregates compared to DMSO control, quantified in (B). Scale bar = 4 pm. N=3 biological replicates *p<0.05 p<0.01 p<0.001

[0020] Figure 4 shows compound of Formula II prevents / reverses aggregation of TDP- 25 in cells. (A) Immunofluorescence image of HeLa cells expressing GFP-TDP-25 treated with 15pM compound of Formula II (referred to as JRMS-44) showing decreased aggregates (arrow) compared to DMSO control, quantified in (B). Scale bar = 50pm. N=3 biological replicates 'p<0.05 "p<0.01 *"p<0.001

[0021] Figure 5 shows that exemplary compound JRMS prevents / reverses TDP-25 aggregation in mouse primary cortical neurons. (A) Mouse primary cortical neurons treated with up to 30pM JRMS exhibit no significant neurotoxicity. (B) Immunofluorescence image of mouse primary cortical neurons transduced to express GFP-TDP-25 treated with 15pM JRMS showing decreased aggregates (arrow) compared to DMSO control. Scale bar = 21 m. (C) Treatment with 15pM JRMS reduces the percentage of mouse primary cortical neurons with TDP-25 aggregates compared to DMSO control. (D) Treatment with 15pM JRMS of mouse primary cortical neurons reduces the size of TDP-25 aggregates compared to DMSO control. (E) Immunoblot analysis of total lysates of mouse primary cortical neurons transduced to express GFP, GFP-TDP-25 and GFP-TDP-43 treated with 15pM JRMS or DMSO, immunolabeled for Kpn|31 , GFP, pTDP (phosphorylated TDP-43 at S409 / 410) and GAPDH. JRMS treatment reduces the amount of phosphorylated TDP-25 without affecting the total level of expression of the GFP-tagged proteins. (F) Quantification of Kpn|31 levels shows no changes upon JRMS and DMSO treatments. (G) Quantification of pTDP levels in GFP-TDP- 25 expressing primary cortical neurons showing a reduction upon JRMS treatment compared to DMSO control. N=3 biological replicates 'p<0.05 "p<0.01 "p<0.001

[0022] Figure 6 shows that exemplary compound JRMS prevents / reverses TDP-25 aggregation in human iPSCs-derived cortical neurons. (A) Immunofluorescence of DMSO or JRMS treated human iPSCs-derived primary cortical neurons transduced to express GFP- TDP-25 treated with 7.5pM JRMS labelled with phosphorylated TDP-43 S409 / 410 (pTDP) and neuronal marker (NeuN). JRMS treatment reduces aggregates (arrow) compared to DMSO control. Scale bar = 20pm. (B) Quantification of number of aggregates per neuron is significantly decreases upon JRMS treatment. N=3 biological replicates 'p<0.05 "p<0.01mp<0.001

[0023] Figure 7 shows Kpn|31 interacts with TDP-25 and modulates its aggregation. (A) Immunoblot of solubility fractionation of cell lysates expressing TDP-25 treated with siScram or siKpnp, then further treated with DMSO control or 15pM of exemplary compound JRMS. Compared to DMSO controls, JRMS treatments leads to a reduction in insoluble phosphorylated S490 / 410 TDP-25 (pTDP) in siScram condition, but leads to an increased insoluble phosphorylated S490 / 410 TDP-25 (pTDP)in siKpn|31 condition, quantified in (B), indicating that JRMS effect is dependent on Kpn|31. (C) Immunofluorescence image of cells expressing GFP or Kpn|31-GFP treated with 15pM JRMS or DMSO control. JRMS treatment leads to increased cytoplasmic Kpn|31 as quantified in (D) as a nuclear:cytoplasmic ratio. Scale bar = 15pm. (E) Immunofluorescence showing no changes in endogenous TDP-43 nuclear localization in cells treated with DMSO or JRMS (Scale bar = 4pm), quantified in (F). (G) Kpn|31 levels modulate TDP-25 aggregation. Overexpression of Kpn|31 reduces insolubleTDP-25, while Kpn|31 knockdown with siRNA increases insoluble TDP-25, quantified in (H). (I) Immunofluorescence image of cells co-expressing GFP with FLAG-mTB-TDP-25 or Kpnpi- GFP with FLAG-mTB-TDP-25, showing reduction in cells with TDP-25 aggregates upon Kpnpi overexpression (Scale bar = 15pm), quantified in (J). (K) Co-immunoprecipitation of GFP-TDP-25 pulling down Kpn|31 in cells treated with both JRMS and DMSO control. JRMS treatment results in an overall less immunoprecipitated Kpn|31 (L) compared to DMSO control. JRMS treatment results in an increased immunoprecipitated Kpn|31 normalized to amount of phosphorylated aggregated TDP-25 (pTDP) compared to DMSO control (M). N=3 biological replicates 'p<0.05 "p<0.01 *"p<0.001

[0024] Figure 8 shows that exemplary compound JRMS prevents / reverses TDP-25 aggregation in vivo. (A) Immunohistochemistry of DMSO or JRMS treated AAV9-GFP-TDP- 25 mouse brains labelled with neuronal marker (NeuN) and phosphorylated TDP-43 S409 / 410 (pTDP). There is lower abundance of phosphorylated TDP-25 aggregates (arros) with JRMS treatment compared to DMSO control. Scale bar = 57pm. TDP-25 aggregates are smaller in JRMS treated mice as quantified in (B), and show a trend toward decreased abundance as quantified in (C). (D) There is no detectable change in neuronal density in the posterior cortex in DMSO and JRMS treated mice, supporting no toxicity associated with JRMS. (E) Immunoblot of DMSO and JRMS treated mice showing equivalent expression levels of Kpn|31 and GFP-TDP-25, but a ~30% decrease in levels of phosphorylated TDP-25, an indicator of reduced aggregation, as quantified in (F). (G) JRMS treatment affects Kpn|31 subcellular localization, increasing its cytoplasmic concentration, quantified in (H). Scale bar = 10pm. N=4 mice per treatment condition for immunohistochemical analysis and N=8 mice per treatment condition for biochemical analysis. 'p<0.05 "p<0.01mp<0.001

[0025] Figure 9 shows that exemplary compound JRMS prevents / reverses TDP-25 aggregation in organotypic slices. (A) Live cell imaging of organotypic slices demonstrating GFP-TDP-25 aggregates (bright round inclusions) abundance over 7 days. Inset in DMSO at day 7 panel is immunohistochemistry with antibody to phosphorylated TDP-43 serine 409 / 410 confirming that bright GFP signal is indeed aggregated TDP-25. Scale bar = 114pm (B) Plot showing percentage of change in number of TDP-25 aggregate over 7 days of treatment. There is a clearance of TDP-25 aggregates in JRMS treated mice compared to DMSO control. N=4 slices per treatment condition. 'p<0.05 "p<0.01mp<0.001

[0026] Figure 10 shows that exemplary compound JRMS prevents / reverses other disease associated aggregation-prone proteins. (A) Immunofluorescence image of HeLa cells expressing SOD1 G93A-FLAG treated with 15pM JRMS showing decreased cytoplasmic inclusions compared to DMSO control, quantified in (B). Scale bar = 15pm. (C) Immunofluorescence image of HeLa cells expressing HA-a-Synuclein-6xHis treated with15pM JRMS showing decreased cytoplasmic aggregates compared to DMSO control, quantified in (D). N=3 biological replicates. p<0.05 p<0.01 p<0.001

[0027] Figure 11 shows the sequence of Kpn|31 [SEQ ID NO:1],

[0028] Figure 12 shows the sequence of TDP-43 [SEQ ID NO:2],

[0029] Figure 13 shows the sequence of TDP-25 [SEQ ID NO:3],

[0030] Figure 14 shows the sequence of TDP-35 [SEQ ID NO:4],

[0031] Figure 15 shows the sequence of TDP-43 ANLS [SEQ ID NO:5],

[0032] Figure 16 shows the sequence of TDP-C-spl-272 [SEQ ID NO:6],

[0033] Figure 17 shows the sequence of SOD1 G93A [SEQ ID NO:7],

[0034] Figure 18 shows the sequence of alpha-synuclein [SEQ ID NO:8],DETAILED DESCRIPTIONI. Definitions

[0035] Unless otherwise indicated, the definitions and embodiments described in this and other sections are intended to be applicable to all embodiments and aspects of the present disclosure herein described for which they are suitable as would be understood by a person skilled in the art. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting.

[0036] In understanding the scope of the present disclosure, the term "comprising" and its derivatives, as used herein, are intended to be open ended terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The foregoing also applies to words having similar meanings such as the terms, "including", "having" and their derivatives. The term "consisting" and its derivatives, as used herein, are intended to be closed terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The term "consisting essentially of, as used herein, is intended to specify the presence of the stated features, elements, components, groups, integers, and / or steps as well as those that do not materially affect the basic and novel characteristic(s) of features, elements, components, groups, integers, and / or steps.

[0037] Terms of degree such as "substantially", "about” and "approximately" as used herein mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms of degree should be construed as including adeviation of at least ±5% of the modified term if this deviation would not negate the meaning of the word it modifies. In addition, all ranges given herein include the end of the ranges and also any intermediate range points, whether explicitly stated or not.

[0038] As used in this disclosure, the singular forms "a", "an" and "the" include plural references unless the content clearly dictates otherwise.

[0039] In embodiments comprising an "additional" or "second" component, the second component as used herein is chemically different from the other components or first component. A "third" component is different from the other, first, and second components, and further enumerated or "additional" components are similarly different.

[0040] The term "and / or" as used herein means that the listed items are present, or used, individually or in combination. In effect, this term means that "at least one of” or "one or more" of the listed items is used or present.

[0041] The term “compound of the application” or “active ingredient” as used herein refers to a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof.

[0042] The term “pharmaceutically acceptable” means compatible with the treatment of subjects, for example humans.

[0043] The term “pharmaceutically acceptable carrier” means a non-toxic solvent, dispersant, excipient, adjuvant or other material which is mixed with the active ingredient in order to permit the formation of a pharmaceutical composition, i.e., a dosage form capable of administration to a subject.

[0044] The term “pharmaceutically acceptable salt” means either an acid addition salt or a base addition salt which is suitable for, or compatible with the treatment of subjects.

[0045] The term “solvate” as used herein means a compound, or a salt, and / or prodrug of a compound, wherein molecules of a suitable solvent are incorporated in the crystal lattice. A suitable solvent is physiologically tolerable at the dosage administered. When the solvent is water, the solvate is referred to as a “hydrate”.

[0046] The term “prodrug” as used herein means a compound, or salt and / or solvate of a compound, that, after administration, is converted into an active drug.

[0044] The term "subject" as used herein includes all members of the animal kingdom including mammals, and suitably refers to humans. Thus the methods of the present application are applicable to both human therapy and veterinary applications.

[0047] The term “treating” or “treatment” as used herein and as is well understood in the art, means an approach for obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission (whether partial or total), whether detectable or undetectable. “Treating” and “treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment. “Treating” and “treatment” as used herein also include prophylactic treatment. Treatment methods comprise administering to a subject a therapeutically effective amount of one or more of the compounds of the application and optionally consist of a single administration, or alternatively comprise a series of administrations.

[0048] As used herein, the term “effective amount” or “therapeutically effective amount” means an amount of a compound, or one or more compounds, of the application that is effective, at dosages and for periods of time necessary to achieve the desired result.

[0049] By “reducing” it is meant any detectable reduction of a stated parameter in the presence of a compound, or one or more compounds, of the application compared to otherwise the same conditions, except for in the absence of the compound, or one or more compounds.

[0050] By “increasing” it is meant any detectable increase of a stated parameter in the presence of a compound, or one or more compounds, of the application compared to otherwise the same conditions, except for in the absence of the compound, or one or more compounds.

[0051] The term “administered” as used herein means administration of a therapeutically effective amount of a compound, or one or more compounds, of the application, or a composition of the application, to a cell either in cell culture or in a subject.

[0052] The term “proteinopathy” as used herein refers to the acquisition of abnormal conformations by proteins (i.e. misfolding of proteins) and in particular, their tendency to aggregate losing their functionality and inducing toxicity, wherein such abnormal conformation of proteins in the brain is associated with a neurodegenerative disease.

[0053] The term “isoform” as used herein refers to a protein that is highly similar to a reference protein and that originates from a single gene or gene family, and are the result of genetic differences. A protein isoform is also known as a protein variant.

[0054] It will be understood that any component defined herein as being included may be explicitly excluded by way of proviso or negative limitation, such as any specific compounds or method steps, whether implicitly or explicitly defined herein.II. Methods and Uses of the Application

[0055] The present application includes a method of increasing Kpn[31 ’s chaperoning function in a subject comprising administering to a subject in need thereof, an effective amount of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof:wherein:R1is selected from Ci.6alkyl, Ci.4alkyleneN(Ci.4alkyl)2, Ci.4alkyleneOH,R2is selected from H and Ci_4alkyl;

[0056] The present application includes a use of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for increasing Kpn[31 ’s chaperoning function in a subject and a use of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for preparation of a medicament for increasing Kpnpi ’s chaperoning function in a subject. Also included is a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for use to increase Kpnpi ’s chaperoning function in a subject.

[0057] In accordance with an aspect of the present application, there is provided a method of treating or preventing a proteinopathy comprising administering to a subject in need thereof an effective amount of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, wherein the proteinopathy comprises an abnormal configuration of one or more proteins that are chaperoned by Kpn[31 .

[0058] The present application includes a use of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for treating or preventing a proteinopathy wherein the proteinopathy comprises an abnormal configuration of one or more proteins thatare chaperoned by Kpn|31 and a use of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for preparation of a medicament for treating or preventing a proteinopathy wherein the proteinopathy comprises an abnormal configuration of one or more proteins that are chaperoned by Kpn|31. Also included is a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for use to treat or prevent a proteinopathy wherein the proteinopathy comprises an abnormal configuration of one or more proteins that are chaperoned by Kpn|31 .

[0059] In some embodiments, Kpn|31 is a protein that in humans has a sequence as shown in [SEQ ID NO:1] in Figure 11.

[0060] In some embodiments, the protein that is chaperoned by Kpn|31 is transactive response DNA-binding protein 43 (TDP-43), Superoxide dismutase 1 (SOD1), Fused in sarcoma (FUS), Chromosome 9 open reading frame 72 (C9orf72), C9orf72-dipetide repeat proteins (GA, GR, PA, PR, GP), Heterogeneous nuclear ribonucleoproteins (hnRNPs), Tubulin associated unit (Tau), alpha-synuclein, Annexin A11 (ANXA11), CHCHD10 (CHC10), prion proteins (PrP), transthyretin (TTR) or huntingtin (HTT), or a fragment or isoform thereof. In some embodiments, the protein that is chaperoned by Kpn|31 is transactive response DNA- binding protein 43 (TDP-43), Superoxide dismutase 1 (SOD1), Fused in sarcoma (FUS), Chromosome 9 open reading frame 72 (C9orf72), C9orf72-dipetide repeat proteins (GA, GR, PA, PR, GP), Heterogeneous nuclear ribonucleoproteins (hnRNPs), Tubulin associated unit (Tau), or alpha-synuclein, or a fragment and / or isoform thereof. In some embodiments, the protein that is chaperoned by Kpn|31 is transactive response DNA-binding protein 43 (TDP- 43), Superoxide dismutase 1 (SOD1) or alpha-synuclein, and / or a fragment or isoform thereof.

[0061] In some embodiments, the protein that is chaperoned by Kpn|31 is TDP-43, a fragment and / or isoform thereof, and the chaperoning prevents or reverses aggregation of TDP-43, a fragment and / or isoform thereof. Accordingly, the present application also includes a method of preventing or reducing the accumulation of pathological aggregates of TDP-43, a fragment and / or isoform thereof, or preventing or reducing pathological distribution of TDP- 43, a fragment and / or isoform thereof, in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above.

[0062] The present application includes a use of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt,solvate and / or prodrug thereof, as defined above, for preventing or reducing the accumulation of pathological aggregates of TDP-43, a fragment and / or isoform thereof, or for preventing or reducing pathological distribution of TDP-43, a fragment and / or isoform thereof, in a subject in need thereof, and a use of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for preparation of a medicament for preventing or reducing the accumulation of pathological aggregates of TDP-43, a fragment and / or isoform thereof, or for preventing or reducing pathological distribution of TDP-43, a fragment and / or isoform thereof, in a subject in need thereof. Also included is a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for preventing or reducing the accumulation of pathological aggregates of TDP-43, a fragment and / or isoform thereof, or for preventing or reducing pathological distribution of TDP-43, a fragment and / or isoform thereof, in a subject in need thereof.

[0063] In some embodiments, the protein that is chaperoned by Kpn[31 is TDP-43. In some embodiments, the protein that is chaperoned by Kpn|31 is a fragment and / or isoform of TDP-43. In some embodiments, the fragment and / or isoform of TDP-43 is TDP-25. In some embodiments, the fragment and / or isoform of TDP-43 is TDP-35. In some embodiments, the fragment and / or isoform of TDP-43 is TDP-43ANLS. In some embodiments, the fragment and / or isoform of TDP-43 is TDP-C-spl-272.

[0064] In some embodiments, TDP-43 is a protein that in humans is encoded by the TARDBP gene and has a sequence as shown in [SEQ ID NO:2] in Figure 12.

[0065] In some embodiments, TDP-25 is a fragment of TDP-43 that has a sequence as shown in [SEQ ID NO:3] in Figure 13. It is of note that there are multiple start sites for TDP- 25, with the exemplary one described herein arising from caspase 3 site at amino acid 219 (Arai et al., 2006; Neumann et al., 2006, Dormann et al., 2009).

[0066] In some embodiments, TDP-35 is a fragment of TDP-43 that has a sequence as shown in [SEQ ID NO:4] in Figure 14. It is of note that there are multiple start sites for TDP- 35, with the exemplary one described herein arising from alternative splicing in exon 2 leading to the start site at amino acid 85 (Xiao, et al., 2015a).

[0067] In some embodiments, TDP-43ANLS is a fragment of TDP-43 that has a sequence as shown in [SEQ ID NO:5] in Figure 15. TDP-43ANLS is the full length protein TDP-43 with three point mutations K82A, R83A, K84A compromising the nuclear localization sequence (NLS) recognition by importin-a (Kpna) (Winton et al., 2008).

[0068] In some embodiments, TDP-C-spl-272 is a fragment of TDP-43 that has a sequence as shown in [SEQ ID NO:6] in Figure 16. It is of note that there are multiple N- terminal fragments of TDP-43, herein the exemplary TDP-C-spl-272 arising from alternative splicing in exon 6 is referenced (Shenouda, et al., 2022).

[0069] In some embodiments, the protein that is chaperoned by Kpn|31 is SOD1 , a fragment and / or isoform thereof, and the chaperoning prevents or reverses aggregation of SOD1 , a fragment and / or isoform thereof. Accordingly, the present application also includes a method of preventing or reducing the accumulation of pathological aggregates of SOD1 , a fragment and / or isoform thereof, or preventing or reducing pathological distribution of SOD1 , a fragment and / or isoform thereof, in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above.

[0070] The present application includes a use of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for preventing or reducing the accumulation of pathological aggregates of SOD1 , a fragment and / or isoform thereof, or for preventing or reducing pathological distribution of SOD1 , a fragment and / or isoform thereof, in a subject in need thereof, and a use of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for preparation of a medicament for preventing or reducing the accumulation of pathological aggregates of SOD1 , a fragment and / or isoform thereof, or for preventing or reducing pathological distribution of SOD1 , a fragment and / or isoform thereof, in a subject in need thereof. Also included is a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for preventing or reducing the accumulation of pathological aggregates of SOD1 , a fragment and / or isoform thereof, or for preventing or reducing pathological distribution of SOD1 , a fragment and / or isoform thereof, in a subject in need thereof.

[0071] In some embodiments, the protein that is chaperoned by Kpn|31 is SOD1. In some embodiments, the protein that is chaperoned by Kpn|31 is a fragment and / or isoform of SOD1 . In some embodiments, the fragment and / or isoform of SOD1 is SOD1 G93A [SEQ ID NO:7] in Figure 17.

[0072] In some embodiments, the protein that is chaperoned by Kpn|31 is alpha- synuclein, a fragment and / or isoform thereof, and the chaperoning prevents or reverses aggregation of alpha-synuclein, a fragment and / or isoform thereof. Accordingly, the presentapplication also includes a method of preventing or reducing the accumulation of pathological aggregates of alpha-synuclein, a fragment and / or isoform thereof, or preventing or reducing pathological distribution of alpha-synuclein, a fragment and / or isoform thereof, in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above.

[0073] The present application includes a use of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for preventing or reducing the accumulation of pathological aggregates of alpha-synuclein, a fragment and / or isoform thereof, or for preventing or reducing pathological distribution of alpha-synuclein, a fragment and / or isoform thereof, in a subject in need thereof, and a use of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for preparation of a medicament for preventing or reducing the accumulation of pathological aggregates of alpha-synuclein, a fragment and / or isoform thereof, or for preventing or reducing pathological distribution of alpha-synuclein, a fragment and / or isoform thereof, in a subject in need thereof. Also included is a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for preventing or reducing the accumulation of pathological aggregates of alpha-synuclein, a fragment and / or isoform thereof, or for preventing or reducing pathological distribution of alpha-synuclein, a fragment and / or isoform thereof, in a subject in need thereof.

[0074] In some embodiments, the protein that is chaperoned by Kpn|31 is alpha- synuclein [SEQ ID NO:8] in Figure 18. In some embodiments, the protein that is chaperoned by Kpn|31 is a fragment and / or isoform of alpha-synuclein.

[0075] In the above method and use embodiments, references to “a fragment” or “an isoform” mean a one fragment or isoform or a mixture of two or more different fragments or isoforms.

[0076] As used in the art and in one aspect of its use and meaning herein, a protein isoform refers to variants of any of the polypeptides referenced herein that are encoded by the same gene, and that may differ in their pl or MW, or both. Such isoforms can differ in their amino acid composition (e.g. as a result of mutation, alternative mRNA or premRNA processing, e.g. alternative splicing, cleavage or limited proteolysis) and in addition, or in the alternative, may arise from differential post-translational modification (e.g., glycosylation, acylation, phosphorylation). It should be noted, that the term “protein isoform” as used hereinincludes both the expected / wild type polypeptide and any variants thereof. The protein isoform is any that may exist in a subject and for which it is beneficial for the aggregation of which to be prevented or reduced.

[0077] A fragment of a protein or protein isoform refers to any sized fragment of a protein or protein isoform referenced herein that may exist in a subject and for which it is beneficial for the aggregation of which to be prevented or reduced.

[0078] In some embodiments, the subject is suffering from a neurodegenerative disorder. Therefore the present application also includes a method of treating or preventing a neurodegenerative disorder in a subject in need thereof comprising administering to the subject, an effective amount of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above.

[0079] The present application includes a use of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for treating or preventing a neurodegenerative disorder and a use of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for preparation of a medicament for treating or preventing a neurodegenerative disorder. Also included is a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined above, for treating or preventing a neurodegenerative disorder. In some embodiments, the neurodegenerative disorder is argyrophilic grain disease, Alzheimer’s disease, amyotrophic lateral sclerosis (ALS), ALS-Parkinsonism dementia complex of Guam, corticobasal degeneration, Dementia with Lewy bodies, Huntington’s disease, Lewy body disease, motor neuron disease, frontotemporal lobar degeneration (FTLD), frontotemporal dementia, frontotemporal lobar degeneration with ubiquitin-positive inclusions, hippocampal sclerosis, inclusion body myopathy, inclusion body myositis, Parkinson’s disease, Parkinson’s disease dementia, Parkinson-dementia complex in Kii peninsula, chronic traumatic encephalopathy, Limbic-predominant age-related TDP-43 encephalopathy (LATE), Pick's disease or Creutzfeldt- Jakob disease, or a combination thereof. In some embodiments, the neurodegenerative disorder is ALS, frontotemporal dementia, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, chronic traumatic encephalopathy or LATE.

[0080] In some embodiments, neurodegenerative disorder is REM sleep behavior disorder (RBD), multiple systems atrophy (MSA) or pure autonomic failure (PAF), or a combination thereof, or a combination with any of the above listed disorders).

[0081] In some embodiments, the methods and uses of the application reduce the number or frequency of neurons in the brain or spinal cord in the subject in need thereof that include mislocalized and / or misfolded TDP-43, or a fragment and / or isoform thereof. In some embodiments, the methods and uses of the application reduce the amount or concentration of misfolded neuronal TDP-43, or a fragment and / or isoform thereof, in the brain or spinal cord of the subject in need thereof.

[0082] In some embodiments, the methods and uses of the application reduce the number or frequency of TDP-43 positive inclusion-positive neurons in the brain or spinal cord in the subject in need thereof. In some embodiments, the methods and uses of the application reduce the amount or concentration of neuronal TDP-43 misfolded protein in the brain or spinal cord the subject in need thereof. In some embodiments, the methods and uses of the application reduce the amount or concentration of neuronal mislocalizaed TDP-43 protein in the brain or spinal cord the subject in need thereof.

[0083] In some embodiments, the methods and uses of the application reduce the number or frequency of neurons in the brain or spinal cord in the subject in need thereof that include mislocalized and / or misfolded SOD1 , or a fragment and / or isoform thereof. In some embodiments, the methods and uses of the application reduce the amount or concentration of misfolded neuronal SOD1 , or a fragment and / or isoform thereof, in the brain or spinal cord of the subject in need thereof.

[0084] In some embodiments, the methods and uses of the application reduce the number or frequency of SOD1 positive inclusion-positive neurons in the brain or spinal cord in the subject in need thereof. In some embodiments, the methods and uses of the application reduce the amount or concentration of neuronal SOD1 misfolded protein in the brain or spinal cord the subject in need thereof. In some embodiments, the methods and uses of the application reduce the amount or concentration of neuronal mislocalizaed SOD1 protein in the brain or spinal cord the subject in need thereof.

[0085] In some embodiments, the methods and uses of the application reduce the number or frequency of neurons in the brain or spinal cord in the subject in need thereof that include mislocalized and / or misfolded alpha-synuclein, or a fragment and / or isoform thereof. In some embodiments, the methods and uses of the application reduce the amount or concentration of misfolded neuronal alpha-synuclein, or a fragment and / or isoform thereof, in the brain or spinal cord of the subject in need thereof.

[0086] In some embodiments, the methods and uses of the application reduce the number or frequency of alpha-synuclein positive inclusion-positive neurons in the brain or spinal cord in the subject in need thereof. In some embodiments, the methods and uses of the application reduce the amount or concentration of neuronal alpha-synuclein misfolded protein in the brain or spinal cord the subject in need thereof. In some embodiments, the methods and uses of the application reduce the amount or concentration of neuronal mislocalizaed alpha-synuclein protein in the brain or spinal cord the subject in need thereof.

[0087] In some embodiments, the subject is a mammal. In some embodiments, the subject is human.

[0088] In some embodiments, the compound is a compound of Formula I, or pharmaceutically acceptable salt, solvate and / or prodrug thereof:(I) wherein: R1is selected from Ci.6alkyl, Ci.4alkyleneN(Ci.4alkyl)2, Ci.4alkyleneOH,R2is selected from H and Ci_4alkyl.

[0089] In some embodiments, the compound of Formula I, or pharmaceutically acceptable salt, solvate and / or prodrug thereof is:solvate and / or prodrug thereof.

[0090] In some embodiments, the compound of Formula I, or pharmaceutically acceptable salt, solvate and / or prodrug thereof ispharmaceutically acceptable salt, solvate and / or prodrug thereof. This compound also has the chemical name (3-(1 H-benzimidazol-2-yl)-1-(3- dimethylaminopropyl)pyrrolo[5,4-b]quinoxalin-2-amine) and is also referred to herein as JRMS.

[0091] In some embodiments, the compound of the application is a compound of Formula II, or pharmaceutically acceptable salt, solvate and / or prodrug thereof:This compound has the chemical name 9-((1- methylpiperidin-3-yl)methoxy)-4-((6-methylpyridin-2-yl)methyl)-7-(5-methylthiophen-2-yl)-2,3,4,5-tetrahydrobenzo[f][1 ,4]oxazepine and is also referred to herein as JRMS-44.

[0092] In some embodiments the pharmaceutically acceptable salt is an acid addition salt or a base addition salt. The selection of a suitable salt may be made by a person skilled in the art (see, for example, S. M. Berge, et al., "Pharmaceutical Salts," J. Pharm. Sci. 1977, 66, 1-19).

[0093] In some embodiments the pharmaceutically acceptable salt is an acid addition salt. An acid addition salt suitable for, or compatible with, the treatment of subjects is any nontoxic organic or inorganic acid addition salt of any basic compound. Basic compounds that form an acid addition salt include, for example, compounds comprising an amine group. Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulfuric, nitric and phosphoric acids, as well as acidic metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids which form suitable salts include mono-, di- and tricarboxylic acids. Illustrative of such organic acids are, for example, acetic, formic, trifluoroacetic, propionic, glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, hydroxymaleic,benzoic, hydroxybenzoic, phenylacetic, cinnamic, mandelic, salicylic, 2-phenoxybenzoic, p- toluenesulfonic acid and other sulfonic acids such as methanesulfonic acid, ethanesulfonic acid and 2-hydroxyethanesulfonic acid. In an embodiment, the mono- or di-acid salts are formed, and such salts exist in either a hydrated, solvated or substantially anhydrous form. In general, acid addition salts are more soluble in water and various hydrophilic organic solvents, and generally demonstrate higher melting points in comparison to their free base forms. The selection criteria for the appropriate salt will be known to one skilled in the art. Other non- pharmaceutically acceptable salts such as but not limited to oxalates may be used, for example in the isolation of compounds of the application for laboratory use, or for subsequent conversion to a pharmaceutically acceptable acid addition salt. In some embodiments, the acid addition salt is a hydrochloric or formic acid addition salt.

[0094] Solvates of compounds of the application include, for example, those made with solvents that are pharmaceutically acceptable. Examples of such solvents include water (resulting solvate is called a hydrate) and ethanol and the like. Suitable solvents are physiologically tolerable at the dosage administered.

[0095] Prodrugs of the compounds of the present application include, for example, conventional esters formed with available hydroxy, thiol, amino or carboxyl groups. Some common esters which have been utilized as prodrugs are phenyl esters, aliphatic (C1-C24) esters, acyloxymethyl esters, carbamates and amino acid esters.

[0096] In some embodiments of the present application, the compounds described herein may have at least one asymmetric center. Where compounds possess more than one asymmetric center, they may exist as diastereomers. It is to be understood that all such isomers and mixtures thereof in any proportion are encompassed within the scope of the present application. It is to be further understood that while the stereochemistry of the compounds may be as shown in any given compound listed herein, such compounds may also contain certain amounts (for example, less than 20%, suitably less than 10%, more suitably less than 5%) of compounds of the present application having an alternate stereochemistry. It is intended that any optical isomers, as separated, pure or partially purified optical isomers or racemic mixtures thereof are included within the scope of the present application.

[0097] The compounds of the present application may also exist in different tautomeric forms and it is intended that any tautomeric forms which the compounds form, as well as mixtures thereof, are included within the scope of the present application.

[0098] The compounds of the present application may further exist in varying polymorphic forms and it is contemplated that any polymorphs, or mixtures thereof, which form are included within the scope of the present application.

[0099] The compounds of the present application may further be radiolabeled and accordingly all radiolabeled versions of the compounds of the application are included within the scope of the present application. The compounds of the application also include those in which one or more radioactive atoms are incorporated within their structure.

[0100] Compounds of the present application are commercially available or may be prepared by various synthetic processes or may be extracted from cells, plants, animals or fungi. The selection of a particular process to prepare a given compound of the application is within the purview of the person of skill in the art. Some starting materials for preparing compounds of the present application are available from commercial chemical sources or may be extracted from cells, plants, animals or fungi.

[0101] The formation of a desired compound salt is achieved using standard techniques. For example, the neutral compound is treated with an acid or base in a suitable solvent and the formed salt is isolated by filtration, extraction or any other suitable method.

[0102] The formation of solvates of the compounds of the application will vary depending on the compound and the solvate. In general, solvates are formed by dissolving the compound in the appropriate solvent and isolating the solvate by cooling or using an antisolvent. The solvate is typically dried or azeotroped under ambient conditions. The selection of suitable conditions to form a particular solvate can be made by a person skilled in the art.

[0103] Prodrugs of the compounds of the present application may be, for example, conventional esters formed with available hydroxy, thiol, amino or carboxyl groups. For example, available hydroxy or amino groups may be acylated using an activated acid in the presence of a base, and optionally, in inert solvent (e.g. an acid chloride in pyridine).

[0104] Treatment methods and uses of the present application comprise administering to a subject a therapeutically effective amount of a compound of the application and optionally consist of a single administration, or alternatively comprise a series of administrations, and optionally comprise concurrent administration or use of one or more other therapeutic agents. For example, in some embodiments, the compound of the application is administered at least once a week. In some embodiments, the compound of the application is administered to the subject from about one time per two or three weeks, or about one time per week to about once daily for a given treatment. In another embodiment, the compound of the application is administered 2, 3, 4, 5 or 6 times daily. The length of the treatment period depends on a varietyof factors, such as the severity of the disease, disorder or condition, the age of the subject, the concentration and / or the activity of the compound of the application, and / or a combination thereof. It will also be appreciated that the effective dosage of the compound of the application used for the treatment may increase or decrease over the course of a particular treatment regime. Changes in dosage may result and become apparent by standard diagnostic assays known in the art. In some instances, chronic administration may be required. For example, the compound of the application is administered to the subject in an amount and for duration sufficient to treat the subject.

[0105] In the treatment methods and uses of the present application, the compounds of the application are suitably formulated in a conventional manner into compositions using one or more carriers. The compounds of the application are suitably formulated into pharmaceutical compositions for administration to subjects in a biologically compatible form suitable for administration in vivo.

[0106] The compounds of the application are administered to a subject in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. For example, compounds of the application are administered by oral, inhalation, parenteral, buccal, sublingual, nasal, rectal, vaginal, patch, pump, minipump, topical or transdermal administration and the pharmaceutical compositions formulated accordingly. In some embodiments, administration is by means of a pump for periodic or continuous delivery. Conventional procedures and ingredients for the selection and preparation of suitable compositions are described, for example, in Remington’s Pharmaceutical Sciences (2000 - 20th edition) and in The United States Pharmacopeia: The National Formulary (USP 24 NF 19) published in 1999.

[0107] Parenteral administration includes systemic delivery routes other than the gastrointestinal (Gl) tract, and includes, for example intravenous, intra-arterial, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary (for example, by use of an aerosol), intrathecal, rectal and topical (including the use of a patch or other transdermal delivery device) modes of administration. Parenteral administration may be by continuous infusion over a selected period of time.

[0108] In some embodiments, compounds of the application are orally administered, for example, with an inert diluent or with an assimilable edible carrier, or are enclosed in hard or soft shell gelatin capsules, or are compressed into tablets, or are incorporated directly with the food of the diet. In some embodiments, the compounds are incorporated with excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, caplets, pellets, granules, lozenges, chewing gum, powders, syrups, elixirs, wafers, aqueous solutions andsuspensions, and the like. In the case of tablets, carriers that are used include lactose, corn starch, sodium citrate and salts of phosphoric acid. Pharmaceutically acceptable excipients include binding agents (e.g., pregelatinized maize starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers (e.g., lactose, microcrystalline cellulose or calcium phosphate); lubricants (e.g., magnesium stearate, talc or silica); disintegrants (e.g., potato starch or sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulphate). In embodiments, the tablets are coated by methods well known in the art. In the case of tablets, capsules, caplets, pellets or granules for oral administration, pH sensitive enteric coatings, such as Eudragits™ designed to control the release of active ingredients are optionally used. Oral dosage forms also include modified release, for example immediate release and timed- release, formulations. Examples of modified-release formulations include, for example, sustained-release (SR), extended-release (ER, XR, or XL), time-release or timed-release, controlled-release (CR), or continuous-release (CR or Contin), employed, for example, in the form of a coated tablet, an osmotic delivery device, a coated capsule, a microencapsulated microsphere, an agglomerated particle, e.g., as of molecular sieving type particles, or, a fine hollow permeable fiber bundle, or chopped hollow permeable fibers, agglomerated or held in a fibrous packet. Timed-release compositions are formulated, for example as liposomes or those wherein the active compound is protected with differentially degradable coatings, such as by microencapsulation, multiple coatings, etc. Liposome delivery systems include, for example, small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. In some embodiments, liposomes are formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines. For oral administration in a capsule form, useful carriers or diluents include lactose and dried corn starch.

[0109] In some embodiments, liquid preparations for oral administration take the form of, for example, solutions, syrups or suspensions, or they are suitably presented as a dry product for constitution with water or other suitable vehicle before use. When aqueous suspensions and / or emulsions are administered orally, the compounds of the application are suitably suspended or dissolved in an oily phase that is combined with emulsifying and / or suspending agents. If desired, certain sweetening and / or flavoring and / or coloring agents are added. Such liquid preparations for oral administration are prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, methyl cellulose or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters or ethyl alcohol); and preservatives (e.g., methyl or propyl p-hydroxybenzoates or sorbic acid). Useful diluents include lactose and high molecular weight polyethylene glycols.

[0110] It is also possible to freeze-dry the compounds of the application and use the lyophilizates obtained, for example, for the preparation of products for injection.

[0111] In some embodiments, compounds of the application are administered parenterally. For example, solutions of compounds of the application are prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose. In some embodiments, dispersions are prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. A person skilled in the art would know how to prepare suitable formulations. For parenteral administration, sterile solutions of the compounds of the application are usually prepared, and the pH’s of the solutions are suitably adjusted and buffered. For intravenous use, the total concentration of solutes should be controlled to render the preparation isotonic. For ocular administration, ointments or droppable liquids are delivered, for example, by ocular delivery systems known to the art such as applicators or eye droppers. In some embodiments, such compositions include mucomimetics such as hyaluronic acid, chondroitin sulfate, hydroxypropyl methylcellulose or polyvinyl alcohol, preservatives such as sorbic acid, EDTA or benzyl chromium chloride, and the usual quantities of diluents or carriers. For pulmonary administration, diluents or carriers will be selected to be appropriate to allow the formation of an aerosol.

[0112] In some embodiments, compounds of the application are formulated for parenteral administration by injection, including using conventional catheterization techniques or infusion. Formulations for injection are, for example, presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. In some embodiments, the compositions take such forms as sterile suspensions, solutions or emulsions in oily or aqueous vehicles, and contain formulating agents such as suspending, stabilizing and / or dispersing agents. In all cases, the form is sterile fluid to the extent that easy syringability exists. Alternatively, the compounds of the application are suitably in a sterile powder form for reconstitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

[0113] In some embodiments, compositions for nasal administration are conveniently formulated as aerosols, drops, gels and powders. For intranasal administration or administration by inhalation, the compounds of the application are conveniently delivered in the form of a solution, dry powder formulation or suspension from a pump spray container that is squeezed or pumped by the patient or as an aerosol spray presentation from a pressurized container or a nebulizer. Aerosol formulations typically comprise a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container,which, for example, take the form of a cartridge or refill for use with an atomising device. Alternatively, the sealed container is a unitary dispensing device such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve which is intended for disposal after use. Where the dosage form comprises an aerosol dispenser, it will contain a propellant which is, for example, a compressed gas such as compressed air or an organic propellant such as fluorochlorohydrocarbon. Suitable propellants include but are not limited to dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, heptafluoroalkanes, carbon dioxide or another suitable gas. In the case of a pressurized aerosol, the dosage unit is suitably determined by providing a valve to deliver a metered amount. In some embodiments, the pressurized container or nebulizer contains a solution or suspension of the active compound. Capsules and cartridges (made, for example, from gelatin) for use in an inhaler or insufflator are, for example, formulated containing a powder mix of a compound of the application and a suitable powder base such as lactose or starch. The aerosol dosage forms can also take the form of a pump-atomizer.

[0114] Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, wherein compounds of the application are formulated with a carrier such as sugar, acacia, tragacanth, or gelatin and glycerine. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base such as cocoa butter.

[0115] Suppository forms of the compounds of the application are useful for vaginal, urethral and rectal administrations. Such suppositories will generally be constructed of a mixture of substances that is solid at room temperature but melts at body temperature. The substances commonly used to create such vehicles include but are not limited to theobroma oil (also known as cocoa butter), glycerinated gelatin, other glycerides, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights and fatty acid esters of polyethylene glycol. See, for example: Remington's Pharmaceutical Sciences, 16th Ed., Mack Publishing, Easton, PA, 1980, pp. 1530-1533 for further discussion of suppository dosage forms.

[0116] In some embodiments compounds of the application are coupled with soluble polymers as targetable drug carriers. Such polymers include, for example, polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxy-ethylaspartamide-phenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues. Furthermore, in some embodiments, compounds of the application are coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals,polydihydropyrans, polycyanoacrylates and crosslinked or amphipathic block copolymers of hydrogels.

[0117] In some embodiments, compounds of the application may be coupled with viral, non-viral or other vectors. Viral vectors may include retrovirus, lentivirus, adenovirus, herpesvirus, poxvirus, alphavirus, vaccinia virus or adeno-associated viruses. Non-viral vectors may include nanoparticles, cationic lipids, cationic polymers, metallic nanoparticles, nanorods, liposomes, micelles, microbubbles, cell-penetrating peptides, or lipospheres. Nanoparticles may include silica, lipid, carbohydrate, or other pharmaceutically acceptable polymers.

[0118] In some embodiments, compounds of the application are formulated using methods to increase blood brain barrier (BBB) penetration. Such methods are known in the art and include, for example, passive transcytosis, intranasal administration, ligand conjugation, membrane coating, and stimuli-triggered BBB disruption, as described in Wi, D. et al. Signal Transduction and Targeted Therapy, 8; Article Number 217 (2023).

[0119] A compound of the application including pharmaceutically acceptable salts, solvates and / or prodrugs thereof, is suitably used on their own but will generally be administered in the form of a pharmaceutical composition in which the one or more compounds of the application (the active ingredient) is in association with a pharmaceutically acceptable carrier. Depending on the mode of administration, the pharmaceutical composition will comprise from about 0.05 wt% to about 99 wt% or about 0.10 wt% to about 70 wt%, of the active ingredient, and from about 1 wt% to about 99.95 wt% or about 30 wt% to about 99.90 wt% of a pharmaceutically acceptable carrier, all percentages by weight being based on the total composition.

[0120] For clarity, the expression “compounds of the application” as used above refers to a single compound of the application or two or more compounds of the application.

[0121] In some embodiments, the method and uses of the present application comprising administering a compound of the application in combination with a second compound of the application and / or another therapeutic agent or any other treatments for the diseases, disorders and conditions described herein, such as treating a TDP-43 proteinopathy or treating a neurodegenerative disorder.EXAMPLES

[0122] The following non-limiting examples are illustrative of the present disclosure:Example 1:Materials & MethodsCell Culture, Transfection and Treatment

[0123] Cell culture. HEK293T and HeLa cells were grown in DMEM with 10% Hi-FBS and 1 x Pen-Strep in 37°C humidified atmosphere with 5% CO2. 24 hours prior to transfection, cells were seeded on a poly-D-lysine (Sigma) coated glass coverslips in 24-well plates for subsequent immunocytochemistry or on 6 well plates for biochemical analysis of cell lysates. Thirty minutes prior to transfection, media was changed to Opti-MEM (Gibco). Cells were transfected with 1 :2 ratio of plasmid DNA and transfection reagent EndoFectin™ Max (Genecopoeia) according to manufacturer instructions. Cells were transfected with GFP, GFP- TDP-25, GFP-TDP-35, GFP-TDP-43ANLS, GFP-TDP-C-spl-272, RFP-TDP-25, Kpn|31-GFP (Addgene plasmid # 106941), 3xFLAG-mTB-TDP-25, SOD1 (G93A)-FLAG or HA-alpha- synuclein-6xHis (Addgene plasmid # 40824). Three hours post-transfection, media was supplemented with Opti-MEM media containing 20% Hi-FBS, for a final concentration of 10% Hi-FBS. 21-hours later, the cells were utilized for either immunocytochemistry or immunoblotting.

[0124] siRNA Knockdown. HEK293T cells were grown and seeded in 6 well plates. After 24 hours, media was changed to Opti-MEM, and cells were transfected with either ON- TARGETplus non-targeting siRNA pool or ON-TARGETplus siRNA targeting Kpn|31 (Horizon) using DharmaFECT transfection reagent (Horizon) according to manufacturer instructions. After 24 hours (day 2), cells media was changed to DMEM with 10% Hi-FBS and 1 x Pen- Strep. On day 3, cells were transfected as above.

[0125] Compound Treatment. Cells were treated with 5, 10, 15, 20 and 25pM of exemplary compound for 90 minutes, 24 hours post-transfection. Alternatively, cells were treated with 2.5 or 5pM of an exemplary compound 3 hours post-transfection for 21 hours.

[0126] Immunoprecipitation. 24-hours post-transfection with GFP-TDP-25, cells grown on 6-well plates were washed two times with cold PBS (pH 7.4; Gibco) before being scrapped in RIPA buffer (150mM NaCI, 50mM Tris pH7.4, 1% NP-40 and 1 x Halt protease and phosphatase inhibitor). Samples were incubated on ice for 20 minutes before being sonicated with a Misonix Microson™ XL 2000 Ultrasonic Cell Disruptor (output wattage 3) for 10 seconds. Cells were subsequently subjected to immunoprecipitation utilizing magnetic agarose control beads and GFP-Trap ® Agarose beads (ChromoTek) according to manufacturer instructions. Given that Kpn|31 interaction with GFP-TDP-25 is seeminglytransient and / or weak, 100% of immunoprecipitated material was loaded on the 10% SDS- PAGE gel.

[0127] Immunocytochemistry. Cells on coverslips were washed two times with PBS (pH 7.4; Gibco) and fixed with 4% paraformaldehyde (PFA) for 15 min at room temperature. Cells were permeabilized and blocked with blocking solution (10% donkey serum, 3% bovine serum albumin (BSA) with 0.3% Trition X-100) for 30 minutes. Primary antibodies were added to coverslips and incubated for 1 hour at room temperature: anti-FLAG (Rabbit, 1 :1000, ProteinTech, #20543- 1-1AP), anti-TDP-43 (Rabbit, 1 :1000, ProteinTech, #10782-2-AP) and anti-HA (Rabbit, 1 :50, Origene, #TA150034). Subsequently, cells were washed three times with PBS+0.1% Tween-20 (PBST) for 5 minutes, prior to the addition of the secondary antibodies anti-rabbit Alexa Fluor™ 594 (1 :1000, Invitrogen, #A21207) in blocking solution for 30 min at room temperature. Cells were then washed three times with PBST for 5 minutes, and mounted with Prolong™ Gold Antifade Mountant with DNA Stain DAPI (ThermoFisher). Cells were imaged on a Leica DM6000 microscope using Velocity Imaging software (PerkinElmer). Cells were counted manually and divided into three groups: non-transfected cells, cells with diffused GFP / RFP signal, and cells with punctuated GFP / RFP signal. The nuclear:cytoplasmic (N:C) ratio of GFP, GFP-TDP-25, GFP-TDP-35, GFP-TDP-43ANLS, GFP-TDP-C-spl-272, RFP-TDP-25, Kpn|31-GFP was performed using Z-stack images by manually drawing the cell body and nucleus boundaries and N:C fluorescence intensities analyzed by Velocity analysis suite. Statistical t-test of two-samples assuming equal variance and graphing of the data were performed on Microsoft Excel.

[0128] Immunoblotting. Cells in wells were washed two times with cold PBS (pH 7.4; Gibco) before being scraped into RIPA buffer (150mM NaCI, 50mM Tris pH7.4, 1% NP-40, 1 mM EDTA and 1 x Halt protease and phosphatase inhibitor). Samples were incubated on ice for 20 minutes before being sonicated with a Misonix Microson™ XL 2000 Ultrasonic Cell Disruptor (output wattage 3) for 10 seconds. To an aliquot of the cell suspension SDS was added to 2%, and labeled “Total Fraction”. For fractionation of soluble vs insoluble proteins, the remaining total cell lysates were centrifuged for 15 mins at 15 OOOxg at 4°C. The supernatant was removed and labeled “Soluble Fraction”. RIPA with 2% SDS was added to the resulting pellet and the sample was sonicated for 20 seconds, and labeled “Insoluble Fraction”. PierceTM BCA assay was performed according to manufacturer instructions to quantify protein concentration of the samples. 4xSample buffer (200mM Tris-CI pH6.8, 40% glycerol, 0.02% bromophenol blue, 8% SDS and 20% |3-mercaptoethanol) was added to samples, and the samples were boiled at 95°C for 5 minutes. Equivalent amounts of proteins were loaded onto 10% SDS-PAGE gels, and then transferred to polyvinyldiflouride (PVDF)membranes. The membranes were then blocked with 5% BSA or 5% skim milk in TBST for 1 hour at room temperature prior to the addition of primary antibodies anti-pTDP S409 / 410 (Mouse, 1 :1000, CosmoBio, #TIP-PTD-M01), anti-Kpn|31 (Mouse, 1 :1000, Santa Cruz, #SC137016), anti-GFP (Mouse, 1 :5,000, Clontech, #632380) or anti-GAPDH (Mouse, 1 :10,000, Abeam, #Ab8245), diluted in blocking solution overnight at 4°C. The membranes were then washed three times with TBS+0.1% Tween-20 (TBST) for 5 minutes, prior to the addition of the secondary antibody, anti-mouse horseradish peroxidase conjugated (1 :5000, VWR, #NA931) diluted in blocking solution for 1 hour at room temperature. The membranes were then washed three times with TBS+0.1% Tween-20 (TBST) for 5 minutes prior to the addition of Western Lighting ® Plus ECL (Perkin Elmer) for developing. The signal was quantified from densitometric scans of autoradiography film (Denville scientific) using Imaged. Statistical t-test of two-samples assuming equal variance and graphing of the data were performed on Microsoft Excel.Primary Cortical Neuron Culture

[0129] Cell culture. Primary cortical neurons were isolated from brains of CD1 mouse embryos on day 15. Following trypsin digestion and dissociation, 250,000 cells / well were plated onto coverslips precoated overnight with poly-d-lysine (Sigma) in 0.1 M borate buffer in a 24 well plate, or 1 million cells / well in a 6-well plate. Cells were seeded in plating media composed of DMEM, 1 x Pen-Strep, 1 x Glutamax and 10% horse serum. After4 hours, plating media was replaced with Neurobasal Media composed of Neurobasal (Thermo Fisher Scientific) supplemented with 1 x Pen-Strep, I x Glutamax (Gibco) and 1 x B27 (Thermo Fisher Scientific). After 4 days, 1 x FDU was added for two days. Subsequently, in-house generated lentivirus expressing GFP-TDP-25 were added to cells for two days. Half media changes were performed every two days. On Day 12, cells were treated with 15pM JRMS or 0.23% DMSO for 90 minutes, and then utilized for immunocytochemistry or immunoblotting, as described earlier.

[0130] Lentivirus Generation. The constructs of GFP, GFP-TDP-43 and GFP-TDP-25 were subcloned into pLM-CMV-R-Cre vector (Gift from Michel Sadelain; Addgene plasmid # 27546) replacing mCherry and Cre sequences, referred to as pLM-GFP, pLM-GFP-TDP-43 and pLM-GFP-TDP-25. To generate the lentivirus, HEK-293 cells were maintained in DMEM supplemented with 10% Hi-FBS and 1 x Pen-Strep in 37°C humidified atmosphere with 5% CO2. 24 hours prior to transfection, cells were seeded on 15 cm dishes. The cells’ media was changed to Opti-MEM supplemented with 10% FBS, and cells were transfected with 4:3:1 ratio of pLM plasmid : psPAX2 plasmid (Gift from Didier Trono; Addgene plasmid # 12259) : pMD2.G (Gift from Didier Trono; Addgene plasmid # 12259) using transfection reagent Lipofectamine 3000 (Invitrogen) according to manufacturer instructions. Two days followingtransfection, the cells’ media was centrifuged at 1000xg for 5 minutes at 4°C, the supernatant filtered through a 0.42pM filter to remove any cell debris, and mixed with Lenti-X concentrator (Takara) and incubated at 4°C for three days. The solution was then spun at 1500xg for 45 minutes at 4°C, and the pellet was resuspended in complete neurobasal media, aliquoted and stored at -80°C.Human iPSCs-derived Cortical Neurons

[0131] Cell culture. Human iPSCs were differentiated into neural progenitor cells (NPCs) using embryoid body-based neural induction methods and were gifted as frozen stocks by Dr. Liliana Attisano (University of Toronto). NPCs were dissociated with 1 mL 0.25% T rypsin-EDTA (Gibco # 25200072) and plated on 0.07% PEI / Laminin coated wells at a density of ~65,00 cells / cm2for subsequent terminal differentiation into cortical neurons. Cells were maintained in neural media with 0.1 pM Compound-E for 4 days. On Day 4, 1 x FDU is added to the cells and removed on day 6. On Day 7, cells were maintained in neural media supplemented with 20ng / ml of both hBDNF and hGDNF. After three weeks, cells transduced with in-house generated lentivirus expressing GFP-TDP-25 were added to cells for three days before being treated with 7.5pM JRMS or 0.12% DMSO for 90 minutes, and then utilized for immunocytochemistry (with an additional primary antibody anti-NeuN (Mouse, 1 :500, Millipore, #MAB377) and secondary antibody anti-mouse Alexa Fluor™ 647 (1 :1000, Invitrogen, # A21235)), as described earlier.In Vivo Testing

[0132] At 1 month of age, mice were i.c.v. injected at Bregma +0.7 mm with 190 pM JRMS or vehicle (3% DMSO) 3 times per week for 2 weeks. Dosage was determined given that a mouse has ~35 pl of cerebrospinal fluid (CSF), signifying that an injection of 3 pl of 190 pM JRMS will have a final CSF concentration of 15 pM JRMS, the concertation at which positive results were observed in cells and primary cortical neurons. Mice were then euthanized.

[0133] Immunohistochemistry. Mouse brains were paraffin embedded and cut into 6pm sections at Bregma ~-3.00 mm using a microtome onto positively charged slides. Bregma ~-3.00 mm allows for quantification of the entorhinal cortex, the region harboring most TDP- 25 aggregates. The slides were deparaffinized and rehydrated through multiple washes with xylene, ethanol and water as outlined in (Xiao et al., 2015b). The antigen retrieval was performed by boiling the slides in TE9 buffer (10 mM Tris, 1 mM EDTA, 0.2% Tween 20 pH 9) at 110°C in a pressure cooker for 15 minutes, followed by cooling for 20 minutes and washing in running tap water for 5 minutes. Slides were then washed for 5 minutes in TBST, then in TBS + 0.3% Triton-X100 for 20 minutes to permeabilize the tissue, and then washed for 5minutes in TBST. Subsequently, the tissue was incubated for 1 hour at room temperature with blocking solution (0.3% Triton-X100, 3% BSA, 5% donkey serum), and then slides incubated overnight at 4°C with primary antibodies diluted in blocking solution. The primary antibodies used were anti-pTDP S409 / 410 (Rat, 1 :500, BioLegend, #829901), anti-GFP (Chicken, 1 :1000; Aves Labs, #GFP1020 anti-NeuN (Mouse, 1 :500, Millipore, #MAB377), and anti- Kpnpi (Mouse, 1 :50, Santa Cruz, #SC137016). The next day, slides were washed three times for 10 minutes each with TBST, followed by secondary antibody incubation for 1 hour at room temperature. The appropriate secondary antibodies Alexa Fluor™ 488, 568 and 647 against rat, chicken, and mouse were used. The slides were washed three times for 10 minutes each with TBST, and mounted with ProLong™ Gold Antifade Mountant with DNA Stain DAPI (ThermoFisher). Analysis of number of neurons and number and size of aggregates were performed in an automated manner by Velocity Analysis Suite (PerkinElmer). The nuclear:cytoplasmic ratio of Kpn[31 was performed using Z-stack images by manually drawing the cell body and nucleus boundaries and analyzed by Velocity analysis suite.

[0134] Tissue iysis. Whole mouse brains (without the cerebellum) were lysed in low salt buffer (50 mM Tris pH 7.5, 150 mM NaCI and 5mM EDTA) with a Dounce tissue grinder (Kimble) with Halt protease and phosphatase inhibitors (ThermoFisher). Following the addition of 2% SDS, the samples were sonicated with Misonix Microson™ XL 2000 Ultrasonic Cell Disruptor (output wattage 3) for 20 seconds. PierceTM BCA assay was performed according to manufacturer instructions to quantify protein concentration of the samples. 4xSample buffer (200mM Tris-CI pH6.8, 40% glycerol, 0.02% bromophenol blue, 8% SDS and 20% |3- mercaptoethanol) was added to samples, and the samples were boiled at 95°C for 5 minutes, then stored at -80 °C until use.Organotypic Slices

[0135] C57BL / 6 neonatal pups were intracerebroventricular (i.e.v.) injected withAAV9-GFP-TDP- 25 as outlined above. Mouse brains were dissected at 10 days of age and posterior cortex (which has the highest abundance of TDP-25 aggregates) was cut using a vibratome into 274 pM sections (2 sections / brain) in dissection media (1 x HBSS pH 7.3, 2.7mM NHCO3, 33mM Glucose, 1 mM Kynurenic acid and 0.2xPen-Strep) as outlined in (Falsig & Aguzzi, 2008). Organotypic slices were aged to 2 weeks in culture in in 37°C humidified atmosphere with 5% CO2, with culture media (50% MEM, 25% BME, 25% horse serum, 0.65% glucose, 1mM Glutamax, and 1 x Pen-Strep) changed every two days. On day 14 in vitro, per brain, one slide was treated with 15pM JRMS and the other with 0.23% DMSO. The slices were treated on day 14, 17 and 19. The brain slices were continuously live-imaged up to day 21 in vitro using Leica DM6000 microscope using Velocity Imaging software (PerkinElmer). Analysis of number of aggregates was performed in an automated manner byVelocity Analysis Suite (PerkinElmer). Statistical t-test of two-samples assuming equal variance and graphing of the data were performed on Microsoft Excel.ResultsJRMS prevents / reverses TDP-43 aggregation in cells and mouse primary cortical neurons in a dose dependent manner

[0136] Through investigation of mechanisms regulating the nuclear import of TDP-43, JRMS, a small molecule, that blocks nuclear import through Kpn|31 was identified. JRMS, binds Kpnpi at amino acids 331-361 , a region that is involved in its NCT function (Moroianu, Blobel, & Radu, 1996). By binding at this region, JRMS blocks Kpn|31 ’s interaction with RanGTP and Importin-a. Treatment of HeLa cells overexpressing TDP-25, TDP-35, TDP- 43ANLS and TDP-C-spl-272 with 15pM JRMS for 90 minutes led to a reduction in the number of cells with aggregates by 45%, 34%, 57% and 65%, respectively (Figure 1 A & E). Treatment with JRMS also resulted in a significant increase of nuclear localization of TDP-35 and TDP- C-spl-272, and an upward trend for TDP-25 and TDP-43-ANLS (Figure 1A & F-l). Given that TDP-25 forms phosphorylated cytoplasmic aggregates and most closely mimics TDP-43 pathology seen in patients (Shenouda, et al., 2018), further validation of JRMS and related compound of formulas (i.e. Compound II or JRMS-44) were done in models of TDP-25.

[0137] Treatment of HeLa cells expressing TDP-25 with 15pM JRMS for 90 minutes led to a ~40% reduction in size of TDP-25 aggregates (Figure 2 A & B). By immunoblotting, JRMS treatment reduced insoluble TDP-25 by ~55% (Figure 2 C & D). Acute (90 mins) and chronic (24-hours) treatment of cells with various concentration of JRMS showed a clear doseresponse curve when assessing the percentage of cells with TDP-25 aggregates and aggregate size (Figure 2 E-G). A previous study has shown that TDP-25 is able to recruit full- length TDP-43 from the nucleus leading to its cytoplasmic localization and incorporation into toxic aggregates in cells (Y.-J. Zhang et al., 2009). Given that this recruitment of the full-length protein is hard to achieve in cells, it was the aim to assess the effect of JRMS effect on this interaction of the full-length by utilizing a mutated nuclear localization sequence fragment of TDP-43 (i.e. TDP-ANLS) and the C-terminal fragment TDP-25 (Figure 3A). Indeed, JRMS treatment resulted in a 33% reduction in the number of cells with GFP-TDP-ANLS and RFP- TDP-25 co-aggregates (Figure 3). It is noteworthy that 15pM compound of formula II (referred herein to as JRMS-44) showed comparable efficacy by reducing TDP-25 aggregates by 34% after 90 mins of treatment in HeLa cells (Figure 4). While not wishing to be limited by theory, this observation may be due to the chaperoning function of Kpn|31 (Guo et al., 2019; Jakel et al., 2002; Khalil et al., 2022; Springhower et al., 2020).

[0138] In primary mouse cortical neurons transduced to express TDP-25 through lentivirus, treatment with JRMS up to 30pM showed no neurotoxicity (Figure 5A). JRMS treatment reduced TDP-25 aggregation by ~45% compared to DMSO control (Figure 5 B-D). By immunoblotting, JRMS reduced level of phosphorylated TDP-25, indicative of aggregated TDP-25, by ~50% without affecting the overall level of TDP-25 or Kpn|31 expression (Figure 5 B-G). To further assess the efficacy of JRMS in a human cellular context, human NPCs were terminally differentiated into cortical neurons and transduced with a lentivirus to express and induce TDP-25 aggregates. Treatment of TDP-35 expressing human iPSCs-derived cortical neurons with 7.5 pM JRMS for 90 mins resulted in a reduction of number of TDP-25 aggregates per neuron by 47% (Figure 6). Thus, small molecule JRMS is effective in preventing / reversing TDP-25 aggregation in cells, primary cortical neurons and human iPSCs- derived cortical neurons.JRMS prevention / reversion of TDP-43 is dependent on lmportin-131

[0139] In seeking to gain more understanding of JRMS’s mechanism of action on reducing TDP-25 aggregation, it was demonstrated that the efficacy of JRMS is dependent on the presence of Kpn|31 (Figure 7 A & B). That is, siRNA knockdown of Kpn|31 led to increased insoluble TDP-25 that was not reduced by JRMS treatment, while JRMS treatment of siRNA scrambled control cells showed a reduction of insoluble TDP-25 (Figure 7A). The subcellular localization of Kpn|31 upon JRMS treatment was then investigated revealing that JRMS treatment increases cytoplasmic levels of Kpn|31 , without affecting nuclear levels of endogenous TDP-43 (Figure 7 C-F). It is therefore proposed that increasing cytoplasmic levels of Kpn|31 promotes its chaperoning function of cytoplasmic TDP-25.

[0140] To further characterize JRMS mechanism of action, a mechanistic understanding of Kpn|31 interactions with TDP-25 was investigated. Overexpression of Kpn|31 reduced insoluble phosphorylated TDP-25 and increased its nuclear localization, while its knockdown increased insoluble phosphorylated TDP-25 (Figure 7 G-J). Taken together, it is apparent that levels of Kpn|31 modulate TDP-25 aggregation. By performing coimmunoprecipitation using TDP-25 as bait, a direct interaction of Kpn|31 with TDP-25 was shown (Figure73K). Interestingly, Kpn|31 co-immunoprecipitated with TDP-25 was shown to be reduced upon JRMS treatment, correlating with decreased phosphorylated TDP-25 (Figure 7 K & L), indicating that Kpn|31 mostly interacts with aggregated TDP-25. Indeed, upon normalizing immunoprecipitated Kpn|31 levels to phosphorylated / aggregated TDP-25, an increased interaction of Kpn|31 to TDP-25 is observed upon JRMS treatment (Figure 7M). Taken together, JRMS harnesses Kpn|31 intrinsic chaperoning function to prevent / reverse TDP-25 aggregation.JRMS prevents / reverses TDP-43 aggregation in mouse model

[0141] To further understand the therapeutic potential of JRMS, JRMS was tested in a TDP-43 proteinopathy mouse model. Assessment of JRMS characteristics against Lipinski’s modified rule of 5 for blood-brain barrier (BBB) permeability was inconclusive suggesting that JRMS is borderline permeable and requires empirical testing (Banks, 2009; Mikitsh & Chacko, 2014). Thus, to bypass the potential BBB hurdle, JRMS treatment was delivered intracerebroventricularly (I.C.V.) in AAV9-GFP-TDP-25 mice at 1 month of age for 2 weeks. Compared to control, JRMS treatment reduced the abundance and size of phosphorylated TDP-25 aggregates in the posterior cortex of mice, the region in which aggregates are most abundant. (Figure 8 A-C). Importantly, treatment with JRMS did not induce any toxicity as demonstrated by similar neuronal density in DMSO and JRMS treated mice (Figure 8D). Biochemically, JRMS treatment reduced levels of phosphorylated TDP-25 by ~35% without affecting TDP-25 expression level (Figure 8 E & F). Moreover, JRMS target engagement with Kpnpi was demonstrated with increased cytoplasmic levels of Kpn|31 in posterior cortex neurons of JRMS treated mice compared to DMSO control (Figure 8 G & H). Thus, JRMS treatment through I.C.V. injections showed therapeutic benefit in reducing TDP-25 aggregates within 2 weeks of treatment.

[0142] Organotypic slices from AAV9-GFP-TDP-25 mice were used to shed light on pharmacodynamics of JRMS. Two organotypic slices from each mouse were harvested and one treated with DMSO and one with JRMS every two days for a week. Over this timeframe, live cell imaging was performed to track the emergence and disappearance of TDP-25 aggregates. As shown in Figure 9, over 7 days, TDP-25 aggregates increased in the DMSO treatment at a rate of 0.64 and decreased with JRMS treatment at a rate of 0.18 compared to baseline levels. Overall, a new small molecule JRMS has been identified which harnesses the chaperone function of endogenous Kpn|31 to p re vent / re verse TDP-43 aggregation in cells, in mouse primary cortical neurons, in vivo and ex-vivo.JRMS prevents / reverses aggregation of other neurodegenerative diseases relevant proteins

[0143] To further explore the therapeutic potential of JRMS in the context of other proteinopathies, cellular models of aggregation of SOD1 and a-synculein were developed. SOD1 was the first gene associated with both familial and sporadic ALS, and mutations in the gene lead to it protein misfolding and accumulation (Redler & Dokholyan, 2012). The G93A mutation in SOD1 was one of the first identified in ALS patients, and was the first used to create a transgenic mouse model (Gurney et al., 1994). The G93A SOD1 mouse model exhibits neuronal cytoplasmic aggregates of misfolded SOD1 protein, causing motor neuron degeneration and paralysis by 4 months of age (Gurney et al., 1994). HeLa cellsoverexpressing SOD1 G93A-FLAG formed cytoplasmic puncta which is reduced by 28% upon 15pM JRMS treatment for 90 minutes (Figure 10 A & B). a-Synuclein is found aggregated as Lewy bodies (LB), Lewy neurites and glial cell inclusions in idiopathic Parkinson’s disease, Dementia with Lewy bodies (DLB), Multiple systems atrophy (MSA), Pure autonomic failure (PAF) and REM sleep behavior disorder (RBD), all of which are neurodegenerative diseases termed a-synucleinopathies (Calabresi, et al., 2023). Overexpression of wild-type a-synuclein in HeLa cells is sufficient to induce aggregate formation, which was reduced by 30% upon 15pM JRMS treatment for 90 minutes (Figure 10 C & D). Taken together, JRMS harnesses Kpnpi intrinsic chaperoning function to prevent / reverse aggregation of different proteins independent of sequences or structures.Discussion

[0144] In the present study, further evidence that Kpn|31 directly interacts with TDP- 43 isoforms (TDP-25) and modulates aggregation is provided. Moreover, a small molecule, JRMS, able to increase Kpn[31 cytoplasmic concentration, and thus elevating its non-canonical chaperoning function to reduce TDP-43 fragments’, SODI ’s and a-synuclein’s aggregation, was identified. Moreover, JRMS was shown to promote TDP-43 isoforms nuclear relocalization, hence addressing both disease mechanisms of loss of function from the nucleus and gain of function of aggregates in the cytoplasm. JRMS’ therapeutic potential was demonstrated in cells, in primary cortical cultures, in human iPSCs-derived cortical neurons, in organotypic slices and in vivo.

[0145] Recently, there has been a growing body of evidence that NOT proteins can modulate the aggregation propensity of aggregation-prone proteins across a wide range of neurodegenerative diseases (Darling & Shorter, 2021 ; Guo et al., 2019; Odeh, Fare, & Shorter, 2022; Springhower et al., 2020). In regard to ALS / FTLD, Kpn|32 was shown to inhibit and reverse aggregation of FUS, TAF15, EWSR1 , hnRNPAI , hnRNPA2 and C9orf72’s GR- DPR, while Kpna and Kpn|31 were shown to inhibit and reverse C9orf72’s GR / PR-DPR and TDP-43 aggregation (Cicardi et al., 2023; Doll et al., 2022; Fare, Rhine, Lam, Myong, & Shorter, 2023; Guo et al., 2019; Hofweber et al., 2018; Hutten et al., 2020; Khalil et al., 2022; Yoshizawa et al., 2018). It is noteworthy that aside from Kpn|31 , other [3-importins, such as IPO3 / TNPO2, IPO4, IPO9 and IPO13, are also able to reduce TDP-43 aggregation, independent of TDP-43 NLS domain (Khalil et al., 2022), indicating a potential conserved chaperoning mechanism within this family.

[0146] Kpn|31 is the best characterized nuclear import receptor with 19 tandem a- helical HEAT repeats (Cingolani, Petosa, Weis, & Muller, 1999; Neuwald & Hirano, 2000; Yoshimura & Hirano, 2016). Khalil et al (2022) has previously showed that Kpn|31 N-terminalis required and sufficient for reducing TDP-43 aggregation and rescuing associated toxicity. Specifically, HEAT repeats 1-8 reduce TDP-43 aggregation by ~70%, while HEAT repeats 1- 6 and 1-7 reduce TDP-43 aggregation by ~50%. In accord, JRMS binds Kpn|31 at residues 331-361 , which is estimated to be near the end of eighth HEAT repeat (Cingolani et al., 1999), and thus does not interfere with its chaperoning domains.

[0147] A series of protein domain interactions revealed that Kpn [31 interacts with TDP- 43 through a sequence in RRM2 (230-240aa) and through the low complexity domain (LCD), with either being sufficient for Kpn|31 chaperoning activity (Khalil et al., 2022). Kpn|31-mediated reduction of TDP-43 aggregation is dependent on its nucleoporin-interacting site in its N- terminal domain, which mediates FG-Nup interactions (Khalil et al., 2022). Specifically, Nup62 was shown to recruit Kpn|31 to TDP-43 to prevent / disassemble its aggregates (Khalil et al., 2022).

[0148] In its canonical function in NCT, Kpn|31 binds RanGTP in the nucleus, after which this complex is transported to the cytoplasm, where RanGTP is hydrolyzed to RanGDP, releasing Kpn|31 which binds Kpna already bound to NLS-cargo-protein and is then relocalized to the nucleus (Tziortzouda, Van Den Bosch, & Hirth, 2021). While not wishing to be limited by theory, it is proposed that JRMS prevents only a subset of Kpn|31 from binding its canonical interactors, RanGTP and Kpna, and thus preventing its nuclear localization, and increasing its cytoplasmic concentration. It is of note, that at the concentrations tested, JRMS did not impact endogenous TDP-43 nuclear localization, nor did it induce any toxicity, suggesting that these are sublethal concentrations not significantly impacting normal NCT. In addition to Kpnpi ’s ability to reduce TDP-25, TDP-35, TDP-43ANLS, and TDP-C-spl-272 aggregation, others have also shown that overexpression of Kpn|31 attenuates aggregation of TDP-43ANLS, TDP-43 Q331 K, TDP-43 M337V, TDP-43 A382T, and sTDP (N-terminal fragment of TDP-43 amino acids 1-269 + unique 18 amino acid sequence tail), and significantly reduced associated cell death (Khalil et al., 2022). This aggregation reduction effect was correlated with overexpressed Kpn|31 (Khalil et al., 2022), highlighting that it is optimal for Kpn|31 to be in proximity of the aggregation-prone proteins. Moreover, while overexpression of Kpn|31 reduces overall levels of TDP-25, it does not affect nuclear full-length TDP-43 levels, indicating a specific affinity to insoluble protein (Khalil et al., 2022). This increased affinity for insoluble TDP-43 isoforms was also observed when a decreased amount of Kpn|31 was co-immunoprecipitated with TDP-25 upon JRMS treatment compared to DMSO control, indicating that Kpn|31 increasingly interacts with aggregated TDP-25. In addition provided evidence that Kpn|31 through JRMS can reduce mutant SOD1 and a-synuclein’s aggregation. Thus, Kpn[31 cytoplasmic localization is optimal for its chaperoning activity, which demonstrates increased affinity towards various aggregated proteins.

[0149] Taken together, it has been demonstrated that Kpn|31 possesses a chaperoning ability, and its levels modulate TDP-43 aggregation and nuclear localization. Exemplary compounds of the application, including the small molecule JRMS (compound l(a), targeting Kpn|31 , has been identified which increases Kpn|31 cytoplasmic concentration, harnessing its chaperone activity to prevent and reverse TDP-43, SOD1 , a-synuclein and potentially other pathological proteins’ aggregates. JRMS reduces TDP-43 aggregation in a dose-dependent manner in cells, primary cortical neurons, human iPSCs-derived cortical neurons, organotypic slices and in a TDP-43 proteinopathy mouse model. Exemplary compound of formula II (referred herein to as JRMS-44) showed comparable efficacy by reducing TDP-25 aggregates by 34% after 90 mins of treatment in HeLa cells. As this therapeutic mechanism is being increasingly shown to alleviate different proteins aggregation, compounds of the application have the potential to possess therapeutic potential beyond TDP- 43-centric diseases.

[0150] While the present disclosure has been described with reference to examples, it is to be understood that the scope of the claims should not be limited by the embodiments set forth in the examples, but should be given the broadest interpretation consistent with the description as a whole.

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Claims

CLAIMS:I . A method of increasing Kpnpi ’s chaperoning function in a subject comprising administering to a subject in need thereof, an effective amount of a compound of Formula I,II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof:wherein:R1is selected from Ci-salkyl, Ci-4alkyleneN(Ci-4alkyl)2, Ci-4alkyleneOH,R2is selected from H and Ci_4alkyl;2. A method of treating or preventing a proteinopathy comprising administering to a subject in need thereof an effective amount of a compound of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV and / or XVI, or pharmaceutically acceptable salt, solvate and / or prodrug thereof, as defined in claim 1 , wherein the proteinopathy comprises an abnormal configuration of one or more proteins that are chaperoned by Kpn|31.

3. The method of claim 3, wherein the protein that is chaperoned by Kpn|31 is TAR-DNA- binding protein 43 kDa (TDP-43), SOD1 , FUS, C9orf72, C9orf72-dipetide repeat proteins (GA, GR, PA, PR, GP), hnRNPs, Tau, alpha-synuclein, Annexin A11 (ANXA11), CHCHD10 (CHC10), prion proteins (PrP), transthyretin (TTR) or huntingtin (HTT), or a fragment or isoform thereof.

4. The method of claim 3, wherein the protein that is chaperoned by Kpn|31 is TAR-DNA- binding protein 43 kDa (TDP-43), or a fragment and / or isoform thereof, and the chaperoning prevents or reverses aggregation of the TDP-43, or a fragment and / or isoform thereof.

5. The method of claim 4, wherein the fragment and / or isoform of TDP-43 is TDP-25, TDP-35, TDP-43ANLS or TDP-C-spl-272.

6. The method of claim 3, wherein the protein that is that is chaperoned by Kpn|31 is SOD1 or a fragment and / or isoform thereof.

7. The method of claim 6, wherein the isoform of SOD1 is SOD1 G93A.

8. The method of claim 3, wherein the protein that is that is chaperoned by Kpn|31 is alpha-synuclein or a fragment and / or isoform thereof.

9. The method of any one of claims 1 to 8, wherein the subject is suffering from a neurodegenerative disorder.

10. The method of claim 9, wherein the neurodegenerative disorder is argyrophilic grain disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), ALS-Parkinsonism dementia complex of Guam, corticobasal degeneration, Dementia with Lewy bodies, Huntington's disease, Lewy body disease, motor neuron disease, frontotemporal lobar degeneration (FTLD), frontotemporal dementia, frontotemporal lobar degeneration with ubiquitin-positive inclusions, hippocampal sclerosis, inclusion body myopathy, inclusion body myositis, Parkinson's disease, Parkinson's disease dementia, Parkinson-dementia complex in Kii peninsula, chronic traumatic encephalopathy, Limbic-predominant age-related TDP- 43 encephalopathy (LATE), Pick's disease, Creutzfeldt-Jakob disease, REM sleep behavior disorder (RBD), multiple systems atrophy (MSA) or pure autonomic failure (PAF), or a combination thereof.

11. The method of claim 10, wherein the neurodegenerative disorder is ALS, frontotemporal dementia, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, chronic traumatic encephalopathy or LATE.

12. The method of any one of claims 1 to 11 , wherein the number or frequency of neurons that include mislocalized and / or misfolded protein that is chaperoned by Kpn[31 , or a fragment and / or isoform thereof, in the brain or spinal cord of the subject are reduced.

13. The method of any one of claims 1 to 11 , wherein the amount or concentration of neuronal misfolded protein that is chaperoned by Kpn[31 , or a fragment and / or isoform thereof, in the brain or spinal cord in the subject are reduced.

14. The method of any one of claims 1 to 13, wherein the subject is human.

15. The method of any one of claims 1 to 14, wherein the compound is a compound of Formula I, or pharmaceutically acceptable salt, solvate and / or prodrug thereof:wherein:R1is selected from Ci_6alkyl, Ci.4alkyleneN(Ci.4alkyl)2, Ci.4alkyleneOH,R2is selected from H and Ci_4alkyl.

16. The method of claim 15, wherein the compound of Formula I, or pharmaceutically acceptable salt, solvate and / or prodrug thereof is:pharmaceutically acceptable salt, solvate and / or prodrug thereof.

17. The method of claim 16, wherein the compound of Formula I, or pharmaceutically acceptable salt, solvate and / or prodrug thereof is:or a pharmaceutically acceptable salt, solvate and / or prodrug thereof.

18. The method of any one of claims 1 to 14, wherein the compound is a compound of Formula II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV or XVI, or a pharmaceutically acceptable salt, solvate and / or prodrug thereof.

19. The method of claim 18, wherein the compound is a compound of Formula II, or a pharmaceutically acceptable salt, solvate and / or prodrug thereof.

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